Breathing new life into T-cell receptor-engineered T-cell therapy in solid tumors: enhancing strategies to expand the universality of precision therapy.

Introduction

For patients with advanced, recurrent, or refractory cancer, the treatment options available at this stage are highly limited. To extend survival time and improve quality of life, research on cancer immunotherapies has achieved substantial progress in recent years, among which adoptive T-cell transfer therapy (ACT) has shown remarkable promise. Initially, ACT demonstrated significant efficacy in clinical application for hematological malignancies, but its application in solid tumors has been hindered by complex tumor immune microenvironment (TME) and various constraints in production process [ 1 , 2 ].

Current ACT technologies under development mainly include TILs, CAR-T and TCR-T cell therapies (Table 1 ) [ 3 – 5 ]. All three approaches enhance anti-tumor effects of T cells by expanding and modifying them in vitro before reinfusion. TILs, composed of tumor-specific T cells, were first clinically tested for treating in metastatic melanoma [ 6 , 7 ]. Owing to their rich TCR heterogeneity, TILs can target and recognize multiple tumor-associated antigens (TAAs). As a population of T cells containing patient-derived, tumor-specific TCRs, TILs possess theoretical efficient tumor-clearing capabilities [ 6 – 8 ]. However, due to their long production cycle and highly specialized manufacturing requirements, their clinical application remains limited. Moreover, numerous bystander T cells and exhausted phenotypes are present during treatment, resulting in limited efficacy [ 9 , 10 ]. Concurrently, CAR-T cells emerged as an alternative effective cellular therapeutic modality. Following the U.S. Food and Drug Administration (FDA) approval of the first CAR-T product in 2017, CAR-T cell therapies have achieved remarkable clinical outcomes in hematologic malignancies and heralded a new era in tumor immunotherapy [ 11 – 14 ]. Genetically engineered CAR structures can precisely recognize tumor surface antigens independent of MHC molecules, providing long-term disease-free survival in hematological malignancies with significantly superior efficacy compared to traditional chemotherapy and radiotherapy [ 15 , 16 ]. However, for solid tumors, CAR-T therapies have not yet demonstrated comparable and stable therapeutic effects, which can be attributed to inherent limitations [ 12 , 17 , 18 ]. CAR-T cells can only recognize target molecules on cell surface, greatly restricting the range of available antigens. Their capacity for tumor cell targeting and tissue infiltration is limited, leading to T-cell exhaustion due to the lack of engagement with targets [ 19 ]. The core functional component of CAR is the single-chain variable fragment (scFv) which functions as the antigen-recognition domain. The immunogenicity of scFv can trigger attacks by immune system and insufficient persistence of CAR-T cells in the body. Additionally, the incidence of adverse effects such as cytokine release syndrome (CRS) is relatively high [ 20 , 21 ].

In this context, TCR-T cell therapies have emerged to further consolidate and optimize the efficacy of engineered T-cell therapies, achieving groundbreaking progress in the clinical treatment of solid tumors [ 22 – 24 ]. TCR-T cells are regarded as among the most effective cytolytic effectors in adoptive cell therapies [ 25 ]. Patient-derived T cells are engineered in vitro to express specific exogenous TCR complexes, including αβ-chain and γδ-chain TCRs. The vast majority of T cells express αβ chains, which are multi-subunit receptor complexes composed of antigen-recognizing TCRαβ subunits and signal-transducing subunits CD3γ, CD3δ, CD3ε, and CD3ζ. Each signal-transducing subunit contains intracellular immunoreceptor tyrosine-based activation motifs (ITAMs). CD4 + or CD8 + TCR-T cells recognize tumor-associated antigen peptides presented by human leukocyte antigen (HLA) Class-I and HLA Class-II molecules without promiscuous binding, respectively. When TCR binds to its homologous peptide-major histocompatibility complex (pMHC) compound, associated CD247 (CD3ζ) chains dimerize to initiate downstream signaling, leading to rapid gene expression driven by transcription factors NF-κB, AP-1, and NFAT, exerting direct cytotoxic effects on target cells. Ideally, TCR-T cells can specifically and tightly bind to pMHC, exerting efficient anti-tumor effects (Fig. 1 ).

These mechanisms confer unique therapeutic benefits upon TCR-T cells. Compared to TILs, TCR-T cells are less affected by patient tumor stage, exhibit superior targeting accuracy and manufacturability trends, and carry a lower risk of CRS. However, TIL can serve as a screening tool to provide rich resources for the development of tumor antigen-specific TCRs and improve the accuracy of antigen prediction [ 26 , 27 ]. Polyclonal TCRs that recognize both tumor-specific antigens (TSAs) and TAAs can be cloned from such naturally occurring tumor-specific T cells to improve the therapeutic and targeting range of TCR-T cells [ 28 , 29 ]. Furthermore, relative to CAR-T cells, TCR-T cell therapy offers three critical advantages for solid tumor treatment: expanded antigen repertoire, enhanced sensitivity and superior tissue infiltration and safety. First of all, the distinct recognition pattern of TCR provides the theoretical foundation for the superior efficacy. TCR-T cells are capable of recognizing a variety of TAAs restricted by HLA on the cell surface, within the cytoplasm, and even in the nucleus, greatly increasing the richness of the antigen spectrum. What’s more, TCRs possess the innate sensitivity to extremely low antigen densities on tumors, allowing them to mount robust responses against target antigens [ 30 ]. The antigenic epitopes recognized by TCRs are typically 8–13 amino acids after processing, rather than the entire peptide segment, which allows for differential expression of target proteins among homologous cells. In hepatitis B virus -related hepatocellular carcinoma (HBV-HCC), specific TCRs can be produced by acquiring short translated mRNA-encoded epitopes, for treatments of most patients who do not express the complete HBV antigen [ 31 ]. Intriguingly, CAR-T cells engineered with scFv targeting pMHC, incorporating TCR-inspired design principles, exhibit significantly enhanced tumor-killing capabilities in vitro and in vivo, highlighting the superiority of TCR-based recognition [ 32 ]. In addition, the natural origin of TCRs endows TCR-T cells with enhanced tumor tissue infiltration ability and lower risk of treatment-related serious adverse events (AEs) compared to CAR-T cells.

TCR-T cell therapies have a killing and eliminating effect on tumors across primary, recurrent, post-treatment pre-metastatic window periods, and metastatic stages. In 1999, human TCR-T cells were first generated, expressing a foreign TCR gene targeting melanoma-associated antigen recognized by T cells (MART-1) [ 33 , 34 ]. To date, the clinical efficacy of TCR-T cell therapies has been demonstrated across diverse solid tumors, including melanoma, synovial sarcoma (SS), metastatic human papillomavirus (HPV)-associated carcinoma, HBV-HCC, ovarian cancer, and myxoid/round cell liposarcoma (MRCLS) [ 35 – 37 ]. On August 1, 2024, afamitresgene autoleucel (afami-cel) was approved by the FDA for treatment of advanced synovial sarcoma, marking a milestone as the first approved TCR-T cell therapy. Additionally, TCR-T cell therapies can achieve potent elimination of disseminated tumor cells (DTCs) and hold potential to prevent recurrence and metastasis after radical treatment of local breast cancer [ 38 ].

However, despite the regulatory approval of several TCR-T cell therapy products, their limited clinical application in solid tumor patients underscores that substantial obstacles persist before the unique therapeutic potential and inherent advantages of this modality can be fully harnessed. The fundamental challenge stems from thymic negative selection, which eliminates high-affinity TCRs that recognize tumor-associated pMHC on self HLA [ 39 ]. Consequently, most tumor antigen-specific TCR-T cells exhibit weak TCR-mediated activation. In vitro efforts focus on identifying TCRs with naturally high affinity or improving the binding affinity between TCR and pMHC through various methods to enhance the therapeutic effect of TCR-T cells. Besides, long production cycles, high costs, and patient-to-patient variability in TAAs and TSAs confine the application of TCR-T cell therapies to a narrow patient population. Expanding the pool of patients eligible for TCR-T cell therapies is a key challenge that needs to be addressed before clinical translation. In this review, we summarize limitations encountered by TCR-T cell therapies in the treatment of solid tumors and corresponding strategies to enhance efficacy, evaluating current progress of TCR-T cell therapies from production processes to clinical applications. We provide insights and references for future paths of TCR-T cell therapies in solid tumor treatment, with the aim of expanding benefited patient populations of the precision therapies.

Deficiencies and limitations of TCR-T cell therapies

Although the natural affinity of TCRs and the broad antigen-presentation capabilities of HLA molecules confer potent anti-tumor efficacy upon TCR-T cell therapies, suboptimal TCR activation signaling and HLA restriction pose unique challenges that are inherent to this therapeutic modality in clinical practice. Additionally, TCR-T cell therapies, analogous to other autologous T-cell modalities, confront parallel obstacles, including insufficient in vivo persistence and significant heterogeneity in patient clinical responses [ 40 , 41 ]. These differential patient responses are related to the imperfection of the product development process in vitro and the inhibition of TME in vivo. It is these defects that result in poor or suboptimal therapeutic responses in patients with solid tumors and thereby limit the application of TCR-T cell therapies (Fig. 2 ).

The in vivo and in vitro function and durability of engineered T cells are suboptimal and insufficient

The exploitation and production of engineered T cells in vitro are still suboptimal. The transduction efficiency, high-affinity TCR screening methods, and function validation models all require refinement through practice and exploration. The traditional methods for identifying functional antigen-specific T-cell clones based on target antigen identification typically take several months and are resource-intensive and costly [ 42 ]. The T-cell clonal expansion method involving pulsing with unprocessed peptides may generate TCR-T cells that become ineffective in vivo due to their inability to recognize endogenously processed mutant targets [ 43 ]. Additionally, the lack of appropriate, clinically relevant animal models is also a major deficiency in the systematic development of TCR-T cell therapies [ 44 ]. In the target antigen screening process, although neoantigens have the advantages of tumor cell-specific and extensive intracellular expression, the fact that most selected neoantigen targets are individual-specific poses another significant challenge [ 23 ]. Moreover, the requirement for precise HLA matching poses a significant barrier to TCR-T cell therapy, restricting the eligible patient population due to the risk of insufficient T-cell activation upon mismatch. Therefore, the identification of a broad spectrum of target tumor antigens and HLA alleles through more efficient research methods is critical for advancing the clinical application of TCR-T cell therapies.

Furthermore, the in vivo persistence of engineered T cells is influenced by the intrinsic condition of patient-derived cells. T cells used for TCR-T cell therapies are typically activated and amplified in vitro and induced to a terminally differentiated phenotype, which results in limited viability after being infused back into the body [ 45 ]. Most cancer patients are of advanced age and have undergone multiple cycles of chemotherapy and radiation therapy, leading to widespread T cells dysfunction. Consequently, the corresponding production of TCR-T cells results in impaired autophagic flux and compromised antitumor activity [ 46 ]. At the same time, the endogenous TCRs in vector-transduced T cells are thought to impair the expression of the transduced target TCRs on the cell surface through dilution and competition. The heterogeneous expression and distribution of TCRs on the surface of T cells may also lead to mispairing of endogenous and transduced α and β TCR chains. This further reduces the expression of the intended TCR, weakening the downstream signal transduction and the anti-tumor activity of the engineered cells in the body [ 25 , 47 , 48 ]. Accordingly, the selection of appropriate transduction cells and combination therapeutic regimens warrants careful consideration in clinical practice.

