Cellular immunotherapy in melanoma: the next frontier in cancer treatment.

Background

Melanoma is among the fastest-rising cancers and poses a disproportionate clinical burden. In 2022, an estimated 331,722 new cases occurred worldwide (age-standardized incidence 3.2 per 100,000) and 58,667 deaths [ 1 ], with the highest rates in fair-skinned populations and a decades-long upward trend in incidence, especially in Australia/New Zealand and North America [ 2 ] with an incidence rising of over 320% in the US since 1975. Arising from neural-crest-derived melanocytes, melanoma can develop in skin and at extra-cutaneous sites where melanocytes normally reside resulting in the epidermal-based cutaneous melanoma and the non-cutaneous melanomas, which occur in the uveal tract of the eye and on mucosal surfaces [ 3 , 4 ].

Cutaneous melanoma is largely driven by ultraviolet (UV) exposure, resulting in a characteristic C > T mutational signature, high mutational burden, and frequent mitogen-activated protein kinase (MAPK) pathway alterations like mutations in B-Raf proto-oncogene (BRAF) V600 or Neuroblastoma RAS viral oncogene homolog (NRAS) [ 5 ]. In line with the central etiological role of UV exposure, more than 80% of global cutaneous melanoma cases in 2022 were estimated to be attributable to UV exposure [ 6 ]. By contrast, non-cutaneous melanomas usually lack UV signatures and arise through distinct genomic alterations [ 7 , 8 ].

Treatment regimens

These biologic distinctions have direct implications for clinical management and therefore shape both the initial work-up and subsequent therapy selection. The pathway begins with histologic confirmation and American Joint Committee on Cancer staging (I–IV) based on tumor features (T), lymph nodes (N), and metastases plus lactate dehydrogenase (LDH; M). In advanced cases, brain magnetic resonance imaging (MRI), LDH, and BRAF V600 testing guide prognosis and treatment selection [ 9 ]. Current international guidelines prioritize programmed cell death protein 1 (PD-1)-based immunotherapy as first-line for most patients regardless of BRAF status [ 10 ].

PD-1, cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), and lymphocyte-activation gene 3 (LAG-3) are inhibitory immune checkpoint receptors on activated T cells that normally limit immune responses and maintain self-tolerance. Tumors exploit this system, for example by expressing Programmed death-ligand 1 (PD-L1), to suppress effector T-cell function and evade immune destruction. Monoclonal antibodies blocking PD-1, other checkpoint receptors, or their ligands release this brake, enhance tumor-specific T-cell activity, and can induce durable responses in some patients with advanced cancers, including melanoma [ 11 – 13 ].

In clinical practice, anti-PD-1 antibodies are established standard monotherapy options for advanced melanoma. This is supported by KEYNOTE-006, in which pembrolizumab significantly improved median overall survival (mOS) compared with ipilimumab (anti-CTLA-4; 32.7 vs. 15.9 months) [ 12 ]. Treatment therefore consists of pembrolizumab (anti-PD-1) or nivolumab (anti-PD-1) monotherapy, or dual-checkpoint blockade with nivolumab (anti-PD-1) plus ipilimumab or relatlimab (anti-LAG-3) [ 10 ]. Combination regimens further extend survival, reaching an mOS of 71.9 months with nivolumab plus ipilimumab in CheckMate-067 [ 14 ] and about 51.0 months with nivolumab plus relatlimab in RELATIVITY-047 [ 15 ].

For tumors harboring BRAF V600 , combined BRAF inhibitor (BRAFi) and mitogen-activated protein kinase (MEK) inhibitor (MEKi) therapy remains a standard, rapidly acting option, achieving a median overall survival (mOS) of 25.9 months with dabrafenib (BRAFi) plus trametinib (MEKi) [ 16 ] and a five-year overall survival of about 35% with encorafenib (BRAFi) plus binimetinib (MEKi), rising to about 45% in patients with normal LDH levels [ 17 ]. Treatment selection between targeted therapy and immunotherapy is individualized [ 18 ], although randomized sequencing studies support starting with immunotherapy when clinically feasible [ 19 ].

Despite these advances, many patients with advanced metastatic melanoma (stage IV) still derive limited or no durable benefit from current systemic therapies. Approximately 40–55% show primary resistance to PD-1- or CTLA-4-based immune checkpoint inhibition (ICI), and about 25% of initial responders later develop acquired resistance [ 20 , 21 ]. In addition, intensified regimens such as nivolumab plus ipilimumab cause grade 3–4 adverse events (AE) in around 60% of patients, often limiting treatment use or causing early discontinuation [ 22 ]. Together, these findings highlight persistent limitations of current checkpoint inhibition and targeted therapy, as many patients do not respond durably, relapse, or cannot tolerate full-intensity treatment [ 23 ].

Studies show this ceiling arises as chronic antigen exposure in melanoma drives CD8⁺ T cells into exhausted states with stable transcriptional and epigenetic changes and persistent inhibitory receptor expression [ 24 ]. This exhaustion is initially permissive to reversal by ICI. However, tumor-intrinsic alterations such as.

(I) loss of antigen presentation, (II) defects in interferon-γ/Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling, (III) oncogenic pathway activation, (IV) tumor-extrinsic factors including T-cell exclusion, (V) expansion of immunosuppressive immune cells including regulatory T cells (T reg ) and myeloid-derived suppressor cells (MDSCs) as well as. (VI) extensive dysfunction of the CD8⁺ compartment,

collectively drive primary and acquired resistance to PD-1, CTLA-4 and BRAF/MEK inhibition in melanoma [ 25 – 27 ]. Because ICI primarily reinvigorates pre-existing antitumor T-cell responses, its efficacy is favored by the presence of tumor-reactive lymphocytes and intact antigen presentation at baseline [ 13 , 24 , 28 ]. In cold or heavily pretreated tumors, however, low antigenicity and a myeloid-dominated, T cell-hostile microenvironment often leave too few potent tumor-specific T cells to be rescued by checkpoint blockade [ 24 , 28 ]. Together, these findings identify a key biological and clinical gap: resistance to ICI may reflect not only inhibitory receptor signaling, but also insufficient quantity, specificity, or functional fitness of tumor-reactive lymphocytes within the tumor microenvironment [ 13 , 23 , 24 , 28 ].

This unmet need provides a strong rationale for cellular therapies that directly supply highly tumor-reactive T cells or generate them ex vivo. Owing to its high mutational burden, frequent T-cell infiltration, and long-standing role as a model disease in immunotherapy, melanoma has been at the forefront of adoptive cell therapy development. Accordingly, a broader range of cellular approaches is now being pursued to exploit melanoma’s immunogenicity while overcoming key resistance mechanisms that limit antibody-based immunotherapy.

Main Text

Cellular immunotherapies for melanoma

While immune checkpoint inhibitors and targeted therapies have transformed the prognosis of metastatic melanoma, many patients still develop resistance or cannot tolerate full-intensity regimens, creating a need for additional immune-based strategies [ 22 , 25 , 29 ]. Cellular immunotherapies seek to fill this gap by inducing de novo tumor-specific immunity or by supplying expanded and engineered effector cells able to persist and function in the tumor microenvironment. In melanoma, the most promising platforms include dendritic cell-based vaccines, tumor-infiltrating lymphocyte transfer, genetically engineered T-cell therapies such as T-cell receptor-engineered T cells (TCR-T) and chimeric antigen receptor (CAR) T cells, and natural killer (NK) cell-based approaches including CAR NK products [ 30 , 31 ] (Graphical Abstract).

Dendritic-cell-based vaccines in melanoma

Dendritic-cell-based vaccination (DCV) in melanoma is a personalized immunotherapy in which autologous dendritic cells (DCs) are loaded ex vivo with tumor-associated antigens (TAAs), such as defined peptides or whole tumor lysates, to prime or boost tumor-specific CD4⁺ and CD8⁺ T-cell responses in vivo [ 32 , 33 ]. As professional antigen-presenting cells (APCs), DCs efficiently process and present antigens on human leukocyte antigen (HLA) class I and II molecules and provide the co-stimulatory and cytokine signals required for effective T-cell activation and differentiation [ 34 , 35 ].

Principle and manufacturing of DCV

In most melanoma protocols, dendritic cells (DCs) are generated from leukapheresis-derived monocytes cultured with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). They are then matured using pro-inflammatory cytokine cocktails (containing tumor necrosis factor α (TNF-α), IL-1β, IL-6 and prostaglandin E2 (PGE2)) or toll-like receptor (TLR) agonists and loaded with melanoma-associated peptides, such as Melanoma antigen recognized by T cells 1 (MART-1), Glycoprotein 100 (gp100), tyrosinase, or Melanoma antigen gene (MAGE), or with autologous whole tumor lysate. The final product is typically administered repeatedly by intradermal, subcutaneous, or intranodal injection [ 34 – 36 ].

These products act as in vivo stimulators to elicit endogenous tumor-specific T cells and can be repeatedly administered in adjuvant and metastatic settings without lymphodepleting conditioning [ 33 , 37 ]. This distinguishes DCV from adoptive T-cell transfer, as it aims to reshape the host T-cell repertoire rather than infuse large numbers of pre-expanded effector cells.

Clinical activity and safety

The clinical development of DC-based therapies in melanoma was pioneered by Nestle et al., who vaccinated metastatic melanoma patients with autologous monocyte-derived DCs (moDC) pulsed with defined melanoma peptides or autologous tumor lysate. This induced antigen-specific cytotoxic T lymphocytes, delayed-type hypersensitivity (DTH) responses, and objective tumor regressions in 5 of 16 evaluable patients, including 2 complete responses (CR) and 3 partial responses (PR), indicating clinical activity in a subset of patients. However, this small, non-randomized phase I study was not designed to assess OS or definitive clinical efficacy [ 32 ].

Subsequent phase I and phase II trials of peptide- or tumor lysate-pulsed mature DCs confirmed consistent immunogenicity, with vaccine-induced tumor-specific CD8⁺ T-cell responses detected in most evaluated patients. Akiyama et al. reported enzyme-linked immunospot (ELISPOT) responses against ≥ 2 melanoma peptides in 6 of 6 immunologically evaluable patients [ 38 ], while Ribas et al. detected tumor antigen-specific CD8⁺ T cells in 26 of 29 patients, with boosted or induced responses in 19 of 29 patients [ 36 ]. Despite this, objective responses remained modest, occurring in about 10% of patients with advanced melanoma, while 11–18% achieved prolonged stable disease [ 36 , 38 , 39 ]. These findings highlight the persistent gap between immunogenicity and clinical benefit, likely due to tumor burden, microenvironmental immunosuppression, and T-cell exhaustion [ 33 , 35 ].

The toxicity profile of DCV is generally favorable and dominated by low-grade local and systemic AE. Most studies reported grade 1–2 flu-like symptoms, low-grade fever, fatigue, and transient injection-site reactions, whereas grade 3 toxicities were rare and treatment-related deaths were not observed [ 33 , 36 , 37 ]. Importantly, severe immune-mediated toxicities characteristic for ICI have not been documented for DCV, which makes these therapies particularly attractive for the adjuvant setting and for combination strategies in patients who have already received multiple lines of systemic treatment and allows a prolonged vaccination schedule [ 33 , 37 , 40 ].

Antigen formats and peptide-based strategies

Optimization of DC-based T-cell therapies in melanoma focuses on antigen selection and formulation to improve the quality, breadth, and durability of vaccine-primed T-cell responses [ 35 , 41 ].

One approach ues synthetic long peptides (SLPs, Fig. 1 ), which require intracellular processing and cross-presentation and contain epitopes presented via HLA class I and class II, thereby engaging both CD8⁺ and CD4⁺ T cells more effectively than minimal epitope vaccines [ 42 , 43 ]. Preclinical studies show that long-peptide-pulsed DCs achieve superior cross-presentation and enhanced activation of cytotoxic T cells (CTL) than short-peptide-loaded DCs [ 43 , 44 ]. Personalized DCV incorporating patient-specific neoantigen peptides identified by tumor sequencing can broaden the targeted epitope repertoire and, in small melanoma cohorts, increase the breadth and diversity of circulating neoantigen-specific T-cell responses, although larger trials are still needed to establish clinical efficacy [ 45 ].

Fig. 1 Advances in manufacturing of dendritic cell vaccination in melanoma. Autologous monocytes obtained from leukapheresis are differentiated ex vivo into monocyte-derived DCs. The resulting immature DCs are then matured and loaded in parallel with tumor-associated antigens, defined melanoma peptides, synthetic long peptides and patient-specific neoantigen peptides identified by tumor sequencing, or alternatively as autologous whole tumor lysate. Antigen formulation refinements, including conjugation to virus-like particles, encapsulation in nanoparticles, or the use of multi-peptide pools, can enhance antigen uptake and support efficient cross-presentation. Maturation is induced with TNF-α, IL-1β, IL-6, and PGE2, optionally complemented by toll-like receptor agonist and type I interferons. Third-generation approaches such as TriMix further potentiate maturation by electroporating moDCs with mRNA encoding CD40L, CD70, and constitutively active TLR4. The final mature, antigen-loaded DC product is administered by injection to stimulate endogenous tumor-specific CD4⁺ and CD8⁺ T-cell responses through antigen presentation on HLA class I and II, co-stimulation, and cytokine signaling

An alternative antigen-agnostic strategy is loading DCs with autologous whole tumor lysate, which includes shared differentiation antigens, cancer-testis antigens, and private or unknown neoantigens without predefined epitope selection [ 46 ]. In the phase I/II study by Salcedo et al. in stage III/IV melanoma, lysate-loaded autologous moDCs were feasible, safe, and immunogenic, with no grade 3–4 vaccine-related toxicities and enhanced T-cell responses to tumor lysate and TAA-derived peptides; among nine patients completing all vaccinations, four survived > 20 months, including one complete remission and one case of stable metastatic disease for ca. 10 months [ 46 ]. In the randomized phase III MIND-DC trial after complete resection of stage IIIB/C melanoma, intranodal vaccination induced functional tumor-specific T cells in about 67% of vaccinated patients versus 3.8% in the placebo arm, but did not significantly improve clinical outcome despite numerically longer 2-year progression-free survival (PFS) and OS [ 47 ]. Overall, tumor-lysate-loaded DC vaccines show biological activity and possible clinical benefit in selected patients, but the evidence remains limited, heterogeneous, and modest compared with contemporary adjuvant PD-1-based therapy.

Formulation refinements, including conjugation of tumor lysates to virus-like particles (Fig. 1 ) and encapsulation of tumor antigens with TLR agonists in nanoparticles, improve antigen uptake and cross-presentation. In preclinical melanoma and other solid-tumor models, these approaches elicit stronger CD8⁺ T-cell priming and better tumor control than conventional lysate-pulsed DCs [ 48 , 49 ]. Likewise, TLR3 agonists such as poly I: C can generate more potent clinical-grade DCs; poly I: C-activated moDCs induce greater expansion of antigen-specific T cells and a lower Forkhead box P3 (FoxP3)⁺ T reg /T effector ratio than DCs matured with lipopolysaccharide or cytokine cocktails [ 50 ]. However, clinical validation in melanoma remains at an early stage.

Optimization of DC maturation and delivery

A second axis of optimization targets DC maturation and delivery route, as immature or incompletely matured DCs may induce tolerance, whereas fully matured DCs support effector and memory differentiation [ 51 ]. Maturation cocktails combining TLR agonists, pro-inflammatory cytokines, and type I interferons upregulate co-stimulatory molecules such as CD80, CD86, and CD40, enhance IL-12 production, and promote chemokine receptor expression favoring lymph node migration [ 52 , 53 ]. In preclinical models and early human studies, these DCs induce stronger T helper type 1 (T H 1) and cytotoxic T-cell (CTL) polarization than conventional GM-CSF/IL-4-derived DCs [ 53 , 54 ].

The TriMix platform (Fig. 1 ) is a third-generation maturation strategy in which moDCs are electroporated with mRNA encoding CD40 ligand, CD70, and constitutively active TLR4, generating highly mature cells with enhanced T-cell stimulatory capacity and reduced donor-to-donor variability [ 55 ]. In pretreated advanced melanoma, TriMixDC-MEL was feasible and well tolerated and induced objective responses in about one quarter of patients, including durable complete responses [ 56 ]. In phase II, combination with the CTLA-4 antibody ipilimumab achieved an overall response rate of about 38%, with complete responses in about 20% and several responses lasting beyond two to three years [ 57 ]. These data suggest that optimized DC maturation can improve durable benefit, particularly in combination with checkpoint inhibition.

Alternative delivery routes for DC vaccines in melanoma should be viewed as route-optimization strategies rather than intrinsically superior approaches. Intradermal injection depends on active DC migration and is inefficient, with in vivo tracking showing that only up to 4% of injected DCs reach draining lymph nodes. Nevertheless, even small numbers of viable DCs in T-cell areas can induce antigen-specific responses [ 58 ]. Intranodal delivery is mechanistically attractive because it bypasses this migratory bottleneck, and an early randomized phase I melanoma trial reported superior CD8⁺ T-cell function after intranodal administration compared with intravenous or intradermal routes [ 59 ]. However, later comparative data did not confirm a general advantage: in 43 advanced melanoma patients, intradermal vaccination induced functional tumor-antigen-specific T cells more frequently than intranodal vaccination, despite greater nodal DC delivery [ 60 ]. Combined intradermal/intravenous approaches such as TriMixDC-MEL were safe and immunogenic in small phase Ib cohorts, but remain non-definitive [ 56 ]. Accordingly, the optimal route likely depends on DC maturation state, migratory capacity, antigen loading, and disease setting. This is underscored by the phase III MIND-DC trial, where intranodal natural DC vaccination induced functional antigen-specific T-cell responses but did not improve recurrence-free survival [ 47 ].

Defined melanoma peptides in DC and non-DC vaccines

A broad range of melanoma-associated peptides has been used to prime or boost T-cell responses in DC-based and conventional peptide vaccine trials. These include differentiation antigens such as Melan-A/MART-1, gp100, and tyrosinase, as well as cancer-testis antigens such as MAGE family members and New York esophageal squamous cell carcinoma 1 (NY-ESO-1), administered either as single epitopes or in peptide pools [ 61 ].

Single-epitope approaches, exemplified by the MAGE-3.A1 peptide DC vaccine, show that a shared tumor peptide can expand tumor-specific cytotoxic T cells and induce regression of individual metastases in a subset of patients, but responses are infrequent, often partial, and limited by antigen-loss escape [ 62 ]. By contrast, multi-peptide DC vaccines, including CD34-progenitor-derived DCs loaded with 4 HLA-A2-restricted melanoma peptides [ 63 ] moDCs pulsed with 5-peptide cocktails tailored to HLA-A2 or HLA-A24, consistently induce broader T-cell repertoires [ 64 ]. In these studies, most patients responded to at least one melanoma antigen and many to multiple epitopes, whereas no single peptide emerged as a dominant target across patients. Objective responses or disease stabilization occurred in a subset of patients, and broader epitope recognition often correlated with improved short-term disease control [ 63 , 64 ].

Non-DC peptide vaccines provide complementary insight. In a randomized phase III trial in HLA-A2-positive advanced melanoma, addition of the gp100:209–217(210 M) peptide to high-dose IL-2 increased objective response rates (ORR) and prolonged PFS and OS compared with IL-2 alone, although absolute response rates remained modest [ 65 ]. Likewise, the NY-ESO-1 79–108 long-peptide vaccine with CpG-B induced integrated NY-ESO-1-specific CD8⁺ and CD4⁺ T-cell and antibody responses in virtually all evaluable patients, with durable polyfunctional cytokine production and epitope spreading, while clinical benefit mainly consisted of stable disease and occasional partial responses [ 66 ].

In the adjuvant setting, multi-peptide vaccines evaluated in the Mel39 and Mel44 programs were tested in completely resected high-risk melanoma (AJCC stage IIB-IV) arising from cutaneous, mucosal, or unknown primary sites [ 67 , 68 ]. In Mel39, 4- and 12-peptide class I HLA-restricted vaccines induced high peripheral immunogenicity and were associated with long-term OS, with a non-significant trend favoring the 12-peptide arm [ 67 ] In the larger randomized phase II Mel44 trial, a 12-peptide vaccine plus melanoma-specific helper peptides (12MP+6MHP) showed more favorable long-term OS than the same vaccine plus tetanus helper peptide (12MP + tet) [ 68 ].

Overall, defined melanoma peptides such as MART-1, gp100, tyrosinase, MAGE-3, and NY-ESO-1 can induce strong antigen-specific T-cell responses in both DC-based and conventional peptide vaccines. Peptide pools more reliably broaden the cytotoxic T-cell repertoire and may improve outcomes within individual cohorts, but a clear survival advantage over single-peptide strategies has not been demonstrated in randomized melanoma trials (Table 1 ).

Table 1 Representative peptide formulations and their immunological and clinical readouts Peptide or peptide set Platform and clinical setting Immunogenicity readouts Clinical activity Single-peptide vs. peptide-pool MAGE-3.A1 minimal epitope (74) Mature moDC + recall antigen in stage IV melanoma ( n = 11). MAGE-3.A1-specific CTL expansion in 8/11; recall responses boosted. Metastatic lesion regression in 6/11; regressing lesions showed CD8⁺ infiltrates, whereas some non-regressing lesions lost MAGE-3. A single shared peptide can induce tumor-specific CTLs and clinical regression, but responses were infrequent/partial; antigen-loss escape supports multi-epitope strategies. Four-peptide HLA-A2 pool (MART-1, gp100, tyrosinase, MAGE-3) (81) CD34⁺-progenitor-derived DCs pulsed with 4 HLA-A2-restricted melanoma peptides + Flu-MP/KLH in metastatic melanoma ( n = 18). Immune responses to KLH/Flu and ≥ 1 melanoma peptide in 16/18; 10/18 responded to > 2 antigens with T H 1-polarized profiles. Among 17 evaluable patients, ≥ 1 metastasis regressed in 7; responses to > 2 antigens were linked to less early progression. Compared with a single peptide, a 4-peptide pool broadened immunity and was associated with better early disease control, although overall activity remained modest. HLA-A2/A24 five-peptide DC cocktail (49) Autologous moDC pulsed with 5 HLA-matched melanoma peptides + KLH in metastatic melanoma (phase I, n = 9). All 6 immunologically evaluable patients developed CTL responses to > 2 epitopes; tetramer⁺ CD8 T cells were detected with a T H 1-biased pattern. Best responses: 1 CR, 1 PR, 1 SD, 6 PD. A focused mid-sized peptide pool induced multi-specific CD8 responses and a small but notable objective response rate. gp100:209–217(210 M) minimal epitope (77) Soluble peptide vaccine + high-dose IL-2 versus IL-2 alone in unresectable stage III/IV melanoma (randomized phase III, n = 185). Earlier studies showed circulating gp100-specific CD8 + T cells in most vaccinated patients; phase III immunomonitoring was limited but confirmatory. gp100 increased ORR from 6% to 16% and mPFS from 1.6 to 2.2 months, with a trend toward longer OS; toxicity was similar between arms and driven by IL-2. A single optimized shared antigen can augment cytokine therapy, but absolute efficacy remained modest. NY-ESO-1 79–108 long peptide (78) Long synthetic NY-ESO-1 peptide + CpG-B in Montanide, ± low-dose IL-2, in resected stage III/IV melanoma (phase I; 18 evaluable). Integrated NY-ESO-1-specific CD8 + and CD4 + T-cell plus antibody responses in 18/18, with polyfunctionality, epitope spreading, and persistence for ≥ 1 year. Clinical benefit consisted mainly of prolonged no evidence of disease (NED) or stable disease (SD), with only occasional PR. The long-peptide format is a strong priming strategy but, as monotherapy, mainly produced durable immune activation and disease stabilization rather than high ORRs. Four-peptide vs. twelve-peptide multiepitope vaccines (Mel39) (80) Adjuvant 4- versus 12-peptide shared-antigen vaccines for resected high-risk melanoma (stage IIB-IV; n = 51). Both formulations were highly immunogenic; broader peripheral immune responses were seen with 12 peptides. Post-hoc 20-year analysis showed OS of about 65% at 10 years and 49% at 20 years overall; HR for OS favored 12 peptides but was not significant, and RFS did not differ significantly. Increasing from 4 to 12 epitopes broadened induced CD8 + responses but did not significantly improve long-term survival. Ten-peptide moDC vaccine (82) Cocktail-matured moDC loaded with 10 defined class I/II melanoma peptides, with DC-licensing strategies, in stage III/IV cutaneous melanoma (phase I/II; n = 62). Multi-epitope T-cell responses occurred in nearly all patients; most responses were induced de novo, and memory persisted for years. With ≥ 12-year follow-up, about 19% of patients with non-resectable metastatic melanoma remained alive, but no WHO-defined CR/PR occurred. This large mixed class I/II peptide pool was highly immunogenic and associated with long-term survival in a subset, but peptide breadth did not correlate with OS, suggesting diminishing returns from simply adding more shared peptides.

Combination strategies of DCV with ICIs and adoptive t-cell transfer

Combination strategies integrate DCV with ICI or adoptive T-cell transfer to overcome tumor-induced immunosuppression and enhance the persistence and function of vaccine-primed T cells. In preclinical melanoma models, combining DCV with PD-1-targeting antibodies increases the density and function of tumor-infiltrating effector T cells, reduces regulatory T-cell and myeloid-derived suppressor-cell populations, and improves tumor control and survival compared with either monotherapy [ 69 , 70 ].

Adoptive T-cell transfer (ATT) programs have incorporated DC-based vaccination before or after infusion of tumor-infiltrating lymphocytes (TIL) or other ex vivo expanded T-cell products. In stage IV melanoma, first clinical trials showed that adding a tumor-antigen-pulsed DC vaccine to TIL therapy was feasible and safe and enhanced antigen-specific T-cell responses, although ORRs did not clearly exceed those expected with TIL therapy alone [ 71 , 72 ]. Other combinations, including DCV with metronomic cyclophosphamide, low-dose IL-2, or cyclo-oxygenase-2 inhibition, achieved disease control in about one third to one half of metastatic melanoma patients in single-arm phase II studies, but did not meaningfully improve OS [ 39 , 73 ].

Overall, available data suggest that optimized DCVs are most effective as part of rational combination strategies with ICI or ATT, in which DCs provide antigen-specific priming while partner therapies relieve inhibitory pathways or increase effector-cell numbers. However, randomized trials are still needed to define the magnitude of clinical benefit in melanoma.

Tumor-infiltrating lymphocytes

ATT with TILs is a personalized immunotherapy that exploits the pre-existing, tumor-experienced T-cell repertoire of individual patients to mediate antitumor activity in solid tumors [ 74 ]. The standard procedure involves resection of a tumor lesion, ex vivo expansion of resident TILs under high-dose IL-2, and reinfusion after non-myeloablative lymphodepletion with short-course high-dose IL-2 support [ 74 , 75 ]. TIL products are typically polyclonal, enriched for CD8 + T cells, and contain clonally expanded populations recognizing multiple autologous tumor epitopes, including neoantigens, which likely underlies the depth and durability of clinical responses [ 74 , 76 ]. Unlike monoclonal ICI, TIL therapy transfers a high-frequency tumor-reactive T-cell population into a host environment optimized for expansion.

The clinical development of TIL therapy was pioneered in metastatic melanoma, whose high mutational burden and frequent baseline T-cell infiltration made it an ideal proof-of-concept setting [ 75 , 77 ]. Early single-center studies reported ORR of about 40–50% in heavily pretreated melanoma, including durable complete responses [ 75 , 78 , 79 ], establishing TIL transfer as one of the most active immunotherapies in melanoma before the checkpoint inhibitor era. These data supported the development of standardized TIL products and randomized comparisons with established ICI.

Lifileucel is a centrally manufactured, cryopreserved, autologous TIL product and currently the most advanced clinical implementation of TIL therapy in melanoma [ 80 , 81 ]. In the phase II C-144-01 trial in unresectable or metastatic melanoma previously treated with PD-1 blockade and, if BRAF V600 -mutated, targeted therapy, a single lifileucel infusion after standard lymphodepletion and high-dose IL-2 achieved an ORR of about 31–36%, with complete responses in about 5–7% and disease control in roughly 70–80% of patients [ 80 , 82 ]. These results led to accelerated approval of lifileucel (AMTAGVI™) by the United States Food and Drug Administration (FDA) in 2024 for adults with unresectable or metastatic melanoma previously treated with a PD-1 inhibitor and, if BRAF V600 -mutated, an appropriate BRAF-targeted regimen, making it the first approved TIL therapy for a solid tumor [ 81 ].

Randomized evidence further established the clinical relevance of TIL therapy. In the phase III trial by Rohaan et al., patients with advanced melanoma progressing after anti-PD-1 therapy were randomized to TIL therapy or ipilimumab; PFS was significantly longer and ORR substantially higher with TIL therapy, supporting its role as an active option for checkpoint-refractory melanoma [ 83 ].

The toxicity profile of TIL therapy is driven mainly by lymphodepleting chemotherapy and high-dose IL-2 rather than antigen off-target effects of the infused cells [ 80 , 84 ]. Lymphodepleting chemotherapy induces in nearly all patients grade 3–4 AE, predominantly hematologic toxicities, which were usually transient and manageable with standard supportive care [ 82 – 84 ]. High-dose IL-2 further contributes to capillary leak syndrome, hypotension, tachycardia, renal and hepatic dysfunction, and the need for intensive monitoring, although these events are generally reversible and limited to the acute treatment period [ 78 , 84 , 85 ]. Treatment-related mortality across modern TIL studies, including lifileucel, has remained in a low range [ 82 , 83 ]. Importantly, severe immune-mediated toxicities characteristic of checkpoint inhibitors, appear less frequent with TIL therapy [ 83 , 84 , 86 ]. Thus, although TIL therapy is intensive and should be delivered in specialized centers, its safety profile is predictable and acceptable given its potential for durable disease control.