Tumors could develop immune evasion and drug resistance

Tumor cells may evade TCR-T cell-mediated cytotoxicity through multiple mechanisms. Within the TME, tumor cells exploit diverse immunosuppressive components to spatially sequester and functionally inhibit TCR-T cells [ 49 ]. The upregulation of β-catenin was observed in synovial sarcoma that recurs after TCR-T cell therapy, which may be involved in T-cell exclusion and resistance to TCR-T treatment [ 50 ]. Melanoma effectively evades killing by New York esophageal squamous cell antigen 1 (NY-ESO-1)-specific TCR-T cells through fusion with melanoma-II type macrophages [ 51 ]. Epstein-Barr virus (EBV)-infected tumors induce CD163 + M2 macrophages to secrete large amounts of matrix metalloprotein 9 (MMP9), which may cause T cell exhaustion and resistance to the TCR-T cell therapies in EBV-positive tumors [ 52 ]. Additionally, tumor cells can employ multifaceted strategies to impair antigen presentation and evade T-cell activation, such as loss of target antigens or abnormal processing, post-translational modifications (PTMs) of epitopes, downregulation the expression or change the structure of HLA molecule, and affecting HLA-pMHC binding, consequently resulting in ineffective treatment or tumor recurrence [ 53 , 54 ]. In a patient with metastatic colon cancer who was treated with KRAS G12D-targeted TCR-T cells, heterozygous loss of the copy of chromosome 6 encoding HLA-C* 08:02 was detected nine months later in one of her progressive colon cancer lung metastasis lesions, demonstrating immune escape by down-regulating HLA expression in tumor cells [ 9 ]. Such disruptions in antigen presentation mean that after infused TCR-T cells enter the body, they cannot recognize the target sites, and thus fail to effectively generate a memory response and then become exhausted.

Abnormal immune system responses

Although derived from patient autologous cells, TCR-T cells, having undergone in vitro genetic modification, retain the potential to elicit aberrant immune responses, including CRS, off-target effects, and immune rejection.

The ability of TCRs to recognize MHC-presented antigens from both surface and intracellular sources provides a diverse target repertoire that enhances antitumor efficacy, yet simultaneously increases the risk of cross-reactivity with unintended targets. Cross-reactivity and CRS are common adverse reactions in TCR-T cell therapies, typically associated with the high affinity of TCRs and the level of tumor antigen expression in patients. In patients with tumors that highly express NY-ESO-1, infusion of TCR-T cells induced significant antitumor responses while concurrently precipitating early-onset CRS [ 48 ]. In some cases, affinity-matured TCRs are more prone to induce severe off-target toxicity [ 55 – 57 ]. A TCR targeting melanoma-associated antigen (MAGE)-A3 through “affinity maturation” has been found to cross-react with a cardiac peptide derived from the TITIN protein, resulting in fatal off-target toxicity. This highlights that high-affinity receptors may coexist with off-target risks, so it is essential to conduct cross-reactivity tests before treatment [ 58 ]. Moreover, the inappropriate selection of target antigens can also lead to the occurrence of off-target effects. For example, clinical trials of MART-1-targeted TCR-T cell therapy were terminated following severe multi-organ toxicity and patient deaths, attributable to MART-1 expression in normal melanocytes [ 22 ]. The safety and feasibility of TCR-T cell therapies targeting other lineage specific antigens such as mesothelin and alpha-fetoprotein (AFP) are now being tested in clinical trials ( NCT04809766 , NCT03132792 ). Other rare side effects have also been documented, including a case of sudden-onset left knee pseudogout arthritis occurring seven days after MAGE-A4-targeted TCR-T cell infusion in a fallopian tube cancer patient, possibly linked to Th17 cell involvement [ 59 ].

Autoimmune reactions also occur as adverse events associated with TCR-T cell therapies. The mispairing between the α and β chains of the introduced TCRs and the endogenous TCRs may result in unpredictable and uncontrollable new hybrid receptors [ 60 – 62 ]. This could increase the risk of autoimmune diseases in addition to resulting in suboptimal function of TCR-T cells [ 5 , 63 ]. It has been proven that in mouse models infusion of mismatched TCR-T cells leads to fatal graft versus-host disease (GVHD) and disruption of the hematopoietic system [ 60 ]. These adverse events can compromise treatment efficacy and, in severe cases, jeopardize patient survival, necessitating rigorous and iterative safety assessments in preclinical studies.

Optimizing strategies to enhance the efficacy of TCR-T cell therapies

To enhance the efficacy of TCR-T cell therapy in patients with solid tumors, investigators have pursued multiple strategies, including optimizing manufacturing protocols (Fig. 3 ), integrating it with complementary therapeutic approaches, screening for high-affinity TCRs and various tumor-specific antigens, and reducing adverse reactions. While improving the tumoricidal efficacy of TCR-T cells, it is also expected that more patients can benefit from this therapy.

Optimizing production platforms and methods

The growing clinical promise of TCR-T therapies has underscored the urgent need to optimize their development pipeline and production efficiency. During TCR-T cell product manufacturing, critical methodological improvements—such as authentic in vitro tumor microenvironment modeling, efficient low-toxicity transduction, rapid screening of tumor antigens and their specific TCRs, and high-affinity TCR-pMHC selection platforms—provide a solid foundation for preclinical exploration, enabling the rational design of next-generation TCR-T cell products with superior therapeutic profiles.

First, refined in vitro modeling platforms are essential to the advancement of TCR-T cell products. It is widely recognized that the TME limits the effectiveness of cell therapy, and its complexity and diversity pose challenges for the development of efficient engineered T cells [ 64 ]. This factor is frequently overlooked in preclinical studies, resulting in suboptimal therapeutic outcomes upon adoptive transfer in patients. To address this, 3D microenvironment models offer a new approach. Andrea Pavesi et al. have developed a simple microfluidic device-based system aggregating tumor cells in a 3D collagen gel area, enabling investigators to observe their interactions with different engineered T cells under diverse oxygen levels and inflammatory cytokine conditions [ 65 ]. Sharon Wei Ling Lee et al. also developed a 3D microfluidic platform to simulate the TME associated with HBV-related hepatocellular carcinoma for the development of HBV-specific TCR-T cells [ 66 ]. These in vitro models recapitulate the patient TME, offering a physiologically relevant platform to assess the antitumor efficacy and migratory competence of TCR-T cells within tumor stroma following adoptive transfer.

Second, the selection of an efficient and low-toxicity transduction method is an integral component in the development of TCR-T cells. Conventional viral transduction methods, including retroviral vectors and lentiviruses, may compromise the anti-tumor efficacy of TCR-T cells due to the risk of random integration and insertional mutagenesis. When vector integration occurs within proto-oncogene enhancers or promoters, or disrupts tumor suppressor genes [ 67 ], it can trigger uncontrolled proliferation of engineered T cells and elevate the risk of malignant transformation [ 68 – 70 ]. The killing efficiency of these T cells may also be weakened. Furthermore, viral transduction methods may increase the susceptibility of TCR-T cells to immunosuppression. Studies have demonstrated that in the microenvironment of HBV-HCC, monocytes could suppress the cytotoxicity of retrovirus-transduced TCR-T cells against cancer cells via the PD-L1/PD-1 axis, whereas the cytotoxicity of mRNA-electroporated TCR-T cells remained unaffected [ 66 ]. Therefore, selection of an optimal transduction methodology is instrumental in enhancing the in vivo cytotoxic efficacy of TCR-T cells. Researchers have made innovative advances in this area. CRISPR-Cas9-mediated gene editing combined with electroporation has been shown to substantially shorten TCR-T cell production cycles while maintaining high editing efficiency [ 71 ]. The synthetic AAV Ark313, characterized by high transduction efficiency, has served as a robust tool for preclinical modeling of transgenic TCR-T cells [ 72 ]. Additionally, non-traditional T cells have also emerged as promising vehicles for TCR transduction. Cytokine-activated T cells have demonstrated superior activation profiles and cytotoxicity compared to equivalent TCR-T cells when used as transduced cells [ 73 ]. Mucosal-associated invariant T cells, which are the most abundant innate-like T cell population in the liver, have exhibited heightened sensitivity to HBV targets and enhanced liver tropism upon engineering with HBV-specific TCRs, outperforming conventional engineered T cells [ 74 ]. These innovations in transduction methods and cellular vehicles enhance both the production efficiency and cytotoxic efficacy of TCR-T cells, thereby establishing favorable conditions for their activity within the TME.

Third, establishing rapid and efficient systems for screening tumor antigens and their specific TCRs is a key step in advancing TCR-T cell therapies. Researchers have continuously refined the selection system by integrating diverse methodologies, including gene library construction, microfluidic platforms, in vitro artificial antigen-presenting cell (APC) systems, single-cell transcriptomics and TCR V (D)J sequencing, and machine learning methods [ 75 ]. Among these, screening TCRs based on TCR-pMHC interactions through in vitro antigen presentation systems represents a well-established approach. For instance, Pu Chen et al. developed the single-plasmid Co-HA system, which co-expresses patient HLA and antigens, enabling effective detection of antigen immunogenicity and verification of T-cell epitope recognition [ 76 ]. Similarly, Teng Wei et al. utilized in vitro amplified TILs and B cell-derived HLA-matched allogeneic C1R cells as APCs to screen for antigen-specific TCRs. Loading these cells with a minigene-encoded antigen library enabled rapid target candidate prioritization and enhanced screening throughput [ 26 ]. The integration of TCR library construction with advanced sequencing technologies has further streamlined the screening process. Jinxing Jiang et al. combined TCR library-based validation with small gene library-based epitope screening to enable large-scale epitope-TCR discovery [ 77 ]. Ken-ichi Hanada et al. employed CITE-seq (Cellular Indexing of Transcriptomes and Epitopes by Sequencing) and TCR-seq to identify antigen-reactive T cells infiltrating in samples of non-small cell lung cancer (NSCLC) at single-cell resolution [ 78 ]. Additionally, Yijian Li et al. and Jingyu Qi et al. used a high-throughput approach based on a droplet microfluidic platform and molecular markers to advance the construction of a tumor antigen-specific TCR database [ 79 , 80 ]. Xu Wang et al. integrated ultradeep single-cell TCR/RNA sequencing with autologous organoid technology to rapidly characterize neoantigen-reactive TCRs [ 81 ]. Collectively, these methodological advances have substantially shortened the production timeline for TCR-T cells, thereby expanding the clinical reach of personalized TCR-T cell therapies—exemplified by neoantigen-specific approaches—to a broader patient population. Other methods, such as immunological approaches, have also contributed uniquely to TCR library construction. By employing the lentivector (lv)-prime and peptide-boost immune methods, a large number of AFP 158 -specific CD8 + T cells generated in mice without repeated in vitro restimulation should keep their TCR diversity and enabled researchers to study their antitumor effects prior to TCR identification [ 82 ]. Yulin Xu et al. established humanized mice using 3D-cultured hematopoietic stem and progenitor cells derived from peripheral blood mononuclear cells (PBMCs), allowing selected TCRs to bypass central tolerance while preserving peptide specificity during affinity maturation [ 83 ]. Notably, a vaccine-induced human TCR for cell therapies has demonstrated clinical feasibility for the first time in patients with glioblastoma [ 84 ]. More recently, Adham S. Bear et al. successfully isolated HLA-A03:01- or HLA-A11:01-restricted TCRs specifically targeting KRAS G12V from patients enrolled in a Phase I mutant KRAS vaccine trial [ 85 ]. Furthermore, the tumor organoids and autologous TILs co-culture systems have effectively amplified and isolated TCR-T cells, addressing challenges associated with imprecise antigen prediction and time-consuming synthesis [ 27 ]. While efficiency remains paramount, safety considerations are equally critical in TCR screening methodology development. Sergey Sennikov and his colleagues proposed an approach that mimics the natural induction of T-cell clones in vivo using peptide-loaded dendritic cells, aiming to minimize off-target toxicity posed by excessively high-affinity TCRs [ 86 ]. However, TCRs identified through these approaches may exhibit suboptimal clinical efficacy. An alternative strategy involves cloning tumor-specific TCRs from long-term surviving patients who respond to immunotherapy, which may yield more potent candidates [ 87 ]. Importantly, Franziska Fuchsl et al. highlighted that the strongly activated T-cell response patterns correlate with the subsequent functional exhaustion. Under repeated stimulation of antigens in vivo, the initially moderately activated TCR can maintain a more durable and stable anti-tumor effect. This is what researchers should pay attention to when screening neoantigen-specific TCRs [ 88 ].

Last, TCR-pMHC interaction affinity is a determinant of in vivo antitumor activity of TCR-T cells. Consequently, relative screening methods are undergoing continual refinement. For instance, the yeast T-cell co-culture system developed by Lihui Wang et al. enables rapid screening of high-affinity TCR-pMHC interactions [ 89 ]. Additionally, a new method detecting activated T cell-tumor interactions based on microfluidic shear stress has been developed to facilitate efficient T-cell cloning and collection [ 90 ]. However, while high-affinity TCR-pMHC binding was historically thought to maximize antitumor efficacy of engineered T cells in vivo, emerging evidence indicates that excessive affinity paradoxically increases off-target toxicity and cross-reactive potential. In this context, “catch bond fishing” has been developed to separate TCR mutants that bind pMHC with high activation signals yet low affinity through acquisition of catch bonds. This method changed the traditional concept of using TCR affinity maturation as the sole screening standard and reduced the tendency of off-target cross reactivity [ 58 ]. Moreover, while functional avidity assays for HLA Class-I restricted TCRs have been well developed [ 91 ], the CD4-2D3 cell line has been established as a valuable tool for evaluating that of HLA Class-II restricted TCRs [ 92 ]. Given that the interaction between TCR and pMHC directly dictates the signal intensity and activation state of TCR-T cells upon in vivo engagement, screening methods with greater physiological fidelity and efficiency warrant further research and exploration.

These methodological innovations across the TCR-T cell development pipeline have reinvigorated this therapeutic modality, shortened production cycles, and provided technical safeguards for efficient cell product screening.

Increased cytotoxic response and in vivo persistence (Fig. 4 )

Researchers have explored diverse strategies to enhance the TCR-T killing response and in vivo persistence—including TCR modification, co-expression of immunostimulatory cytokines, combined immunotherapy, and immunometabolic modulation—with the aim of augmenting solid tumor clearance efficacy.

In engineered T cells, modifying TCRs to couple with distinct co-stimulatory domains represents a powerful approach to augment T-cell activation. Kotaro Miyao et al. inserted the CD28 or 4-1BB intracellular domain (ICD) into CD3ζ, constituting the artificial T cell-activating adapter molecules (ATAMs). The ATAMs enhanced signal transduction after TCR binding to specific epitopes without additional modifications [ 93 ]. However, this method failed to achieve satisfactory synergy in vivo. To further improve the signal transduction near the membrane, researchers then integrated the minimal tumor necrosis factor receptor-associated factor (TRAF)-binding motifs of 4-1BB into CD3ζ, demonstrating excellent antitumor efficacy in a mouse xenotransplantation model [ 94 ]. Wenhua Liang et al. found that the introduction of an I/A point mutation in the intracellular motifs of the CD3ε subunit also enhanced signaling of the TCR/CD3 complex [ 95 ]. Beyond canonical co-stimulatory domains, the engagement of inhibitory signaling domains can, in some cases, produce unexpected effects on TCR signal activation. Nadja Sailer et al. found that the co-stimulatory signal can transform the inhibitory PD-1 signal into an activation signal in T cells, when the PD-1 extracellular domain and 4-1BB signaling domain are fused with TCR [ 96 ]. These co-stimulatory domains amplify TCR signaling, enhancing the activation status of TCR-T cells in vivo. Nevertheless, supraphysiological TCR signal activation poses significant risks, including off-target reactions and compromised efficacy. This challenge can be addressed by 7-alpha-hydroxycholesterol (7a-HC), an effective inhibitor of TCR signaling, which precisely modulates TCR signal intensity to prevent deleterious overactivation and preserve long-term TCR-T cell efficacy [ 97 ].

Moreover, distinct genetic engineering strategies beyond co-stimulatory domain incorporation can further optimize TCR-T cell therapies. Co-delivery of TCRs and T cell activating transcription factor c-Jun can enhance the proliferative capacity and in vivo persistence of TCR-T cells [ 98 ]. Additionally, targeted knockout of endogenous TCRαβ chains in TCR-T cells can improve transgenic TCR expression efficiency and its epitope recognition efficiency [ 99 ]. Given allelic exclusion and thymic positive selection, in vitro reprogrammed hematopoietic stem cells (HSCs) or induced pluripotent stem cells (iPSCs) have emerged as more suitable sources than PBMC-derived T cells for generating gene-edited T cells. Knockout of endogenous TCR genes ( TRAC/TRBC ) and inhibition of gene rearrangement at the HSC/iPSC stage significantly reduce TCR mispairing while preserving antigen specificity [ 100 – 102 ]. Alternative approaches, including engineering transgenic TCRs with murine constant regions or introducing disulfide bonds to stabilize transgenic TCRs, also effectively mitigate mispairing risk [ 5 ] ,which is critical for accurate target antigen recognition and optimal in vivo activation of TCR-T cells.

Co-expression of immunostimulatory cytokines represents another strategy to overcome TME-mediated inhibition and enhance the cytotoxicity response. For instance, the combination of p38 inhibitors, IL-7 and IL-15 promotes T cell proliferation [ 103 ]. P1A tumor antigen-specific TCR-T cells engineered to express IL7/CCL19 exhibit enhanced antitumor efficacy and durable memory responses [ 104 ]. IL-21 prevents T cell differentiation and maintains a naive phenotype of T cells. The mutant IL-21 receptor that can provide constitutive IL-21 signaling in AFP-specific TCR-T cells enhances their proliferative capacity [ 105 ]. Nevertheless, systemic administration of certain cytokines, such as recombinant human IL-12, carries significant risks of severe toxicity and adverse events. To address this, Ling Zhang et al. engineered therapeutic T cells to constitutive express anchored IL-12 (aIL-12) in their membranes, thereby improving the effectiveness of T cells while reducing circulating IL-12 and IFN-γ levels [ 106 ]. Building upon this approach, Jiemiao Hu et al. further enhanced the tumor-killing effects by fusing a cell-surface vimentin (CSV)-targeted peptide VNTANST to the IL-12 p40 subunit, generating membrane anchoring tumor targeting IL-12 (attIL12) [ 107 ]. For tighter control of IL-12 expression, Segi Kim et al. inserted the IL-12 gene into the PDCD1 locus of NY-ESO-1-specific TCR-T cells, simultaneously disrupting endogenous PD-1 expression and enabling endogenous PDCD1 regulatory elements to control recombinant IL-12 secretion [ 108 ]. Similarly, optimal co-expression and membrane anchoring of IL-15 and IL-21 limit the emergence of dysfunctional natural killer (NK)-like T cells and prevent functional T-cell exhaustion under tumor burden in vitro and in vivo [ 109 ]. These strategies promot the in vivo expansion and anti-tumor function of TCR-T cells while avoiding systemic cytokine exposure-related toxicities.

The combined treatment with immune agents clears the obstacles for the function of T cells and exerts a significant synergistic effect. For example, spermidine restores the autophagy flux of aging TCR-T cells, reduces expression of inhibitory immune receptors (PD-1, TIM-3, or LAG-3), and enhances the proliferation and antitumor activity of TCR-T cells [ 46 ]. PCSK9 inhibitors downregulate the PD-1 expression on AFP-specific TCR-T cells, thereby significantly enhancing their efficacy against HCC [ 110 ]. Combining the TRPV2 inhibitor tranilast with TCR-T cell therapy for esophageal cancer enhances cytotoxicity compared with monotherapy [ 111 ]. MMP9 inhibitors improve T-cell function in the presence of M2 macrophages, consequently augmenting the antitumor efficacy of TCR-T cells against EBV-positive tumors [ 52 ]. Prior to activation of artificial TCR-T cells in vitro, Cbl-b inhibitors can maintain TCR-T cells in a less differentiated state and significantly improve cell survival in vivo after antigen secondary exposure [ 112 ]. Collectively, these combination strategies enable TCR-T cells to overcome TME-mediated immunosuppression and achieve potent antitumor efficacy.

Metabolic reprogramming within the TME drives CD8 + T-cell exhaustion, characterized by impaired proliferation and loss of effector function [ 113 , 114 ]. In contrast to effector T cells, which are in a high-energy state, exhausted T cells display low glycolysis, mitochondrial dysfunction, and an inefficient reliance on fatty acid oxidation to adapt to nutrient-deprived environments [ 115 ]. Upon TME infiltration, TCR-T cells undergo detrimental metabolic reprogramming that impairs their function. Therefore, metabolic intervention, such as increasing glycolysis or inhibiting fatty acid oxidation, has gained prominence as a requisite strategy to regulate T cell differentiation and enhance immune responses within the TME. For instance, hypoxia-induced lactate accumulation directly binds to the intracellular motif of glucose transporter GLUT10, inhibiting glucose uptake and activation of CD8 + T cells, thereby attenuating their antitumor efficacy [ 116 ]. Conditional knockout of monocarboxylate transporter 11 (MCT11) specifically reduces lactate uptake by exhausted T cells and improves their effector functions [ 117 ]. Moreover, metabolic interventions—including D-mannose supplementation [ 118 ], N-acetylcysteine (NAC) administration [ 119 ], lysine-specific histone demethylase 1A (LSD1) inhibition [ 120 ], and prolyl 4-hydroxylase (P4HA1) targeting [ 121 ]—promote differentiation of CD8 + T cells into a more durable phenotype, mitigate hypoxia-induced stress and mitochondrial failure, and enhance immune responses. Modulating metabolic targets is key to overcoming the suppressive effects of the TME on TCR-T cells, enabling them to exert potent and sustained effector functions. Despite its therapeutic potential, optimal combination strategies require further preclinical validation before clinical implementation.

Additional combination approaches, including oncolytic viruses and tumor vaccines, are currently under investigation [ 122 ]. Notably, oncolytic virus therapy demonstrates potent synergistic antitumor effects when combined with TCR-T cell therapy. Specifically, TCR-T cells can serve as delivery vehicles for myxoma virus (MYXV). TCR-T MYXV cells induce tumor-cell autosis and mediate effective bystander killing of antigen-negative tumor cells, thereby achieving tumor clearance while circumventing antigen escape [ 123 ]. Amphiphile-bound vaccination enabled TCR-T cells to colocalize to lymph nodes with a group of APCs presenting highly co-stimulated homologous peptides, delivering to them endogenous costimulatory signals and cytokine-supported TCR stimulation [ 124 ]. Furthermore, other immune components within the TME can also potentiate TCR-T cell function. TCR-T cells engineered to secrete a high-affinity signal regulatory protein α decoy (CV1) that blocks CD47 exhibit enhanced function and persistence, particularly with the assistance of macrophages [ 125 ]. NK cells recognize and eliminate cancer cells that evade T-cell recognition via HLA downregulation. When combined with NY-ESO-1-specific 1G4 TCR-T cells, this innate capacity synergistically enhances tumor lysis [ 126 ]. Nevertheless, these combination strategies remain in the preclinical stage, and further evidence is required to establish effective protocols for achieving robust efficacy against solid tumors.

Overall, researchers have implemented synergistic combination strategies that enable TCR-T cells to achieve maximal tumor-killing efficacy within the immunosuppressive TME, thereby unlocking greater potential for future clinical applications in solid tumor therapy.

Promoting antigen presentation and expanding target ranges (Fig. 5 )

The application scope of personalized TCR-T cell therapies is limited by tumor types, targeted organs, HLA alleles, and types and numbers of target antigens expressed in solid tumor patients. While a growing repertoire of antigens, epitopes, and TCRs has been cataloged, the translation of these discoveries into tumor-agnostic and broadly applicable TCR-T cell therapies faces persistent, substantial challenges [ 127 ]. Therefore, the development of novel therapeutic targets and cell products, as well as the improvement of antigen presentation processes, represents a significant component of preclinical research. These studies focus on screening diverse novel targets and HLA types while developing adaptable TCR-T products to broaden patient eligibility and expand clinical trial access [ 128 ].