Optimization strategies for TIL therapy

Optimization of TIL-product composition and manufacturing

Several strategies aim to optimize TIL therapy in melanoma and other solid tumors by improving response rates and durability while simplifying logistics and reducing toxicity (Fig. 2 ). A central focus is TIL composition, especially enrichment of neoantigen-specific or stem-like T-cell subsets and avoidance of terminally exhausted or regulatory populations, which correlates with superior clinical responses in retrospective analyses [ 76 , 87 ]. Antigen-specific enrichment can be achieved by cytokine capture, in which activated T cells are isolated by magnetic-activated cell sorting (MACS) or fluorescence-activated cell sorting (FACS) after retaining secreted cytokines on a surface-bound matrix [ 88 ]. Interferon-γ (IFN-γ) is the most widely used functional readout for tumor recognition in CD8⁺ and CD4⁺ T cells, with Good Manufacturing Practice (GMP)-grade and automated workflows reported [ 89 ]. However, cytokine production is confined to particular T-cell subsets, so enrichment based solely on IFN-γ or related readouts may fail to recover all antigen-specific populations [ 90 ].

Fig. 2 Advances in TIL immunotherapy through optimization of product composition, functional fitness, and tumor microenvironment resistance. Multiple strategies aim to improve TIL therapy in melanoma and other solid tumors by enhancing response durability and simplifying manufacturing. Tumor-reactive and neoantigen-specific TILs can be enriched using cytokine capture assays, activation-induced markers, PD-1, CD39, CD103, CD137, or peptide-HLA multimers. Long-term persistence is promoted by preserving stem-like TIL states, shortening expansion, and using IL-7/IL-15 instead of exclusive high-dose IL-2, with optional IL-21 and 4-1BB co-stimulation. Efficacy may be further enhanced by combining TILs with PD-1 blockade and by engineering resistance to tumor-mediated suppression through approaches such as CISH knockout, TGF-β or adenosine pathway targeting, chemokine receptor retargeting, and membrane-bound IL-15 expression

Activation-induced marker capture is an alternative based on marker upregulation after brief co-culture with autologous tumor or personalized neoantigen pools presented by professional APCs [ 90 , 91 ]. Antigen-agnostic enrichment using PD-1 can recover patient-specific tumor-reactive CD8⁺ T cells, but exhausted and bystander cells remain a limitation [ 92 , 93 ]. CD39 and CD103 co-expression likewise identifies tissue-resident memory CD8⁺ T cells enriched for tumor reactivity, although these cells often display an exhausted phenotype that may impair persistence [ 94 , 95 ]. Among available markers, CD137 (4-1BB) appears particularly selective, as depletion of CD137⁺ cells markedly reduces tumor reactivity within PD-1-, CD39-, or CD103-enriched TIL populations [ 91 ].

An alternative strategy uses fluorochrome-conjugated peptide-HLA (pHLA) multimers to identify antigen-specific T-cell receptor (TCR) specificities independent of functional readouts. Although highly specific for predicted neoantigens, this method is constrained by HLA restriction, imperfect epitope prediction, and substantial labor and cost [ 96 , 97 ].

Stem-like phenotypes can be enriched by selecting CD39⁻ CD69⁻ CD8⁺ TILs, which mark a TCF1⁺ progenitor population associated with complete regression and long-term persistence after ATT [ 87 ]. Shortening pre-expansion preserves younger, less differentiated TILs [ 98 , 99 ], whereas replacing exclusive high-dose IL-2 with IL-7 and IL-15, optionally with IL-21, and adding 4-1BB co-stimulation during outgrowth or rapid expansion can further improve CD8⁺ yield and fitness [ 100 – 102 ].

Overall, enrichment of neoantigen-specific and TCF1⁺/stem-like subsets while avoiding terminally exhausted or regulatory populations is a promising strategy to improve TIL durability. However, prospective evidence that such selection improves ORR over unselected bulk TIL products remains limited [ 76 , 87 ].

Combination strategies with immune checkpoint inhibition

Combining TIL therapy with PD-1 inhibition is conceptually attractive because TIL infusion replenishes tumor-reactive lymphocytes, whereas PD-1 blockade may sustain their function and limit re-induction of hypofunctional or exhausted states in the tumor microenvironment. Consistent with the model proposed by Datsi and Garg, TIL plus PD-1 strategies aim to increase both the number and functional quality of tumor-specific T cells beyond a critical threshold, rather than merely releasing inhibitory brakes on an insufficient endogenous repertoire [ 24 ]. Early clinical data support this rationale: in the phase II IOV-COM-202 trial in previously untreated unresectable or metastatic melanoma, lifileucel plus pembrolizumab achieved an ORR of 65.2%, including a CR of 30.4%, with regression of target lesions in all evaluable patients and expected regimen-related toxicity [ 103 ]. These findings support biologically plausible synergy and justify randomized first-line trials comparing TIL therapy with combined ICI plus TIL approaches.

Genetic-engineering strategies

Genetic modification of TILs is a key strategy to improve their efficacy and durability. Cytokine-inducible SH2-containing protein (CISH) is an intracellular negative regulator of T-cell receptor (TCR)- and cytokine signaling, and preclinical models show that CISH knockout (CISH KO ) enhances T-cell proliferation, cytokine production, metabolic fitness, and tumor recognition without broad non-specific autoreactivity [ 104 , 105 ]. In the first-in-human phase I trial of CRISPR-Cas9-edited CISH KO TILs in metastatic gastrointestinal epithelial cancers, large-scale manufacturing, engraftment, and an acceptable safety profile were demonstrated, without dose-limiting gene-editing-specific toxicities. Preliminary efficacy signals included stable disease in several patients and at least one durable complete response in microsatellite instability-high colorectal cancer, providing clinical proof-of-concept that intracellular checkpoint deletion can enhance TIL function in humans [ 106 ].

Additional engineering strategies aim to increase the therapeutic index of TIL therapy, including deletion of inhibitory receptors such as PD-1 [ 107 ], disruption of suppressive cytokine pathways such as TGF-β signaling [ 108 ], introduction of autocrine cytokine support circuits [ 109 ], and chemokine-receptor retargeting to improve homing [ 110 ]. A related approach, termed cytokine engineering, seeks to overcome dependence on systemic high-dose IL-2. For example, OBX-115 expresses membrane-bound human IL-15 under pharmacological control to provide cell-intrinsic survival and proliferation signals and potentially reduce the need for exogenous IL-2 [ 109 , 111 ].

TIL therapy provides clinically meaningful durable responses in patients with advanced melanoma after failure of ICI and targeted therapies, with a predictable and manageable short-term toxicity profile [ 82 ]. Long-term lifileucel data, superiority over ipilimumab [ 83 ], and FDA approval as Amtagvi [ 81 ] have established TIL therapy as a key treatment for advanced melanoma. Ongoing engineered and combination approaches, including CISH KO and other gene-edited products, may further improve efficacy, safety, and applicability across cancers. However, unlike CAR T cells, TIL production requires resectable tumor tissue.

CAR T cell therapy

CAR T cells combine an antibody-derived binding domain with intracellular activation and costimulatory modules such as CD28 or 4-1BB, enabling HLA-independent recognition of tumor-associated surface antigens and rapid T-cell activation [ 112 ]. Manufacturing involves isolation of autologous T cells from leukapheresis, ex vivo activation, gene transfer of the CAR transgene, usually by γ-retroviral or lentiviral vectors, and large-scale expansion under GMP conditions before lymphodepleting chemotherapy and reinfusion [ 113 , 114 ]. Process variables such as cytokine composition and culture duration critically influence phenotype, function, and in vivo kinetics of the final product [ 115 , 116 ].

Clinically, CAR T cells show high efficacy in hematologic malignancies such as diffuse large B-cell lymphoma, other B-cell lymphomas, and multiple myeloma, where pivotal or randomized trials have demonstrated superior disease control compared with standard of care [ 117 , 118 ]. The principal toxicities are cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity (ICANS), which result from on-target immune activation and range from mild to life-threatening [ 119 ]. Rare post-therapy T-cell neoplasms have also been reported, leading to increased pharmacovigilance and label warnings in 2024, although the absolute risk appears low relative to clinical benefit [ 120 ]. To date, no CAR T-cell therapy has been approved for melanoma or other solid tumors, and all CAR-based strategies in melanoma remain investigational.

Melanoma Targets for designing the CARs

Despite successes in hematologic malignancies, translating CAR T-cell therapy to melanoma remains difficult because of antigen heterogeneity, shared expression with normal melanocytic and neural tissues, and a profoundly immunosuppressive tumor microenvironment (TIME) [ 121 ]. The following section therefore focuses on melanoma-relevant target antigens that form the basis for current and emerging CAR T-cell strategies in this disease (Table 2 ).

Table 2 Expression of Melanoma Targets Target Alias / Synonyms Expression in Melanoma Potential Off-Tumor Risk (examples) Evidence Level Clinical Status / Activity Example Studies / IDs GD2 Disialoganglioside GD2 Frequent/variable in cutaneous & metastatic melanoma Peripheral nervous system, pain fibers; occasional skin/mesenchymal expression Early clinical Phase I: safe/feasible; biological activity, limited clinical efficacy JITC 2024 B7-H3 CD276 Broad across solid tumors incl. melanoma; low in normal tissues Low basal expression in healthy tissues (widely distributed, mostly weak) Recruiting Dose-finding (safety/MTD) NCT04897321 IL13Rα2 IL-13 receptor α2 Subsets of melanoma (heterogeneous); elevated in some brain mets Limited normal expression (e.g., testis, smooth muscle, variable) Recruiting Phase I (safety/dose) NCT04119024 CSPG4 MCSP, HMW-MAA High and frequent on melanoma cells Pericytes, melanocyte precursors; limited normal expression Strong preclinical CAR T active preclinically DOI:10.3390/cancers11081198 (131) TYRP1 gp75 Melanosomal with surface pools shown (cutaneous, acral, uveal models) Pigmented tissues (melanocytes); vitiligo-like effects possible Strong preclinical Efficacy in PDX/models; clinical pending DOI: 10.1038/s41467-024-45221-2 (135)

B7 homolog 3 (B7-H3) is broadly expressed across solid tumors, including melanoma, and on tumor vasculature and stroma, whereas normal tissues show mostly low protein expression, supporting a potential therapeutic window [ 122 , 123 ]. In the first-in-human STRIvE-02 phase I trial of systemic B7-H3 CAR Ts in relapsed or refractory solid tumors, 1 of 9 treated patients achieved a partial response, and toxicity was mainly limited to low-grade CRS and transient transaminitis, without clear acute organ injury attributable to B7-H3 expression in healthy tissues [ 124 ]. Several additional B7-H3 CAR T studies allow enrollment of B7-H3 positive melanoma, yet no melanoma specific safety or response data have been reported [ 125 , 126 ]. Preclinical data further suggest that epigenetic modulators may pharmacologically increase B7-H3 density, although this has not been tested in melanoma and could increase on-target, off-tumor risk [ 127 ].

Chondroitin sulfate proteoglycan 4 (CSPG4), also referred to as NG2, is frequently expressed on primary and metastatic melanoma and other aggressive tumors, whereas normal expression is largely restricted to pericytes and selected stromal subsets, implying possible vascular off-tumor toxicity [ 128 – 130 ]. CSPG4-directed CAR-engineered lymphocytes show potent melanoma killing in preclinical models, and GMP-compliant transient CSPG4 CAR T production has been established. However, the dedicated melanoma trial is still recruiting, and no human efficacy or safety data are yet available [ 131 , 132 ].

Interleukin-13 receptor alpha 2 (IL13Rα2) is overexpressed in glioblastoma and subsets of melanoma and other solid tumors, with minimal to absent protein in most normal tissues [ 133 , 134 ]. In a phase I trial of locoregional IL13Rα2 CAR Ts in recurrent high-grade glioma, treatment was feasible, toxicities were mostly low grade, and objective responses, including durable regressions, occurred without a consistent off-tumor injury signal [ 134 ] An ongoing phase I trial is evaluating IL13Rα2 CAR Ts in stage IIIC/IV melanoma and other IL13Rα2-positive solid tumors, but melanoma-specific efficacy data have not yet been reported [ 133 ].

Tyrosinase-related protein 1 (TYRP1) is a melanocytic differentiation antigen involved in melanin synthesis and is expressed in cutaneous melanocytes and retinal pigment epithelium as well as multiple melanoma subtypes [ 135 ]. High-affinity TYRP1 CAR Ts show selective recognition of TYRP1-high melanoma cells and robust antitumor activity in preclinical models, apparently driven by antigen-density-dependent activation, but no clinical data are available and toxicity to skin or ocular melanocytes remains theoretical [ 135 – 137 ].

Disialoganglioside (GD2) is highly expressed on neuroblastoma, other neuroectodermal tumors, and a substantial subset of metastatic melanoma, whereas normal expression is largely restricted to central nervous system (CNS) neurons, peripheral nerves, and melanocytes [ 138 ]. Clinical experience in neuroblastoma and diffuse midline glioma shows that GD2 CAR T cells can induce meaningful responses with manageable toxicities under close CNS monitoring [ 139 – 141 ]. In the first phase I trial of third-generation GD2 CAR Ts in adults with GD2-positive metastatic melanoma and other solid cancers, manufacture was feasible and no dose-limiting toxicities or severe unexpected neuropathic events occurred, but clinical responses were limited, suggesting that melanoma efficacy will likely require combination strategies or next-generation constructs [ 142 ].

Taken together, B7-H3, CSPG4, IL13Rα2, TYRP1, and GD2 are biologically plausible but imperfect melanoma targets. Their therapeutic windows are constrained by heterogeneous antigen density, residual expression in normal tissues, and resistance mechanisms within the TIME, highlighting the need for next-generation CAR T-cell designs that improve both safety and efficacy.

CAR T Upgrades

The limited efficacy of first-generation CAR T-cell therapies in melanoma reflects persistent barriers including heterogeneous and plastic antigen expression, shared targets on normal melanocytic and neural tissues, a profoundly immunosuppressive TIME, and insufficient trafficking into metastatic lesions [ 143 – 145 ]. Next-generation CAR T designs therefore aim to broaden on-tumor recognition, improve localization, enhance resistance to immune suppression, and increase safety (Fig. 3 ).

Fig. 3 Modular next-generation CAR T-cell engineering strategies to overcome key barriers in melanoma. First-generation CAR T cells show limited activity in melanoma due to heterogeneous and plastic antigen expression, shared targets on normal melanocytes or neural tissues, a suppressive and metabolically hostile tumor immune microenvironment, and inefficient trafficking into metastatic lesions. Next-generation designs combine improved on-tumor recognition with tighter spatial control and enhanced safety. Multi-antigen targeting broadens coverage and limits antigen escape using dual or pan-CAR concepts that recognize more than one melanoma-relevant antigen. AND-gate CAR architectures restrict activity by requiring two inputs for full function, thereby reducing off-tumor activation. Logic-gated circuits also include synNotch IF-THEN programs that induce CAR expression only after a priming signal and OR-gates that maintain activity across heterogeneous target expression. Tumor localization can be increased by locoregional delivery and by trafficking enhancement through engineered chemokine receptors matched to tumor chemokine gradients. In the CAR–IL-12 concept shown, CAR–IL-12 is expressed after an upstream activation, recognizes the tumor antigen via its CAR-domain and simultaneously engages the T-cell IL-12 receptor via its IL-12 domain, driving downstream signaling that induces chemokine receptor expression and thereby improves migration into the tumor milieu. Resistance to suppression can be strengthened through disruption or decoying of inhibitory axes including TGF-β signaling via dominant-negative TGFβRII and adenosine signaling via A2A receptor targeting. Safety is reinforced through clinically controllable fail-safes such as inducible caspase-9 suicide switches activated by a small molecule CID (chemical inducer of dimerization) enabling rapid pharmacologic ablation of infused cells if severe toxicity occurs

A first area of optimization is antigen recognition. Dual, tandem, and conditional CAR constructs targeting more than one antigen can reduce antigen escape and improve control of heterogeneous tumors in preclinical models without introducing dominant new safety signals when targets are carefully selected [ 146 , 147 ]. Supporting this concept, a pan-CAR T approach co-targeting B7-H3 and CSPG4 achieved broad antigen coverage and effective tumor control in heterogeneous triple-negative breast cancer models, suggesting relevance for melanoma, although clinical validation is still lacking [ 148 ].

Logic-gated CAR T cells, including synthetic notch (synNotch)-based and AND/NOT-gate architectures, further refine specificity by requiring defined antigen combinations before activation. In murine models, these systems reduce off-tumor toxicity while preserving antitumor activity, which is particularly relevant for melanoma targets such as TYRP1 and GD2 that are shared with essential normal tissues [ 146 , 149 , 150 ].

A second area is enhancement of CAR T-cell fitness within the TIME. Armored CAR T cells can conditionally deliver immunomodulators at the tumor site and thereby preserve proliferation, persistence, and cytotoxicity under chronically inflamed conditions [ 151 , 152 ]. Constructs incorporating inducible IL-12, IL-18, IL-15, or αPD-L1/IL-12 fusion proteins have shown superior expansion, IFN-γ production, and durable tumor rejection in solid tumor models while limiting systemic toxicity when expression remains tightly linked to antigen engagement [ 153 – 156 ]. Early clinical experience, such as huCART19 IL-18 CARs in refractory lymphoma, supports the translatability of armored approaches [ 157 ]. In parallel, genetic disruption of inhibitory pathways can reduce susceptibility to soluble suppressive mediators. CRISPR-mediated deletion of the adenosine A2A receptor (A2AR) improved cytokine production and tumor control in Human epidermal growth factor receptor 2 (HER2) + and mesothelin + solid tumor models [ 158 – 160 ], whereas co-expression of dominant-negative transforming growth factor-β receptor type II (dnTGFβRII) or transforming growth factor-β traps improved proliferation and cytotoxicity under high TGF-β conditions [ 161 , 162 ].

A third area is trafficking. Engineering CAR T cells to express chemokine receptors such as C-X-C chemokine receptor (CXCR) type 2 or C-C chemokine receptor (CCR) type 2, matched to melanoma-associated chemokine patterns, increased intratumoral accumulation and tumor control in xenograft models, although off-tumor chemokine gradients remain a concern [ 163 – 166 ]. Microenvironment-responsive designs that couple CAR signaling to hypoxia or P-selectin can further focus activity to tumor sites and reduce systemic exposure in murine models [ 167 – 169 ]. Locoregional administration, already explored for IL13Rα2 and B7-H3 CAR T cells in brain tumors, may also be relevant for melanoma with dominant CNS involvement, although dedicated melanoma trials are lacking [ 134 , 170 ].

A fourth area is safety. Inducible suicide switches such as inducible caspase-9 (iCas9) allow rapid elimination of infused cells and have shown proof of principle in preclinical and early clinical studies [ 171 – 173 ]. Additional synthetic-biology approaches, including tunable ON switches, split CAR designs, and small-molecule-responsive domains, may allow in vivo control of CAR T-cell activity with improved safety [ 174 , 175 ]. Such safeguards are especially important for targets such as GD2 and CSPG4, where low-level expression in normal tissues raises concern for neurotoxicity or skin toxicity. This concept is already reflected in melanoma-directed platforms: a B7-H3-targeted CAR T-cell strategy for metastatic uveal melanoma incorporates iCas9, whereas CSPG4-specific melanoma CAR T cells are being developed with transient mRNA electroporation to limit CAR expression duration [ 131 , 176 ]. Fourth-generation platforms such as 4SCAR, combining a conventional CAR backbone with iCas9 and additional co-stimulatory modules, have entered early-phase solid-tumor trials with encouraging preliminary safety and antitumor activity [ 177 – 179 ].

Collectively, these upgrade strategies provide a modular toolkit to address the central obstacles identified for melanoma CAR T-cell therapy:

(I) suboptimal target specificity. (II) antigen heterogeneity, (III) a suppressive microenvironment, (IV) poor trafficking. (V) safety concerns.

The available data suggest that melanoma targets are unlikely to succeed as simple single-antigen, first-generation CARs, but may become more effective when integrated into logic-gated, armored, trafficking-enhanced, and safety-switch-equipped platforms (Table 3 ). The critical next step is rational clinical evaluation of such composite designs in biomarker-selected melanoma populations, with careful pharmacovigilance and systematic correlation of antigen expression, CAR T-cell dynamics, and clinical outcomes.

Table 3 Advantages and limitations of the therapeutic options for melanoma treatment Therapy modality Key advantages (pros) Key disadvantages / limitations (cons) Immune checkpoint inhibitors (ICI) (PD-1 and CTLA-4–based) Can induce meaningful and sometimes durable responses by rescuing pre-existing antitumor T cells. Primary and acquired resistance are common; efficacy is limited in cold or myeloid-inflamed TIME, and intensified regimens may be poorly tolerated. Targeted therapy Provides rapid disease control in appropriate molecular subsets and can be integrated into sequencing strategies. Durable benefit is often limited in therapy-resistant disease, and repeated treatment lines add cumulative toxicity. Dendritic cell (DC) vaccines Excellent safety profile; can prime broad antigen-specific immunity and support T-cell, B-cell, and NK-cell responses with memory potential. Objective responses as monotherapy are modest in advanced melanoma; best viewed as priming or backbone partners for combinations and dependent on a permissive TIME. Tumor-infiltrating lymphocyte (TIL) therapy Replaces the missing tumor-reactive repertoire; recognizes diverse targets including neoantigens; achieves ~ 30–40% ORR in checkpoint-refractory melanoma with durable complete responses in a subset. Requires tumor resection, complex manufacturing, lymphodepletion, and high-dose IL-2; intensive treatment best delivered in specialized centers. TCR-engineered T cells (TCR-T) Can target intracellular antigens and neoantigens presented by HLA and may support long-lived memory when manufactured in a stem-like state. HLA-restricted and usually narrow in specificity, with risk of immune escape; safety depends on strict control of off-target and cross-reactive toxicity. CAR-T / CAR-NK cells Not HLA-restricted, do not usually require patient-specific tumor material for target discovery, and allow programmable, potentially off-the-shelf development. In melanoma and other solid tumors, efficacy is limited by scarce tumor-selective surface antigens, antigen heterogeneity, CRS/neurotoxicity, and limited persistence in TIME. NK-cell–based immunotherapy (including adoptive NK, CAR-NK, engagers; exploratory) HLA-independent, potentially off-the-shelf, often with a more favorable toxicity profile; may target antigen-loss and HLA-deficient clones and cooperate with DCs and T cells. Still exploratory; limited by poor in vivo persistence, incomplete trafficking, suppressive TME factors, and the need for better targets, stronger platforms, and larger controlled studies.

Engineered T-cell receptor T-cell therapy in melanoma

Engineered TCR T-cell therapy redirects autologous T cells to tumor cells by introducing a defined, high-affinity TCR that recognizes a peptide-HLA complex derived from a tumor antigen. Unlike CAR T cells, which bind intact surface proteins independently of HLA, TCR-engineered T cells recognize intracellular proteins presented on HLA class I and, in some designs, class II, thereby expanding the target space to cancer-testis antigens, lineage antigens, and neoantigens inaccessible to conventional antibodies or CARs [ 180 ].

Manufacturing broadly parallels CAR T-cell production: T cells are collected by leukapheresis, activated ex vivo, transduced with a γ-retroviral or lentiviral vector encoding the tumor-specific TCR, expanded under controlled culture conditions, and infused after non-myeloablative lymphodepletion to promote engraftment and in vivo expansion [ 180 , 181 ].

A key distinction is the emphasis on preserving an early memory, stem-like phenotype, which is associated with superior engraftment, self-renewal, and long-term persistence [ 180 , 182 ]. Clinical studies of NY-ESO-1- and other tumor-specific TCR T cells have shown multi-year persistence and durable antitumor responses, consistent with formation of a functional memory pool [ 183 ]. In contrast, conventional CAR T-cell products are often skewed toward more differentiated effector phenotypes and are more prone to exhaustion, which can limit long-term persistence. Accordingly, process design for TCR T cells must balance efficient viral transduction with maintenance of a less differentiated, memory-like state [ 181 , 184 , 185 ].

Clinical activity and safety in melanoma and related solid tumors

Early proof-of-concept for TCR-engineered T cells in melanoma came from a landmark trial using an HLA-A*02:01-restricted NY-ESO-1-specific TCR. In patients with metastatic melanoma or synovial sarcoma refractory to standard therapy, NY-ESO-1 TCR-transduced T cells plus high-dose IL-2 induced objective responses in 5 of 11 melanoma patients and 4 of 6 synovial sarcoma patients, including durable complete responses in melanoma, establishing that TCR-engineered T cells can mediate deep and lasting regressions when a suitable target antigen is present [ 186 ].

More recently, MART-1-specific TCR-engineered peripheral-blood T cells were evaluated in metastatic cutaneous and uveal melanoma. Although the study achieved an ORR of 18%, it was terminated early because of dose-dependent, on-target/off-tumor toxicity affecting skin, eyes, and ears, including one treatment-related death at the highest dose level, illustrating the narrow therapeutic window of differentiation antigens shared with normal melanocytes [ 187 ].

Preferentially Expressed Antigen in Melanoma (PRAME)-directed TCR-T therapy is currently among the most advanced TCR-T programs in melanoma. PRAME is broadly expressed across melanoma subtypes and other solid tumors while being largely absent from healthy adult tissues [ 188 , 189 ]. In the phase I ACTengine IMA203 trial, an autologous PRAME-specific TCR product achieved as presented at the 2025 ASCO Annual Meeting an ORR of 54% in heavily pretreated melanoma at the recommended phase II dose, with tumor shrinkage in most patients, a median duration of response of about 12 months, activity across cutaneous, mucosal, and uveal melanoma, low rates of high-grade CRS, and no consistent signal of off-target organ toxicity [ 190 ]. These findings prompted the randomized phase III SUPRAME trial comparing IMA203 with investigator’s choice in previously treated cutaneous melanoma [ 191 ].

Overall, current data support TCR-engineered T cells as a clinically active modality in melanoma when suitable antigens such as NY-ESO-1 or PRAME are targeted. Safety reflects both regimen-related toxicity from lymphodepletion and, where used, IL-2, and antigen-dependent risks. Cytopenias, infections, and transient systemic symptoms are common but usually manageable [ 182 ]. A distinct risk is unexpected cross-reactivity with structurally related self-peptides: affinity-enhanced MAGE-A3-specific TCRs caused fatal cardiac toxicity through off-target recognition of a titin-derived peptide, and other MAGE- and MART-1-directed products have produced severe neuro- and skin toxicities in melanoma [ 187 , 192 – 194 ]. These events have driven increasingly stringent preclinical specificity testing and the development of additional genetic safety strategies.

Genetic-engineering strategies

Many engineering concepts described for CAR T cells and genetically modified TILs, including checkpoint disruption, cytokine armoring, chemokine-receptor retargeting, and safety switches, also apply to TCR-engineered T cells (Fig. 4 ).

Fig. 4 Advances in TCR-engineered T-cell immunotherapy for melanoma and other solid tumors. Engineered TCR T-cell therapy redirects autologous T cells by introducing a defined, tumor-reactive TCR that recognizes peptide–HLA complexes, thereby enabling targeting of intracellular antigens such as cancer-testis antigens, lineage antigens, and neoantigens that are not accessible to CAR T cells. Next-generation design features include structure-guided optimization to improve antigen discrimination and reduce off-target recognition, optimized TCR α/β pairing to favor correct chain assembly and limit mispairing and prevention of endogenous/transgenic TCR mispairing through silencing or deletion of native TCRα/β chains to generate monospecific products. Additional engineering modules aim to enhance function within the tumor microenvironment, including checkpoint pathway modulation (PD-1, CTLA-4), resistance to soluble suppression via adenosine (A2AR) and TGF-β (dnTGFβRII) axis interference, improved trafficking through chemokine receptor retargeting (e.g., CXCR1/2/3), and improved persistence through cytokine support circuits such as membrane-bound IL-15 and IL-15Rα

Central to TCR-T therapy is the engineered receptor itself. Current clinical programs mainly target cancer-testis antigens (NY-ESO-1, MAGE-A3/A4, PRAME), lineage-restricted differentiation antigens (MART-1, gp100), and self-proteins that are expressed at low levels in some normal tissues but upregulated in multiple tumors, such as GD2 and CSPG4 [ 180 ]. Compared with CARs, TCRs naturally recognize peptide-HLA complexes at very low antigen density, but affinity enhancement by mutagenesis can improve tumor recognition at the cost of increased cross-reactivity. The MAGE-A3/titin experience showed that even modest affinity enhancement can cause severe organ toxicity, underscoring the narrow therapeutic window of some targets [ 192 , 193 ]. Current strategies therefore balance affinity optimization with extensive in vitro specificity testing and, where possible, structure-guided design to improve discrimination between target and off-target peptide-HLA complexes [ 189 , 193 ].

Unlike CAR T cells, TCR-T products retain the native TCR repertoire in addition to the introduced tumor-specific TCR, creating a risk of mispairing between endogenous and transgenic α and β chains. This can reduce expression of the intended receptor and generate mixed TCRs with unpredictable specificities. One mitigation strategy is pairing optimization of the transgenic TCR to favor correct α/β assembly. In the PRAME-directed product IMA203, two charge-complementary amino acid substitutions at the α/β interface were introduced to promote preferential pairing and improve receptor stability and binding properties [ 189 ]. More stringent approaches combine TCR insertion with silencing or deletion of endogenous TCRα/β chains using shRNA, zinc-finger nucleases, or CRISPR-Cas9 to generate monospecific T cells expressing only the desired transgenic TCR [ 195 , 196 ].

Positioning of engineered TCR-Ts within melanoma cell therapy

Within melanoma cell therapy, engineered TCR-T cells occupy an intermediate position between CAR T cells and TIL therapy. Like CAR T cells, they are generated from peripheral blood and allow modular genetic engineering [ 197 ]. Like TILs, they are HLA-restricted and rely on physiological TCR signaling, but they can target intracellular antigens inaccessible to CARs [ 180 ]. In melanoma, TCR-T has shown deep and sometimes durable responses, particularly against cancer-testis antigens such as NY-ESO-1 [ 198 ] and PRAME [ 189 ], while also illustrating that antigen selection and receptor design are central to safety.

As PRAME-targeted TCR-T programs progress into phase III trials [ 191 ] and multiplex-edited, armored, and safety-switch-equipped products emerge, TCR-T is likely to become an important complement to DC vaccines, CAR T cells, and TIL therapy in advanced melanoma. Its ultimate role will depend on whether ongoing trials translate response rates into durable survival benefit and reliably control off-target risks.

Natural killer cell-based therapy in melanoma

Natural killer (NK) cells are innate lymphocytes that detect and kill transformed or infected cells through a balance of activating and inhibitory receptors without prior antigen sensitization, enabling recognition of tumor cells with altered or absent HLA class I expression [ 199 ]. Their antitumoral activity is mediated through direct cytotoxicity, engagement of death receptors such as Fas and TNF-related apoptosis-inducing ligand (TRAIL), antibody-dependent cellular cytotoxicity via CD16, and secretion of IFN-γ and TNF that support DC activation and T-cell priming [ 200 , 201 ].

In melanoma, NK cells recognize tumor targets through activating receptors such as natural killer group 2, member D (NKG2D), DNAX accessory molecule-1 (DNAM-1), and the natural cytotoxicity receptors NKp30, NKp44, and NKp46, which interact with induced self-ligands including HLA class I chain-related protein A (MICA), HLA class I chain-related protein B (MICB), UL16-binding protein (ULBP) family members, and the DNAM-1 ligands poliovirus receptor (PVR, also known as CD155) and Nectin-2 [ 202 ]. Human NK cells comprise cytotoxic CD56 dim CD16 + cells and more immunoregulatory CD56 bright CD16 −/low cells, with additional tissue-resident populations in skin and tumors [ 203 ]. In advanced melanoma, higher frequencies of CD56 bright NK cells are associated with reduced degranulation and shorter OS, whereas predominance of mature CD56 dim NK cells and high intratumoral NK cell density or NK gene signatures correlate with favorable prognosis and better responses to ICI [ 204 , 205 ].

Melanoma evades NK surveillance through upregulation of HLA class I and inhibitory ligands, expression of HLA-E and HLA-G, and shedding or downregulation of NKG2D ligands, thereby reducing activating input and restoring dominant inhibition [ 206 ]. In addition, tumor-derived mediators such as TGFβ, PGE2, and indoleamine 2,3-dioxygenase (IDO) suppress NK activating receptors and promote poorly cytotoxic or regulatory NK subsets [ 206 , 207 ]. The TIME further impairs NK function through hypoxia [ 208 ], nutrient deprivation [ 209 ], acidosis [ 210 ] and accumulation of MDSCs and T regs [ 211 ], driving an exhausted phenotype with reduced cytotoxicity and cytokine production. Preclinical melanoma studies indicate that intact NK activity supports the efficacy of targeted therapy [ 212 ] and ICI [ 213 , 214 ], whereas NK-cell loss or dysfunction promotes resistance, metastasis, and immune escape [ 214 , 215 ], , providing a strong rationale for NK cell-based therapy in melanoma [ 200 , 214 ].

Conventional and memory-like adoptive NK cell transfer

Adoptive NK cell therapy aims to restore or augment tumor-reactive NK cell function through ex vivo-expanded and activated autologous or allogeneic NK cell products [ 199 ]. Early trials with autologous NK cells in metastatic melanoma demonstrated feasibility and safety but only minimal clinical activity, likely because of dominant inhibition by self HLA ligands and limited in vivo persistence [ 199 , 216 ]. This led to development of allogeneic NK strategies exploiting killer-cell immunoglobulin-like receptor (KIR) and HLA mismatch to reduce inhibitory signaling and enhance antitumor activity, an approach with proven benefit in acute myeloid leukemia that is now being translated to solid tumors including melanoma [ 216 – 219 ].

Allogeneic NK manufacture usually starts from leukapheresis products of healthy donors or umbilical cord blood (CB), followed by cytokine-based expansion with IL-2 or IL-15 and feeder-cell-based systems or artificial APCs to support large-scale expansion and a cytotoxic phenotype [ 220 ]. Process optimization has generated clinically relevant NK doses with preserved activating receptor expression, potent melanoma cell killing in vitro, and the option of cryopreservation for off-the-shelf use [ 221 , 222 ]. In a phase I trial of haploidentical NK transfer in cancer patients including metastatic melanoma patients, treatment was feasible and generally well tolerated, but clinical activity remained limited: 4 of 10 melanoma patients achieved only transient stable disease and all subsequently progressed [ 218 ].

Cytokine-induced memory-like (CIML) NK cells represent a key refinement. Brief ex vivo stimulation with IL-12, IL-15, and IL-18 induces a memory-like state with enhanced IFN-γ production, cytotoxicity, and persistence upon restimulation [ 223 , 224 ]. CIML NK cells can be generated from healthy donors and melanoma patients, retain cytotoxicity against autologous and allogeneic melanoma targets, and outperform conventional IL-2-activated NK cells in preclinical models [ 225 ]. Building on preclinical data demonstrating that cytokine-induced memory-like NK cells from healthy donors and melanoma patients efficiently control autologous and allogeneic melanoma targets in vitro and in vivo [ 225 ], early phase I trials of allogeneic CIML NK cells in other heavily pretreated solid tumors have shown acceptable safety, transient disease control, and occasional regressions [ 226 ]. However, robust melanoma-specific efficacy data are still lacking, and a dedicated phase I trial in advanced melanoma is ongoing ( NCT05629546 ).

Engineered NK cells and CAR NK strategies in melanoma

Genetic engineering can further enhance NK-cell recognition and persistence by introducing CARs, cytokine support, or resistance to inhibitory pathways [ 227 ] (Fig. 5 ). Compared with CAR T cells, CAR NK products can be generated from allogeneic donors with low risk of Graft-versus-host disease, and early-phase trials across tumor types have shown favorable safety, with low rates of severe CRS or neurotoxicity and minimal long-term toxicity [ 228 , 229 ]. Preclinical studies show that CAR NK cells targeting GD2 [ 230 ] or B7-H3 [ 231 ] mediate potent killing of melanoma cell lines and xenografts while retaining lower alloreactive risk than CAR T cells, supporting their development as off-the-shelf products.

Fig. 5 Advances in adoptive NK cell immunotherapy and engineered NK platforms for melanoma. Adoptive NK cell therapy uses ex vivo expanded and activated NK cells and is increasingly pursued with allogeneic products to mitigate self HLA mediated inhibition and limited persistence observed with early autologous approaches. Cytokine-induced memory-like (CIML) NK cells are generated by brief priming with IL-12, IL-15, and IL-18, imprinting enhanced IFN-γ production, increased cytotoxicity, and improved persistence upon restimulation. Genetic engineering further extends NK functionality through CAR expression and modular targeting formats, with the advantage of low graft-versus-host disease risk and generally favorable early safety signals across tumor types. In melanoma-relevant designs, tri-specific NK cell engagers link a tumor-associated antigen to CD16 to trigger NK activation while simultaneously delivering IL-15 as an intrinsic support signal for survival and expansion, and, in parallel, the UniCAR NK-92 platform uses a switchable adaptor module to redirect a universal CAR to different tumor antigens, decoupling the effector cell product from antigen specificity and adding a pharmacologically controllable safety layer. Additional engineering modules improve persistence through membrane-bound or secreted IL-15 and IL-15Rα support, enhance recruitment into inflamed lesions via chemokine receptor retargeting such as CXCR1, CXCR2, or CXCR3, and strengthen resistance to tumor microenvironment suppression by disrupting TGF-β signaling or targeting adenosine A2A receptor pathways. Combination strategies further aim to integrate NK therapies with checkpoint inhibition, targeted therapy, chemotherapy, or radiation to counteract tumor-induced suppression and exploit synergistic effects, although melanoma-specific clinical outcome data remain limited

NK-specific engineering strategies include membrane-bound or secreted IL-15 to support survival [ 232 ], expression of chemokine receptors such as CXCR1, CXCR2, or CXCR3 to improve trafficking to inflamed melanoma lesions [ 233 – 235 ] and disruption of TGF-β signaling [ 236 , 237 ] or targeting of A2ARs to overcome microenvironmental suppression [ 238 ]. Early clinical trials of engineered NK cells outside melanoma already provide encouraging safety signals, including CD19 CAR NK cells in hematologic malignancies [ 239 ] and trispecific NK-cell engagers combining tumor targeting, CD16 engagement, and IL-15 delivery, which have shown objective responses with limited severe toxicity [ 240 , 241 ]. These findings support cautious optimism for translation to melanoma.

Melanoma-specific CAR NK programs remain early but are progressing. Preclinical and translational studies describe CAR NK constructs against GD2, CSPG4, MCSP, and other melanoma-associated antigens that achieve robust tumor control in xenograft models and can be produced from CB or induced pluripotent stem cell-derived NK cells at scale [ 242 , 243 ]. A universal CAR NK concept has also been developed with the UniCAR NK-92 platform, in which NK-92 cells expressing a universal CAR are redirected to different antigens via soluble target modules, adding flexibility and pharmacologic safety control [ 230 ]. In parallel, supercharged NK cells generated through combined cytokine and co-stimulatory receptor signaling have shown strong melanoma control in humanized mice, suggesting an additional route to enhance NK potency with or without CARs [ 244 , 245 ].

Combinatorial approaches

Combination strategies integrate NK cell-based therapies with immune checkpoint inhibition, targeted therapy, or radiation to counteract tumor-induced suppression and exploit synergy [ 246 , 247 ]. Preclinical studies show that PD-1 and CTLA-4 blockade can enhance NK-cell activity by reshaping the TIME, and that tumor-infiltrating NK cells may themselves express checkpoint receptors such as PD-1 and T cell immunoreceptor with Ig and ITIM domains (TIGIT), providing a rationale for direct checkpoint blockade on NK cells [ 248 ]. In BRAF V600E melanoma models, NK cells are required for sustained tumor control during BRAF inhibition, and loss of NK function accelerates relapse and metastatic outgrowth [ 212 ]. Clinically, early-phase trials are evaluating ex vivo-expanded or engineered NK-cell products with anti-PD-1 antibodies [ 249 ] or chemotherapy [ 250 ] in advanced solid tumors. In biliary tract cancer, allogeneic SMT-NK cells plus pembrolizumab achieved an overall response rate of 17.4% in the full-analysis set and 50.0% in the per-protocol population without severe combination-related AEs [ 249 ]. In locally advanced colon carcinoma, adjuvant NK-cell infusions added to chemotherapy improved 5-year PFS and OS without new safety signals [ 250 ]. Melanoma-specific clinical outcome data, however, remain limited.

Current limitations and future directions

Despite conceptual advantages, including HLA-independent recognition, off-the-shelf applicability, and a more favorable toxicity profile than many T-cell products [ 251 – 253 ], NK cell-based therapies for melanoma remain at an exploratory stage [ 216 , 254 ]. Major obstacles include limited in vivo persistence and expansion, insufficient trafficking into metastatic sites, strong suppression by TGF-β, adenosine, and other microenvironmental factors, and the need to identify melanoma antigens with sufficient selectivity for engineered NK targeting [ 255 , 256 ]. Systematic reviews in solid tumors conclude that current evidence, derived mainly from early-phase small trials, supports favorable safety and signs of clinical and biological activity, but also underscores the need for more potent and standardized NK platforms, rational combination strategies, and larger controlled studies to define their therapeutic potential [ 253 , 255 , 257 ].

Conclusion

Despite major advances with ICI and targeted therapy, therapy-resistant melanoma remains a major clinical and socioeconomic challenge, as primary and acquired resistance to PD-1- and CTLA-4-based regimens occurs in a substantial proportion of patients [ 21 , 26 ]. Many patients never respond, relapse after an initial response, or cannot tolerate intensified checkpoint-based regimens and therefore receive multiple lines of systemic treatment with cumulative toxicity but limited durable benefit [ 20 – 22 , 25 ]. These patients require strategies that not only modulate pre-existing immunity but rebuild and sustain a competent tumor-reactive immune compartment [ 31 ].

Resistance to ICI in melanoma is not explained by T-cell hypofunction alone, but reflects multiple T-cell-intrinsic, microenvironmental, and tumor-intrinsic mechanisms within the TIME. Chronic antigen exposure, metabolic stress, and inhibitory cytokine signaling promote exhausted or pre-exhausted T-cell states with reduced proliferative capacity and impaired cytokine production [ 24 , 28 ]. Beyond this, resistance may result from insufficient numbers or quality of tumor-reactive lymphocytes, impaired antigen presentation, myeloid- and T reg -mediated immunosuppression, and tumor-intrinsic resistance programs [ 21 , 24 , 26 , 28 ].

Cellular immunotherapies directly address this problem by supplying tumor-reactive lymphocytes or by priming them de novo, while also offering opportunities to remodel the TIME and re-establish immune memory [ 30 , 31 ]. DC vaccines exemplify this approach by introducing new T-cell specificities through shared antigens, neoantigens, or whole tumor lysates and priming polyfunctional CD4⁺ and CD8⁺ T cells [ 33 , 45 ]. Their major strengths are excellent safety, outpatient feasibility, and the potential to induce broad and durable immunity, including immune memory [ 33 , 36 , 37 , 258 ]. However, objective response rates as monotherapy in advanced melanoma remain modest, so DC vaccines are increasingly viewed as priming or backbone components of combination regimens rather than stand-alone treatments in late-line disease [ 33 , 35 , 69 , 70 ].

TIL therapy directly replaces the missing tumor-reactive repertoire. TIL products are enriched for clonotypes that have already infiltrated the tumor and can target a broad antigen spectrum, including patient-specific neoantigens, making them less vulnerable to single-target escape than engineered products [ 74 , 76 , 259 ]. Clinically, TIL therapy achieves response rates that exceed those of most salvage therapies in checkpoint-refractory melanoma, with lifileucel and related products showing objective response rates of about 30–40% and durable complete responses in a meaningful subset of patients [ 82 , 83 ]. However, TIL therapy requires tumor resection, complex manufacturing, and lymphodepleting conditioning with high-dose IL-2, and is therefore best delivered in specialized centers to patients able to tolerate an intensive but time-limited regimen [ 74 , 78 , 86 , 259 ].

Genetically engineered TCR-T cells and CAR T/CAR NK cells offer complementary advantages and limitations. TCR-T cells can target intracellular cancer-testis antigens and neoantigens presented by defined HLA molecules and, when manufactured in a stem-like state, may form long-lived memory populations [ 87 , 180 , 182 , 184 ]. Their limitations are HLA restriction, narrow antigen specificity, and safety risks related to off-target or cross-reactive peptide recognition [ 180 , 182 , 192 , 193 ]. CAR-based strategies overcome HLA restriction and usually do not require autologous tumor material for target discovery, enabling off-the-shelf development and rapid retargeting of both T and NK cells [ 112 , 242 , 243 , 252 ]. In melanoma and other solid tumors, however, efficacy is constrained by the scarcity of truly tumor-selective surface antigens, antigen heterogeneity and plasticity, and acute toxicities for CAR Ts such as cytokine release syndrome and neurotoxicity [ 119 , 155 ]. Many conventional CAR T products also acquire terminal effector differentiation and exhaustion, limiting persistence and memory formation in the TIME [ 145 , 184 ].

NK-cell-based immunotherapy remains exploratory in melanoma. Its potential advantages include HLA-independent recognition, off-the-shelf manufacturability, and a comparatively favorable toxicity profile, but these are counterbalanced by limited in vivo persistence and expansion, incomplete trafficking into metastatic sites, strong suppression by TGF-β, adenosine, and other microenvironmental factors, and the need for sufficiently selective melanoma targets for engineered NK approaches [ 206 , 214 , 216 , 229 , 242 , 255 ]. Current early-phase studies mainly support excellent safety and encouraging biological activity, but more potent and standardized NK platforms, optimized combinations, and larger controlled trials are needed before NK-cell therapies can claim a defined position alongside TIL, TCR-T, and CAR-based approaches in advanced melanoma [ 216 , 242 , 253 ].

Taken together, no single cellular therapy fully resolves the multifactorial problem of therapy-resistant melanoma. DC vaccines excel at safe, antigen-diverse priming and memory induction but depend on a minimally permissive TIME [ 33 , 35 , 258 ]. TIL therapy provides a broad tumor-experienced repertoire with preserved homing and broad target recognition, but at the cost of individualized and intensive treatment [ 74 , 83 , 259 ]. TCR-T and CAR T/CAR NK products offer potent and programmable effector cells, yet remain limited by antigen specificity, toxicity risks, and incomplete persistence [ 112 , 155 , 180 , 182 , 229 , 242 ]. NK-cell-based strategies may complement these platforms by targeting antigen-loss and HLA-deficient clones and by cooperating with DCs and T cells, but they still require further optimization for persistence, trafficking, and resistance to the suppressive TME [ 206 , 251 – 253 ] (Fig. 5 ).

The most plausible future direction is rational combination and sequencing of cellular platforms with checkpoint modulation and TIME-directed strategies. DC vaccines may be used to prime de novo tumor-specific T- and NK-cell responses before or after ATT, thereby enriching the effector pool that can then be expanded or sustained by TIL, TCR-T, or CAR-based approaches [ 33 , 35 , 71 , 260 ]. In heavily pretreated, checkpoint-refractory melanoma, strategies such as TIL plus PD-1 blockade or DC vaccination followed by TIL infusion and delayed ICI are particularly attractive and warrant further clinical evaluation [ 72 , 103 , 259 ]. The therapeutic options are summarized in Table 3 .

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Acknowledgements

The authors thank their team for support and input.

Authors’ contributions

Conceptualization, A.W. and A.D. writing—original draft preparation, A.W.; writing—review and editing, A.W. and A.D.; supervision, A.D.; funding acquisition. All authors have read and agreed to the published version of the manuscript.

Funding

Open Access funding enabled and organized by Projekt DEAL. This research was supported by the intramural Research Foundation FOKO2025 via the networking research grant (ID 2025-18; A.D.).

Data availability

No datasets were generated or analysed during the current study.

Declarations

Ethics approval and consent to participate

not applicable.

Consent for publication

not applicable.

Competing interests

The authors declare no competing interests.

Abbreviations

12-peptide vaccine plus melanoma-specific helper peptides

CD137

Adenosine A2A Receptor

Adverse Events

Adoptive T-cell transfer

B7 Homolog 3

B-Raf proto-oncogene

Chimeric Antigen Receptor

Cord Blood

Chemokine Receptor

Chemical Inducer of Dimerization

Cytokine-induced memory-like

Cytokine-inducible SH2-containing protein

CISH knockout

Central Nervous System

Complete Responses

Cytokine Release Syndrome

Chondroitin Sulfate Proteoglycan 4

Cytotoxic T cells

Cytotoxic T-Lymphocyte-Associated Protein 4

C-X-C Chemokine Receptor

Dendritic-cell-based Vaccination

DNAX Accessory Molecule-1

Delayed-type Hypersensitivity

Enzyme-Linked Immunospot

Fluorescence-Activated Cell Sorting

Food and Drug Administration

Disialoganglioside

Granulocyte-Macrophage Colony-Stimulating Factor

Good Manufacturing Practice

Human epidermal growth factor receptor 2

Human Leukocyte Antigen

Immune-effector Cell-Associated Neurotoxicity

Immune Checkpoint Inhibition

3-Idolamine-Dioxygenase

Interferon-γ

Interleukin-4

Interleukin-13 receptor alpha 2

Janus kinase-signal transducer and activator of transcription

Killer-Cell Immunoglobulin-like Receptor

Lymphocyte-Activation Gene 3

Magnetic-Activated Cell Sorting

Melanoma Antigen Gene

Mitogen-Activated Protein Kinase

Melanoma Antigen Recognized by T cells 1

Mitogen-activated protein kinase

induced self-ligands including HLA class I chain-related protein A

HLA class I chain-related protein B

Magnetic Resonance Imaging

Natural Killer

Neuroblastoma RAS viral oncogene homolog

New York Esophageal Squamous Cell Carcinoma 1

Objective Response Rates

Forkhead box P3

Programmed Cell Death Protein 1

Programmed Death Ligand 1

Progression-Free Survival

Prostaglandin E2

Partial Responses

Preferentially Expressed Antigen in Melanoma

T-Cell Receptor

T-Cell Receptor-engineered T cells

T helper type 1

T cell immunoreceptor with Ig and ITIM domains

Tumor-Infiltrating Lymphocyte

Tumor Microenvironment

Toll-Like Receptor

Tumor Necrosis Factor α

TNF-Related Apoptosis-Inducing Ligand

Tyrosinase-Related Rrotein 1

UL16-Binding Protein

Ultraviolet

References

1. Ferlay J, Ervik M, Lam F, Laversanne M, Colombet M, Mery L, Piñeros M, Znaor A, Soerjomataram I, Bray F. Global Cancer Observatory: Cancer Today.; 2024 [cited 2025 Dec 27]. Available from: URL: https://gco.iarc.who.int/today .

2. Wang M Gao X Zhang L Recent global patterns in skin cancer incidence, mortality, and prevalence Chin Med J (Engl) 2025 138 2 185 92 10.1097/CM9.0000000000003416 39682020 PMC11745855 Wang M, Gao X, Zhang L. Recent global patterns in skin cancer incidence, mortality, and prevalence. Chin Med J (Engl). 2025;138(2):185–92. 10.1097/CM9.0000000000003416. 39682020 10.1097/CM9.0000000000003416 PMC11745855

3. Saginala K, Barsouk A, Aluru JS, Rawla P, Barsouk A. Epidemiol Melanoma Med Sci (Basel) 2021;9(4):63. 10.3390/medsci904006.

4. Mihajlovic M Vlajkovic S Jovanovic P Stefanovic V Primary mucosal melanomas: a comprehensive review Int J Clin Exp Pathol 2012 5 8 739 53 23071856 PMC3466987 Mihajlovic M, Vlajkovic S, Jovanovic P, Stefanovic V. Primary mucosal melanomas: a comprehensive review. Int J Clin Exp Pathol. 2012;5(8):739–53. 23071856 PMC3466987

5. Akbani R Akdemir KC Aksoy BA Albert M Ally A Amin SB Genomic Classif Cutan Melanoma Cell 2015 161 7 1681 96 10.1016/j.cell.2015.05.044 PMC4580370 26091043 Akbani R, Akdemir KC, Aksoy BA, Albert M, Ally A, Amin SB, et al. Genomic Classif Cutan Melanoma Cell. 2015;161(7):1681–96. 10.1016/j.cell.2015.05.044. 10.1016/j.cell.2015.05.044 PMC4580370 26091043

6. Langselius O Rumgay H de Vries E Whiteman DC Jemal A Parkin DM Global burden of cutaneous melanoma incidence attributable to ultraviolet radiation in 2022 Int J Cancer 2025 157 6 1110 9 10.1002/ijc.35463 40421619 Langselius O, Rumgay H, de Vries E, Whiteman DC, Jemal A, Parkin DM, et al. Global burden of cutaneous melanoma incidence attributable to ultraviolet radiation in 2022. Int J Cancer. 2025;157(6):1110–9. 10.1002/ijc.35463. 40421619 10.1002/ijc.35463

7. Robertson AG Shih J Yau C Gibb EA Oba J Mungall KL Integrative Analysis Identifies Four Molecular and Clinical Subsets in Uveal Melanoma Cancer Cell 2017 32 2 204 e22015 10.1016/j.ccell.2017.07.003 28810145 PMC5619925 Robertson AG, Shih J, Yau C, Gibb EA, Oba J, Mungall KL, et al. Integrative Analysis Identifies Four Molecular and Clinical Subsets in Uveal Melanoma. Cancer Cell. 2017;32(2):204–e22015. 10.1016/j.ccell.2017.07.003. 28810145 10.1016/j.ccell.2017.07.003 PMC5619925

8. Newell F Kong Y Wilmott JS Johansson PA Ferguson PM Cui C Whole-genome landscape of mucosal melanoma reveals diverse drivers and therapeutic targets Nat Commun 2019 10 1 3163 10.1038/s41467-019-11107-x 31320640 PMC6639323 Newell F, Kong Y, Wilmott JS, Johansson PA, Ferguson PM, Cui C, et al. Whole-genome landscape of mucosal melanoma reveals diverse drivers and therapeutic targets. Nat Commun. 2019;10(1):3163. 10.1038/s41467-019-11107-x. 31320640 10.1038/s41467-019-11107-x PMC6639323

9. Keung EZ Gershenwald JE The eighth edition American Joint Committee on Cancer (AJCC) melanoma staging system: implications for melanoma treatment and care Expert Rev Anticancer Ther 2018 18 8 775 84 10.1080/14737140.2018.1489246 29923435 PMC7652033 Keung EZ, Gershenwald JE. The eighth edition American Joint Committee on Cancer (AJCC) melanoma staging system: implications for melanoma treatment and care. Expert Rev Anticancer Ther. 2018;18(8):775–84. 10.1080/14737140.2018.1489246. 29923435 10.1080/14737140.2018.1489246 PMC7652033

10. Amaral T, Ottaviano M, Arance A, Blank C, Chiarion-Sileni V, Donia M et al. Cutaneous melanoma: ESMO Clinical Practice Guideline for diagnosis, treatment and follow-up. Annals of oncology: official journal of the European Society for Medical Oncology. 2025; 36(1):10–30. Available from: URL: https://pubmed.ncbi.nlm.nih.gov/39550033/ . 10.1016/j.annonc.2024.11.006 PMC7618628 39550033

11. Qin S Xu L Yi M Yu S Wu K Luo S Novel immune checkpoint targets: moving beyond PD-1 and CTLA-4 Mol Cancer 2019 18 1 155 10.1186/s12943-019-1091-2 31690319 PMC6833286 Qin S, Xu L, Yi M, Yu S, Wu K, Luo S. Novel immune checkpoint targets: moving beyond PD-1 and CTLA-4. Mol Cancer. 2019;18(1):155. 10.1186/s12943-019-1091-2. 31690319 10.1186/s12943-019-1091-2 PMC6833286

12. Long GV Carlino MS McNeil C Ribas A Gaudy-Marqueste C Schachter J Pembrolizumab versus ipilimumab for advanced melanoma: 10-year follow-up of the phase III KEYNOTE-006 study Annals oncology: official J Eur Soc Med Oncol 2024 35 12 1191 9 10.1016/j.annonc.2024.08.2330 39306585 Long GV, Carlino MS, McNeil C, Ribas A, Gaudy-Marqueste C, Schachter J, et al. Pembrolizumab versus ipilimumab for advanced melanoma: 10-year follow-up of the phase III KEYNOTE-006 study. Annals oncology: official J Eur Soc Med Oncol. 2024;35(12):1191–9. 10.1016/j.annonc.2024.08.2330. 10.1016/j.annonc.2024.08.2330 39306585

13. Ribas A Wolchok JD Cancer immunotherapy using checkpoint blockade Science 2018 359 6382 1350 5 10.1126/science.aar4060 29567705 PMC7391259 Ribas A, Wolchok JD. Cancer immunotherapy using checkpoint blockade. Science. 2018;359(6382):1350–5. 10.1126/science.aar4060. 29567705 10.1126/science.aar4060 PMC7391259

14. Wolchok JD Chiarion-Sileni V Rutkowski P Cowey CL Schadendorf D Wagstaff J Final, 10-Year Outcomes with Nivolumab plus Ipilimumab in Advanced Melanoma N Engl J Med 2025 392 1 11 22 10.1056/NEJMoa2407417 39282897 PMC12080919 Wolchok JD, Chiarion-Sileni V, Rutkowski P, Cowey CL, Schadendorf D, Wagstaff J, et al. Final, 10-Year Outcomes with Nivolumab plus Ipilimumab in Advanced Melanoma. N Engl J Med. 2025;392(1):11–22. 10.1056/NEJMoa2407417. 39282897 10.1056/NEJMoa2407417 PMC12080919

15. Tawbi HA Hodi FS Lipson EJ Schadendorf D Ascierto PA Matamala L Three-Year Overall Survival With Nivolumab Plus Relatlimab in Advanced Melanoma From RELATIVITY-047 J Clin Oncol 2025 43 13 1546 52 10.1200/JCO.24.01124 39671533 PMC12054981 Tawbi HA, Hodi FS, Lipson EJ, Schadendorf D, Ascierto PA, Matamala L, et al. Three-Year Overall Survival With Nivolumab Plus Relatlimab in Advanced Melanoma From RELATIVITY-047. J Clin Oncol. 2025;43(13):1546–52. 10.1200/JCO.24.01124. 39671533 10.1200/JCO.24.01124 PMC12054981