Initially, a specific tumor antigen can generate numerous distinct epitopes for presentation. Thus, identifying highly immunogenic epitopes and corresponding high-affinity TCRs represents a valuable research avenue. In the case of HPV E7, Benjamin Y. Jin et al. identified an HPV-16 E7-specific HLA-A* 02:01-restricted TCR in cervical biopsies from patients, notable for its high functional affinity and CD8 co-receptor-independent tumor targeting [ 129 ]. Subsequently, by single cell sorting and sequencing, Xiaowen Li et al. identified two TCRs that specifically bind E7 89-97 in the context of HLA-A*11:01 [ 130 ]. The screening of different HLA allele-restricted TCRs expands the HLA repertoire for this antigen target, broadening patient eligibility and providing more therapeutic options. Moreover, such screening facilitates the identification of TCRs with enhanced recognition specificity, thereby improving therapeutic efficacy. NY-ESO-1-specific novel functional HZ6 and HZ8 TCRs exhibited a different recognition pattern from the widely used 1G4 TCR [ 131 ]. Furthermore, target antigens with broader expression profiles in tumor cells can potentiate more substantial therapeutic efficacy of TCR-T cell therapies. For example, the cancer germline antigen (CGA) Ropporin-1 (ROPN1) is expressed in 90% of patients with primary and metastatic Triple-negative breast cancer (TNBC). Highly specific TCR-mediated killing of tumor cells targeting ROPN1-derived FLYTYIAKV (FLY-1A) epitopes exhibits superior efficacy compared with standard therapeutics [ 127 ]. To enable tumor cells to “express” the target antigens more generally, Ruihan Xu et al. developed HLA-peptide-addressing universal (HAUL) TCR-T cell therapy, deploying membrane fusion nanoparticles to install HLA-peptide complexes (pHLAs) on tumor cell membranes. In this way, TCR-T cells can recognize and kill tumor cells regardless of original target antigen expression level in tumors [ 132 ]. Similarly, Qin Wang et al. achieved this goal by delivering and loading exogenous antigens onto tumor cell membranes through DSPE-PEG-NY-ESO-1 157–165 liposomes (NY-ESO-1 Lips) [ 133 ]. Despite the unique innovation and therapeutic prospects of these methods, treatment-related risks such as cross-reactivity require thorough assessment prior to clinical trials. Importantly, owing to their high immunogenicity, strict tumor specificity, and potential safety advantages, tumor-specific antigens, including tumor hotspot mutant peptides [ 26 , 99 , 134 ] and tumor-associated viral proteins [ 135 ], have emerged as promising therapeutic targets. Jianxin Wang first identified non-mutated epitope LAMC2 203-211 on naturally occurring laminin subunit gamma 2 (LAMC2) in PDAC [ 136 ]. Adham S. Bear et al. confirmed that G12V/A03:01 and G12V/A11:01 serve as shared tumor neoantigen targets, which can be utilized in development of adoptive cell therapies for patients with KRAS G12V-mutated tumors [ 85 ]. Furthermore, in addition to mutation-derived neoantigens, tumor-associated viral proteins offer significant therapeutic potential for certain tumor types. Qi Liu et al. found that mutant TCR-transduced engineered T-cells (Ai-TCR-T) with improved affinity demonstrated enhanced cytotoxicity and the same in vivo safety as wild-type TCR-T cells in HBV-associated HCC therapies [ 137 ]. As research on TSA-specific TCR-T cell therapy advances, an expanding portfolio of cellular therapy products is entering clinical evaluation. However, targeting TSAs with TCR-T cell therapies inherently entails profound personalization and substantial treatment costs. Enhancing manufacturing efficiency of relevant cellular products and expanding eligible patient population are therefore integral components for realizing the full therapeutic potential of TCR-T cell therapies. Moreover, discovery of expanded repertoire of tumor-associated antigens propels advancement of TCR-T cell therapies. Inspired by CAR-T cell therapy targeting Glypican-3 (GPC3), Enric Vercher and colleagues successfully cloned TCR-T cells that recognize GPC3 (522–530) epitope. These TCR-T cells demonstrated superior effector functions and lower exhaustion characteristics compared to CAR-T cells in xenograft liver cancer models [ 138 ]. Pengju Yao et al. found that bacterial peptides are more likely to bypass central immune tolerance in colorectal cancer (CRC) and are more immunogenic than cancer-testis antigens (CTA), potentially serving as universal anti-CRC targets [ 139 ]. Furthermore, it is believed that TCRs of naturally occurring tumor antigen-specific T cells (Tas) are advantageous for application in TCR-T cell therapies. Selected by surface markers CD200 and ENTPD1 from patients’ tumors, TCRs derived from Tas cells are utilized for the production of numerous polyclonal TCR-T cells recognizing both TSAs and TAAs simultaneously for clinical therapies [ 28 ].

CD4 + TCR-T cells exhibit distinct cytolytic pathways and killing characteristics compared with CD8 + TCR-T cells. In vitro studies have demonstrated significant therapeutic effects of HLA Class II-restricted TCRs, exemplified by T helper cell type 1 (Th1) cell-derived TCR Radium-4 targeting human telomerase reverse transcriptase (hTERT) presented by HLA-DP04 and HLA-DP03 [ 140 ]. In nasopharyngeal carcinoma, TCR135 exhibits high functional activity and affinity as a CD4 TCR recognizing EBV nuclear antigen 1 (EBNA1) presented by HLA-DP5, and the transduced T cells can effectively infiltrate the tumor microenvironment and significantly inhibit tumor growth [ 141 ]. Similarly, the discovery of an HLA-DRA/DRB1*09:01-restricted HPV18 E7 84-98 -specific TCR (10F04) with excellent antitumor activity in preclinical models highlights the potential of HLA Class-II-restricted TCR-T cell therapies [ 87 ]. Nevertheless, rare expression of HLA Class-II molecules on most tumor cells has limited the clinical application of CD4 + TCR-T cell therapy. To overcome this limitation, HLA Class-I-restricted CD4 + TCR-T cell therapies have been proposed as a promising alternative [ 142 ]. In addition, other T-cell subsets, such as NKT cells, may represent a novel direction for TCR-T cell therapies. CD1d-restricted invariant natural killer T (iNKT) cells possess intrinsic tumor immunosurveillance capabilities. It has been reported that iNKT cells transduced with TAA-targeting TCRs demonstrate superior antitumor efficacy against solid tumors compared to conventional TCR-T cells in both in vitro and in vivo experiments [ 143 ]. Recent studies have revealed that genetically engineered iNKT cells exhibit robust tumor-killing effects and excellent tumor-homing capacity in solid tumor treatment, suggesting their potential as a powerful modality for ACT [ 144 , 145 ]. However, the extremely low frequency of NKT cells in human blood poses a substantial challenge for large-scale therapeutic production and clinical translation, thereby warranting further investigation.

Augmenting expression of target antigens and HLA molecules in solid tumors promotes antigen presentation process and drives the transformation of the tumor immune microenvironment from “cold” to “hot”, creating favorable conditions for T cell-mediated tumor elimination. Synergistic improvement of efficiency of NY-ESO-1 TCR-T cell therapies by decitabine exemplifies this approach [ 146 ]. Similarly, eribulin, used for the treatment of recurrent metastatic breast cancer, up-regulates expression of HLA Class-I molecules by increasing expression of NOD-like family CARD domain-containing 5 (NLRC5), and enhances response of NY-ESO-1-specific TCR-T cells to breast cancer cells in combination therapy [ 147 ]. Atovaquone (ATO) induces ROS accumulation and up-regulation of HLA Class-I expression in tumor cells, and when combined with AFP-specific TCR-T cell therapy, not only improves cytotoxicity of engineered T cells but also potentiates ferroptosis of tumor cells [ 148 ]. Additionally, STING agonist diABZI enhances TCR signaling and augments antigen presentation in Mel526 tumor cells, consequently boosting TCR-T cell therapy efficacy [ 149 ]. Beyond directly increasing tumor antigen or HLA expression, enhancing antigen capture efficiency and presentation capacity of TCR-T cells can improve their antitumor effects as well. For instance, by combining the extracellular domain and the intracellular signaling domain of TIM-4 (a phagocytic receptor that recognizes “eat me” signal phosphatidylserine) in the TCR, Brandon Cieniewicz et al. designed chimeric phagocytic receptor (CER)-1236 T cells with target-dependent phagocytic function, demonstrating enhanced antigen capture efficiency and presentation capacity relative to conventional TCR-T cells [ 150 ].

Collectively, these findings underscore that optimizing antigen presentation and recognition constitutes the most critical step in the development of TCR-T cell therapies. Expanding the spectrum of target antigens, TCR repertoires, and HLA typing facilitates the transition from limited to broad clinical applications, whereas promoting tumor cell antigen presentation elevates the therapeutic ceiling of TCR-T cell therapy.

Reducing toxic side effects

Allogeneic T-cell engineering offers a solution to the dual challenges of insufficient quantity and compromised quality in patient-derived T cells, but carries the risk of immune rejection. In HBV-HCC patients treated with liver transplantation (LT), immunosuppressive drug-resistant armored (IDRA) HBV-redirected TCR-T cells, developed through mRNA electroporation, have been shown to mitigate immunosuppression of engineered T cells. Electroporated mRNA allows drug resistance to last only 3–5 days, preventing the persistence of potential IDRA alloreactive T cells in transplant patients, thus significantly reducing the risk of potential organ rejection. Moreover, this approach reduces the risk of serious liver inflammatory events caused by off-target effects in HBV-infected normal liver cells [ 151 , 152 ]. IDRA HBV-TCRs can dissolve free-floating HBV-HCC cells in whole blood to prevent relapse after LT [ 153 ]. Furthermore, generating transgenic TCR-T (TgTCR-T) cells from engineered PSCs addresses the problem of inadequate activity of patient-derived T cells, while reducing the risk of autoreaction [ 47 ].

To increase TCR affinity and signal strength after activation, TCRs are usually modified or synergized with other immune agents. However, excessive TCR signaling activation and high affinity may lead to CRS, cross-reactivity and off-target effects. Abdullah Degirmencay et al. found that altering a single amino acid in the framework region of TCR variable β domain can lead to improved surface expression of engineered TCRs, enhanced target cell-specific killing, and reduced off-target effects [ 154 ]. The introduction of the suicide gene, inducible caspase 9 (iC9) gene, into the SLL TCR framework can induce rapid apoptosis and clearance of transgenic T cells upon administration of the small molecule dimerization drug AP1903, thereby enabling control of serious adverse reactions caused by TCR-T cells in vivo [ 155 ].

Clinical research progress and experience

TCR-T cell therapies are currently in the early stages of clinical exploration for patients with solid tumors. The focus is primarily on two fronts: enhancing efficacy and improving safety. Investigations encompass the validation of established and emerging antigens, and concurrently exploring combination regimens to augment therapeutic efficacy (Tables 2 and 3 ).

Verifying safety and efficacy in vivo

Clinically, target antigens for TCR-T cell therapy are primarily categorized into two classes: TAAs comprising classic targets, and TSAs mainly consisting of tumor hotspot mutation peptides. Accordingly, current therapeutic improvements largely center on three priorities: enhancing the cytotoxic efficacy of established approaches, reducing treatment-associated toxicities, and exploring next-generation target antigens with superior therapeutic potential.