16. Robert C Grob JJ Stroyakovskiy D Karaszewska B Hauschild A Levchenko E Five-Year Outcomes with Dabrafenib plus Trametinib in Metastatic Melanoma N Engl J Med 2019 381 7 626 36 10.1056/NEJMoa1904059 31166680 Robert C, Grob JJ, Stroyakovskiy D, Karaszewska B, Hauschild A, Levchenko E, et al. Five-Year Outcomes with Dabrafenib plus Trametinib in Metastatic Melanoma. N Engl J Med. 2019;381(7):626–36. 10.1056/NEJMoa1904059. 31166680 10.1056/NEJMoa1904059

17. Dummer R Flaherty KT Robert C Arance A de Groot JWB Garbe C COLUMBUS 5-Year Update: A Randomized, Open-Label, Phase III Trial of Encorafenib Plus Binimetinib Versus Vemurafenib or Encorafenib in Patients With BRAF V600-Mutant Melanoma J Clin Oncol 2022 40 36 4178 88 10.1200/JCO.21.02659 35862871 PMC9916040 Dummer R, Flaherty KT, Robert C, Arance A, de Groot JWB, Garbe C, et al. COLUMBUS 5-Year Update: A Randomized, Open-Label, Phase III Trial of Encorafenib Plus Binimetinib Versus Vemurafenib or Encorafenib in Patients With BRAF V600-Mutant Melanoma. J Clin Oncol. 2022;40(36):4178–88. 10.1200/JCO.23.00385. 35862871 10.1200/JCO.21.02659 PMC9916040

18. Garbe C Amaral T Peris K Hauschild A Arenberger P Basset-Seguin N European consensus-based interdisciplinary guideline for melanoma. Part 2: Treatment - Update 2024 Eur J Cancer 2025 215 115153 10.1016/j.ejca.2024.115153 39709737 Garbe C, Amaral T, Peris K, Hauschild A, Arenberger P, Basset-Seguin N, et al. European consensus-based interdisciplinary guideline for melanoma. Part 2: Treatment - Update 2024. Eur J Cancer. 2025;215:115153. 10.1016/j.ejca.2024.115153. 39709737 10.1016/j.ejca.2024.115153

19. Atkins MB Lee SJ Chmielowski B Tarhini AA Cohen GI Truong T-G Combination Dabrafenib and Trametinib Versus Combination Nivolumab and Ipilimumab for Patients With Advanced BRAF-Mutant Melanoma: The DREAMseq Trial-ECOG-ACRIN EA6134 J Clin Oncol 2023 41 2 186 97 10.1200/JCO.22.01763 36166727 PMC9839305 Atkins MB, Lee SJ, Chmielowski B, Tarhini AA, Cohen GI, Truong T-G, et al. Combination Dabrafenib and Trametinib Versus Combination Nivolumab and Ipilimumab for Patients With Advanced BRAF-Mutant Melanoma: The DREAMseq Trial-ECOG-ACRIN EA6134. J Clin Oncol. 2023;41(2):186–97. 10.1200/JCO.22.01763. 36166727 10.1200/JCO.22.01763 PMC9839305

20. Amaral T, Seeber O, Mersi E, Sanchez S, Thomas I, Meiwes A et al. Primary Resistance to PD-1-Based Immunotherapy-A Study in 319 Patients with Stage IV Melanoma. Cancers (Basel). 2020;12(4):1027. 10.3390/cancers12041027. 10.3390/cancers12041027 PMC7226601 32331243

21. Lim SY Shklovskaya E Lee JH Pedersen B Stewart A Ming Z The molecular and functional landscape of resistance to immune checkpoint blockade in melanoma Nat Commun 2023 14 1 1516 10.1038/s41467-023-36979-y 36934113 PMC10024679 Lim SY, Shklovskaya E, Lee JH, Pedersen B, Stewart A, Ming Z, et al. The molecular and functional landscape of resistance to immune checkpoint blockade in melanoma. Nat Commun. 2023;14(1):1516. 10.1038/s41467-023-36979-y. 36934113 10.1038/s41467-023-36979-y PMC10024679

22. Wolchok JD Chiarion-Sileni V Gonzalez R Rutkowski P Grob J-J Cowey CL Overall Survival with Combined Nivolumab and Ipilimumab in Advanced Melanoma N Engl J Med 2017 377 14 1345 56 10.1056/NEJMoa1709684 28889792 PMC5706778 Wolchok JD, Chiarion-Sileni V, Gonzalez R, Rutkowski P, Grob J-J, Cowey CL, et al. Overall Survival with Combined Nivolumab and Ipilimumab in Advanced Melanoma. N Engl J Med. 2017;377(14):1345–56. 10.1056/NEJMoa1709684. 28889792 10.1056/NEJMoa1709684 PMC5706778

23. Fateeva A, Eddy K, Chen S. Current State of Melanoma Therapy and Next Steps: Battling Therapeutic Resistance. Cancers (Basel). 2024;16(8):1571. 10.3390/cancers16081571. 10.3390/cancers16081571 PMC11049326 38672652

24. Datsi A Sorg RV Garg AD The conundrum of CD8 + T cell trajectories in low antigenic tumors: How to overcome a hypofunctional state distinct from antigen-driven exhaustion? Genes Immun 2024 25 5 353 5 10.1038/s41435-024-00299-y 39384954 Datsi A, Sorg RV, Garg AD. The conundrum of CD8 + T cell trajectories in low antigenic tumors: How to overcome a hypofunctional state distinct from antigen-driven exhaustion? Genes Immun. 2024;25(5):353–5. 10.1038/s41435-024-00299-y. 39384954 10.1038/s41435-024-00299-y

25. Gide TN Wilmott JS Scolyer RA Long GV Primary and Acquired Resistance to Immune Checkpoint Inhibitors in Metastatic Melanoma Clin Cancer Res 2018 24 6 1260 70 10.1158/1078-0432.CCR-17-2267 29127120 Gide TN, Wilmott JS, Scolyer RA, Long GV. Primary and Acquired Resistance to Immune Checkpoint Inhibitors in Metastatic Melanoma. Clin Cancer Res. 2018;24(6):1260–70. 10.1158/1078-0432.CCR-17-2267. 29127120 10.1158/1078-0432.CCR-17-2267

26. Alsaafeen BH Ali BR Elkord E Resistance mechanisms to immune checkpoint inhibitors: updated insights Mol Cancer 2025 24 1 20 10.1186/s12943-024-02212-7 39815294 PMC11734352 Alsaafeen BH, Ali BR, Elkord E. Resistance mechanisms to immune checkpoint inhibitors: updated insights. Mol Cancer. 2025;24(1):20. 10.1186/s12943-024-02212-7. 39815294 10.1186/s12943-024-02212-7 PMC11734352

27. Zielińska MK, Ciążyńska M, Sulejczak D, Rutkowski P, Czarnecka AM. Mechanisms of Resistance to Anti-PD-1 Immunotherapy in Melanoma and Strategies to Overcome It. Biomolecules. 2025;15(2):269. 10.3390/biom15020269. 10.3390/biom15020269 PMC11853485 40001572

28. Ziogas DC Theocharopoulos C Koutouratsas T Haanen J Gogas H Mechanisms of resistance to immune checkpoint inhibitors in melanoma: What we have to overcome? Cancer Treat Rev 2023 113 102499 10.1016/j.ctrv.2022.102499 36542945 Ziogas DC, Theocharopoulos C, Koutouratsas T, Haanen J, Gogas H. Mechanisms of resistance to immune checkpoint inhibitors in melanoma: What we have to overcome? Cancer Treat Rev. 2023;113:102499. 10.1016/j.ctrv.2022.102499. 36542945 10.1016/j.ctrv.2022.102499

29. Dummer R Ascierto PA Nathan P Robert C Schadendorf D Rationale for Immune Checkpoint Inhibitors Plus Targeted Therapy in Metastatic Melanoma: A Review JAMA Oncol 2020 6 12 1957 66 10.1001/jamaoncol.2020.4401 32970096 Dummer R, Ascierto PA, Nathan P, Robert C, Schadendorf D. Rationale for Immune Checkpoint Inhibitors Plus Targeted Therapy in Metastatic Melanoma: A Review. JAMA Oncol. 2020;6(12):1957–66. 10.1001/jamaoncol.2020.4401. 32970096 10.1001/jamaoncol.2020.4401

30. Qian J Liu Y Recent advances in adoptive cell therapy for cancer immunotherapy Front Immunol 2025 16 1665488 10.3389/fimmu.2025.1665488 41058692 PMC12497837 Qian J, Liu Y. Recent advances in adoptive cell therapy for cancer immunotherapy. Front Immunol. 2025;16:1665488. 10.3389/fimmu.2025.1665488. 41058692 10.3389/fimmu.2025.1665488 PMC12497837

31. Filin IY, Mayasin YP, Kharisova CB, Gorodilova AV, Kitaeva KV, Chulpanova DS, et al. Cell Immunotherapy against Melanoma: Clinical Trials Review. Int J Mol Sci. 2023;24(3):2413. 10.3390/ijms24032413. 10.3390/ijms24032413 PMC9916554 36768737

32. Nestle FO Alijagic S Gilliet M Sun Y Grabbe S Dummer R Vaccination of melanoma patients with peptide- or tumor lysate-pulsed dendritic cells Nat Med 1998 4 3 328 32 10.1038/nm0398-328 9500607 Nestle FO, Alijagic S, Gilliet M, Sun Y, Grabbe S, Dummer R, et al. Vaccination of melanoma patients with peptide- or tumor lysate-pulsed dendritic cells. Nat Med. 1998;4(3):328–32. 10.1038/nm0398-328. 9500607 10.1038/nm0398-328

33. Dillman RO Nistor GI Cornforth AN Dendritic cell vaccines for melanoma: past, present and future Melanoma Manag 2016 3 4 273 89 10.2217/mmt-2016-0014 30190899 PMC6094661 Dillman RO, Nistor GI, Cornforth AN. Dendritic cell vaccines for melanoma: past, present and future. Melanoma Manag. 2016;3(4):273–89. 10.2217/mmt-2016-0014. 30190899 10.2217/mmt-2016-0014 PMC6094661

34. Datsi A Falkowski L Sorg RV Generation and quality control of mature monocyte-derived dendritic cells for immunotherapy Methods Cell Biol 2024 183 1 31 38548408 10.1016/bs.mcb.2023.05.007 Datsi A, Falkowski L, Sorg RV. Generation and quality control of mature monocyte-derived dendritic cells for immunotherapy. Methods Cell Biol. 2024;183:1–31. 10.1016/bs.mcb.2023.05.007. 38548408 10.1016/bs.mcb.2023.05.007

35. Zhang S, Mo S, Huang W, Zhong D, Yang X, Xie S, et al. Dendritic cell vaccines: Current research progress, challenges, and opportunities. Genes Dis. 2025;13(4):101913. 10.1016/j.gendis.2025.101913. 10.1016/j.gendis.2025.101913 PMC13050070 41938709

36. Ribas A Camacho LH Lee SM Hersh EM Brown CK Richards JM Multicenter phase II study of matured dendritic cells pulsed with melanoma cell line lysates in patients with advanced melanoma J Transl Med 2010 8 89 10.1186/1479-5876-8-89 20875102 PMC2954849 Ribas A, Camacho LH, Lee SM, Hersh EM, Brown CK, Richards JM, et al. Multicenter phase II study of matured dendritic cells pulsed with melanoma cell line lysates in patients with advanced melanoma. J Transl Med. 2010;8:89. 10.1186/1479-5876-8-89. 20875102 10.1186/1479-5876-8-89 PMC2954849

37. Bulgarelli J Piccinini C Scarpi E Gentili G Renzi L Carloni S Adjuvant dendritic cell-based immunotherapy in melanoma: insights into immune cell dynamics and clinical evidence from a phase II trial J Transl Med 2025 23 1 455 10.1186/s12967-025-06403-8 40251644 PMC12007200 Bulgarelli J, Piccinini C, Scarpi E, Gentili G, Renzi L, Carloni S, et al. Adjuvant dendritic cell-based immunotherapy in melanoma: insights into immune cell dynamics and clinical evidence from a phase II trial. J Transl Med. 2025;23(1):455. 10.1186/s12967-025-06403-8. 40251644 10.1186/s12967-025-06403-8 PMC12007200

38. Akiyama Y Tanosaki R Inoue N Shimada M Hotate Y Yamamoto A Clinical response in Japanese metastatic melanoma patients treated with peptide cocktail-pulsed dendritic cells J Transl Med 2005 3 1 4 10.1186/1479-5876-3-4 15676080 PMC549033 Akiyama Y, Tanosaki R, Inoue N, Shimada M, Hotate Y, Yamamoto A, et al. Clinical response in Japanese metastatic melanoma patients treated with peptide cocktail-pulsed dendritic cells. J Transl Med. 2005;3(1):4. 10.1186/1479-5876-3-4. 15676080 10.1186/1479-5876-3-4 PMC549033

39. Hersey P Halliday GM Farrelly ML DeSilva C Lett M Menzies SW Phase I/II study of treatment with matured dendritic cells with or without low dose IL-2 in patients with disseminated melanoma Cancer Immunol Immunother 2008 57 7 1039 51 10.1007/s00262-007-0435-8 18157724 PMC11030839 Hersey P, Halliday GM, Farrelly ML, DeSilva C, Lett M, Menzies SW. Phase I/II study of treatment with matured dendritic cells with or without low dose IL-2 in patients with disseminated melanoma. Cancer Immunol Immunother. 2008;57(7):1039–51. 10.1007/s00262-007-0435-8. 18157724 10.1007/s00262-007-0435-8 PMC11030839

40. Schadendorf D Ugurel S Schuler-Thurner B Nestle FO Enk A Bröcker E-B Dacarbazine (DTIC) versus vaccination with autologous peptide-pulsed dendritic cells (DC) in first-line treatment of patients with metastatic melanoma: a randomized phase III trial of the DC study group of the DeCOG Annals oncology: official J Eur Soc Med Oncol 2006 17 4 563 70 10.1093/annonc/mdj138 16418308 Schadendorf D, Ugurel S, Schuler-Thurner B, Nestle FO, Enk A, Bröcker E-B, et al. Dacarbazine (DTIC) versus vaccination with autologous peptide-pulsed dendritic cells (DC) in first-line treatment of patients with metastatic melanoma: a randomized phase III trial of the DC study group of the DeCOG. Annals oncology: official J Eur Soc Med Oncol. 2006;17(4):563–70. 10.1093/annonc/mdj138. 10.1093/annonc/mdj138 16418308

41. Hargadon KM Strategies to Improve the Efficacy of Dendritic Cell-Based Immunotherapy for Melanoma Front Immunol 2017 8 1594 10.3389/fimmu.2017.01594 29209327 PMC5702020 Hargadon KM. Strategies to Improve the Efficacy of Dendritic Cell-Based Immunotherapy for Melanoma. Front Immunol. 2017;8:1594. 10.3389/fimmu.2017.01594. 29209327 10.3389/fimmu.2017.01594 PMC5702020

42. Rabu C Rangan L Florenceau L Fortun A Charpentier M Dupré E Cancer vaccines: designing artificial synthetic long peptides to improve presentation of class I and class II T cell epitopes by dendritic cells OncoImmunology 2019 8 4 e1560919 10.1080/2162402X.2018.1560919 30906653 PMC6422379 Rabu C, Rangan L, Florenceau L, Fortun A, Charpentier M, Dupré E, et al. Cancer vaccines: designing artificial synthetic long peptides to improve presentation of class I and class II T cell epitopes by dendritic cells. OncoImmunology. 2019;8(4):e1560919. 10.1080/2162402X.2018.1560919. 30906653 10.1080/2162402X.2018.1560919 PMC6422379

43. Chen X Yang J Wang L Liu B Personalized neoantigen vaccination with synthetic long peptides: recent advances and future perspectives Theranostics 2020 10 13 6011 23 10.7150/thno.38742 32483434 PMC7255011 Chen X, Yang J, Wang L, Liu B. Personalized neoantigen vaccination with synthetic long peptides: recent advances and future perspectives. Theranostics. 2020;10(13):6011–23. 10.7150/thno.38742. 32483434 10.7150/thno.38742 PMC7255011

44. Rosalia RA Quakkelaar ED Redeker A Khan S Camps M Drijfhout JW Dendritic cells process synthetic long peptides better than whole protein, improving antigen presentation and T-cell activation Eur J Immunol 2013 43 10 2554 65 10.1002/eji.201343324 23836147 Rosalia RA, Quakkelaar ED, Redeker A, Khan S, Camps M, Drijfhout JW, et al. Dendritic cells process synthetic long peptides better than whole protein, improving antigen presentation and T-cell activation. Eur J Immunol. 2013;43(10):2554–65. 10.1002/eji.201343324. 23836147 10.1002/eji.201343324

45. Carreno BM Magrini V Becker-Hapak M Kaabinejadian S Hundal J Petti AA A dendritic cell vaccine increases the breadth and diversity of melanoma neoantigen-specific T cells Science 2015 348 6236 803 8 10.1126/science.aaa3828 25837513 PMC4549796 Carreno BM, Magrini V, Becker-Hapak M, Kaabinejadian S, Hundal J, Petti AA, et al. A dendritic cell vaccine increases the breadth and diversity of melanoma neoantigen-specific T cells. Science. 2015;348(6236):803–8. 10.1126/science.aaa3828. 25837513 10.1126/science.aaa3828 PMC4549796

46. Salcedo M Bercovici N Taylor R Vereecken P Massicard S Duriau D Vaccination of melanoma patients using dendritic cells loaded with an allogeneic tumor cell lysate Cancer Immunol Immunother 2006 55 7 819 29 10.1007/s00262-005-0078-6 16187085 PMC11030805 Salcedo M, Bercovici N, Taylor R, Vereecken P, Massicard S, Duriau D, et al. Vaccination of melanoma patients using dendritic cells loaded with an allogeneic tumor cell lysate. Cancer Immunol Immunother. 2006;55(7):819–29. 10.1007/s00262-005-0078-6. 16187085 10.1007/s00262-005-0078-6 PMC11030805

47. Bol KF Schreibelt G Bloemendal M van Willigen WW Hins-de Bree S de Goede AL Adjuvant dendritic cell therapy in stage IIIB/C melanoma: the MIND-DC randomized phase III trial Nat Commun 2024 15 1 1632 10.1038/s41467-024-45358-0 38395969 PMC10891118 Bol KF, Schreibelt G, Bloemendal M, van Willigen WW, Hins-de Bree S, de Goede AL, et al. Adjuvant dendritic cell therapy in stage IIIB/C melanoma: the MIND-DC randomized phase III trial. Nat Commun. 2024;15(1):1632. 10.1038/s41467-024-45358-0. 38395969 10.1038/s41467-024-45358-0 PMC10891118

48. Win SJ McMillan DGG Errington-Mais F Ward VK Young SL Baird MA Enhancing the immunogenicity of tumour lysate-loaded dendritic cell vaccines by conjugation to virus-like particles Br J Cancer 2012 106 1 92 8 10.1038/bjc.2011.538 22134504 PMC3251873 Win SJ, McMillan DGG, Errington-Mais F, Ward VK, Young SL, Baird MA, et al. Enhancing the immunogenicity of tumour lysate-loaded dendritic cell vaccines by conjugation to virus-like particles. Br J Cancer. 2012;106(1):92–8. 10.1038/bjc.2011.538. 22134504 10.1038/bjc.2011.538 PMC3251873

49. Zhu M Ding X Zhao R Liu X Shen H Cai C Co-delivery of tumor antigen and dual toll-like receptor ligands into dendritic cell by silicon microparticle enables efficient immunotherapy against melanoma J Control Release 2018 272 72 82 10.1016/j.jconrel.2018.01.004 29325699 PMC5825289 Zhu M, Ding X, Zhao R, Liu X, Shen H, Cai C, et al. Co-delivery of tumor antigen and dual toll-like receptor ligands into dendritic cell by silicon microparticle enables efficient immunotherapy against melanoma. J Control Release. 2018;272:72–82. 10.1016/j.jconrel.2018.01.004. 29325699 10.1016/j.jconrel.2018.01.004 PMC5825289

50. Fučíková J Rožková D Ulčová H Budinský V Sochorová K Pokorná K Poly I: C-activated dendritic cells that were generated in CellGro for use in cancer immunotherapy trials J Transl Med 2011 9 1 223 10.1186/1479-5876-9-223 22208910 PMC3259090 Fučíková J, Rožková D, Ulčová H, Budinský V, Sochorová K, Pokorná K, et al. Poly I: C-activated dendritic cells that were generated in CellGro for use in cancer immunotherapy trials. J Transl Med. 2011;9(1):223. 10.1186/1479-5876-9-223. 22208910 10.1186/1479-5876-9-223 PMC3259090

51. Lutz MB Schuler G Immature, semi-mature and fully mature dendritic cells: which signals induce tolerance or immunity? Trends Immunol 2002 23 9 445 9 10.1016/S1471-4906(02)02281-0 12200066 Lutz MB, Schuler G. Immature, semi-mature and fully mature dendritic cells: which signals induce tolerance or immunity? Trends Immunol. 2002;23(9):445–9. 10.1016/s1471-4906(02)02281-0. 12200066 10.1016/s1471-4906(02)02281-0

52. Boullart ACI Aarntzen EHJG Verdijk P Jacobs JFM Schuurhuis DH Benitez-Ribas D Maturation of monocyte-derived dendritic cells with Toll-like receptor 3 and 7/8 ligands combined with prostaglandin E2 results in high interleukin-12 production and cell migration Cancer Immunol Immunother 2008 57 11 1589 97 10.1007/s00262-008-0489-2 18322684 PMC2522299 Boullart ACI, Aarntzen EHJG, Verdijk P, Jacobs JFM, Schuurhuis DH, Benitez-Ribas D, et al. Maturation of monocyte-derived dendritic cells with Toll-like receptor 3 and 7/8 ligands combined with prostaglandin E2 results in high interleukin-12 production and cell migration. Cancer Immunol Immunother. 2008;57(11):1589–97. 10.1007/s00262-008-0489-2. 18322684 10.1007/s00262-008-0489-2 PMC2522299

53. Mailliard RB Wankowicz-Kalinska A Cai Q Wesa A Hilkens CM Kapsenberg ML alpha-type-1 polarized dendritic cells: a novel immunization tool with optimized CTL-inducing activity Cancer Res 2004 64 17 5934 7 10.1158/0008-5472.CAN-04-1261 15342370 Mailliard RB, Wankowicz-Kalinska A, Cai Q, Wesa A, Hilkens CM, Kapsenberg ML, et al. alpha-type-1 polarized dendritic cells: a novel immunization tool with optimized CTL-inducing activity. Cancer Res. 2004;64(17):5934–7. 10.1158/0008-5472.CAN-04-1261. 15342370 10.1158/0008-5472.CAN-04-1261

54. Giermasz AS Urban JA Nakamura Y Watchmaker P Cumberland RL Gooding W Type-1 polarized dendritic cells primed for high IL-12 production show enhanced activity as cancer vaccines Cancer Immunol Immunother 2009 58 8 1329 36 10.1007/s00262-008-0648-5 19156413 PMC2907477 Giermasz AS, Urban JA, Nakamura Y, Watchmaker P, Cumberland RL, Gooding W, et al. Type-1 polarized dendritic cells primed for high IL-12 production show enhanced activity as cancer vaccines. Cancer Immunol Immunother. 2009;58(8):1329–36. 10.1007/s00262-008-0648-5. 19156413 10.1007/s00262-008-0648-5 PMC2907477

55. Bonehill A Tuyaerts S van Nuffel AMT Heirman C Bos TJ Fostier K Enhancing the T-cell stimulatory capacity of human dendritic cells by co-electroporation with CD40L, CD70 and constitutively active TLR4 encoding mRNA Mol Ther 2008 16 6 1170 80 10.1038/mt.2008.77 18431362 Bonehill A, Tuyaerts S, van Nuffel AMT, Heirman C, Bos TJ, Fostier K, et al. Enhancing the T-cell stimulatory capacity of human dendritic cells by co-electroporation with CD40L, CD70 and constitutively active TLR4 encoding mRNA. Mol Ther. 2008;16(6):1170–80. 10.1038/mt.2008.77. 18431362 10.1038/mt.2008.77

56. Wilgenhof S van Nuffel AMT Benteyn D Corthals J Aerts C Heirman C A phase IB study on intravenous synthetic mRNA electroporated dendritic cell immunotherapy in pretreated advanced melanoma patients Annals oncology: official J Eur Soc Med Oncol 2013 24 10 2686 93 10.1093/annonc/mdt245 23904461 Wilgenhof S, van Nuffel AMT, Benteyn D, Corthals J, Aerts C, Heirman C, et al. A phase IB study on intravenous synthetic mRNA electroporated dendritic cell immunotherapy in pretreated advanced melanoma patients. Annals oncology: official J Eur Soc Med Oncol. 2013;24(10):2686–93. 10.1093/annonc/mdt245. 10.1093/annonc/mdt245 23904461

57. Wilgenhof S Corthals J Heirman C van Baren N Lucas S Kvistborg P Phase II Study of Autologous Monocyte-Derived mRNA Electroporated Dendritic Cells (TriMixDC-MEL) Plus Ipilimumab in Patients With Pretreated Advanced Melanoma JCO 2016 34 12 1330 8 10.1200/JCO.2015.63.4121 26926680 Wilgenhof S, Corthals J, Heirman C, van Baren N, Lucas S, Kvistborg P, et al. Phase II Study of Autologous Monocyte-Derived mRNA Electroporated Dendritic Cells (TriMixDC-MEL) Plus Ipilimumab in Patients With Pretreated Advanced Melanoma. JCO. 2016;34(12):1330–8. 10.1200/JCO.2015.63.4121. 10.1200/JCO.2015.63.4121 26926680

58. Verdijk P Aarntzen EHJG Lesterhuis WJ Boullart ACI Kok E van Rossum MM Limited amounts of dendritic cells migrate into the T-cell area of lymph nodes but have high immune activating potential in melanoma patients Clin Cancer Res 2009 15 7 2531 40 10.1158/1078-0432.CCR-08-2729 19318472 Verdijk P, Aarntzen EHJG, Lesterhuis WJ, Boullart ACI, Kok E, van Rossum MM, et al. Limited amounts of dendritic cells migrate into the T-cell area of lymph nodes but have high immune activating potential in melanoma patients. Clin Cancer Res. 2009;15(7):2531–40. 10.1158/1078-0432.CCR-08-2729. 19318472 10.1158/1078-0432.CCR-08-2729

59. Bedrosian I Mick R Xu S Nisenbaum H Faries M Zhang P Intranodal administration of peptide-pulsed mature dendritic cell vaccines results in superior CD8 + T-cell function in melanoma patients JCO 2003 21 20 3826 35 10.1200/JCO.2003.04.042 14551301 Bedrosian I, Mick R, Xu S, Nisenbaum H, Faries M, Zhang P, et al. Intranodal administration of peptide-pulsed mature dendritic cell vaccines results in superior CD8 + T-cell function in melanoma patients. JCO. 2003;21(20):3826–35. 10.1200/JCO.2003.04.042. 10.1200/JCO.2003.04.042 14551301

60. Lesterhuis WJ de Vries IJM Schreibelt G Lambeck AJA Aarntzen EHJG Jacobs JFM Route of administration modulates the induction of dendritic cell vaccine-induced antigen-specific T cells in advanced melanoma patients Clin Cancer Res 2011 17 17 5725 35 10.1158/1078-0432.CCR-11-1261 21771874 Lesterhuis WJ, de Vries IJM, Schreibelt G, Lambeck AJA, Aarntzen EHJG, Jacobs JFM, et al. Route of administration modulates the induction of dendritic cell vaccine-induced antigen-specific T cells in advanced melanoma patients. Clin Cancer Res. 2011;17(17):5725–35. 10.1158/1078-0432.CCR-11-1261. 21771874 10.1158/1078-0432.CCR-11-1261

61. Engell-Noerregaard L Hansen TH Andersen MH thor Straten P Svane IM Review of clinical studies on dendritic cell-based vaccination of patients with malignant melanoma: assessment of correlation between clinical response and vaccine parameters Cancer Immunol Immunother 2009 58 1 1 14 10.1007/s00262-008-0568-4 18719915 PMC11030652 Engell-Noerregaard L, Hansen TH, Andersen MH, thor Straten P, Svane IM. Review of clinical studies on dendritic cell-based vaccination of patients with malignant melanoma: assessment of correlation between clinical response and vaccine parameters. Cancer Immunol Immunother. 2009;58(1):1–14. 10.1007/s00262-008-0568-4. 18719915 10.1007/s00262-008-0568-4 PMC11030652

62. Thurner B Haendle I Röder C Dieckmann D Keikavoussi P Jonuleit H Vaccination with mage-3A1 peptide-pulsed mature, monocyte-derived dendritic cells expands specific cytotoxic T cells and induces regression of some metastases in advanced stage IV melanoma J Exp Med 1999 190 11 1669 78 10.1084/jem.190.11.1669 10587357 PMC2195739 Thurner B, Haendle I, Röder C, Dieckmann D, Keikavoussi P, Jonuleit H, et al. Vaccination with mage-3A1 peptide-pulsed mature, monocyte-derived dendritic cells expands specific cytotoxic T cells and induces regression of some metastases in advanced stage IV melanoma. J Exp Med. 1999;190(11):1669–78. 10.1084/jem.190.11.1669. 10587357 10.1084/jem.190.11.1669 PMC2195739

63. Fay JW Palucka AK Paczesny S Dhodapkar M Johnston DA Burkeholder S Long-term outcomes in patients with metastatic melanoma vaccinated with melanoma peptide-pulsed CD34(+) progenitor-derived dendritic cells Cancer Immunol Immunother 2006 55 10 1209 18 10.1007/s00262-005-0106-6 16331519 PMC11030596 Fay JW, Palucka AK, Paczesny S, Dhodapkar M, Johnston DA, Burkeholder S, et al. Long-term outcomes in patients with metastatic melanoma vaccinated with melanoma peptide-pulsed CD34(+) progenitor-derived dendritic cells. Cancer Immunol Immunother. 2006;55(10):1209–18. 10.1007/s00262-005-0106-6. 16331519 10.1007/s00262-005-0106-6 PMC11030596

64. Oshita C Takikawa M Kume A Miyata H Ashizawa T Iizuka A Dendritic cell-based vaccination in metastatic melanoma patients: phase II clinical trial Oncol Rep 2012 28 4 1131 8 10.3892/or.2012.1956 22895835 PMC3583475 Oshita C, Takikawa M, Kume A, Miyata H, Ashizawa T, Iizuka A, et al. Dendritic cell-based vaccination in metastatic melanoma patients: phase II clinical trial. Oncol Rep. 2012;28(4):1131–8. 10.3892/or.2012.1956. 22895835 10.3892/or.2012.1956 PMC3583475

65. Schwartzentruber DJ Lawson DH Richards JM Conry RM Miller DM Treisman J gp100 peptide vaccine and interleukin-2 in patients with advanced melanoma N Engl J Med 2011 364 22 2119 27 10.1056/NEJMoa1012863 21631324 PMC3517182 Schwartzentruber DJ, Lawson DH, Richards JM, Conry RM, Miller DM, Treisman J, et al. gp100 peptide vaccine and interleukin-2 in patients with advanced melanoma. N Engl J Med. 2011;364(22):2119–27. 10.1056/NEJMoa1012863. 21631324 10.1056/NEJMoa1012863 PMC3517182

66. Baumgaertner P Costa Nunes C Cachot A Maby-El Hajjami H Cagnon L Braun M Vaccination of stage III/IV melanoma patients with long NY-ESO-1 peptide and CpG-B elicits robust CD8 + and CD4 + T-cell responses with multiple specificities including a novel DR7-restricted epitope OncoImmunology 2016 5 10 e1216290 10.1080/2162402X.2016.1216290 27853637 PMC5087303 Baumgaertner P, Costa Nunes C, Cachot A, Maby-El Hajjami H, Cagnon L, Braun M, et al. Vaccination of stage III/IV melanoma patients with long NY-ESO-1 peptide and CpG-B elicits robust CD8 + and CD4 + T-cell responses with multiple specificities including a novel DR7-restricted epitope. OncoImmunology. 2016;5(10):e1216290. 10.1080/2162402X.2016.1216290. 27853637 10.1080/2162402X.2016.1216290 PMC5087303

67. Ninmer EK Zhu H Chianese-Bullock KA Slingluff CL Twenty-year survival outcomes after multipeptide vaccination for resected high-risk melanoma: A post-hoc analysis of a randomized clinical trial Int J Cancer 2025 157 9 1912 23 10.1002/ijc.70006 40536492 PMC12407048 Ninmer EK, Zhu H, Chianese-Bullock KA, Slingluff CL. Twenty-year survival outcomes after multipeptide vaccination for resected high-risk melanoma: A post-hoc analysis of a randomized clinical trial. Int J Cancer. 2025;157(9):1912–23. 10.1002/ijc.70006. 40536492 10.1002/ijc.70006 PMC12407048

68. Ninmer EK Zhu H Chianese-Bullock KA von Mehren M Haas NB Ross MI Multipeptide vaccines for melanoma in the adjuvant setting: long-term survival outcomes and post-hoc analysis of a randomized phase II trial Nat Commun 2024 15 1 2570 10.1038/s41467-024-46877-6 38519525 PMC10959948 Ninmer EK, Zhu H, Chianese-Bullock KA, von Mehren M, Haas NB, Ross MI, et al. Multipeptide vaccines for melanoma in the adjuvant setting: long-term survival outcomes and post-hoc analysis of a randomized phase II trial. Nat Commun. 2024;15(1):2570. 10.1038/s41467-024-46877-6. 38519525 10.1038/s41467-024-46877-6 PMC10959948

69. Yazdani M Gholizadeh Z Nikpoor AR Mohamadian Roshan N Jaafari MR Badiee A Ex vivo dendritic cell-based (DC) vaccine pulsed with a low dose of liposomal antigen and CpG-ODN improved PD-1 blockade immunotherapy Sci Rep 2021 11 1 14661 10.1038/s41598-021-94250-0 34282215 PMC8290007 Yazdani M, Gholizadeh Z, Nikpoor AR, Mohamadian Roshan N, Jaafari MR, Badiee A. Ex vivo dendritic cell-based (DC) vaccine pulsed with a low dose of liposomal antigen and CpG-ODN improved PD-1 blockade immunotherapy. Sci Rep. 2021;11(1):14661. 10.1038/s41598-021-94250-0. 34282215 10.1038/s41598-021-94250-0 PMC8290007

70. Grees M Sharbi-Yunger A Evangelou C Baumann D Cafri G Tzehoval E Optimized dendritic cell vaccination induces potent CD8 T cell responses and anti-tumor effects in transgenic mouse melanoma models OncoImmunology 2018 7 7 e1445457 10.1080/2162402X.2018.1445457 29900058 PMC5993502 Grees M, Sharbi-Yunger A, Evangelou C, Baumann D, Cafri G, Tzehoval E, et al. Optimized dendritic cell vaccination induces potent CD8 T cell responses and anti-tumor effects in transgenic mouse melanoma models. OncoImmunology. 2018;7(7):e1445457. 10.1080/2162402X.2018.1445457. 29900058 10.1080/2162402X.2018.1445457 PMC5993502

71. Poschke I Lövgren T Adamson L Nyström M Andersson E Hansson J A phase I clinical trial combining dendritic cell vaccination with adoptive T cell transfer in patients with stage IV melanoma Cancer Immunol Immunother 2014 63 10 1061 71 10.1007/s00262-014-1575-2 24993563 PMC11028804 Poschke I, Lövgren T, Adamson L, Nyström M, Andersson E, Hansson J, et al. A phase I clinical trial combining dendritic cell vaccination with adoptive T cell transfer in patients with stage IV melanoma. Cancer Immunol Immunother. 2014;63(10):1061–71. 10.1007/s00262-014-1575-2. 24993563 10.1007/s00262-014-1575-2 PMC11028804

72. Saberian C, Amaria RN, Najjar AM, Radvanyi LG, Haymaker CL, Forget M-A et al. Randomized phase II trial of lymphodepletion plus adoptive cell transfer of tumor-infiltrating lymphocytes, with or without dendritic cell vaccination, in patients with metastatic melanoma. J Immunother Cancer. 2021;9(5):e002449. 10.1136/jitc-2021-002449. 10.1136/jitc-2021-002449 PMC8144048 34021033

73. Ellebaek E Engell-Noerregaard L Iversen TZ Froesig TM Munir S Hadrup SR Metastatic melanoma patients treated with dendritic cell vaccination, Interleukin-2 and metronomic cyclophosphamide: results from a phase II trial Cancer Immunol Immunother 2012 61 10 1791 804 10.1007/s00262-012-1242-4 22426890 PMC11029126 Ellebaek E, Engell-Noerregaard L, Iversen TZ, Froesig TM, Munir S, Hadrup SR, et al. Metastatic melanoma patients treated with dendritic cell vaccination, Interleukin-2 and metronomic cyclophosphamide: results from a phase II trial. Cancer Immunol Immunother. 2012;61(10):1791–804. 10.1007/s00262-012-1242-4. 22426890 10.1007/s00262-012-1242-4 PMC11029126

74. Rohaan MW van den Berg JH Kvistborg P Haanen JBAG Adoptive transfer of tumor-infiltrating lymphocytes in melanoma: a viable treatment option J Immunother Cancer 2018 6 1 102 10.1186/s40425-018-0391-1 30285902 PMC6171186 Rohaan MW, van den Berg JH, Kvistborg P, Haanen JBAG. Adoptive transfer of tumor-infiltrating lymphocytes in melanoma: a viable treatment option. J Immunother Cancer. 2018;36(1):102. 10.1186/s40425-018-0391-1. 30285902 10.1186/s40425-018-0391-1 PMC6171186

75. Rosenberg SA Dudley ME Adoptive cell therapy for the treatment of patients with metastatic melanoma Curr Opin Immunol 2009 21 2 233 40 10.1016/j.coi.2009.03.002 19304471 PMC3459355 Rosenberg SA, Dudley ME. Adoptive cell therapy for the treatment of patients with metastatic melanoma. Curr Opin Immunol. 2009;21(2):233–40. 10.1016/j.coi.2009.03.002. 19304471 10.1016/j.coi.2009.03.002 PMC3459355

76. Kristensen NP, Heeke C, Tvingsholm SA, Borch A, Draghi A, Crowther MD et al. Neoantigen-reactive CD8 + T cells affect clinical outcome of adoptive cell therapy with tumor-infiltrating lymphocytes in melanoma. J Clin Invest. 2022;18;132(2):e150535. 10.1172/JCI150535.132(2). 10.1172/JCI150535 PMC8759789 34813506

77. Rosenberg SA Packard BS Aebersold PM Solomon D Topalian SL Toy ST Use of tumor-infiltrating lymphocytes and interleukin-2 in the immunotherapy of patients with metastatic melanoma. A preliminary report N Engl J Med 1988 319 25 1676 80 10.1056/NEJM198812223192527 3264384 Rosenberg SA, Packard BS, Aebersold PM, Solomon D, Topalian SL, Toy ST, et al. Use of tumor-infiltrating lymphocytes and interleukin-2 in the immunotherapy of patients with metastatic melanoma. A preliminary report. N Engl J Med. 1988;319(25):1676–80. 10.1056/NEJM198812223192527. 3264384 10.1056/NEJM198812223192527

78. Dafni U Michielin O Lluesma SM Tsourti Z Polydoropoulou V Karlis D Efficacy of adoptive therapy with tumor-infiltrating lymphocytes and recombinant interleukin-2 in advanced cutaneous melanoma: a systematic review and meta-analysis Annals oncology: official J Eur Soc Med Oncol 2019 30 12 1902 13 10.1093/annonc/mdz398 31566658 Dafni U, Michielin O, Lluesma SM, Tsourti Z, Polydoropoulou V, Karlis D, et al. Efficacy of adoptive therapy with tumor-infiltrating lymphocytes and recombinant interleukin-2 in advanced cutaneous melanoma: a systematic review and meta-analysis. Annals oncology: official J Eur Soc Med Oncol. 2019;30(12):1902–13. 10.1093/annonc/mdz398. 10.1093/annonc/mdz398 31566658

79. Rosenberg SA Yang JC Sherry RM Kammula US Hughes MS Phan GQ Durable complete responses in heavily pretreated patients with metastatic melanoma using T-cell transfer immunotherapy Clin Cancer Res 2011 17 13 4550 7 10.1158/1078-0432.CCR-11-0116 21498393 PMC3131487 Rosenberg SA, Yang JC, Sherry RM, Kammula US, Hughes MS, Phan GQ, et al. Durable complete responses in heavily pretreated patients with metastatic melanoma using T-cell transfer immunotherapy. Clin Cancer Res. 2011;17(13):4550–7. 10.1158/1078-0432.CCR-11-0116. 21498393 10.1158/1078-0432.CCR-11-0116 PMC3131487

80. Chesney J, Lewis KD, Kluger H, Hamid O, Whitman E, Thomas S et al. Efficacy and safety of lifileucel, a one-time autologous tumor-infiltrating lymphocyte (TIL) cell therapy, in patients with advanced melanoma after progression on immune checkpoint inhibitors and targeted therapies: pooled analysis of consecutive cohorts of the C-144-01 study. J Immunother Cancer. 2022;Dec;10(12):e005755. 10.1136/jitc-2022-005755.10(12). 10.1136/jitc-2022-005755 PMC9748991 36600653

81. Hu L Fan C Bross P Das A Cho ES Knudson KM FDA Approval Summary: Lifileucel for Unresectable or Metastatic Melanoma Previously Treated with an Anti-PD-1-Based Immunotherapy Clin Cancer Res 2025 31 19 4004 9 10.1158/1078-0432.CCR-25-0880 40699950 PMC12313226 Hu L, Fan C, Bross P, Das A, Cho ES, Knudson KM, et al. FDA Approval Summary: Lifileucel for Unresectable or Metastatic Melanoma Previously Treated with an Anti-PD-1-Based Immunotherapy. Clin Cancer Res. 2025;31(19):4004–9. 10.1158/1078-0432.CCR-25-0880. 40699950 10.1158/1078-0432.CCR-25-0880 PMC12313226

82. Medina T Chesney JA Kluger HM Hamid O Whitman ED Cusnir M Long-Term Efficacy and Safety of Lifileucel Tumor-Infiltrating Lymphocyte Cell Therapy in Patients With Advanced Melanoma: A 5-Year Analysis of the C-144-01 Study JCO 2025 43 33 3565 72 10.1200/JCO-25-00765 PMC12622280 40454684 Medina T, Chesney JA, Kluger HM, Hamid O, Whitman ED, Cusnir M, et al. Long-Term Efficacy and Safety of Lifileucel Tumor-Infiltrating Lymphocyte Cell Therapy in Patients With Advanced Melanoma: A 5-Year Analysis of the C-144-01 Study. JCO. 2025;43(33):3565–72. 10.1200/JCO-25-00765. 10.1200/JCO-25-00765 PMC12622280 40454684

83. Rohaan MW Borch TH van den Berg JH Met Ö Kessels R Geukes Foppen MH Tumor-Infiltrating Lymphocyte Therapy or Ipilimumab in Advanced Melanoma N Engl J Med 2022 387 23 2113 25 10.1056/NEJMoa2210233 36477031 Rohaan MW, Borch TH, van den Berg JH, Met Ö, Kessels R, Geukes Foppen MH, et al. Tumor-Infiltrating Lymphocyte Therapy or Ipilimumab in Advanced Melanoma. N Engl J Med. 2022;387(23):2113–25. 10.1056/NEJMoa2210233. 36477031 10.1056/NEJMoa2210233

84. Martín-Lluesma S Dafni U Vervita K Karlis D Dimopoulou G Tsourti Z Safety of adoptive therapy with tumor-infiltrating lymphocytes and high-dose recombinant interleukin 2 in advanced cutaneous melanoma: a systematic review and meta-analysis Annals oncology: official J Eur Soc Med Oncol 2025 36 8 909 19 10.1016/j.annonc.2025.04.001 40210086 Martín-Lluesma S, Dafni U, Vervita K, Karlis D, Dimopoulou G, Tsourti Z, et al. Safety of adoptive therapy with tumor-infiltrating lymphocytes and high-dose recombinant interleukin 2 in advanced cutaneous melanoma: a systematic review and meta-analysis. Annals oncology: official J Eur Soc Med Oncol. 2025;36(8):909–19. 10.1016/j.annonc.2025.04.001. 10.1016/j.annonc.2025.04.001 40210086

85. Davar D Ding F Saul M Sander C Tarhini AA Kirkwood JM High-dose interleukin-2 (HD IL-2) for advanced melanoma: a single center experience from the University of Pittsburgh Cancer Institute J Immunother Cancer 2017 5 1 74 10.1186/s40425-017-0279-5 28923120 PMC5604296 Davar D, Ding F, Saul M, Sander C, Tarhini AA, Kirkwood JM, et al. High-dose interleukin-2 (HD IL-2) for advanced melanoma: a single center experience from the University of Pittsburgh Cancer Institute. J Immunother Cancer. 2017;5(1):74. 10.1186/s40425-017-0279-5. 28923120 10.1186/s40425-017-0279-5 PMC5604296

86. Naidoo J, Murphy C, Atkins MB, Brahmer JR, Champiat S, Feltquate D et al. Society for Immunotherapy of Cancer (SITC) consensus definitions for immune checkpoint inhibitor-associated immune-related adverse events (irAEs) terminology. J Immunother Cancer. 2023;11(3):e006398. 10.1136/jitc-2022-006398. 10.1136/jitc-2022-006398 PMC10069596 37001909

87. Krishna S Lowery FJ Copeland AR Bahadiroglu E Mukherjee R Jia L Stem-like CD8 T cells mediate response of adoptive cell immunotherapy against human cancer Science 2020 370 6522 1328 34 10.1126/science.abb9847 33303615 PMC8883579 Krishna S, Lowery FJ, Copeland AR, Bahadiroglu E, Mukherjee R, Jia L, et al. Stem-like CD8 T cells mediate response of adoptive cell immunotherapy against human cancer. Science. 2020;370(6522):1328–34. 10.1126/science.abb9847. 33303615 10.1126/science.abb9847 PMC8883579

88. Campbell JDM Detection and enrichment of antigen-specific CD4 + and CD8 + T cells based on cytokine secretion Methods 2003 31 2 150 9 10.1016/S1046-2023(03)00125-7 12957573 Campbell JDM. Detection and enrichment of antigen-specific CD4 + and CD8 + T cells based on cytokine secretion. Methods. 2003;31(2):150–9. 10.1016/s1046-2023(03)00125-7. 12957573 10.1016/s1046-2023(03)00125-7

89. Kayser S Boβ C Feucht J Witte K-E Scheu A Bülow H-J Rapid generation of NY-ESO-1-specific CD4 + THELPER1 cells for adoptive T-cell therapy OncoImmunology 2015 4 5 e1002723 10.1080/2162402X.2014.1002723 26155389 PMC4485840 Kayser S, Boβ C, Feucht J, Witte K-E, Scheu A, Bülow H-J, et al. Rapid generation of NY-ESO-1-specific CD4 + THELPER1 cells for adoptive T-cell therapy. OncoImmunology. 2015;4(5):e1002723. 10.1080/2162402X.2014.1002723. 26155389 10.1080/2162402X.2014.1002723 PMC4485840

90. Bianchi V Harari A Coukos G Neoantigen-Specific Adoptive Cell Therapies for Cancer: Making T-Cell Products More Personal Front Immunol 2020 11 1215 10.3389/fimmu.2020.01215 32695101 PMC7333784 Bianchi V, Harari A, Coukos G. Neoantigen-Specific Adoptive Cell Therapies for Cancer: Making T-Cell Products More Personal. Front Immunol. 2020;11:1215. 10.3389/fimmu.2020.01215. 32695101 10.3389/fimmu.2020.01215 PMC7333784

91. Völzke C Ehrhardt L Fischer L Maul P Wenzel C Riabinska A Clinical-scale, modular manufacturing of tumor-reactive TILs using a closed and automated culture system Front Immunol 2024 15 1483254 10.3389/fimmu.2024.1483254 39717766 PMC11664263 Völzke C, Ehrhardt L, Fischer L, Maul P, Wenzel C, Riabinska A, et al. Clinical-scale, modular manufacturing of tumor-reactive TILs using a closed and automated culture system. Front Immunol. 2024;15:1483254. 10.3389/fimmu.2024.1483254. 39717766 10.3389/fimmu.2024.1483254 PMC11664263

92. Simon S Labarriere N PD-1 expression on tumor-specific T cells: Friend or foe for immunotherapy? OncoImmunology 2017 7 1 e1364828 10.1080/2162402X.2017.1364828 29296515 PMC5739549 Simon S, Labarriere N. PD-1 expression on tumor-specific T cells: Friend or foe for immunotherapy? OncoImmunology. 2017;7(1):e1364828. 10.1080/2162402X.2017.1364828. 29296515 10.1080/2162402X.2017.1364828 PMC5739549

93. Gros A Robbins PF Yao X Li YF Turcotte S Tran E PD-1 identifies the patient-specific CD8⁺ tumor-reactive repertoire infiltrating human tumors J Clin Invest 2014 124 5 2246 59 10.1172/JCI73639 24667641 PMC4001555 Gros A, Robbins PF, Yao X, Li YF, Turcotte S, Tran E, et al. PD-1 identifies the patient-specific CD8⁺ tumor-reactive repertoire infiltrating human tumors. J Clin Invest. 2014;124(5):2246–59. 10.1172/JCI73639. 24667641 10.1172/JCI73639 PMC4001555

94. Duhen T Duhen R Montler R Moses J Moudgil T de Miranda NF Co-expression of CD39 and CD103 identifies tumor-reactive CD8 T cells in human solid tumors Nat Commun 2018 9 1 2724 10.1038/s41467-018-05072-0 30006565 PMC6045647 Duhen T, Duhen R, Montler R, Moses J, Moudgil T, de Miranda NF, et al. Co-expression of CD39 and CD103 identifies tumor-reactive CD8 T cells in human solid tumors. Nat Commun. 2018;9(1):2724. 10.1038/s41467-018-05072-0. 30006565 10.1038/s41467-018-05072-0 PMC6045647

95. Chow A Uddin FZ Liu M Dobrin A Nabet BY Mangarin L The ectonucleotidase CD39 identifies tumor-reactive CD8 + T cells predictive of immune checkpoint blockade efficacy in human lung cancer Immunity 2023 56 1 93 e1066 10.1016/j.immuni.2022.12.001 36574773 PMC9887636 Chow A, Uddin FZ, Liu M, Dobrin A, Nabet BY, Mangarin L, et al. The ectonucleotidase CD39 identifies tumor-reactive CD8 + T cells predictive of immune checkpoint blockade efficacy in human lung cancer. Immunity. 2023;56(1):93–e1066. 10.1016/j.immuni.2022.12.001. 36574773 10.1016/j.immuni.2022.12.001 PMC9887636

96. Dolton G Tungatt K Lloyd A Bianchi V Theaker SM Trimby A More tricks with tetramers: a practical guide to staining T cells with peptide-MHC multimers Immunology 2015 146 1 11 22 10.1111/imm.12499 26076649 PMC4552497 Dolton G, Tungatt K, Lloyd A, Bianchi V, Theaker SM, Trimby A, et al. More tricks with tetramers: a practical guide to staining T cells with peptide-MHC multimers. Immunology. 2015;146(1):11–22. 10.1111/imm.12499. 26076649 10.1111/imm.12499 PMC4552497

97. van Buuren MM Dijkgraaf FE Linnemann C Toebes M Chang CXL Mok JY HLA micropolymorphisms strongly affect peptide-MHC multimer-based monitoring of antigen-specific CD8 + T cell responses J Immunol 2014 192 2 641 8 10.4049/jimmunol.1301770 24342804 van Buuren MM, Dijkgraaf FE, Linnemann C, Toebes M, Chang CXL, Mok JY, et al. HLA micropolymorphisms strongly affect peptide-MHC multimer-based monitoring of antigen-specific CD8 + T cell responses. J Immunol. 2014;192(2):641–8. 10.4049/jimmunol.1301770. 24342804 10.4049/jimmunol.1301770

98. Dudley ME Gross CA Langhan MM Garcia MR Sherry RM Yang JC CD8 + enriched young tumor infiltrating lymphocytes can mediate regression of metastatic melanoma Clin Cancer Res 2010 16 24 6122 31 10.1158/1078-0432.CCR-10-1297 20668005 PMC2978753 Dudley ME, Gross CA, Langhan MM, Garcia MR, Sherry RM, Yang JC, et al. CD8 + enriched young tumor infiltrating lymphocytes can mediate regression of metastatic melanoma. Clin Cancer Res. 2010;16(24):6122–31. 10.1158/1078-0432.CCR-10-1297. 20668005 10.1158/1078-0432.CCR-10-1297 PMC2978753

99. Tran KQ Zhou J Durflinger KH Langhan MM Shelton TE Wunderlich JR Minimally cultured tumor-infiltrating lymphocytes display optimal characteristics for adoptive cell therapy J Immunother 2008 31 8 742 51 10.1097/CJI.0b013e31818403d5 18779745 PMC2614999 Tran KQ, Zhou J, Durflinger KH, Langhan MM, Shelton TE, Wunderlich JR, et al. Minimally cultured tumor-infiltrating lymphocytes display optimal characteristics for adoptive cell therapy. J Immunother. 2008;31(8):742–51. 10.1097/CJI.0b013e31818403d5. 18779745 10.1097/CJI.0b013e31818403d5 PMC2614999

100. Gong W Hoffmann J-M Stock S Wang L Liu Y Schubert M-L Comparison of IL-2 vs IL-7/IL-15 for the generation of NY-ESO-1-specific T cells Cancer Immunol Immunother 2019 68 7 1195 209 10.1007/s00262-019-02354-4 31177329 PMC11028180 Gong W, Hoffmann J-M, Stock S, Wang L, Liu Y, Schubert M-L, et al. Comparison of IL-2 vs IL-7/IL-15 for the generation of NY-ESO-1-specific T cells. Cancer Immunol Immunother. 2019;68(7):1195–209. 10.1007/s00262-019-02354-4. 31177329 10.1007/s00262-019-02354-4 PMC11028180

101. Santegoets SJAM Turksma AW Suhoski MM Stam AGM Albelda SM Hooijberg E IL-21 promotes the expansion of CD27 + CD28+ tumor infiltrating lymphocytes with high cytotoxic potential and low collateral expansion of regulatory T cells J Transl Med 2013 11 37 10.1186/1479-5876-11-37 23402380 PMC3626797 Santegoets SJAM, Turksma AW, Suhoski MM, Stam AGM, Albelda SM, Hooijberg E, et al. IL-21 promotes the expansion of CD27 + CD28+ tumor infiltrating lymphocytes with high cytotoxic potential and low collateral expansion of regulatory T cells. J Transl Med. 2013;11:37. 10.1186/1479-5876-11-37. 23402380 10.1186/1479-5876-11-37 PMC3626797

102. Chacon JA Wu RC Sukhumalchandra P Molldrem JJ Sarnaik A Pilon-Thomas S Co-stimulation through 4-1BB/CD137 improves the expansion and function of CD8(+) melanoma tumor-infiltrating lymphocytes for adoptive T-cell therapy PLoS ONE 2013 8 4 e60031 10.1371/journal.pone.0060031 23560068 PMC3613355 Chacon JA, Wu RC, Sukhumalchandra P, Molldrem JJ, Sarnaik A, Pilon-Thomas S, et al. Co-stimulation through 4-1BB/CD137 improves the expansion and function of CD8(+) melanoma tumor-infiltrating lymphocytes for adoptive T-cell therapy. PLoS ONE. 2013;8(4):e60031. 10.1371/journal.pone.0060031. 23560068 10.1371/journal.pone.0060031 PMC3613355

103. Thomas SS, Gogas H, Hong YK, In GK, de Doger B, Furness AJ et al. Efficacy and safety of lifileucel, an autologous tumor-infiltrating lymphocyte cell therapy, and pembrolizumab in patients with immune checkpoint inhibitor-naive unresectable or metastatic melanoma: Updated results from IOV-COM-202 cohort 1A. JCO. 2024;42(16_suppl):9505.

104. Palmer DC Guittard GC Franco Z Crompton JG Eil RL Patel SJ Cish actively silences TCR signaling in CD8 + T cells to maintain tumor tolerance J Exp Med 2015 212 12 2095 113 10.1084/jem.20150304 26527801 PMC4647263 Palmer DC, Guittard GC, Franco Z, Crompton JG, Eil RL, Patel SJ, et al. Cish actively silences TCR signaling in CD8 + T cells to maintain tumor tolerance. J Exp Med. 2015;212(12):2095–113. 10.1084/jem.20150304. 26527801 10.1084/jem.20150304 PMC4647263

105. Lv J Le Qin, Zhao R Di Wu, Wu Z Zheng D Disruption of CISH promotes the antitumor activity of human T cells and decreases PD-1 expression levels Mol Ther Oncolytics 2023 28 46 58 10.1016/j.omto.2022.12.003 36654786 PMC9827364 Lv J, Le Qin, Zhao R, Di Wu, Wu Z, Zheng D, et al. Disruption of CISH promotes the antitumor activity of human T cells and decreases PD-1 expression levels. Mol Ther Oncolytics. 2023;28:46–58. 10.1016/j.omto.2022.12.003. 36654786 10.1016/j.omto.2022.12.003 PMC9827364

106. Lou E Choudhry MS Starr TK Folsom TD Bell J Rathmann B Targeting the intracellular immune checkpoint CISH with CRISPR-Cas9-edited T cells in patients with metastatic colorectal cancer: a first-in-human, single-centre, phase 1 trial Lancet Oncol 2025 26 5 559 70 10.1016/S1470-2045(25)00083-X 40315882 Lou E, Choudhry MS, Starr TK, Folsom TD, Bell J, Rathmann B, et al. Targeting the intracellular immune checkpoint CISH with CRISPR-Cas9-edited T cells in patients with metastatic colorectal cancer: a first-in-human, single-centre, phase 1 trial. Lancet Oncol. 2025;26(5):559–70. 10.1016/S1470-2045(25)00083-X. 40315882 10.1016/S1470-2045(25)00083-X

107. Chamberlain CA Bennett EP Kverneland AH Svane IM Donia M Met Ö Highly efficient PD-1-targeted CRISPR-Cas9 for tumor-infiltrating lymphocyte-based adoptive T cell therapy Mol Ther Oncolytics 2022 24 417 28 10.1016/j.omto.2022.01.004 35141398 PMC8807971 Chamberlain CA, Bennett EP, Kverneland AH, Svane IM, Donia M, Met Ö. Highly efficient PD-1-targeted CRISPR-Cas9 for tumor-infiltrating lymphocyte-based adoptive T cell therapy. Mol Ther Oncolytics. 2022;24:417–28. 10.1016/j.omto.2022.01.004. 35141398 10.1016/j.omto.2022.01.004 PMC8807971

108. Fix SM, Forget M-A, Sakellariou-Thompson D, Wang Y, Griffiths TM, Lee M et al. CRISPR-mediated TGFBR2 knockout renders human ovarian cancer tumor-infiltrating lymphocytes resistant to TGF-β signaling. J Immunother Cancer 2022Jul;10(7):e003750. 10.1136/jitc-2021-003750. 10.1136/jitc-2021-003750 PMC9330322 35882447

109. Amaria RN Diab A Tawbi HA Glitza IC McQuade JL Davies MA OBX-115 engineered tumor-infiltrating lymphocytes (TIL) with regulatable membrane-bound IL15 (mbIL15): Translational data from a single-center phase 1 trial in patients (pts) with immune checkpoint inhibitor (ICI)–resistant advanced melanoma JCO 2025 43 9519 10.1200/JCO.2025.43.16_suppl.9519 Amaria RN, Diab A, Tawbi HA, Glitza IC, McQuade JL, Davies MA, et al. OBX-115 engineered tumor-infiltrating lymphocytes (TIL) with regulatable membrane-bound IL15 (mbIL15): Translational data from a single-center phase 1 trial in patients (pts) with immune checkpoint inhibitor (ICI)–resistant advanced melanoma. JCO. 2025;43:9519. (16_suppl).