TCR-T cell therapies targeting classical antigens can be improved and enhanced to show new vitality in clinical practice. CTAs represent a kind of classic target antigens with expression in healthy cells typically restricted to immune-privileged testicular tissue, and TCR-T cell therapies targeting CTAs have shown particularly significant efficacy and benefits in melanoma [ 156 ]. Clinical trials aimed at enhancing efficacy of these TCR-T cells are driving the broader clinical adoption of this approach. The MAGE family of tumor antigens represents one of the most prominent and well-characterized CTA families. Currently, the prevailing method involving site-directed mutagenesis of the TCR complementarity-determining regions is commonly employed to generate T-cell products with self-specific peptide-enhanced affinity receptors (SPEAR). The SPEAR T cell product ADP-A2M10, exhibiting higher affinity and specificity for MAGE-A10 antigen peptide, demonstrated a manageable safety profile in two Phase I clinical studies enrolling patients with advanced NSCLC ( NCT02592577 ) and patients with advanced head and neck squamous cell carcinoma (HNSCC), melanoma, and uroepithelial carcinoma (UC) ( NCT 02989064 ) [ 41 , 157 ]. However, due to overlapping expression patterns of MAGE-A10 and MAGE-A4, as well as immunosuppressive effects within the TME that can dampen the efficacy of infused T cells, researchers have developed the next-generation SPEAR T cells targeting MAGE-A4. These cells, known as afami-cel, are engineered to co-express additional immune-modulating molecules such as CD8α or IL-17/CCL19. This product is generated by transducing CD4 + and CD8 + T cells with affinity-optimized TCRs via a lentiviral vector, and has demonstrated early and durable responses in patients with various solid tumors, particularly those with synovial sarcoma ( NCT03132922 ) [ 158 ]. To further investigate its efficacy, researchers conducted an open-label, non-randomized Phase II trial (SPEARHEAD-1, NCT04044768 ) in patients with advanced synovial sarcoma or myxoid/round cell liposarcoma expressing MAGE-A4. Effective targeting ability and durable therapeutic responses were demonstrated in this study, evidenced by an overall response rate of 37% (19 out of 52 patients) and durable responses (median 11.6 months) [ 36 ]. This therapy was approved by the FDA on August 1, 2024, for the treatment of adult patients with unresectable or metastatic synovial sarcoma who have previously received chemotherapy, offering a new therapeutic option for this patient population.

Similarly, NY-ESO-1 also stands as a critical member of the CTA family. Analogous augmentation approaches are presently undergoing clinical assessment. Paul F. Robbins et al. conducted the first-in-human clinical trial utilizing affinity-enhanced NY-ESO-1-reactive TCR-transduced autologous PBMCs for adoptive cell therapy, reporting objective clinical responses in patients with refractory synovial sarcoma and melanoma [ 37 ]. The NY-ESO-1-specific TCR-T cell product TBI-1301 utilizes endogenous TCR silencing by siRNA to enhance TCR affinity, and its first -in-human Phase I clinical trial was conducted in 9 patients with NY-ESO-1-expressing solid tumors. Efficacy and safety of this approach were validated, with the exception of grade 3 lung injury [ 48 ]. The Phase I clinical trial ( NCT04318964 ) of affinity-enhanced TCR-T cells (TAEST16001 cells) specifically recognizing HLA-A* 02:01-restricted NY-ESO-1 157–165 epitope demonstrated safety and promising efficacy in 12 patients with advanced soft tissue sarcoma [ 159 ]. Additionally, letetresgene autoleucel (lete-cel), targeting NY-ESO-1, employs an efficacy-enhancing strategy analogous to afami-cel and has demonstrated significant antitumor efficacy and durable responses in patients with synovial sarcoma and MRCLS [ 160 ]. However, in a single-arm study treating patients with advanced or recurrent NSCLC ( NCT02588612 ), lete-cel demonstrated favorable tolerability but limited efficacy, with only one out of 18 patients showing a durable therapeutic response [ 40 ]. Collectively, these findings indicate that current TCR-T clinical therapies for solid tumors exhibit a relative efficacy in patients with sarcomas, whereas antitumor activity in other solid malignancies requires further enhancement.

Due to the technological innovation, some TCR-T cell therapies, exhibiting obvious application limitations originally, can also show superior clinical efficacy under the premise of ensuring safety. HBV-TCR-T immunotherapy was previously considered to have severe liver toxicity and was not adopted clinically. In recent years, this therapy has improved the safety of in vivo application by shortening the lifespan of engineered T cells, and has shown promising efficacy in advanced HBV-HCC patients. In a proof-of-concept study, Anthony Tanoto Tan et al. obtained short-translated mRNA coding epitopes in 2 HBV-HCC patients with recurrence and metastasis after liver transplantation, and used mRNA electroporation to transiently express TCRs to provide safety assurance. In one of these patients, five of the six lung metastases showed a volume reduction during the 1-year follow-up period [ 31 ]. Subsequently, the team conducted an open-label Phase I dose-escalation study ( NCT03899415 ) to evaluate the safety of transient autologous T cells expressing HBV-specific TCRs in 8 patients with advanced HBV-HCC who did not meet liver transplantation criteria [ 152 , 161 ]. Three of them developed long-term stabilization of tumor lesions with dose-escalation therapy, showing long-term clinical benefit associated with transient immune changes [ 152 ]. They further evaluated the safety of multiple infusions of HBV-specific mRNA electroporation TCR-T cell therapy in six patients with HBV-associated HCC recurrence after liver transplantation ( NCT02719782 ), and demonstrated satisfactory tolerability in patients [ 162 ]. Therefore, addressing cellular product toxicity and adverse events through targeted transduction strategies or alternative interventions constitutes a clinically imperative objective.

Moreover, addressing the limited diversity of target antigens and HLA alleles represents an urgent clinical priority. Different alleles and epitopes show disparate profiles of major tumor type coverage [ 163 ]. Targeting novel effective antigens enables broader patient inclusion in individualized TCR-T cell therapies, thereby enhancing the universal applicability of precision strategies. Martin Wermke et al. conducted the first-in-human study of safety and tolerability of the TCR-T cells IMA202, with a natural high specificity and binding affinity for a nine-amino acid peptide derived from MAGE-A1 in 16 patients with advanced solid tumors positive for HLA-A *02:01 MAGE-A1 [ 30 ]. Certain subtypes of sarcomas and malignant melanomas were found to be particularly sensitive to this therapy, once again demonstrating the potential of targeting cancer-testis antigens in the treatment of solid tumors, particularly sarcomas. Meanwhile, clinical trials are expanding to encompass diverse solid tumors, such as renal cell carcinoma, epithelial cancer, and pancreatic cancer. Preclinical and clinical research on emerging targets, such as HBV16/18 E7 and KK-LC-1, is also being actively pursued, collectively propelling TCR-T cell therapy development forward ( NCT05787535 , NCT05122221 , NCT05035407 ). In recent years, TCR-T cell therapies targeting neoantigens have gradually emerged. Targeting of tumor hotspot mutation antigens has gained traction among researchers owing to its high precision and broad applicability. Preliminary validation of this strategy in a patient with metastatic CRC demonstrated both safety and efficacy. In this case, the patient’s lung metastases showed significant regression after treatment, with six out of seven metastatic lesions completely regressing or continuing to regress nine months post-treatment [ 9 ]. Another study reported a patient with progressive metastatic pancreatic cancer who, following the failure of TIL therapy, received autologous T cells expressing two HLA-C*08:02-restricted TCRs targeting mutant KRAS-G12D. This intervention produced sustained metastatic regression for 6 months and an overall partial response rate of 72% [ 164 ]. Sanghyun P. Ki and colleagues applied TCR gene-edited peripheral blood lymphocytes (PBL) targeting the p53 R175H neoantigen to a patient with chemotherapy-refractory breast cancer. Compared with other patients in the same cohort who received TIL containing low-frequency p53 mutation-reactive cells, the infused cells in this patient exhibited enhanced durability of immune phenotypes and reactivity, with objective tumor regression lasting 6 months [ 165 ]. These findings confirm the therapeutic potential of neoantigen-targeted TCR-T cell therapies. Several cell products targeting neoantigens, such as KRAS-G12D, KRAS-G12V or p53, are currently being evaluated in Phase I clinical trials.

Overall, approaches aimed at reducing adverse events and potentiating TCR-T cell efficacy, grounded in mechanistic insights, hold promise for surmounting clinical obstacles and expanding the therapeutic landscape in the treatment of solid tumors.

Progress of combination therapies

TCR-T cell therapy in combination with other therapeutic regimens represents another important strategy for enhancing efficacy. When TCR-T cell therapy is combined with clinically established cancer treatments such as chemotherapy, tumor vaccines, immunotherapy, and radiation therapy, these modalities may mutually reinforce each other, achieving synergistic solid tumor treatment efficacy that surpasses the cumulative benefit of their individual effects.

In clinical settings, the effectiveness of TCR-T cell therapies is affected by previous chemotherapy and other immunotherapy, with high lymphodepleting (LD) chemotherapy doses increasing the risk of adverse reactions such as febrile neutropenia and grade 3 or higher anemia [ 166 ]. By analyzing the outcomes of the cell therapy in 45 patients with synovial sarcoma ( NCT01343043 ), Alexandra Gyurdieva et al. found that prior to adoptive T-cell infusion, patient response could be predicted based on biomarker levels such as IL-15. Consequently, individualized LD chemotherapy regimens affecting in vivo cell expansion and associated with clinical response could be formulated to improve treatment outcomes and effector memory [ 167 ]. Additionally, the selection of the dosage for the LD regimen is also closely related to the efficacy and prognosis of TCR-T cells. High-dose LD regimen of fludarabine and cyclophosphamide, for example, promotes better infiltration of NY-ESO-1 SPEAR T cells into synovial sarcoma, to exert their functions [ 168 ]. In contrast, a single cycle of cyclophosphamide lymphodepletion yielded minimal enhancement of the therapeutic response of afami-cel for the same malignancy [ 36 ].

Furthermore, the synergistic effect of tumor vaccination on TCR-T cell therapies is still under exploration. In a clinical trial applying lymphocytes engineered with NY-ESO-1-reactive TCR genes ( NCT00670748 ), 11 patients were immunized with recombinant NY-ESO-1 AVIPOX vaccine, but the clinical responses were comparable to those of the monotherapy group [ 37 ]. After innovation and exploration, combining low-dose IL-2 and fresh adoptive cell infusion as synergistic methods, Theodore S. Nowicki et al. demonstrated the safety and feasibility of a dual-cell therapy combining genetically engineered NY-ESO-1 TCR-T cells with NY-ESO-1 peptide-pulsed DC vaccines in patients with NY-ESO-1-positive synovial sarcoma and other solid tumors. Among 6 patients, 4 showed evidence of short-term tumor regression ( NCT02070406 ), but long-term remission was rare and the durability was insufficient [ 169 ]. In addition, the pullulan nanogel drug delivery system containing the long peptide antigen (LPA) vaccine has been shown to increase the number of TCR-T cells in lymph nodes and tumor tissues. In a Phase I trial in 3 patients with advanced refractory soft tissue sarcoma, pullulan nanogel containing LPA targeting NY-ESO-1 157–165 epitope demonstrated potential to enhance long-lasting efficacy of TCR-T cells [ 170 ]. Given the limited patient cohort in clinical trials and uncertain efficacy, vaccine-combined TCR-T cell therapy, despite its theoretical rationale, still requires extensive exploration in larger cohorts to validate its clinical utility.

However, clinical trials combining immune checkpoint inhibitors have not shown significant improvements in completed trials to date. A multi-arm clinical trial ( NCT03709706 ) of evaluating lete-cel combined with PD-1 inhibitor pembrolizumab for advanced or recurrent NSCLC showed that the combination therapy did not increase toxicity, but the efficacy was limited, which may be due to other resistance mechanisms [ 40 ]. Similarly, a trial ( NCT01697527 ) combining NY-ESO-1-specific TCR-T cells with CTLA-4 inhibitor ipilimumab also failed to show significant therapeutic benefits [ 169 ].