110. Amaria RN Haymaker CL Bernatchez C Forget M-A Patel V Hwu W-J A phase I/II study of lymphodepletion plus adoptive cell transfer (ACT) with T cells transduced with CXCR2 and NGFR followed by high dose interleukin-2 (IL-2) in patients with metastatic melanoma (MM) JCO 2016 34 15suppl TPS9594 9594 10.1200/JCO.2016.34.15_suppl.TPS9594 Amaria RN, Haymaker CL, Bernatchez C, Forget M-A, Patel V, Hwu W-J, et al. A phase I/II study of lymphodepletion plus adoptive cell transfer (ACT) with T cells transduced with CXCR2 and NGFR followed by high dose interleukin-2 (IL-2) in patients with metastatic melanoma (MM). JCO. 2016;34(15suppl):TPS9594–9594.

111. Burga RA Aksoy BA Ao Z Tchaicha JH Sethi DK Villasmil Ocando A IL-2-independent expansion, persistence, and antitumor activity in TIL expressing regulatable membrane-bound IL-15 Mol Ther 2025 33 8 3605 23 10.1016/j.ymthe.2025.04.031 40285351 PMC12461655 Burga RA, Aksoy BA, Ao Z, Tchaicha JH, Sethi DK, Villasmil Ocando A, et al. IL-2-independent expansion, persistence, and antitumor activity in TIL expressing regulatable membrane-bound IL-15. Mol Ther. 2025;33(8):3605–23. 10.1016/j.ymthe.2025.04.031. 40285351 10.1016/j.ymthe.2025.04.031 PMC12461655

112. June CH O’Connor RS Kawalekar OU Ghassemi S Milone MC CAR T cell immunotherapy for human cancer Science 2018 359 6382 1361 5 10.1126/science.aar6711 29567707 June CH, O’Connor RS, Kawalekar OU, Ghassemi S, Milone MC. CAR T cell immunotherapy for human cancer. Science. 2018;359(6382):1361–5. 10.1126/science.aar6711. 29567707 10.1126/science.aar6711

113. Piscopo NJ, Mueller KP, Das A, Hematti P, Murphy WL, Palecek SP et al. Bioengineering Solutions for Manufacturing Challenges in CAR T Cells. Biotechnol J. 2018;13(2):10.1002/biot.201700095. 10.1002/biot.201700095. 10.1002/biot.201700095 PMC5796845 28840981

114. Levine BL Miskin J Wonnacott K Keir C Global Manufacturing of CAR T Cell Therapy Mol Ther Methods Clin Dev 2017 4 92 101 10.1016/j.omtm.2016.12.006 28344995 PMC5363291 Levine BL, Miskin J, Wonnacott K, Keir C. Global Manufacturing of CAR T Cell Therapy. Mol Ther Methods Clin Dev. 2017;4:92–101. 10.1016/j.omtm.2016.12.006. 28344995 10.1016/j.omtm.2016.12.006 PMC5363291

115. Watanabe N Mo F McKenna MK Impact of Manufacturing Procedures on CAR T Cell Functionality Front Immunol 2022 13 876339 10.3389/fimmu.2022.876339 35493513 PMC9043864 Watanabe N, Mo F, McKenna MK. Impact of Manufacturing Procedures on CAR T Cell Functionality. Front Immunol. 2022;13:876339. 10.3389/fimmu.2022.876339. 35493513 10.3389/fimmu.2022.876339 PMC9043864

116. Silveira CRF Corveloni AC Caruso SR Macêdo NA Brussolo NM Haddad F Cytokines as an important player in the context of CAR-T cell therapy for cancer: Their role in tumor immunomodulation, manufacture, and clinical implications Front Immunol 2022 13 947648 10.3389/fimmu.2022.947648 36172343 PMC9512053 Silveira CRF, Corveloni AC, Caruso SR, Macêdo NA, Brussolo NM, Haddad F, et al. Cytokines as an important player in the context of CAR-T cell therapy for cancer: Their role in tumor immunomodulation, manufacture, and clinical implications. Front Immunol. 2022;13:947648. 10.3389/fimmu.2022.947648. 36172343 10.3389/fimmu.2022.947648 PMC9512053

117. Schuster SJ Svoboda J Chong EA Nasta SD Mato AR Anak Ö Chimeric Antigen Receptor T Cells in Refractory B-Cell Lymphomas N Engl J Med 2017 377 26 2545 54 10.1056/NEJMoa1708566 29226764 PMC5788566 Schuster SJ, Svoboda J, Chong EA, Nasta SD, Mato AR, Anak Ö, et al. Chimeric Antigen Receptor T Cells in Refractory B-Cell Lymphomas. N Engl J Med. 2017;377(26):2545–54. 10.1056/NEJMoa1708566. 29226764 10.1056/NEJMoa1708566 PMC5788566

118. Locke FL Miklos DB Jacobson CA Perales M-A Kersten M-J Oluwole OO Axicabtagene Ciloleucel as Second-Line Therapy for Large B-Cell Lymphoma N Engl J Med 2022 386 7 640 54 10.1056/NEJMoa2116133 34891224 Locke FL, Miklos DB, Jacobson CA, Perales M-A, Kersten M-J, Oluwole OO, et al. Axicabtagene Ciloleucel as Second-Line Therapy for Large B-Cell Lymphoma. N Engl J Med. 2022;386(7):640–54. 10.1056/NEJMoa2116133. 34891224 10.1056/NEJMoa2116133

119. Lee DW Santomasso BD Locke FL Ghobadi A Turtle CJ Brudno JN ASTCT Consensus Grading for Cytokine Release Syndrome and Neurologic Toxicity Associated with Immune Effector Cells Biol Blood Marrow Transpl 2019 25 4 625 38 10.1016/j.bbmt.2018.12.758 PMC12180426 30592986 Lee DW, Santomasso BD, Locke FL, Ghobadi A, Turtle CJ, Brudno JN, et al. ASTCT Consensus Grading for Cytokine Release Syndrome and Neurologic Toxicity Associated with Immune Effector Cells. Biol Blood Marrow Transpl. 2019;25(4):625–38. 10.1016/j.bbmt.2018.12.758. 10.1016/j.bbmt.2018.12.758 PMC12180426 30592986

120. Verdun N Marks P Secondary Cancers after Chimeric Antigen Receptor T-Cell Therapy N Engl J Med 2024 390 7 584 6 10.1056/NEJMp2400209 38265704 Verdun N, Marks P. Secondary Cancers after Chimeric Antigen Receptor T-Cell Therapy. N Engl J Med. 2024;390(7):584–6. 10.1056/NEJMp2400209. 38265704 10.1056/NEJMp2400209

121. Soltantoyeh T, Akbari B, Karimi A, Mahmoodi Chalbatani G, Ghahri-Saremi N, Hadjati J et al. Chimeric Antigen Receptor (CAR) T Cell Therapy for Metastatic Melanoma: Challenges and Road Ahead. Cells. 2021;10(6):1450. 10.3390/cells10061450 10.3390/cells10061450 PMC8230324 34207884

122. Kontos F Michelakos T Kurokawa T Sadagopan A Schwab JH Ferrone CR B7-H3: An Attractive Target for Antibody-based Immunotherapy Clin Cancer Res 2021 27 5 1227 35 10.1158/1078-0432.CCR-20-2584 33051306 PMC7925343 Kontos F, Michelakos T, Kurokawa T, Sadagopan A, Schwab JH, Ferrone CR, et al. B7-H3: An Attractive Target for Antibody-based Immunotherapy. Clin Cancer Res. 2021;27(5):1227–35. 10.1158/1078-0432.CCR-20-2584. 33051306 10.1158/1078-0432.CCR-20-2584 PMC7925343

123. Seaman S Zhu Z Saha S Zhang XM Yang MY Hilton MB Eradication of Tumors through Simultaneous Ablation of CD276/B7-H3-Positive Tumor Cells and Tumor Vasculature Cancer Cell 2017 31 4 501 e5158 10.1016/j.ccell.2017.03.005 28399408 PMC5458750 Seaman S, Zhu Z, Saha S, Zhang XM, Yang MY, Hilton MB, et al. Eradication of Tumors through Simultaneous Ablation of CD276/B7-H3-Positive Tumor Cells and Tumor Vasculature. Cancer Cell. 2017;31(4):501–e5158. 10.1016/j.ccell.2017.03.005. 28399408 10.1016/j.ccell.2017.03.005 PMC5458750

124. Pinto N Albert CM Taylor MR Ullom HB Wilson AL Huang W STRIvE-02: A First-in-Human Phase I Study of Systemically Administered B7-H3 Chimeric Antigen Receptor T Cells for Patients With Relapsed/Refractory Solid Tumors JCO 2024 42 35 4163 72 10.1200/JCO.23.02229 39255444 Pinto N, Albert CM, Taylor MR, Ullom HB, Wilson AL, Huang W, et al. STRIvE-02: A First-in-Human Phase I Study of Systemically Administered B7-H3 Chimeric Antigen Receptor T Cells for Patients With Relapsed/Refractory Solid Tumors. JCO. 2024;42(35):4163–72. 10.1200/JCO.23.02229. 10.1200/JCO.23.02229 39255444

125. Maher J, Davies DM. CAR Based Immunotherapy of Solid Tumours-A Clinically Based Review of Target Antigens. Biology (Basel). 2023;12(2):287. 10.3390/biology12020287. 10.3390/biology12020287 PMC9953298 36829563

126. Guo Y Wang X Zhang C Chen W Fu Y Yu Y Tumor Immunotherapy Targeting B7-H3: From Mechanisms to Clinical Applications Immunotargets Ther 2025 14 291 320 10.2147/ITT.S507522 40171330 PMC11960814 Guo Y, Wang X, Zhang C, Chen W, Fu Y, Yu Y, et al. Tumor Immunotherapy Targeting B7-H3: From Mechanisms to Clinical Applications. Immunotargets Ther. 2025;14:291–320. 10.2147/ITT.S507522. 40171330 10.2147/ITT.S507522 PMC11960814

127. Yamada Y Venkadakrishnan VB Mizuno K Bakht M Ku S-Y Garcia MM Targeting DNA methylation and B7-H3 in RB1-deficient and neuroendocrine prostate cancer Sci Transl Med 2023 15 722 eadf6732 10.1126/scitranslmed.adf6732 37967200 PMC10954288 Yamada Y, Venkadakrishnan VB, Mizuno K, Bakht M, Ku S-Y, Garcia MM, et al. Targeting DNA methylation and B7-H3 in RB1-deficient and neuroendocrine prostate cancer. Sci Transl Med. 2023;15(722):eadf6732. 10.1126/scitranslmed.adf6732. 37967200 10.1126/scitranslmed.adf6732 PMC10954288

128. Jiang Z Zhou J Li L Liao S He J Zhou S Pericytes in the tumor microenvironment Cancer Lett 2023 556 216074 10.1016/j.canlet.2023.216074 36682706 Jiang Z, Zhou J, Li L, Liao S, He J, Zhou S, et al. Pericytes in the tumor microenvironment. Cancer Lett. 2023;556:216074. 10.1016/j.canlet.2023.216074. 36682706 10.1016/j.canlet.2023.216074

129. Boudin L de Nonneville A Finetti P Mescam L Le Cesne A Italiano A CSPG4 expression in soft tissue sarcomas is associated with poor prognosis and low cytotoxic immune response J Transl Med 2022 20 1 464 10.1186/s12967-022-03679-y 36221119 PMC9552405 Boudin L, de Nonneville A, Finetti P, Mescam L, Le Cesne A, Italiano A, et al. CSPG4 expression in soft tissue sarcomas is associated with poor prognosis and low cytotoxic immune response. J Transl Med. 2022;20(1):464. 36221119 10.1186/s12967-022-03679-y PMC9552405

130. Chen X, Habib S, Alexandru M, Chauhan J, Evan T, Troka JM, et al. Chondroitin Sulfate Proteoglycan 4 (CSPG4) as an Emerging Target for Immunotherapy to Treat Melanoma. Cancers (Basel). 2024;16(19):3260. 10.3390/cancers16193260. 10.3390/cancers16193260 PMC11476251 39409881

131. Wiesinger M, März J, Kummer M, Schuler G, Dörrie J, Schuler-Thurner B, et al. Clinical-Scale Production of CAR-T Cells for the Treatment of Melanoma Patients by mRNA Transfection of a CSPG4-Specific CAR under Full GMP Compliance. Cancers (Basel). 2019;11(8):1198. 10.3390/cancers11081198. 10.3390/cancers11081198 PMC6721485 31426437

132. Giraudo L Cattaneo G Gammaitoni L Iaia I Donini C Massa A CSPG4 CAR-redirected Cytokine Induced Killer lymphocytes (CIK) as effective cellular immunotherapy for HLA class I defective melanoma J Exp Clin Cancer Res 2023 42 1 310 10.1186/s13046-023-02884-x 37993874 PMC10664597 Giraudo L, Cattaneo G, Gammaitoni L, Iaia I, Donini C, Massa A, et al. CSPG4 CAR-redirected Cytokine Induced Killer lymphocytes (CIK) as effective cellular immunotherapy for HLA class I defective melanoma. J Exp Clin Cancer Res. 2023;42(1):310. 10.1186/s13046-023-02884-x. 37993874 10.1186/s13046-023-02884-x PMC10664597

133. Knudson KM Hwang S McCann MS Joshi BH Husain SR Puri RK Recent Advances in IL-13Rα2-Directed Cancer Immunotherapy Front Immunol 2022 13 878365 10.3389/fimmu.2022.878365 35464460 PMC9023787 Knudson KM, Hwang S, McCann MS, Joshi BH, Husain SR, Puri RK. Recent Advances in IL-13Rα2-Directed Cancer Immunotherapy. Front Immunol. 2022;13:878365. 10.3389/fimmu.2022.878365. 35464460 10.3389/fimmu.2022.878365 PMC9023787

134. Brown CE Hibbard JC Alizadeh D Blanchard MS Natri HM Wang D Locoregional delivery of IL-13Rα2-targeting CAR-T cells in recurrent high-grade glioma: a phase 1 trial Nat Med 2024 30 4 1001 12 10.1038/s41591-024-02875-1 38454126 PMC11031404 Brown CE, Hibbard JC, Alizadeh D, Blanchard MS, Natri HM, Wang D, et al. Locoregional delivery of IL-13Rα2-targeting CAR-T cells in recurrent high-grade glioma: a phase 1 trial. Nat Med. 2024;30(4):1001–12. 10.1038/s41591-024-02875-1. 38454126 10.1038/s41591-024-02875-1 PMC11031404

135. Jilani S Saco JD Mugarza E Pujol-Morcillo A Chokry J Ng C CAR-T cell therapy targeting surface expression of TYRP1 to treat cutaneous and rare melanoma subtypes Nat Commun 2024 15 1 1244 10.1038/s41467-024-45221-2 38336975 PMC10858182 Jilani S, Saco JD, Mugarza E, Pujol-Morcillo A, Chokry J, Ng C, et al. CAR-T cell therapy targeting surface expression of TYRP1 to treat cutaneous and rare melanoma subtypes. Nat Commun. 2024;15(1):1244. 10.1038/s41467-024-45221-2. 38336975 10.1038/s41467-024-45221-2 PMC10858182

136. Zhu EF Gai SA Opel CF Kwan BH Surana R Mihm MC Synergistic innate and adaptive immune response to combination immunotherapy with anti-tumor antigen antibodies and extended serum half-life IL-2 Cancer Cell 2015 27 4 489 501 10.1016/j.ccell.2015.03.004 25873172 PMC4398916 Zhu EF, Gai SA, Opel CF, Kwan BH, Surana R, Mihm MC, et al. Synergistic innate and adaptive immune response to combination immunotherapy with anti-tumor antigen antibodies and extended serum half-life IL-2. Cancer Cell. 2015;27(4):489–501. 10.1016/j.ccell.2015.03.004 25873172 10.1016/j.ccell.2015.03.004 PMC4398916

137. Khalil DN Postow MA Ibrahim N Ludwig DL Cosaert J Kambhampati SRP An Open-Label, Dose-Escalation Phase I Study of Anti-TYRP1 Monoclonal Antibody IMC-20D7S for Patients with Relapsed or Refractory Melanoma Clin Cancer Res 2016 22 21 5204 10 10.1158/1078-0432.CCR-16-1241 27797971 PMC5117650 Khalil DN, Postow MA, Ibrahim N, Ludwig DL, Cosaert J, Kambhampati SRP, et al. An Open-Label, Dose-Escalation Phase I Study of Anti-TYRP1 Monoclonal Antibody IMC-20D7S for Patients with Relapsed or Refractory Melanoma. Clin Cancer Res. 2016;22(21):5204–10. 10.1158/1078-0432.CCR-16-1241. 27797971 10.1158/1078-0432.CCR-16-1241 PMC5117650

138. Machy P Mortier E Birklé S Biology of GD2 ganglioside: implications for cancer immunotherapy Front Pharmacol 2023 14 1249929 10.3389/fphar.2023.1249929 37670947 PMC10475612 Machy P, Mortier E, Birklé S. Biology of GD2 ganglioside: implications for cancer immunotherapy. Front Pharmacol. 2023;14:1249929. 10.3389/fphar.2023.1249929. 37670947 10.3389/fphar.2023.1249929 PMC10475612

139. Del Bufalo F de Angelis B Caruana I Del Baldo G de Ioris MA Serra A GD2-CART01 for Relapsed or Refractory High-Risk Neuroblastoma N Engl J Med 2023 388 14 1284 95 10.1056/NEJMoa2210859 37018492 Del Bufalo F, de Angelis B, Caruana I, Del Baldo G, de Ioris MA, Serra A, et al. GD2-CART01 for Relapsed or Refractory High-Risk Neuroblastoma. N Engl J Med. 2023;388(14):1284–95. 10.1056/NEJMoa2210859. 37018492 10.1056/NEJMoa2210859

140. Locatelli F Pagliara D de Ioris MA Becilli M Del Baldo G Serra A GD2-targeting CAR T cells in high-risk neuroblastoma: a phase 1/2 trial Nat Med 2025 31 11 3689 99 10.1038/s41591-025-03874-6 40841488 Locatelli F, Pagliara D, de Ioris MA, Becilli M, Del Baldo G, Serra A, et al. GD2-targeting CAR T cells in high-risk neuroblastoma: a phase 1/2 trial. Nat Med. 2025;31(11):3689–99. 10.1038/s41591-025-03874-6. 40841488 10.1038/s41591-025-03874-6

141. Monje M Mahdi J Majzner R Yeom KW Schultz LM Richards RM Intravenous and intracranial GD2-CAR T cells for H3K27M+ diffuse midline gliomas Nature 2025 637 8046 708 15 10.1038/s41586-024-08171-9 39537919 PMC11735388 Monje M, Mahdi J, Majzner R, Yeom KW, Schultz LM, Richards RM, et al. Intravenous and intracranial GD2-CAR T cells for H3K27M+ diffuse midline gliomas. Nature. 2025;637(8046):708–15. 10.1038/s41586-024-08171-9. 39537919 10.1038/s41586-024-08171-9 PMC11735388

142. Gargett T, Truong NTH, Gardam B, Yu W, Ebert LM, Johnson A et al. Safety and biological outcomes following a phase 1 trial of GD2-specific CAR-T cells in patients with GD2-positive metastatic melanoma and other solid cancers. J Immunother Cancer. 2024;12(5):e008659. 10.1136/jitc-2023-008659. 10.1136/jitc-2023-008659 PMC11097842 38754916

143. Razavi A Keshavarz-Fathi M Pawelek J Rezaei N Chimeric antigen receptor T-cell therapy for melanoma Expert Rev Clin Immunol 2021 17 3 209 23 10.1080/1744666X.2021.1880895 33481629 Razavi A, Keshavarz-Fathi M, Pawelek J, Rezaei N. Chimeric antigen receptor T-cell therapy for melanoma. Expert Rev Clin Immunol. 2021;17(3):209–23. 10.1080/1744666X.2021.1880895. 33481629 10.1080/1744666X.2021.1880895

144. Yang DD Macmorland W Arnold JN Current strategies for armoring chimeric antigen receptor T-cells to overcome barriers of the solid tumor microenvironment Front Immunol 2025 16 1643941 10.3389/fimmu.2025.1643941 41019052 PMC12460333 Yang DD, Macmorland W, Arnold JN. Current strategies for armoring chimeric antigen receptor T-cells to overcome barriers of the solid tumor microenvironment. Front Immunol. 2025;16:1643941. 10.3389/fimmu.2025.1643941. 41019052 10.3389/fimmu.2025.1643941 PMC12460333

145. Wagner J Wickman E DeRenzo C Gottschalk S CAR T Cell Therapy for Solid Tumors: Bright Future or Dark Reality? Mol Ther 2020 28 11 2320 39 10.1016/j.ymthe.2020.09.015 32979309 PMC7647674 Wagner J, Wickman E, DeRenzo C, Gottschalk S. CAR T Cell Therapy for Solid Tumors: Bright Future or Dark Reality? Mol Ther. 2020;28(11):2320–39. 10.1016/j.ymthe.2020.09.015. 32979309 10.1016/j.ymthe.2020.09.015 PMC7647674

146. Simon S Bugos G Salter AI Riddell SR Synthetic receptors for logic gated T cell recognition and function Curr Opin Immunol 2022 74 9 17 10.1016/j.coi.2021.09.003 34571290 PMC8901444 Simon S, Bugos G, Salter AI, Riddell SR. Synthetic receptors for logic gated T cell recognition and function. Curr Opin Immunol. 2022;74:9–17. 10.1016/j.coi.2021.09.003. 34571290 10.1016/j.coi.2021.09.003 PMC8901444

147. Zhu B Yin H Di Zhang, Zhang M Chao X Scimeca L Synthetic biology approaches for improving the specificity and efficacy of cancer immunotherapy Cell Mol Immunol 2024 21 5 436 47 10.1038/s41423-024-01153-x 38605087 PMC11061174 Zhu B, Yin H, Di Zhang, Zhang M, Chao X, Scimeca L, et al. Synthetic biology approaches for improving the specificity and efficacy of cancer immunotherapy. Cell Mol Immunol. 2024;21(5):436–47. 10.1038/s41423-024-01153-x. 38605087 10.1038/s41423-024-01153-x PMC11061174

148. Stucchi S, Borea R, Garcia-Recio S, Zingarelli M, Rädler PD, Camerini E et al. B7-H3 and CSPG4 co-targeting as Pan-CAR-T cell treatment of triple-negative breast cancer. J Immunother Cancer. 2025;13(5):e011533. 10.1136/jitc-2025-011533. 10.1136/jitc-2025-011533 PMC12107568 40425233

149. Williams JZ Allen GM Shah D Sterin IS Kim KH Garcia VP Precise T cell recognition programs designed by transcriptionally linking multiple receptors Science 2020 370 6520 1099 104 10.1126/science.abc6270 33243890 PMC8054651 Williams JZ, Allen GM, Shah D, Sterin IS, Kim KH, Garcia VP, et al. Precise T cell recognition programs designed by transcriptionally linking multiple receptors. Science. 2020;370(6520):1099–104. 10.1126/science.abc6270. 33243890 10.1126/science.abc6270 PMC8054651

150. Hyrenius-Wittsten A, Su Y, Park M, Garcia JM, Alavi J, Perry N et al. SynNotch CAR circuits enhance solid tumor recognition and promote persistent antitumor activity in mouse models. Sci Transl Med. 2021;13(591):eabd8836. 10.1126/scitranslmed.abd8836. 10.1126/scitranslmed.abd8836 PMC8594452 33910981

151. Andreou T Neophytou C Kalli M Mpekris F Stylianopoulos T Breaking barriers: enhancing CAR-armored T cell therapy for solid tumors through microenvironment remodeling Front Immunol 2025 16 1638186 10.3389/fimmu.2025.1638186 40969762 PMC12441034 Andreou T, Neophytou C, Kalli M, Mpekris F, Stylianopoulos T. Breaking barriers: enhancing CAR-armored T cell therapy for solid tumors through microenvironment remodeling. Front Immunol. 2025;16:1638186. 10.3389/fimmu.2025.1638186. 40969762 10.3389/fimmu.2025.1638186 PMC12441034

152. Kankeu Fonkoua LA Sirpilla O Sakemura R Siegler EL Kenderian SS CAR T cell therapy and the tumor microenvironment: Current challenges and opportunities Mol Ther Oncolytics 2022 25 69 77 10.1016/j.omto.2022.03.009 35434273 PMC8980704 Kankeu Fonkoua LA, Sirpilla O, Sakemura R, Siegler EL, Kenderian SS. CAR T cell therapy and the tumor microenvironment: Current challenges and opportunities. Mol Ther Oncolytics. 2022;25:69–77. 10.1016/j.omto.2022.03.009. 35434273 10.1016/j.omto.2022.03.009 PMC8980704

153. Murad JP, Christian L, Rosa R, Ren Y, Buckley AJ, Lee EHJ, et al. Solid tumour CAR-T cells engineered with fusion proteins targeting PD-L1 for localized IL-12 delivery. Nat Biomed Eng. 2025;10(4):613–29. 10.1038/s41551-025-01509-2. 10.1038/s41551-025-01509-2 PMC13099387 41034514

154. Kessel C, Rossig C, Abken H. Weal and woe of interleukin-18 in the T cell therapy of cancer. J Immunother Cancer. 2025;13(1):e010545. 10.1136/jitc-2024-010545. 10.1136/jitc-2024-010545 PMC11749219 39809514

155. Rafii S, Mukherji D, Komaranchath AS, Khalil C, Iqbal F, Abdelwahab SI, et al. Advancing CAR T-Cell Therapy in Solid Tumors: Current Landscape and Future Directions. Cancers (Basel). 2025;17(17):2898. 10.3390/cancers17172898. 10.3390/cancers17172898 PMC12428560 40940995

156. Chmielewski M, Abken H. TRUCKS, the fourth-generation CAR T cells: Current developments and clinical translation. Adv Cell Gene Ther 2020;3(3).

157. Svoboda J, Landsburg DJ, Nasta SD, Barta SK, Chong EA, Lariviere MJ et al. Safety and efficacy of armored huCART19-IL18 in patients with relapsed/refractory lymphomas that progressed after anti-CD19 CAR T cells. JCO 2024;42(16_suppl):7004.

158. Giuffrida L Sek K Henderson MA Lai J Chen AXY Meyran D CRISPR/Cas9 mediated deletion of the adenosine A2A receptor enhances CAR T cell efficacy Nat Commun 2021 12 1 3236 10.1038/s41467-021-23331-5 34050151 PMC8163771 Giuffrida L, Sek K, Henderson MA, Lai J, Chen AXY, Meyran D, et al. CRISPR/Cas9 mediated deletion of the adenosine A2A receptor enhances CAR T cell efficacy. Nat Commun. 2021;12(1):3236. 10.1038/s41467-021-23331-5. 34050151 10.1038/s41467-021-23331-5 PMC8163771

159. Beavis PA Henderson MA Giuffrida L Mills JK Sek K Cross RS Targeting the adenosine 2A receptor enhances chimeric antigen receptor T cell efficacy J Clin Invest 2017 127 3 929 41 10.1172/JCI89455 28165340 PMC5330718 Beavis PA, Henderson MA, Giuffrida L, Mills JK, Sek K, Cross RS, et al. Targeting the adenosine 2A receptor enhances chimeric antigen receptor T cell efficacy. J Clin Invest. 2017;127(3):929–41. 10.1172/JCI89455. 28165340 10.1172/JCI89455 PMC5330718

160. Masoumi E Jafarzadeh L Mirzaei HR Alishah K Fallah-Mehrjardi K Rostamian H Genetic and pharmacological targeting of A2a receptor improves function of anti-mesothelin CAR T cells J Exp Clin Cancer Res 2020 39 1 49 10.1186/s13046-020-01546-6 32151275 PMC7063771 Masoumi E, Jafarzadeh L, Mirzaei HR, Alishah K, Fallah-Mehrjardi K, Rostamian H, et al. Genetic and pharmacological targeting of A2a receptor improves function of anti-mesothelin CAR T cells. J Exp Clin Cancer Res. 2020;39(1):49. 10.1186/s13046-020-01546-6. 32151275 10.1186/s13046-020-01546-6 PMC7063771

161. Li K Xu J Wang J Lu C Dai Y Dai Q Dominant-negative transforming growth factor-β receptor-armoured mesothelin-targeted chimeric antigen receptor T cells slow tumour growth in a mouse model of ovarian cancer Cancer Immunol Immunother 2023 72 4 917 28 10.1007/s00262-022-03290-6 36166071 PMC10025183 Li K, Xu J, Wang J, Lu C, Dai Y, Dai Q, et al. Dominant-negative transforming growth factor-β receptor-armoured mesothelin-targeted chimeric antigen receptor T cells slow tumour growth in a mouse model of ovarian cancer. Cancer Immunol Immunother. 2023;72(4):917–28. 10.1007/s00262-022-03290-6. 36166071 10.1007/s00262-022-03290-6 PMC10025183

162. Zhu S Hu J Lin J Wang C Wang E Co-Expression of Dominant-Negative TGF-β Receptor 2 Enhances the Therapeutic Efficacy of Novel TREM1/DAP12-BB-Based CAR-T Cells in Solid Tumours Immunology 2025 174 3 310 21 10.1111/imm.13888 39746895 Zhu S, Hu J, Lin J, Wang C, Wang E. Co-Expression of Dominant-Negative TGF-β Receptor 2 Enhances the Therapeutic Efficacy of Novel TREM1/DAP12-BB-Based CAR-T Cells in Solid Tumours. Immunology. 2025;174(3):310–21. 10.1111/imm.13888. 39746895 10.1111/imm.13888

163. Whilding LM, Halim L, Draper B, Parente-Pereira AC, Zabinski T, Davies DM, et al. CAR T-Cells Targeting the Integrin αvβ6 and Co-Expressing the Chemokine Receptor CXCR2 Demonstrate Enhanced Homing and Efficacy against Several Solid Malignancies. Cancers (Basel). 2019;11(5):674. 10.3390/cancers11050674. 10.3390/cancers11050674 PMC6563120 31091832

164. Moon EK Carpenito C Sun J Wang L-CS Kapoor V Predina J Expression of a functional CCR2 receptor enhances tumor localization and tumor eradication by retargeted human T cells expressing a mesothelin-specific chimeric antibody receptor Clin Cancer Res 2011 17 14 4719 30 10.1158/1078-0432.CCR-11-0351 21610146 PMC3612507 Moon EK, Carpenito C, Sun J, Wang L-CS, Kapoor V, Predina J, et al. Expression of a functional CCR2 receptor enhances tumor localization and tumor eradication by retargeted human T cells expressing a mesothelin-specific chimeric antibody receptor. Clin Cancer Res. 2011;17(14):4719–30. 10.1158/1078-0432.CCR-11-0351. 21610146 10.1158/1078-0432.CCR-11-0351 PMC3612507

165. Wang Z Li P Zeng X Guo J Zhang C Fan Z CAR-T therapy dilemma and innovative design strategies for next generation Cell Death Dis 2025 16 1 211 10.1038/s41419-025-07454-x 40148310 PMC11950394 Wang Z, Li P, Zeng X, Guo J, Zhang C, Fan Z, et al. CAR-T therapy dilemma and innovative design strategies for next generation. Cell Death Dis. 2025;16(1):211. 10.1038/s41419-025-07454-x. 40148310 10.1038/s41419-025-07454-x PMC11950394

166. Liu G Rui W Zheng H Huang D Yu F Zhang Y CXCR2-modified CAR-T cells have enhanced trafficking ability that improves treatment of hepatocellular carcinoma Eur J Immunol 2020 50 5 712 24 10.1002/eji.201948457 31981231 Liu G, Rui W, Zheng H, Huang D, Yu F, Zhang Y, et al. CXCR2-modified CAR-T cells have enhanced trafficking ability that improves treatment of hepatocellular carcinoma. Eur J Immunol. 2020;50(5):712–24. 10.1002/eji.201948457. 31981231 10.1002/eji.201948457

167. Vogt KC Silberman PC Lin Q Han JE Laflin A Gellineau HA Microenvironment actuated CAR T cells improve solid tumor efficacy without toxicity Sci Adv 2025 11 4 eads3403 10.1126/sciadv.ads3403 39841845 PMC11753401 Vogt KC, Silberman PC, Lin Q, Han JE, Laflin A, Gellineau HA, et al. Microenvironment actuated CAR T cells improve solid tumor efficacy without toxicity. Sci Adv. 2025;11(4):eads3403. 10.1126/sciadv.ads3403. 39841845 10.1126/sciadv.ads3403 PMC11753401

168. Juillerat A Marechal A Filhol JM Valogne Y Valton J Duclert A An oxygen sensitive self-decision making engineered CAR T-cell Sci Rep 2017 7 39833 10.1038/srep39833 28106050 PMC5247770 Juillerat A, Marechal A, Filhol JM, Valogne Y, Valton J, Duclert A, et al. An oxygen sensitive self-decision making engineered CAR T-cell. Sci Rep. 2017;7:39833. 10.1038/srep39833. 28106050 10.1038/srep39833 PMC5247770

169. Kosti P Opzoomer JW Larios-Martinez KI Henley-Smith R Scudamore CL Okesola M Hypoxia-sensing CAR T cells provide safety and efficacy in treating solid tumors Cell Rep Med 2021 2 4 100227 10.1016/j.xcrm.2021.100227 33948568 PMC8080111 Kosti P, Opzoomer JW, Larios-Martinez KI, Henley-Smith R, Scudamore CL, Okesola M, et al. Hypoxia-sensing CAR T cells provide safety and efficacy in treating solid tumors. Cell Rep Med. 2021;2(4):100227. 10.1016/j.xcrm.2021.100227. 33948568 10.1016/j.xcrm.2021.100227 PMC8080111

170. Vitanza NA Wilson AL Huang W Seidel K Brown C Gustafson JA Intraventricular B7-H3 CAR T Cells for Diffuse Intrinsic Pontine Glioma: Preliminary First-in-Human Bioactivity and Safety Cancer Discov 2023 13 1 114 31 10.1158/2159-8290.CD-22-0750 36259971 PMC9827115 Vitanza NA, Wilson AL, Huang W, Seidel K, Brown C, Gustafson JA, et al. Intraventricular B7-H3 CAR T Cells for Diffuse Intrinsic Pontine Glioma: Preliminary First-in-Human Bioactivity and Safety. Cancer Discov. 2023;13(1):114–31. 10.1158/2159-8290.CD-22-0750. 36259971 10.1158/2159-8290.CD-22-0750 PMC9827115

171. Guercio M Manni S Boffa I Caruso S Di Cecca S Sinibaldi M Inclusion of the Inducible Caspase 9 Suicide Gene in CAR Construct Increases Safety of CAR.CD19 T Cell Therapy in B-Cell Malignancies Front Immunol 2021 12 755639 10.3389/fimmu.2021.755639 34737753 PMC8560965 Guercio M, Manni S, Boffa I, Caruso S, Di Cecca S, Sinibaldi M, et al. Inclusion of the Inducible Caspase 9 Suicide Gene in CAR Construct Increases Safety of CAR.CD19 T Cell Therapy in B-Cell Malignancies. Front Immunol. 2021;12:755639. 10.3389/fimmu.2021.755639. 34737753 10.3389/fimmu.2021.755639 PMC8560965

172. Straathof KC Pulè MA Yotnda P Dotti G Vanin EF Brenner MK An inducible caspase 9 safety switch for T-cell therapy Blood 2005 105 11 4247 54 10.1182/blood-2004-11-4564 15728125 PMC1895037 Straathof KC, Pulè MA, Yotnda P, Dotti G, Vanin EF, Brenner MK, et al. An inducible caspase 9 safety switch for T-cell therapy. Blood. 2005;105(11):4247–54. 10.1182/blood-2004-11-4564. 15728125 10.1182/blood-2004-11-4564 PMC1895037

173. Gargett T Brown MP The inducible caspase-9 suicide gene system as a safety switch to limit on-target, off-tumor toxicities of chimeric antigen receptor T cells Front Pharmacol 2014 5 235 10.3389/fphar.2014.00235 25389405 PMC4211380 Gargett T, Brown MP. The inducible caspase-9 suicide gene system as a safety switch to limit on-target, off-tumor toxicities of chimeric antigen receptor T cells. Front Pharmacol. 2014;5:235. 10.3389/fphar.2014.00235. 25389405 10.3389/fphar.2014.00235 PMC4211380

174. Lu L Xie M Yang B Zhao W-B Cao J Enhancing the safety of CAR-T cell therapy: Synthetic genetic switch for spatiotemporal control Sci Adv 2024 10 8 eadj6251 10.1126/sciadv.adj6251 38394207 PMC10889354 Lu L, Xie M, Yang B, Zhao W-B, Cao J. Enhancing the safety of CAR-T cell therapy: Synthetic genetic switch for spatiotemporal control. Sci Adv. 2024;10(8):eadj6251. 10.1126/sciadv.adj6251. 38394207 10.1126/sciadv.adj6251 PMC10889354

175. Russell GC Hamzaoui Y Rho D Sutrave G Choi JS Missan DS Synthetic biology approaches for enhancing safety and specificity of CAR-T cell therapies for solid cancers Cytotherapy 2024 26 8 842 57 10.1016/j.jcyt.2024.03.484 38639669 Russell GC, Hamzaoui Y, Rho D, Sutrave G, Choi JS, Missan DS, et al. Synthetic biology approaches for enhancing safety and specificity of CAR-T cell therapies for solid cancers. Cytotherapy. 2024;26(8):842–57. 10.1016/j.jcyt.2024.03.484. 38639669 10.1016/j.jcyt.2024.03.484

176. Ventin M Cattaneo G Arya S Jia J Gelmi MC Sun Y Chimeric Antigen Receptor T Cell with an Inducible Caspase-9 Suicide Gene Eradicates Uveal Melanoma Liver Metastases via B7-H3 Targeting Clin Cancer Res 2024 30 15 3243 58 10.1158/1078-0432.CCR-24-0071 38767611 PMC11572477 Ventin M, Cattaneo G, Arya S, Jia J, Gelmi MC, Sun Y, et al. Chimeric Antigen Receptor T Cell with an Inducible Caspase-9 Suicide Gene Eradicates Uveal Melanoma Liver Metastases via B7-H3 Targeting. Clin Cancer Res. 2024;30(15):3243–58. 10.1158/1078-0432.CCR-24-0071. 38767611 10.1158/1078-0432.CCR-24-0071 PMC11572477

177. Jiao C Zvonkov E Lai X Zhang R Liu Y Qin Y SCAR2.0: a multi-CAR-T therapy regimen for the treatment of relapsed/refractory B cell lymphomas Blood Cancer J 2021 4 3 59 10.1038/s41408-021-00455-x PMC7966809 33727525 Jiao C, Zvonkov E, Lai X, Zhang R, Liu Y, Qin Y, et al. SCAR2.0: a multi-CAR-T therapy regimen for the treatment of relapsed/refractory B cell lymphomas. Blood Cancer J. 2021;4(3):59. 10.1038/s41408-021-00455-x. 10.1038/s41408-021-00455-x PMC7966809 33727525

178. Xu X Zhao W Yue Z Qin M Jin M Chang L-J 4SCAR-GD2-modified T-cell therapy in neuroblastoma with MYCN amplification: A case report with over 4-year follow-up data Pediatr Investig 2020 4 1 55 8 10.1002/ped4.12181 32851343 PMC7331334 Xu X, Zhao W, Yue Z, Qin M, Jin M, Chang L-J, et al. 4SCAR-GD2-modified T-cell therapy in neuroblastoma with MYCN amplification: A case report with over 4-year follow-up data. Pediatr Investig. 2020;4(1):55–8. 10.1002/ped4.12181. 32851343 10.1002/ped4.12181 PMC7331334

179. Liu Z Zhou J Yang X Liu Y Zou C Lv W Safety and antitumor activity of GD2-Specific 4SCAR-T cells in patients with glioblastoma Mol Cancer 2023 22 1 3 10.1186/s12943-022-01711-9 36617554 PMC9827625 Liu Z, Zhou J, Yang X, Liu Y, Zou C, Lv W, et al. Safety and antitumor activity of GD2-Specific 4SCAR-T cells in patients with glioblastoma. Mol Cancer. 2023;22(1):3. 10.1186/s12943-022-01711-9. 36617554 10.1186/s12943-022-01711-9 PMC9827625

180. Shafer P Kelly LM Hoyos V Cancer Therapy With TCR-Engineered T Cells: Current Strategies, Challenges, and Prospects Front Immunol 2022 13 835762 10.3389/fimmu.2022.835762 35309357 PMC8928448 Shafer P, Kelly LM, Hoyos V. Cancer Therapy With TCR-Engineered T Cells: Current Strategies, Challenges, and Prospects. Front Immunol. 2022;13:835762. 10.3389/fimmu.2022.835762. 35309357 10.3389/fimmu.2022.835762 PMC8928448

181. Dan L Kang-Zheng L Optimizing viral transduction in immune cell therapy manufacturing: key process design considerations J Transl Med 2025 23 1 501 10.1186/s12967-025-06524-0 40316943 PMC12046913 Dan L, Kang-Zheng L. Optimizing viral transduction in immune cell therapy manufacturing: key process design considerations. J Transl Med. 2025;23(1):501. 10.1186/s12967-025-06524-0. 40316943 10.1186/s12967-025-06524-0 PMC12046913

182. Baulu E Gardet C Chuvin N Depil S TCR-engineered T cell therapy in solid tumors: State of the art and perspectives Sci Adv 2023 9 7 eadf3700 10.1126/sciadv.adf3700 36791198 PMC9931212 Baulu E, Gardet C, Chuvin N, Depil S. TCR-engineered T cell therapy in solid tumors: State of the art and perspectives. Sci Adv. 2023;9(7):eadf3700. 10.1126/sciadv.adf3700. 36791198 10.1126/sciadv.adf3700 PMC9931212

183. Stadtmauer EA Faitg TH Lowther DE Badros AZ Chagin K Dengel K Long-term safety and activity of NY-ESO-1 SPEAR T cells after autologous stem cell transplant for myeloma Blood Adv 2019 3 13 2022 34 10.1182/bloodadvances.2019000194 31289029 PMC6616263 Stadtmauer EA, Faitg TH, Lowther DE, Badros AZ, Chagin K, Dengel K, et al. Long-term safety and activity of NY-ESO-1 SPEAR T cells after autologous stem cell transplant for myeloma. Blood Adv. 2019;3(13):2022–34. 10.1182/bloodadvances.2019000194. 31289029 10.1182/bloodadvances.2019000194 PMC6616263

184. Tao Z Chyra Z Kotulová J Celichowski P Mihályová J Charvátová S Impact of T cell characteristics on CAR-T cell therapy in hematological malignancies Blood Cancer J 2024 14 1 213 10.1038/s41408-024-01193-6 39627220 PMC11615218 Tao Z, Chyra Z, Kotulová J, Celichowski P, Mihályová J, Charvátová S, et al. Impact of T cell characteristics on CAR-T cell therapy in hematological malignancies. Blood Cancer J. 2024;14(1):213. 10.1038/s41408-024-01193-6. 39627220 10.1038/s41408-024-01193-6 PMC11615218

185. Ayala Ceja M, Khericha M, Harris CM, Puig-Saus C, Chen YY. CAR-T cell manufacturing: Major process parameters and next-generation strategies. J Exp Med. 2024;221(2):e20230903. 10.1084/jem.20230903 10.1084/jem.20230903 PMC10791545 38226974

186. Robbins PF Morgan RA Feldman SA Yang JC Sherry RM Dudley ME Tumor regression in patients with metastatic synovial cell sarcoma and melanoma using genetically engineered lymphocytes reactive with NY-ESO-1 JCO 2011 29 7 917 24 10.1200/JCO.2010.32.2537 PMC3068063 21282551 Robbins PF, Morgan RA, Feldman SA, Yang JC, Sherry RM, Dudley ME, et al. Tumor regression in patients with metastatic synovial cell sarcoma and melanoma using genetically engineered lymphocytes reactive with NY-ESO-1. JCO. 2011;29(7):917–24. 10.1200/JCO.2010.32.2537. 10.1200/JCO.2010.32.2537 PMC3068063 21282551

187. Rohaan MW, Gomez-Eerland R, van den Berg JH, Geukes Foppen MH, van Zon M, Raud B et al. MART-1 TCR gene-modified peripheral blood T cells for the treatment of metastatic melanoma: a phase I/IIa clinical trial. Immunooncol Technol. 2022;15:100089. Available from: URL: https://pubmed.ncbi.nlm.nih.gov/35865122/ . 10.1016/j.iotech.2022.100089 PMC9293760 35865122

188. Lezcano C Jungbluth AA Nehal KS Hollmann TJ Busam KJ PRAME Expression in Melanocytic Tumors Am J Surg Pathol 2018 42 11 1456 65 10.1097/PAS.0000000000001134 30045064 PMC6631376 Lezcano C, Jungbluth AA, Nehal KS, Hollmann TJ, Busam KJ. PRAME Expression in Melanocytic Tumors. Am J Surg Pathol. 2018;42(11):1456–65. 10.1097/PAS.0000000000001134. 30045064 10.1097/PAS.0000000000001134 PMC6631376

189. Wermke M Araujo DM Chatterjee M Tsimberidou AM Holderried TAW Jazaeri AA Autologous T cell therapy for PRAME+ advanced solid tumors in HLA-A*02 + patients: a phase 1 trial Nat Med 2025 31 7 2365 74 10.1038/s41591-025-03650-6 40205198 PMC12283372 Wermke M, Araujo DM, Chatterjee M, Tsimberidou AM, Holderried TAW, Jazaeri AA, et al. Autologous T cell therapy for PRAME+ advanced solid tumors in HLA-A*02 + patients: a phase 1 trial. Nat Med. 2025;31(7):2365–74. 10.1038/s41591-025-03650-6. 40205198 10.1038/s41591-025-03650-6 PMC12283372

190. Wermke M Alsdorf W Araujo DM Busse A Chatterjee M Hernandez-Aya LF Phase 1 clinical update of IMA203, an autologous TCR-T targeting PRAME in patients with PD1 refractory metastatic melanoma JCO 2025 43 16suppl 2508 10.1200/JCO.2025.43.16_suppl.2508 Wermke M, Alsdorf W, Araujo DM, Busse A, Chatterjee M, Hernandez-Aya LF, et al. Phase 1 clinical update of IMA203, an autologous TCR-T targeting PRAME in patients with PD1 refractory metastatic melanoma. JCO. 2025;43(16suppl):2508.

191. Luke JJ, Betof Warner A, Chmielowski B, Diab A, Gebhardt C, Hernandez-Aya LF et al. SUPRAME: A phase 3 trial comparing IMA203, an engineered T-cell receptor expressing T cell therapy (TCR-T) vs investigator’s choice in patients with previously treated advanced cutaneous melanoma. JCO 2025;43(16_suppl).

192. Linette GP Stadtmauer EA Maus MV Rapoport AP Levine BL Emery L Cardiovascular toxicity and titin cross-reactivity of affinity-enhanced T cells in myeloma and melanoma Blood 2013 122 6 863 71 10.1182/blood-2013-03-490565 23770775 PMC3743463 Linette GP, Stadtmauer EA, Maus MV, Rapoport AP, Levine BL, Emery L, et al. Cardiovascular toxicity and titin cross-reactivity of affinity-enhanced T cells in myeloma and melanoma. Blood. 2013;122(6):863–71. 10.1182/blood-2013-03-490565. 23770775 10.1182/blood-2013-03-490565 PMC3743463

193. Raman MCC Rizkallah PJ Simmons R Donnellan Z Dukes J Bossi G Direct molecular mimicry enables off-target cardiovascular toxicity by an enhanced affinity TCR designed for cancer immunotherapy Sci Rep 2016 6 18851 10.1038/srep18851 26758806 PMC4725365 Raman MCC, Rizkallah PJ, Simmons R, Donnellan Z, Dukes J, Bossi G, et al. Direct molecular mimicry enables off-target cardiovascular toxicity by an enhanced affinity TCR designed for cancer immunotherapy. Sci Rep. 2016;6:18851. 10.1038/srep18851. 26758806 10.1038/srep18851 PMC4725365

194. Golikova EA Alshevskaya AA Alrhmoun S Sivitskaya NA Sennikov SV TCR-T cell therapy: current development approaches, preclinical evaluation, and perspectives on regulatory challenges J Transl Med 2024 22 1 897 10.1186/s12967-024-05703-9 39367419 PMC11451006 Golikova EA, Alshevskaya AA, Alrhmoun S, Sivitskaya NA, Sennikov SV. TCR-T cell therapy: current development approaches, preclinical evaluation, and perspectives on regulatory challenges. J Transl Med. 2024;22(1):897. 10.1186/s12967-024-05703-9. 39367419 10.1186/s12967-024-05703-9 PMC11451006

195. Sun Q, Zhang X, Wang L, Gao X, Xiong Y, Liu L et al. T-cell receptor gene therapy targeting melanoma-associated antigen-A4 by silencing of endogenous TCR inhibits tumor growth in mice and human. Cell Death Dis. 2019;10(7):475. Available from: URL: https://pubmed.ncbi.nlm.nih.gov/31209257/ . 10.1038/s41419-019-1717-8 PMC6572850 31209257

196. Wei F Cheng X-X Xue JZ Xue S-A Emerging Strategies in TCR-Engineered T Cells Front Immunol 2022 13 850358 10.3389/fimmu.2022.850358 35432319 PMC9006933 Wei F, Cheng X-X, Xue JZ, Xue S-A. Emerging Strategies in TCR-Engineered T Cells. Front Immunol. 2022;13:850358. 10.3389/fimmu.2022.850358. 35432319 10.3389/fimmu.2022.850358 PMC9006933

197. Ellis GI Sheppard NC Riley JL Genetic engineering of T cells for immunotherapy Nat Rev Genet 2021 22 7 427 47 10.1038/s41576-021-00329-9 33603158 PMC8217325 Ellis GI, Sheppard NC, Riley JL. Genetic engineering of T cells for immunotherapy. Nat Rev Genet. 2021;22(7):427–47. 10.1038/s41576-021-00329-9. 33603158 10.1038/s41576-021-00329-9 PMC8217325

198. Robbins PF Kassim SH Tran TLN Crystal JS Morgan RA Feldman SA A pilot trial using lymphocytes genetically engineered with an NY-ESO-1-reactive T-cell receptor: long-term follow-up and correlates with response Clin Cancer Res 2015 21 5 1019 27 10.1158/1078-0432.CCR-14-2708 25538264 PMC4361810 Robbins PF, Kassim SH, Tran TLN, Crystal JS, Morgan RA, Feldman SA, et al. A pilot trial using lymphocytes genetically engineered with an NY-ESO-1-reactive T-cell receptor: long-term follow-up and correlates with response. Clin Cancer Res. 2015;21(5):1019–27. 10.1158/1078-0432.CCR-14-2708. 25538264 10.1158/1078-0432.CCR-14-2708 PMC4361810

199. Ma S Yu J Caligiuri MA Natural killer cell-based immunotherapy for cancer J Immunol 2025 214 7 1444 56 10.1093/jimmun/vkaf036 40246292 PMC12311385 Ma S, Yu J, Caligiuri MA. Natural killer cell-based immunotherapy for cancer. J Immunol. 2025;214(7):1444–56. 10.1093/jimmun/vkaf036. 40246292 10.1093/jimmun/vkaf036 PMC12311385

200. Tarazona R Duran E Solana R Natural Killer Cell Recognition of Melanoma: New Clues for a More Effective Immunotherapy Front Immunol 2015 6 649 26779186 10.3389/fimmu.2015.00649 PMC4703774 Tarazona R, Duran E, Solana R. Natural Killer Cell Recognition of Melanoma: New Clues for a More Effective Immunotherapy. Front Immunol. 2015;6:649. 10.3389/fimmu.2015.00649. 26779186 10.3389/fimmu.2015.00649 PMC4703774

201. Prager I Watzl C Mechanisms of natural killer cell-mediated cellular cytotoxicity J Leukoc Biol 2019 105 6 1319 29 10.1002/JLB.MR0718-269R 31107565 Prager I, Watzl C. Mechanisms of natural killer cell-mediated cellular cytotoxicity. J Leukoc Biol. 2019;105(6):1319–29. 10.1002/JLB.MR0718-269R. 31107565 10.1002/JLB.MR0718-269R

202. Casado JG Pawelec G Morgado S Sanchez-Correa B Delgado E Gayoso I Expression of adhesion molecules and ligands for activating and costimulatory receptors involved in cell-mediated cytotoxicity in a large panel of human melanoma cell lines Cancer Immunol Immunother 2009 58 9 1517 26 10.1007/s00262-009-0682-y 19259667 PMC11030684 Casado JG, Pawelec G, Morgado S, Sanchez-Correa B, Delgado E, Gayoso I, et al. Expression of adhesion molecules and ligands for activating and costimulatory receptors involved in cell-mediated cytotoxicity in a large panel of human melanoma cell lines. Cancer Immunol Immunother. 2009;58(9):1517–26. 10.1007/s00262-009-0682-y. 19259667 10.1007/s00262-009-0682-y PMC11030684

203. Chen S, Zhu H, Jounaidi Y. Comprehensive snapshots of natural killer cells functions, signaling, molecular mechanisms and clinical utilization. Signal Transduct Target Ther. 2024;9(1):302. Available from: URL: https://pubmed.ncbi.nlm.nih.gov/39511139/ . 10.1038/s41392-024-02005-w PMC11544004 39511139

204. Netskar H Pfefferle A Goodridge JP Sohlberg E Dufva O Teichmann SA Pan-cancer profiling of tumor-infiltrating natural killer cells through transcriptional reference mapping Nat Immunol 2024 25 8 1445 59 10.1038/s41590-024-01884-z 38956379 PMC11291284 Netskar H, Pfefferle A, Goodridge JP, Sohlberg E, Dufva O, Teichmann SA, et al. Pan-cancer profiling of tumor-infiltrating natural killer cells through transcriptional reference mapping. Nat Immunol. 2024;25(8):1445–59. 10.1038/s41590-024-01884-z. 38956379 10.1038/s41590-024-01884-z PMC11291284

205. Lee H Quek C Silva I Tasker A Batten M Rizos H Integrated molecular and immunophenotypic analysis of NK cells in anti-PD-1 treated metastatic melanoma patients OncoImmunology 2019 8 2 e1537581 10.1080/2162402X.2018.1537581 30713793 PMC6343795 Lee H, Quek C, Silva I, Tasker A, Batten M, Rizos H, et al. Integrated molecular and immunophenotypic analysis of NK cells in anti-PD-1 treated metastatic melanoma patients. OncoImmunology. 2019;8(2):e1537581. 10.1080/2162402X.2018.1537581. 30713793 10.1080/2162402X.2018.1537581 PMC6343795

206. Lee H, Da Silva IP, Palendira U, Scolyer RA, Long GV, Wilmott JS. Targeting NK Cells to Enhance Melanoma Response to Immunotherapies. Cancers (Basel). 2021;13(6):1363. 10.3390/cancers13061363. 10.3390/cancers13061363 PMC8002669 33802954

207. Pietra G, Manzini C, Rivara S, Vitale M, Cantoni C, Petretto A et al. Melanoma cells inhibit natural killer cell function by modulating the expression of activating receptors and cytolytic activity. Cancer Res. 2012; 72(6):1407–15. Available from: URL: https://pubmed.ncbi.nlm.nih.gov/22258454/ . 10.1158/0008-5472.CAN-11-2544 22258454

208. Hasmim M Messai Y Ziani L Thiery J Bouhris J-H Noman MZ Critical Role of Tumor Microenvironment in Shaping NK Cell Functions: Implication of Hypoxic Stress Front Immunol 2015 6 482 10.3389/fimmu.2015.00482 26441986 PMC4585210 Hasmim M, Messai Y, Ziani L, Thiery J, Bouhris J-H, Noman MZ, et al. Critical Role of Tumor Microenvironment in Shaping NK Cell Functions: Implication of Hypoxic Stress. Front Immunol. 2015;6:482. 10.3389/fimmu.2015.00482. 26441986 10.3389/fimmu.2015.00482 PMC4585210

209. Miao L Lu C Zhang B Li H Zhao X Chen H Advances in metabolic reprogramming of NK cells in the tumor microenvironment on the impact of NK therapy J Transl Med 2024 22 1 229 10.1186/s12967-024-05033-w 38433193 PMC10909296 Miao L, Lu C, Zhang B, Li H, Zhao X, Chen H, et al. Advances in metabolic reprogramming of NK cells in the tumor microenvironment on the impact of NK therapy. J Transl Med. 2024;22(1):229. 10.1186/s12967-024-05033-w. 38433193 10.1186/s12967-024-05033-w PMC10909296