Notably, TCR-T cell therapies combined with radiotherapy may represent a new approach for patients who are intolerant to chemotherapy or with refractory systemic therapy. Low-dose radiotherapy (LDRT) combined with TCR-T cells afami-cel targeting MAGE-A4 tumor antigen showed a more durable response in a patient with metastatic mucosal melanoma, with a PR of 10 weeks after treatment [ 171 ].

In summary, combination therapeutic strategies incorporating TCR-T cells are still in an exploratory phase, with chemotherapeutic regimens currently representing the most established option. Meanwhile, emerging approaches such as tumor vaccines are progressively being integrated into clinical investigations due to compatible therapeutic mechanisms and promising efficacy profiles.

Outlook

The application of TCR-T cell therapies in clinical solid tumor patients has not been widespread, which is largely due to the complex mechanisms underlying TCR-T cell function. TCR-T cells recognize antigen epitopes presented by HLA, and execute their antitumor effects via multiple steps: infiltration into tumor tissue; specific recognition of HLA-presented antigen epitopes; activation of TCR signals and cytotoxic killing of target cells; and establishment of long-term memory responses or exhaustion in vivo. Disruption at any step may compromise efficacy of TCR-T cells. In addition, effective T cells may be lost in the production process of engineered T cells. As a result, the quantity and quality of engineered T cells surviving in the patients’ body may be insufficient to achieve therapeutic efficacy. From foundational antigen and TCR screening through preclinical optimization to synergistic clinical application, global researchers have made transformative innovations and improvements in every aspect of TCR-T cell therapies. This collaborative progress is now translating TCR-T cell therapies from conceptual promise into clinical reality for patients with solid malignancies.

The complexity of the TME may compromise the efficacy of combinatorial regimens or even elicit paradoxical effects. Renpeng Ding et al. found that although elevated tumor PD-L1 expression impaired the effector function of TCR-T cells in vitro, the results of scRNA-seq revealed that high tumor PD-L1 expression could downregulate both pro-inflammatory and anti-inflammatory transcription factors, and blockade of tumor PD-L1 alone may inhibit not only anti-inflammatory response, but also pro-inflammatory response [ 172 ]. Interventions with immune agents may be influenced by dynamic pathway crosstalk in tumor cells, so it is necessary to select the appropriate combination therapies and doses, and the implementation of the multi-pathway and multi-target integrated therapies to kill “cunning and variable” tumor cells.

Current evidence indicates that single-component TCR-T cells exhibit limited cytotoxicity against tumors and can barely achieve the purpose of tumor eradication. The combination of CD4 + and CD8 + TCR-T cells may be able to more fully simulate human immune response and extend patient survival [ 77 , 173 ]. Notably, CD8 + T cells engineered with mismatched HLA Class-II-restricted TCRs exhibit targeted cytotoxicity akin to conventional cytotoxic CD4 + T cells [ 174 ]. This suggests a novel strategy for the combined use of different HLA-restricted T-cell products. Garry Dolton et al. discovered a single TCR targeting multiple HLA-A* 02:01-restricted epitopes in a case of stage IV malignant melanoma after successful TIL treatment [ 29 ]. Such TCRs target and eliminate various tumor cell types, facilitating broader applicability of TCR-T cell therapies. Dual TCR-T cells with strong antitumor activity and tumor tissue retention preference have been found to be abundant in patients with NSCLC, providing a new perspective for the development of TCR-T cell therapies [ 175 ]. The quality of current TCR-T cell products remains to be standardized, and significant challenges persist in identifying optimal antigens, epitopes and TCRs [ 127 ]. TCR-T cell products should be compared with natural CD8 + (or CD4 + ) T cells before clinical application to thoroughly evaluate the targeting efficacy of engineered T cells [ 25 ], thereby enabling the removal of cell products that may be clinically ineffective to improve the efficiency of clinical studies. Recently, a BRAF mutation neoantigen-specific T-cell product for melanoma, BNT221, was obtained through an in vitro induction process known as NEO-STIM, which induces the response of CD8 + and CD4 + T cells in peripheral blood to neoantigen epitopes. In a Phase I clinical trial, significant therapeutic responses were observed, with a 20% reduction in tumor size after just 6 weeks of treatment [ 176 ]. These findings not only validate the superiority of polyclonal T-cell products but also offer new insights into future development of personalized TCR-T cell therapies.

In summary, the current therapeutic efficacy of TCR-T cells depends on the type of tumor and the level of target antigen expression, with significant individual differences. In the clinical trial of the HLA-A*02-restricted MAGE-A4 TCR-T cell therapy, the expression rate of MAGE-A4 was 26%, with synovial sarcoma being the highest (70%) and gastric cancer the lowest (only 9%) [ 163 ]. Patient eligibility is constrained by multiple factors, and patients who meet the treatment conditions do not account for the majority. Current TCR-T cell therapies benefit only a limited subset of patients, and insufficient expression or mutation of the target antigen can result in ineffective treatment [ 9 ]. Future development of TCR-T cell therapies requires not only enhancing the antitumor activity of engineered T cells against solid tumors, but also expanding the eligible patient population. Our team is currently conducting an open-label, single-arm, Phase I clinical trial ( NCT06478251 ) to evaluate the safety and efficacy of KRAS G12V/G12D mutation-specific TCR-T cell therapy in patients with advanced pancreatic cancer and colorectal cancer harboring KRAS G12V/G12D mutations. We aim to provide clinical evidence to support the development of TCR-T cell products targeting hotspot mutations as neoantigens. By establishing a tumor-specific antigen library and inducing tumor cells to express target antigens, researchers are working to include patients with various solid tumors in the treatment cohort from multiple dimensions, so as to achieve the universal application of individualized TCR-T cell therapies in patients with solid tumors. This represents both a critical challenge and a primary objective for future research.

Recently, KSX01-TCR-T, developed by a Chinese biotech firm, leverages AI-driven TCR recognition technology combined with single-cell omics data and tumor-reactive T-cell fingerprinting to rapidly identify personalized TCR sequences suitable for patients in just 10 days. This approach is not restricted by target antigens or HLA types, significantly expanding the patient population eligible for TCR-T cell therapies. The preliminary results of a Phase I clinical trial, reported at the annual meeting of the European Society for Medical Oncology (ESMO), showed objective response rates (ORR) of 33% (1/3) and 67% (2/3) in two different dosing cohorts ( NCT06150365 ). The incorporation of AI technology offers a new perspective for the development of TCR-T cell therapies by greatly accelerating personalized TCR discovery and expanding patients who can potentially receive tailored treatments. This approach enhances feasibility and scalability of individualized TCR-T cell therapy, enabling more patients with advanced solid tumors to be treated efficiently.

Supplementary Information

Supplementary Material 1.

Acknowledgements

Not applicable.

Author contributions

YL: Writing—original draft preparation, review and editing; XZ: Conceptualization, Resources; YC and JL: Writing—review and editing, Investigation; TL: Supervision, Conceptualization; XB: Supervision, Resources. All authors read and approved the final manuscript.

Funding

This work was supported by grants from the National Key Research and Development Program of China [2024YFA1306400 to XLB, China]; the Joint Fund for Regional Innovation and Development of National Natural Science Foundation of China [U23A20462 to XLB, China]; “Ling Yan” Research and Development Program of Department of Zhejiang Province Science and Technology [2024C03167 to XLB, China]; National Natural Science Foundation of China [82403852 and 82573008 to XZZ, China]; Natural Science Foundation of Zhejiang Province [LQN25H160036 to XZZ, China].

Data availability

No datasets were generated or analyzed 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.

Footnotes

Publisher's Note

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

Yawen Li and Xiaozhen Zhang have contributed equally to this work.

Contributor Information

Tingbo Liang, Email: liangtingbo@zju.edu.cn.

Xueli Bai, Email: shirleybai@zju.edu.cn.

References

1. Lin H, Liu C, Hu A, Zhang D, Yang H, Mao Y. Understanding the immunosuppressive microenvironment of glioma: Mechanistic insights and clinical perspectives. J Hematol Oncol. 2024;17(1):31.

2. Yin Y, Feng W, Chen J, Chen X, Wang G, Wang S, et al. Immunosuppressive tumor microenvironment in the progression, metastasis, and therapy of hepatocellular carcinoma: From bench to bedside. Exp Hematol Oncol. 2024;13(1):72.

3. Liu Z, Lei W, Wang H, Liu X, Fu R. Challenges and strategies associated with CAR-T cell therapy in blood malignancies. Exp Hematol Oncol. 2024;13(1):22.

4. Ai K, Liu B, Chen X, Huang C, Yang L, Zhang W, et al. Optimizing CAR-T cell therapy for solid tumors: Current challenges and potential strategies. J Hematol Oncol. 2024;17(1):105.

5. Shao W, Yao Y, Yang L, Li X, Ge T, Zheng Y, et al. Novel insights into TCR-T cell therapy in solid neoplasms: Optimizing adoptive immunotherapy. Exp Hematol Oncol. 2024;13(1):37.

6. Albarrán Fernández V, Ballestín Martínez P, Stoltenborg Granhøj J, Borch TH, Donia M, Marie Svane I. Biomarkers for response to TIL therapy: A comprehensive review. J Immunother Cancer. 2024;12(3):e008640.

7. Sarnaik AA, Hwu P, Mulé JJ, Pilon-Thomas S. Tumor-infiltrating lymphocytes: a new hope. Cancer Cell. 2024;42(8):1315–8.

8. Hu W, Bian Y, Ji H. TIL therapy in lung cancer: current progress and perspectives. Adv Sci. 2024;11(46):e2409356.

9. Tran E, Robbins PF, Lu Y-C, Prickett TD, Gartner JJ, Jia L, et al. T-cell transfer therapy targeting mutant KRAS in cancer. N Engl J Med. 2016;375(23):2255–62.

10. Goff SL, Dudley ME, Citrin DE, Somerville RP, Wunderlich JR, Danforth DN, et al. Randomized, prospective evaluation comparing intensity of lymphodepletion before adoptive transfer of tumor-infiltrating lymphocytes for patients with metastatic melanoma. J Clin Oncol. 2016;34(20):2389–97.

11. Lin H, Yang X, Ye S, Huang L, Mu W. Antigen escape in CAR-T cell therapy: mechanisms and overcoming strategies. Biomed Pharmacother. 2024;178:117252.

12. Liu J, Jiao X, Ma D, Fang Y, Gao Q. CAR-T therapy and targeted treatments: emerging combination strategies in solid tumors. Medicine (Baltimore). 2024;5(6):530–49.

13. Pu J, Liu T, Sharma A, Jiang L, Wei F, Ren X, et al. Advances in adoptive cellular immunotherapy and therapeutic breakthroughs in multiple myeloma. Exp Hematol Oncol. 2024;13(1):105.

14. Hushmandi K, Imani Fooladi AA, Reiter RJ, Farahani N, Liang L, Aref AR, et al. Next-generation immunotherapeutic approaches for blood cancers: exploring the efficacy of CAR-T and cancer vaccines. Exp Hematol Oncol. 2025;14(1):75.

15. Zhao Z, Chen Y, Francisco NM, Zhang Y, Wu M. The application of CAR-T cell therapy in hematological malignancies: advantages and challenges. Acta Pharm Sin B. 2018;8(4):539–51.

16. June CH, Sadelain M. Chimeric antigen receptor therapy. N Engl J Med. 2018;379(1):64–73.

17. Yang C, Liu Y, Huang Z, Liu S, Zhang X, Liu Q, et al. Recent advances and challenges of cellular immunotherapies in lung cancer treatment. Exp Hematol Oncol. 2025;14(1):94.

18. Liu Y, Duan Y, Du Z, Lu B, Liu S, Li L, et al. Current challenges and emerging opportunities of chimeric antigen receptor-engineered cell immunotherapy. Exp Hematol Oncol. 2025;14(1):92.

19. Li X, Chen T, Li X, Zhang H, Li Y, Zhang S, et al. Therapeutic targets of armored chimeric antigen receptor T cells navigating the tumor microenvironment. Exp Hematol Oncol. 2024;13(1):96.

20. 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.

21. Hong M, Talluri S, Chen YY. Advances in promoting chimeric antigen receptor T cell trafficking and infiltration of solid tumors. Curr Opin Biotechnol. 2023;84:103020.

22. 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.

23. Xu R, Du S, Zhu J, Meng F, Liu B. Neoantigen-targeted TCR-T cell therapy for solid tumors: how far from clinical application. Cancer Lett. 2022;546:215840.

24. Ma P, Jiang Y, Zhao G, Wang W, Xing S, Tang Q, et al. Toward a comprehensive solution for treating solid tumors using T-cell receptor therapy: a review. Eur J Cancer. 2024;209:114224.

25. Sykulev Y. Factors contributing to the potency of CD8+ T cells. Trends Immunol. 2023;44(9):693–700.

26. Wei T, Leisegang M, Xia M, Kiyotani K, Li N, Zeng C, et al. Generation of neoantigen-specific T cells for adoptive cell transfer for treating head and neck squamous cell carcinoma. OncoImmunology. 2021;10(1):1929726.

27. Li Z, Ma L, Gao Z, Wang X, Che X, Zhang P, et al. Identification and validation of tumor-specific T cell receptors from tumor infiltrating lymphocytes using tumor organoid co-cultures. Cancer Immunol Immunother. 2024;73(9):164–76.

28. He J, Xiong X, Yang H, Li D, Liu X, Li S, et al. Defined tumor antigen-specific T cells potentiate personalized TCR-T cell therapy and prediction of immunotherapy response. Cell Res. 2022;32(6):530–42.

29. Dolton G, Rius C, Wall A, Szomolay B, Bianchi V, Galloway SAE, et al. Targeting of multiple tumor-associated antigens by individual T cell receptors during successful cancer immunotherapy. Cell. 2023;186(16):3333-49.e27.

30. Wermke M, Holderried TAW, Luke JJ, Morris VK, Alsdorf WH, Wetzko K, et al. First-in-human dose escalation trial to evaluate the clinical safety and efficacy of an anti-MAGEA1 autologous TCR-transgenic T cell therapy in relapsed and refractory solid tumors. J Immunother Cancer. 2024;12(7):e008668.

31. Tan AT, Yang N, Lee Krishnamoorthy T, Oei V, Chua A, Zhao X, et al. Use of expression profiles of HBV-DNA integrated into genomes of hepatocellular carcinoma cells to select T cells for immunotherapy. Gastroenterology. 2019;156(6):1862-76.e9.

32. Liu M, Akahori Y, Imai N, Wang L, Negishi K, Kato T, et al. MAGE-A4 pMHC-targeted CAR-T cells exploiting TCR machinery exhibit significantly improved in vivo function while retaining antigen specificity. J Immunother Cancer. 2024;12(11):e010248.

33. Clay TM, Custer MC, Sachs J, Hwu P, Rosenberg SA, Nishimura MI. Efficient transfer of a tumor antigen-reactive TCR to human peripheral blood lymphocytes confers anti-tumor reactivity. J Immunol. 1999;163(1):507–13.

34. Fetsch PA, Marincola FM, Filie A, Hijazi YM, Kleiner DE, Abati A. Melanoma-associated antigen recognized by T cells (MART-1): the advent of a preferred immunocytochemical antibody for the diagnosis of metastatic malignant melanoma with fine-needle aspiration. Cancer. 1999;87(1):37–42.

35. Nagarsheth NB, Norberg SM, Sinkoe AL, Adhikary S, Meyer TJ, Lack JB, et al. TCR-engineered T cells targeting E7 for patients with metastatic HPV-associated epithelial cancers. Nat Med. 2021;27(3):419–25.

36. D’Angelo SP, Araujo DM, Abdul Razak AR, Agulnik M, Attia S, Blay J-Y, et al. Afamitresgene autoleucel for advanced synovial sarcoma and myxoid round cell liposarcoma (SPEARHEAD-1): an international, open-label, phase 2 trial. Lancet. 2024;403(10435):1460–71.

37. 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.

38. Goddard ET, Linde MH, Srivastava S, Klug G, Shabaneh TB, Iannone S, et al. Immune evasion of dormant disseminated tumor cells is due to their scarcity and can be overcome by T cell immunotherapies. Cancer Cell. 2024;42(1):119-34.e12.

39. Davari K, Holland T, Prassmayer L, Longinotti G, Ganley KP, Pechilis LJ, et al. Development of a CD8 co-receptor independent T-cell receptor specific for tumor-associated antigen MAGE-A4 for next generation T-cell-based immunotherapy. J Immunother Cancer. 2021;9(3):e002035.

40. Altan M, Lopes G, Hiltermann TJN, Govindan R, Villaruz LC, Calvo E, et al. Safety and tolerability of letetresgene autoleucel (GSK3377794): pilot studies in patients with advanced non-small cell lung cancer. Clin Cancer Res. 2025;31(3):529–42.

41. Blumenschein GR, Devarakonda S, Johnson M, Moreno V, Gainor J, Edelman MJ, et al. Phase I clinical trial evaluating the safety and efficacy of ADP-A2M10 SPEAR T cells in patients with MAGE-A10+ advanced non-small cell lung cancer. J Immunother Cancer. 2022;10(1):e003581.

42. Simon P, Omokoko TA, Breitkreuz A, Hebich L, Kreiter S, Attig S, et al. Functional TCR retrieval from single antigen-specific human T cells reveals multiple novel epitopes. Cancer Immunol Res. 2014;2(12):1230–44.

43. Shafer P, Leung WK, Woods M, Choi JM, Rodriguez-Plata CM, Maknojia A, et al. Incongruity between T cell receptor recognition of breast cancer hotspot mutations ESR1 Y537S and D538G following exogenous peptide loading versus endogenous antigen processing. Cytotherapy. 2024;26(3):266–75.

44. Festag J, Festag MM, Asen T, Wettengel JM, Mück-Häusl MA, Abdulhaqq S, et al. Vector-mediated delivery of human major histocompatibility complex-I into hepatocytes enables investigation of T cell receptor-redirected hepatitis B virus-specific T cells in mice, and in macaque cell cultures. Hum Gene Ther. 2023;34(23–24):1204–18.

45. Xia S, Huang J, Yan L, Han J, Zhang W, Shao H, et al. miR-150 promotes progressive T cell differentiation via inhibiting FOXP1 and RC3H1. Hum Immunol. 2022;83(11):778–88.

46. Zhang C, Sun Y, Li S, Shen L, Teng X, Xiao Y, et al. Autophagic flux restoration of senescent T cells improves antitumor activity of TCR‐engineered T cells. Clin Transl Immunol. 2022;11(9):e1419.

47. Wu B, Zhang Q, Hong P, Liu L, Peng H, Xia C, et al. Antigen‐specific TCR‐T cells from Rag2 gene‐deleted pluripotent stem cells impede solid tumour growth in a mouse model. Cell Prolif. 2023;56(4):e13389.

48. Ishihara M, Kitano S, Kageyama S, Miyahara Y, Yamamoto N, Kato H, et al. NY-ESO-1-specific redirected T cells with endogenous TCR knockdown mediate tumor response and cytokine release syndrome. J Immunother Cancer. 2022;10(6):e003811.

49. Zheng S, Wang W, Shen L, Yao Y, Xia W, Ni C. Tumor battlefield within inflamed, excluded or desert immune phenotypes: the mechanisms and strategies. Exp Hematol Oncol. 2024;13(1):80.

50. Asanuma K, Ishihara M, Nakamura T, Uchida K, Hagi T, Kageyama S, et al. Histological assessment of synovial sarcoma before and after TCR-T Cell therapy and cryoablation: a case report. Anticancer Res. 2023;43(12):5737–48.

51. Minowa T, Hirohashi Y, Murata K, Sasaki K, Handa T, Nakatsugawa M, et al. Fusion with type 2 macrophages induces melanoma cell heterogeneity that potentiates immunological escape from cytotoxic T lymphocytes. J Pathol. 2023;260(3):304–16.

52. Chen Y, Ouyang D, Wang Y, Pan Q, Zhao J, Chen H, et al. EBV promotes TCR-T-cell therapy resistance by inducing CD163+M2 macrophage polarization and MMP9 secretion. J Immunother Cancer. 2024;12(6):e008375.

53. Lahman MC, Schmitt TM, Paulson KG, Vigneron N, Buenrostro D, Wagener FD, et al. Targeting an alternate Wilms’ tumor antigen 1 peptide bypasses immunoproteasome dependency. Sci Transl Med. 2022;14(631):eabg8070.

54. Maggs L, Sadagopan A, Moghaddam AS, Ferrone S. HLA class I antigen processing machinery defects in antitumor immunity and immunotherapy. Trends Cancer. 2021;7(12):1089–101.

55. Morgan RA, Dudley ME, Wunderlich JR, Hughes MS, Yang JC, Sherry RM, et al. Cancer regression in patients after transfer of genetically engineered lymphocytes. Science. 2006;314(5796):126–9.

56. Cameron BJ, Gerry AB, Dukes J, Harper JV, Kannan V, Bianchi FC, et al. Identification of a Titin-derived HLA-A1-presented peptide as a cross-reactive target for engineered MAGE A3-directed T cells. Sci Transl Med. 2013;5(197):197ra03.

57. 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.

58. Zhao X, Kolawole EM, Chan W, Feng Y, Yang X, Gee MH, et al. Tuning T cell receptor sensitivity through catch bond engineering. Science. 2022. 10.1126/science.abl5282.

59. Kim ST, Tayar J, Fu S, Ke D, Norry E, Sun A, et al. Newly developed pseudogout arthritis after therapy with MAGE-A4 directed TCR T cells responded to treatment with tocilizumab. J Immunother Cancer. 2021. 10.1136/jitc-2021-002716.

60. Bendle GM, Linnemann C, Hooijkaas AI, Bies L, de Witte MA, Jorritsma A, et al. Lethal graft-versus-host disease in mouse models of T cell receptor gene therapy. Nat Med. 2010;16(5):565–70.

61. Heemskerk MH, Hagedoorn RS, van der Hoorn MA, van der Veken LT, Hoogeboom M, Kester MG, et al. Efficiency of T-cell receptor expression in dual-specific T cells is controlled by the intrinsic qualities of the TCR chains within the TCR-CD3 complex. Blood. 2007;109(1):235–43.

62. Provasi E, Genovese P, Lombardo A, Magnani Z, Liu PQ, Reik A, et al. Editing T cell specificity towards leukemia by zinc finger nucleases and lentiviral gene transfer. Nat Med. 2012;18(5):807–15.

63. Zhang Y, Liu Z, Wei W, Li Y. TCR engineered T cells for solid tumor immunotherapy. Exp Hematol Oncol. 2022;11(1):38.

64. Wang R, Zhuang J, Zhang Q, Wu W, Yu X, Zhang H, et al. Decoding the metabolic dialogue in the tumor microenvironment: from immune suppression to precision cancer therapies. Exp Hematol Oncol. 2025;14(1):99.

65. Pavesi A, Tan AT, Koh S, Chia A, Colombo M, Antonecchia E, et al. A 3D microfluidic model for preclinical evaluation of TCR-engineered T cells against solid tumors. JCI Insight. 2017;2(12):e89762.

66. Lee SWL, Adriani G, Ceccarello E, Pavesi A, Tan AT, Bertoletti A, et al. Characterizing the role of monocytes in T cell cancer immunotherapy using a 3D microfluidic model. Front Immunol. 2018;9:416.