210. Brand A Singer K Koehl GE Kolitzus M Schoenhammer G Thiel A LDHA-Associated Lactic Acid Production Blunts Tumor Immunosurveillance by T and NK Cells Cell Metab 2016 24 5 657 71 10.1016/j.cmet.2016.08.011 27641098 Brand A, Singer K, Koehl GE, Kolitzus M, Schoenhammer G, Thiel A, et al. LDHA-Associated Lactic Acid Production Blunts Tumor Immunosurveillance by T and NK Cells. Cell Metab. 2016;24(5):657–71. 10.1016/j.cmet.2016.08.011. 27641098 10.1016/j.cmet.2016.08.011

211. Bruno A Mortara L Baci D Noonan DM Albini A Myeloid Derived Suppressor Cells Interactions With Natural Killer Cells and Pro-angiogenic Activities: Roles in Tumor Progression Front Immunol 2019 10 771 10.3389/fimmu.2019.00771 31057536 PMC6482162 Bruno A, Mortara L, Baci D, Noonan DM, Albini A. Myeloid Derived Suppressor Cells Interactions With Natural Killer Cells and Pro-angiogenic Activities: Roles in Tumor Progression. Front Immunol. 2019;10:771. 10.3389/fimmu.2019.00771. 31057536 10.3389/fimmu.2019.00771 PMC6482162

212. Ferrari de Andrade L Ngiow SF Stannard K Rusakiewicz S Kalimutho M Khanna KK Natural killer cells are essential for the ability of BRAF inhibitors to control BRAFV600E-mutant metastatic melanoma Cancer Res 2014 74 24 7298 308 10.1158/0008-5472.CAN-14-1339 25351955 Ferrari de Andrade L, Ngiow SF, Stannard K, Rusakiewicz S, Kalimutho M, Khanna KK, et al. Natural killer cells are essential for the ability of BRAF inhibitors to control BRAFV600E-mutant metastatic melanoma. Cancer Res. 2014;74(24):7298–308. 10.1158/0008-5472.CAN-14-1339. 25351955 10.1158/0008-5472.CAN-14-1339

213. Hsu J Hodgins JJ Marathe M Nicolai CJ Bourgeois-Daigneault M-C Trevino TN Contribution of NK cells to immunotherapy mediated by PD-1/PD-L1 blockade J Clin Invest 2018 128 10 4654 68 10.1172/JCI99317 30198904 PMC6159991 Hsu J, Hodgins JJ, Marathe M, Nicolai CJ, Bourgeois-Daigneault M-C, Trevino TN, et al. Contribution of NK cells to immunotherapy mediated by PD-1/PD-L1 blockade. J Clin Invest. 2018;128(10):4654–68. 10.1172/JCI99317. 30198904 10.1172/JCI99317 PMC6159991

214. Rahimi A Malakoutikhah Z Rahimmanesh I Ferns GA Nedaeinia R Ishaghi SMM The nexus of natural killer cells and melanoma tumor microenvironment: crosstalk, chemotherapeutic potential, and innovative NK cell-based therapeutic strategies Cancer Cell Int 2023 23 1 312 10.1186/s12935-023-03134-y 38057843 PMC10702008 Rahimi A, Malakoutikhah Z, Rahimmanesh I, Ferns GA, Nedaeinia R, Ishaghi SMM, et al. The nexus of natural killer cells and melanoma tumor microenvironment: crosstalk, chemotherapeutic potential, and innovative NK cell-based therapeutic strategies. Cancer Cell Int. 2023;23(1):312. 10.1186/s12935-023-03134-y. 38057843 10.1186/s12935-023-03134-y PMC10702008

215. Wang X Cui Y Luo G Wang Q Hu J He W Activated mouse CD4(+)Foxp3(-) T cells facilitate melanoma metastasis via Qa-1-dependent suppression of NK-cell cytotoxicity Cell Res 2012 22 12 1696 706 10.1038/cr.2012.128 22945357 PMC3515753 Wang X, Cui Y, Luo G, Wang Q, Hu J, He W, et al. Activated mouse CD4(+)Foxp3(-) T cells facilitate melanoma metastasis via Qa-1-dependent suppression of NK-cell cytotoxicity. Cell Res. 2012;22(12):1696–706. 10.1038/cr.2012.128. 22945357 10.1038/cr.2012.128 PMC3515753

216. van Vliet AA, Georgoudaki A-M, Raimo M, de Gruijl TD, Spanholtz J. Adoptive NK Cell Therapy: A Promising Treatment Prospect for Metastatic Melanoma. Cancers (Basel). 2021;13(18):4722. 10.3390/cancers13184722. 10.3390/cancers13184722 PMC8471577 34572949

217. Ruggeri L Capanni M Urbani E Perruccio K Shlomchik WD Tosti A Effectiveness of donor natural killer cell alloreactivity in mismatched hematopoietic transplants Science 2002 295 5562 2097 100 10.1126/science.1068440 11896281 Ruggeri L, Capanni M, Urbani E, Perruccio K, Shlomchik WD, Tosti A, et al. Effectiveness of donor natural killer cell alloreactivity in mismatched hematopoietic transplants. Science. 2002;295(5562):2097–100. 10.1126/science.1068440. 11896281 10.1126/science.1068440

218. Miller JS Soignier Y Panoskaltsis-Mortari A McNearney SA Yun GH Fautsch SK Successful adoptive transfer and in vivo expansion of human haploidentical NK cells in patients with cancer Blood 2005 105 8 3051 7 10.1182/blood-2004-07-2974 15632206 Miller JS, Soignier Y, Panoskaltsis-Mortari A, McNearney SA, Yun GH, Fautsch SK, et al. Successful adoptive transfer and in vivo expansion of human haploidentical NK cells in patients with cancer. Blood. 2005;105(8):3051–7. 10.1182/blood-2004-07-2974. 15632206 10.1182/blood-2004-07-2974

219. Zhao X Weinhold S Brands J Hejazi M Degistirici Ö Kögler G NK cell development in a human stem cell niche: KIR expression occurs independently of the presence of HLA class I ligands Blood Adv 2018 2 19 2452 61 10.1182/bloodadvances.2018019059 30266820 PMC6177642 Zhao X, Weinhold S, Brands J, Hejazi M, Degistirici Ö, Kögler G, et al. NK cell development in a human stem cell niche: KIR expression occurs independently of the presence of HLA class I ligands. Blood Adv. 2018;2(19):2452–61. 10.1182/bloodadvances.2018019059. 30266820 10.1182/bloodadvances.2018019059 PMC6177642

220. Becker PSA Suck G Nowakowska P Ullrich E Seifried E Bader P Selection and expansion of natural killer cells for NK cell-based immunotherapy Cancer Immunol Immunother 2016 65 4 477 84 10.1007/s00262-016-1792-y 26810567 PMC4826432 Becker PSA, Suck G, Nowakowska P, Ullrich E, Seifried E, Bader P, et al. Selection and expansion of natural killer cells for NK cell-based immunotherapy. Cancer Immunol Immunother. 2016;65(4):477–84. 10.1007/s00262-016-1792-y. 26810567 10.1007/s00262-016-1792-y PMC4826432

221. Spanholtz J Preijers F Tordoir M Trilsbeek C Paardekooper J de Witte T Clinical-grade generation of active NK cells from cord blood hematopoietic progenitor cells for immunotherapy using a closed-system culture process PLoS ONE 2011 6 6 e20740 10.1371/journal.pone.0020740 21698239 PMC3116834 Spanholtz J, Preijers F, Tordoir M, Trilsbeek C, Paardekooper J, de Witte T, et al. Clinical-grade generation of active NK cells from cord blood hematopoietic progenitor cells for immunotherapy using a closed-system culture process. PLoS ONE. 2011;6(6):e20740. 10.1371/journal.pone.0020740. 21698239 10.1371/journal.pone.0020740 PMC3116834

222. van Vliet AA Peters E Vodegel D Steenmans D Raimo M Gibbs S Early TRAIL-engagement elicits potent multimodal targeting of melanoma by CD34 + progenitor cell-derived NK cells iScience 2023 26 7 107078 10.1016/j.isci.2023.107078 37426355 PMC10329179 van Vliet AA, Peters E, Vodegel D, Steenmans D, Raimo M, Gibbs S, et al. Early TRAIL-engagement elicits potent multimodal targeting of melanoma by CD34 + progenitor cell-derived NK cells. iScience. 2023;26(7):107078. 10.1016/j.isci.2023.107078. 37426355 10.1016/j.isci.2023.107078 PMC10329179

223. Romee R Schneider SE Leong JW Chase JM Keppel CR Sullivan RP Cytokine activation induces human memory-like NK cells Blood 2012 120 24 4751 60 10.1182/blood-2012-04-419283 22983442 PMC3520618 Romee R, Schneider SE, Leong JW, Chase JM, Keppel CR, Sullivan RP, et al. Cytokine activation induces human memory-like NK cells. Blood. 2012;120(24):4751–60. 10.1182/blood-2012-04-419283. 22983442 10.1182/blood-2012-04-419283 PMC3520618

224. Romee R Rosario M Berrien-Elliott MM Wagner JA Jewell BA Schappe T Cytokine-induced memory-like natural killer cells exhibit enhanced responses against myeloid leukemia Sci Transl Med 2016 8 357 357ra123 10.1126/scitranslmed.aaf2341 27655849 PMC5436500 Romee R, Rosario M, Berrien-Elliott MM, Wagner JA, Jewell BA, Schappe T, et al. Cytokine-induced memory-like natural killer cells exhibit enhanced responses against myeloid leukemia. Sci Transl Med. 2016;8(357):357ra123. 10.1126/scitranslmed.aaf2341. 27655849 10.1126/scitranslmed.aaf2341 PMC5436500

225. Marin ND Krasnick BA Becker-Hapak M Conant L Goedegebuure SP Berrien-Elliott MM Memory-like Differentiation Enhances NK Cell Responses to Melanoma Clin Cancer Res 2021 27 17 4859 69 10.1158/1078-0432.CCR-21-0851 34187852 PMC8416927 Marin ND, Krasnick BA, Becker-Hapak M, Conant L, Goedegebuure SP, Berrien-Elliott MM, et al. Memory-like Differentiation Enhances NK Cell Responses to Melanoma. Clin Cancer Res. 2021;27(17):4859–69. 10.1158/1078-0432.CCR-21-0851. 34187852 10.1158/1078-0432.CCR-21-0851 PMC8416927

226. Shapiro RM Sheffer M Booker MA Tolstorukov MY Birch GC Sade-Feldman M First-in-human evaluation of memory-like NK cells with an IL-15 super-agonist and CTLA-4 blockade in advanced head and neck cancer J Hematol Oncol 2025 18 1 17 10.1186/s13045-025-01669-3 39948608 PMC11827236 Shapiro RM, Sheffer M, Booker MA, Tolstorukov MY, Birch GC, Sade-Feldman M, et al. First-in-human evaluation of memory-like NK cells with an IL-15 super-agonist and CTLA-4 blockade in advanced head and neck cancer. J Hematol Oncol. 2025;18(1):17. 10.1186/s13045-025-01669-3. 39948608 10.1186/s13045-025-01669-3 PMC11827236

227. Mantesso S Geerts D Spanholtz J Kučerová L Genetic Engineering of Natural Killer Cells for Enhanced Antitumor Function Front Immunol 2020 11 607131 10.3389/fimmu.2020.607131 33391277 PMC7772419 Mantesso S, Geerts D, Spanholtz J, Kučerová L. Genetic Engineering of Natural Killer Cells for Enhanced Antitumor Function. Front Immunol. 2020;11:607131. 10.3389/fimmu.2020.607131. 33391277 10.3389/fimmu.2020.607131 PMC7772419

228. Mehta RS Rezvani K Chimeric Antigen Receptor Expressing Natural Killer Cells for the Immunotherapy of Cancer Front Immunol 2018 9 283 10.3389/fimmu.2018.00283 29497427 PMC5818392 Mehta RS, Rezvani K. Chimeric Antigen Receptor Expressing Natural Killer Cells for the Immunotherapy of Cancer. Front Immunol. 2018;9:283. 10.3389/fimmu.2018.00283. 29497427 10.3389/fimmu.2018.00283 PMC5818392

229. Kong JC Sa’ad MA Vijayan HM Ravichandran M Balakrishnan V Tham SK Chimeric antigen receptor-natural killer cell therapy: current advancements and strategies to overcome challenges Front Immunol 2024 15 1384039 10.3389/fimmu.2024.1384039 38726000 PMC11079817 Kong JC, Sa’ad MA, Vijayan HM, Ravichandran M, Balakrishnan V, Tham SK, et al. Chimeric antigen receptor-natural killer cell therapy: current advancements and strategies to overcome challenges. Front Immunol. 2024;15:1384039. 10.3389/fimmu.2024.1384039. 38726000 10.3389/fimmu.2024.1384039 PMC11079817

230. Mitwasi N Feldmann A Arndt C Koristka S Berndt N Jureczek J UniCAR-modified off-the-shelf NK-92 cells for targeting of GD2-expressing tumour cells Sci Rep 2020 10 1 2141 10.1038/s41598-020-59082-4 32034289 PMC7005792 Mitwasi N, Feldmann A, Arndt C, Koristka S, Berndt N, Jureczek J, et al. UniCAR-modified off-the-shelf NK-92 cells for targeting of GD2-expressing tumour cells. Sci Rep. 2020;10(1):2141. 10.1038/s41598-020-59082-4. 32034289 10.1038/s41598-020-59082-4 PMC7005792

231. Grote S, Ureña-Bailén G, Chan KC-H, Baden C, Mezger M, Handgretinger R et al. In Vitro Evaluation of CD276-CAR NK-92 Functionality, Migration and Invasion Potential in the Presence of Immune Inhibitory Factors of the Tumor Microenvironment. Cells. 2021;10(5).10.3390/cells10051020 https://pubmed.ncbi.nlm.nih.gov/33925968/ 10.3390/cells10051020 PMC8145105 33925968

232. Xu X Cao P Wang M Wan Y Sun S Chen Y Signaling intact membrane-bound IL-15 enables potent anti-tumor activity and safety of CAR-NK cells Front Immunol 2025 16 1658580 10.3389/fimmu.2025.1658580 41098734 PMC12518233 Xu X, Cao P, Wang M, Wan Y, Sun S, Chen Y, et al. Signaling intact membrane-bound IL-15 enables potent anti-tumor activity and safety of CAR-NK cells. Front Immunol. 2025;16:1658580. 10.3389/fimmu.2025.1658580. 41098734 10.3389/fimmu.2025.1658580 PMC12518233

233. Susek KH Karvouni M Alici E Lundqvist A The Role of CXC Chemokine Receptors 1–4 on Immune Cells in the Tumor Microenvironment Front Immunol 2018 9 2159 10.3389/fimmu.2018.02159 30319622 PMC6167945 Susek KH, Karvouni M, Alici E, Lundqvist A. The Role of CXC Chemokine Receptors 1–4 on Immune Cells in the Tumor Microenvironment. Front Immunol. 2018;9:2159. 10.3389/fimmu.2018.02159. 30319622 10.3389/fimmu.2018.02159 PMC6167945

234. Eguchi S Luo W Zhu H Rosenblum JM Cooper AR Cairo MS Advances in augmenting infiltration of active natural killer cells into pediatric and adult solid tumors Mol Ther Oncol 2025 33 4 201061 10.1016/j.omton.2025.201061 41112978 PMC12528893 Eguchi S, Luo W, Zhu H, Rosenblum JM, Cooper AR, Cairo MS. Advances in augmenting infiltration of active natural killer cells into pediatric and adult solid tumors. Mol Ther Oncol. 2025;33(4):201061. 10.1016/j.omton.2025.201061. 41112978 10.1016/j.omton.2025.201061 PMC12528893

235. Cho S, Choi SH, Maeng E, Park H, Ryu KS, Park K-S. Boosting tumor homing of endogenous natural killer cells via therapeutic secretomes of chemically primed natural killer cells. J Immunother Cancer. 2025;13(3):e010371. 10.1136/jitc-2024-010371. 10.1136/jitc-2024-010371 PMC11883546 40044578

236. Yvon ES Burga R Powell A Cruz CR Fernandes R Barese C Cord blood natural killer cells expressing a dominant negative TGF-β receptor: Implications for adoptive immunotherapy for glioblastoma Cytotherapy 2017 19 3 408 18 10.1016/j.jcyt.2016.12.005 28109751 Yvon ES, Burga R, Powell A, Cruz CR, Fernandes R, Barese C, et al. Cord blood natural killer cells expressing a dominant negative TGF-β receptor: Implications for adoptive immunotherapy for glioblastoma. Cytotherapy. 2017;19(3):408–18. 10.1016/j.jcyt.2016.12.005. 28109751 10.1016/j.jcyt.2016.12.005

237. Thangaraj JL Coffey M Lopez E Kaufman DS Disruption of TGF-β signaling pathway is required to mediate effective killing of hepatocellular carcinoma by human iPSC-derived NK cells Cell Stem Cell 2024 31 9 1327 e13435 10.1016/j.stem.2024.06.009 38986609 PMC11380586 Thangaraj JL, Coffey M, Lopez E, Kaufman DS. Disruption of TGF-β signaling pathway is required to mediate effective killing of hepatocellular carcinoma by human iPSC-derived NK cells. Cell Stem Cell. 2024;31(9):1327–e13435. 10.1016/j.stem.2024.06.009. 38986609 10.1016/j.stem.2024.06.009 PMC11380586

238. Young A Ngiow SF Gao Y Patch A-M Barkauskas DS Messaoudene M A2AR Adenosine Signaling Suppresses Natural Killer Cell Maturation in the Tumor Microenvironment Cancer Res 2018 78 4 1003 16 10.1158/0008-5472.CAN-17-2826 29229601 Young A, Ngiow SF, Gao Y, Patch A-M, Barkauskas DS, Messaoudene M, et al. A2AR Adenosine Signaling Suppresses Natural Killer Cell Maturation in the Tumor Microenvironment. Cancer Res. 2018;78(4):1003–16. 10.1158/0008-5472.CAN-17-2826. 29229601 10.1158/0008-5472.CAN-17-2826

239. Liu E Marin D Banerjee P Macapinlac HA Thompson P Basar R Use of CAR-Transduced Natural Killer Cells in CD19-Positive Lymphoid Tumors N Engl J Med 2020 382 6 545 53 10.1056/NEJMoa1910607 32023374 PMC7101242 Liu E, Marin D, Banerjee P, Macapinlac HA, Thompson P, Basar R, et al. Use of CAR-Transduced Natural Killer Cells in CD19-Positive Lymphoid Tumors. N Engl J Med. 2020;382(6):545–53. 10.1056/NEJMoa1910607. 32023374 10.1056/NEJMoa1910607 PMC7101242

240. Vallera DA Felices M McElmurry R McCullar V Zhou X Schmohl JU IL15 Trispecific Killer Engagers (TriKE) Make Natural Killer Cells Specific to CD33 + Targets While Also Inducing Persistence, In Vivo Expansion, and Enhanced Function Clin Cancer Res 2016 22 14 3440 50 10.1158/1078-0432.CCR-15-2710 26847056 PMC4947440 Vallera DA, Felices M, McElmurry R, McCullar V, Zhou X, Schmohl JU, et al. IL15 Trispecific Killer Engagers (TriKE) Make Natural Killer Cells Specific to CD33 + Targets While Also Inducing Persistence, In Vivo Expansion, and Enhanced Function. Clin Cancer Res. 2016;22(14):3440–50. 10.1158/1078-0432.CCR-15-2710. 26847056 10.1158/1078-0432.CCR-15-2710 PMC4947440

241. Felices M, Warlick E, Juckett M, Weisdorf D, Vallera D, Miller S, et al. 444 GTB-3550 tri-specific killer engager TriKE™ drives NK cells expansion and cytotoxicity in acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS) patients. BMJ Publishing Group Ltd; 2021;A473–473.

242. Hibler W, Merlino G, Yu Y. CAR NK Cell Therapy for the Treatment of Metastatic Melanoma: Potential & Prospects. Cells. 2023;12(23):2750. 10.3390/cells12232750. 10.3390/cells12232750 PMC10706172 38067178

243. Bahmanyar M Vakil MK Al-Awsi GRL Kouhpayeh SA Mansoori Y Mansoori B Anticancer traits of chimeric antigen receptors (CARs)-Natural Killer (NK) cells as novel approaches for melanoma treatment BMC Cancer 2022 22 1 1220 10.1186/s12885-022-10320-0 36434591 PMC9701052 Bahmanyar M, Vakil MK, Al-Awsi GRL, Kouhpayeh SA, Mansoori Y, Mansoori B, et al. Anticancer traits of chimeric antigen receptors (CARs)-Natural Killer (NK) cells as novel approaches for melanoma treatment. BMC Cancer. 2022;22(1):1220. 10.1186/s12885-022-10320-0. 36434591 10.1186/s12885-022-10320-0 PMC9701052

244. Kaur K, Topchyan P, Jewett A. Supercharged Natural Killer (sNK) Cells Inhibit Melanoma Tumor Progression and Restore Endogenous NK Cell Function in Humanized BLT Mice. Cancers (Basel). 2025;17(15). 10.3390/cancers17152430. 10.3390/cancers17152430 PMC12346276 40805135

245. Kaur K Jewett A Supercharged NK Cell-Based Immuotherapy in Humanized Bone Marrow Liver and Thymus (Hu-BLT) Mice Model of Oral, Pancreatic, Glioblastoma, Hepatic, Melanoma and Ovarian Cancers Crit Rev Immunol 2023 43 2 13 25 10.1615/CritRevImmunol.2023050618 37938193 Kaur K, Jewett A, Supercharged NK. Cell-Based Immuotherapy in Humanized Bone Marrow Liver and Thymus (Hu-BLT) Mice Model of Oral, Pancreatic, Glioblastoma, Hepatic, Melanoma and Ovarian Cancers. Crit Rev Immunol. 2023;43(2):13–25. 10.1615/CritRevImmunol.2023050618. 37938193 10.1615/CritRevImmunol.2023050618

246. Shin MH Kim J Lim SA Kim J Kim S-J Lee K-M NK Cell-Based Immunotherapies in Cancer Immune Netw 2020 20 2 e14 10.4110/in.2020.20.e14 32395366 PMC7192832 Shin MH, Kim J, Lim SA, Kim J, Kim S-J, Lee K-M. NK Cell-Based Immunotherapies in Cancer. Immune Netw. 2020;20(2):e14. 10.4110/in.2020.20.e14. 32395366 10.4110/in.2020.20.e14 PMC7192832

247. Zheng W Ling S Cao Y Shao C Sun X Combined use of NK cells and radiotherapy in the treatment of solid tumors Front Immunol 2023 14 1306534 10.3389/fimmu.2023.1306534 38264648 PMC10803658 Zheng W, Ling S, Cao Y, Shao C, Sun X. Combined use of NK cells and radiotherapy in the treatment of solid tumors. Front Immunol. 2023;14:1306534. 10.3389/fimmu.2023.1306534. 38264648 10.3389/fimmu.2023.1306534 PMC10803658

248. Zhang C Liu Y Targeting NK Cell Checkpoint Receptors or Molecules for Cancer Immunotherapy Front Immunol 2020 11 1295 10.3389/fimmu.2020.01295 32714324 PMC7344328 Zhang C, Liu Y. Targeting NK Cell Checkpoint Receptors or Molecules for Cancer Immunotherapy. Front Immunol. 2020;11:1295. 10.3389/fimmu.2020.01295. 32714324 10.3389/fimmu.2020.01295 PMC7344328

249. Leem G, Jang S-I, Cho J-H, Jo JH, Lee HS, Chung MJ et al. Safety and Efficacy of Allogeneic Natural Killer Cells in Combination with Pembrolizumab in Patients with Chemotherapy-Refractory Biliary Tract Cancer: A Multicenter Open-Label Phase 1/2a Trial. Cancers (Basel). 2022;14(17). 10.3390/cancers14174229. 10.3390/cancers14174229 PMC9454779 36077766

250. Li L Li W Wang C Yan X Wang Y Niu C Adoptive transfer of natural killer cells in combination with chemotherapy improves outcomes of patients with locally advanced colon carcinoma Cytotherapy 2018 20 1 134 48 10.1016/j.jcyt.2017.09.009 29056549 Li L, Li W, Wang C, Yan X, Wang Y, Niu C, et al. Adoptive transfer of natural killer cells in combination with chemotherapy improves outcomes of patients with locally advanced colon carcinoma. Cytotherapy. 2018;20(1):134–48. 10.1016/j.jcyt.2017.09.009. 29056549 10.1016/j.jcyt.2017.09.009

251. Laskowski TJ Biederstädt A Rezvani K Natural killer cells in antitumour adoptive cell immunotherapy Nat Rev Cancer 2022 22 10 557 75 10.1038/s41568-022-00491-0 35879429 PMC9309992 Laskowski TJ, Biederstädt A, Rezvani K. Natural killer cells in antitumour adoptive cell immunotherapy. Nat Rev Cancer. 2022;22(10):557–75. 10.1038/s41568-022-00491-0. 35879429 10.1038/s41568-022-00491-0 PMC9309992

252. Peng L Sferruzza G Yang L Zhou L Chen S CAR-T and CAR-NK as cellular cancer immunotherapy for solid tumors Cell Mol Immunol 2024 21 10 1089 108 10.1038/s41423-024-01207-0 39134804 PMC11442786 Peng L, Sferruzza G, Yang L, Zhou L, Chen S. CAR-T and CAR-NK as cellular cancer immunotherapy for solid tumors. Cell Mol Immunol. 2024;21(10):1089–108. 10.1038/s41423-024-01207-0. 39134804 10.1038/s41423-024-01207-0 PMC11442786

253. Park H Kim G Kim N Ha S Yim H Efficacy and safety of natural killer cell therapy in patients with solid tumors: a systematic review and meta-analysis Front Immunol 2024 15 1454427 10.3389/fimmu.2024.1454427 39478866 PMC11522797 Park H, Kim G, Kim N, Ha S, Yim H. Efficacy and safety of natural killer cell therapy in patients with solid tumors: a systematic review and meta-analysis. Front Immunol. 2024;15:1454427. 10.3389/fimmu.2024.1454427. 39478866 10.3389/fimmu.2024.1454427 PMC11522797

254. Nassief G, Anaeme A, Moussa K, Mansour AN, Ansstas G. Recent Advancements in Cell-Based Therapies in Melanoma. Int J Mol Sci. 2024;25(18):9848. 10.3390/ijms25189848. 10.3390/ijms25189848 PMC11432154 39337333

255. Lian G, Mak TS-K, Yu X, Lan H-Y. Challenges and Recent Advances in NK Cell-Targeted Immunotherapies in Solid Tumors. Int J Mol Sci. 2021;23(1):164. 10.3390/ijms23010164. 10.3390/ijms23010164 PMC8745474 35008589

256. Shen Z Meng X Rautela J Chopin M Huntington ND Adjusting the scope of natural killer cells in cancer therapy Cell Mol Immunol 2025 22 7 699 711 10.1038/s41423-025-01297-4 40410571 PMC12206923 Shen Z, Meng X, Rautela J, Chopin M, Huntington ND. Adjusting the scope of natural killer cells in cancer therapy. Cell Mol Immunol. 2025;22(7):699–711. 10.1038/s41423-025-01297-4. 40410571 10.1038/s41423-025-01297-4 PMC12206923

257. Tao J-H Zhang J Li H-S Zhou Y Guan C-X Nature killer cell for solid tumors: Current obstacles and prospective remedies in NK cell therapy and beyond Crit Rev Oncol Hematol 2025 205 104553 10.1016/j.critrevonc.2024.104553 39515404 Tao J-H, Zhang J, Li H-S, Zhou Y, Guan C-X. Nature killer cell for solid tumors: Current obstacles and prospective remedies in NK cell therapy and beyond. Crit Rev Oncol Hematol. 2025;205:104553. 10.1016/j.critrevonc.2024.104553. 39515404 10.1016/j.critrevonc.2024.104553

258. Gross S, Erdmann M, Haendle I, Voland S, Berger T, Schultz E et al. Twelve-year survival and immune correlates in dendritic cell-vaccinated melanoma patients. JCI Insight. 2017;2(8):e91438. 10.1172/jci.insight.91438. 10.1172/jci.insight.91438 PMC5396520 28422751

259. Tsai KK Komanduri KV Tumor-Infiltrating Lymphocyte Therapy for the Treatment of Metastatic Melanoma Am J Clin Dermatol 2025 26 5 733 45 10.1007/s40257-025-00957-5 40549109 PMC12436572 Tsai KK, Komanduri KV. Tumor-Infiltrating Lymphocyte Therapy for the Treatment of Metastatic Melanoma. Am J Clin Dermatol. 2025;26(5):733–45. 10.1007/s40257-025-00957-5. 40549109 10.1007/s40257-025-00957-5 PMC12436572

260. Tu S Zhou L Huang R Zhou X Yang J He Y Dendritic cell vaccines extend CAR T-cell persistence and improve the efficacy of CD19 CAR T-cell therapy in refractory or relapsed adult B-ALL patients Am J Hematol 2024 99 7 1437 40 10.1002/ajh.27349 38712616 Tu S, Zhou L, Huang R, Zhou X, Yang J, He Y, et al. Dendritic cell vaccines extend CAR T-cell persistence and improve the efficacy of CD19 CAR T-cell therapy in refractory or relapsed adult B-ALL patients. Am J Hematol. 2024;99(7):1437–40. 10.1002/ajh.27349. 38712616 10.1002/ajh.27349