67. Bushman FD. Retroviral insertional mutagenesis in humans: evidence for four genetic mechanisms promoting expansion of cell clones. Mol Ther. 2020;28(2):352–6.

68. Hacein-Bey-Abina S, Von Kalle C, Schmidt M, McCormack MP, Wulffraat N, Leboulch P, et al. LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science (New York, NY).302 (5644):415–9.

69. Nobles CL, Sherrill-Mix S, Everett JK, Reddy S, Fraietta JA, Porter DL, et al. CD19-targeting CAR T cell immunotherapy outcomes correlate with genomic modification by vector integration. J Clin Invest. 2019;130(2):673–85.

70. Shah NN, Qin H, Yates B, Su L, Shalabi H, Raffeld M, et al. Clonal expansion of CAR T cells harboring lentivector integration in the CBL gene following anti-CD22 CAR T-cell therapy. Blood Adv. 2019;3(15):2317–22.

71. Foy SP, Jacoby K, Bota DA, Hunter T, Pan Z, Stawiski E, et al. Non-viral precision T cell receptor replacement for personalized cell therapy. Nature. 2022;615(7953):687–96.

72. Nyberg WA, Ark J, To A, Clouden S, Reeder G, Muldoon JJ, et al. An evolved AAV variant enables efficient genetic engineering of murine T cells. Cell. 2023;186(2):446-60.e19.

73. Kang S, Li Y, Bao Y, Li Y. High-affinity T cell receptors redirect cytokine-activated T cells (CAT) to kill cancer cells. Front Med. 2019;13(1):69–82.

74. Healy K, Pavesi A, Parrot T, Sobkowiak MJ, Reinsbach SE, Davanian H, et al. Human MAIT cells endowed with HBV specificity are cytotoxic and migrate towards HBV-HCC while retaining antimicrobial functions. JHEP Rep. 2021;3(4):100318.

75. Zhang B, Ren Z, Zhao J, Zhu Y, Huang B, Xiao C, et al. Global analysis of HLA-A2 restricted MAGE-A3 tumor antigen epitopes and corresponding TCRs in non-small cell lung cancer. Theranostics. 2023;13(13):4449–68.

76. Chen P, Chen D, Bu D, Gao J, Qin W, Deng K, et al. Dominant neoantigen verification in hepatocellular carcinoma by a single-plasmid system coexpressing patient HLA and antigen. J Immunother Cancer. 2023;11(4):e006334.

77. Jiang J, Xia M, Zhang L, Chen X, Zhao Y, Zeng C, et al. Rapid generation of genetically engineered T cells for the treatment of virus-related cancers. Cancer Sci. 2022;113(11):3686–97.

78. Hanada KI, Zhao C, Gil-Hoyos R, Gartner JJ, Chow-Parmer C, Lowery FJ, et al. A phenotypic signature that identifies neoantigen-reactive T cells in fresh human lung cancers. Cancer Cell. 2022;40(5):479–93.

79. Li Y, Qi J, Liu Y, Zheng Y, Zhu H, Zang Y, et al. High-throughput screening of functional neo-antigens and their specific T-cell receptors via the Jurkat reporter system combined with droplet microfluidics. Anal Chem. 2023;95(25):9697–705.

80. Qi J, Zhu H, Li Y, Guan X, He Y, Ren G, et al. Creation of a high‐throughput microfluidic platform for single‐cell transcriptome sequencing of cell‐cell interactions. Small Methods. 2023;7(11):e2300730.

81. Wang X, Dai Z, Lin X, Zou X, Wang R, Tasiheng Y, et al. Antigen/HLA-agnostic strategies for Characterizing Tumor-responsive T cell receptors in PDAC patients via single-cell sequencing and autologous organoid application. Cancer Lett. 2024;588:216741.

82. Zhu W, Peng Y, Wang L, Hong Y, Jiang X, Li Q, et al. Identification of α-fetoprotein-specific T-cell receptors for hepatocellular carcinoma immunotherapy. Hepatology. 2018;68(2):574–89.

83. Xu Y, Shan W, Luo Q, Zhang M, Huo D, Chen Y, et al. Establishment of a humanized mouse model using steady-state peripheral blood-derived hematopoietic stem and progenitor cells facilitates screening of cancer-targeted T-cell repertoires. Cancer Innovation. 2024;3(3):e118.

84. Krämer C, Kilian M, Chih Y-C, Kourtesakis A, Hoffmann DC, Boschert T, et al. NLGN4X TCR transgenic T cells to treat gliomas. Neuro Oncol. 2024;26(2):266–78.

85. Bear AS, Nadler RB, O’Hara MH, Stanton KL, Xu C, Saporito RJ, et al. Natural TCRs targeting KRASG12V display fine specificity and sensitivity to human solid tumors. J Clin Invest. 2024;134(21):e175790.

86. Sennikov S, Volynets M, Alrhmoun S, Perik-Zavodskii R, Perik-Zavodskaia O, Fisher M, et al. Modified dendritic cell-based T-cell expansion protocol and single-cell multi-omics allow for the selection of the most expanded and in vitro-effective clonotype via profiling of thousands of MAGE-A3-specific T-cells. Front Immunol. 2024;15:1470130.

87. Long J, Chen X, He M, Ou S, Zhao Y, Yan Q, et al. HLA-class II restricted TCR targeting human papillomavirus type 18 E7 induces solid tumor remission in mice. Nat Commun. 2024;15(1):2271.

88. Füchsl F, Untch J, Kavaka V, Zuleger G, Braun S, Schwanzer A, et al. High-resolution profile of neoantigen-specific TCR activation links moderate stimulation to increased resilience of engineered TCR-T cells. Nat Commun. 2024;15(1):10520.

89. Wang L, Lan X. Rapid screening of TCR-pMHC interactions by the YAMTAD system. Cell Discov. 2022;8(1):30.

90. Ashby JF, Schmidt J, Kc N, Kurum A, Koch C, Harari A, et al. Microfluidic T cell selection by cellular avidity. Adv Healthcare Mater. 2022;11(16):e2200169.

91. Morimoto S, Fujiki F, Kondo K, Nakajima H, Kobayashi Y, Inatome M, et al. Establishment of a novel platform cell line for efficient and precise evaluation of T cell receptor functional avidity. Oncotarget. 2018;9(75):34132.

92. Fujiki F, Morimoto S, Nishida Y, Tanii S, Aoyama N, Inatome M, et al. Establishment of a novel NFAT-GFP reporter platform useful for the functional avidity maturation of HLA class II-restricted TCRs. Cancer Immunol Immunother. 2023;72(7):2347–56.

93. Miyao K, Terakura S, Okuno S, Julamanee J, Watanabe K, Hamana H, et al. Introduction of genetically modified CD3ζ improves proliferation and persistence of antigen-specific CTLs. Cancer Immunol Res. 2018;6(6):733–44.

94. Lah S, Kim S, Kang I, Kim H, Hupperetz C, Jung H, et al. Engineering second-generation TCR-T cells by site-specific integration of TRAF-binding motifs into theCD247locus. J Immunother Cancer. 2023;11(4):e005519.

95. Liang W, Yi R, Wang W, Shi Y, Zhang J, Xu X, et al. Enhancing the antitumor immunity of T cells by engineering the lipid-regulatory site of the TCR/CD3 complex. Cancer Immunol Res. 2023;11(1):93–108.

96. Sailer N, Fetzer I, Salvermoser M, Braun M, Brechtefeld D, Krendl C, et al. T-cells expressing a highly potent PRAME-specific T-cell receptor in combination with a chimeric PD1-41BB co-stimulatory receptor show a favorable preclinical safety profile and strong anti-tumor reactivity. Cancers. 2022;14(8):1998.

97. Ren Z, Wang K, Zhang Y, Chen H, Zhu Y, Li H, et al. Transient hydroxycholesterol treatment restrains TCR signaling to promote long-term immunity. Cell Chem Biol. 2024;31(5):920-31.e6.

98. Hussein MS, Li Q, Mao R, Peng Y, He Y. TCR T cells overexpressing c-Jun have better functionality with improved tumor infiltration and persistence in hepatocellular carcinoma. Front Immunol. 2023;14:1114770.

99. Shen M, Chen S, Han X, Hao Y, Wang J, Li L, et al. Identification of an HLA-A*11:01-restricted neoepitope of mutant PIK3CA and its specific T cell receptors for cancer immunotherapy targeting hotspot driver mutations. Cancer Immunol Immunother. 2024;73(8):150.

100. Li J, Xiao Z, Wang D, Jia L, Nie S, Zeng X, et al. The screening, identification, design and clinical application of tumor-specific neoantigens for TCR-T cells. Mol Cancer. 2023;22(1):141.

101. Minagawa A, Yoshikawa T, Yasukawa M, Hotta A, Kunitomo M, Iriguchi S, et al. Enhancing T cell receptor stability in rejuvenated iPSC-derived T cells improves their use in cancer immunotherapy. Cell Stem Cell. 2018;23(6):850-8.e4.

102. Saito H, Okita K, Chang AE, Ito F. Adoptive transfer of CD8+ T cells generated from induced pluripotent stem cells triggers regressions of large tumors along with immunological memory. Cancer Res. 2016;76(12):3473–83.

103. Chen S, Zhang J, Shen M, Han X, Li S, Hu C, et al. p38 inhibition enhances TCR-T cell function and antagonizes the immunosuppressive activity of TGF-β. Int Immunopharmacol. 2021;98:107848.

104. Tokunaga Y, Sasaki T, Goto S, Adachi K, Sakoda Y, Tamada K. Enhanced antitumor responses of tumor antigen-specific TCR T cells genetically engineered to produce IL7 and CCL19. Mol Cancer Ther. 2022;21(1):138–48.

105. Zhu W, Zhang Z, Chen J, Chen X, Huang L, Zhang X, et al. A novel engineered IL-21 receptor arms T-cell receptor-engineered T cells (TCR-T cells) against hepatocellular carcinoma. Signal Transduct Target Ther. 2024;9(1):101.

106. Zhang L, Davies JS, Serna C, Yu Z, Restifo NP, Rosenberg SA, et al. Enhanced efficacy and limited systemic cytokine exposure with membrane-anchored interleukin-12 T-cell therapy in murine tumor models. J Immunother Cancer. 2020;8(1):e000210.

107. Hu J, Yang Q, Zhang W, Du H, Chen Y, Zhao Q, et al. Cell membrane-anchored and tumor-targeted IL-12 (attIL12)-T cell therapy for eliminating large and heterogeneous solid tumors. J Immunother Cancer. 2022;10(1):e003633.

108. Kim S, Park CI, Lee S, Choi HR, Kim CH. Reprogramming of IL-12 secretion in the PDCD1 locus improves the anti-tumor activity of NY-ESO-1 TCR-T cells. Front Immunol. 2023;14:1062365.

109. Nguyen R, Doubrovina E, Mousset CM, Jin BY, Okada R, Zhang X, et al. Cooperative armoring of CAR and TCR T cells by T cell-restricted IL15 and IL21 universally enhances solid tumor efficacy. Clin Cancer Res. 2024;30(8):1555–66.

110. Xu W, Hu M, Lu X, Lao Y, Ma N, Wang Y, et al. Inhibition of PCSK9 enhances the anti-hepatocellular carcinoma effects of TCR-T cells and anti-PD-1 immunotherapy. Int J Biol Sci. 2024;20(10):3942–55.

111. Amissah OB, Chen W, de Dieu HJ, Sun Y, Lin L, Liu Y, et al. NY-ESO-1-specific T cell receptor-engineered T cells and Tranilast, a TRPV2 antagonist bivalent treatment enhances the killing of esophageal cancer: a dual-targeted cancer therapeutic route. Cancer Cell Int. 2024;24(1):64.

112. Wang J, Han X, Hao Y, Chen S, Pang B, Zou L, et al. Cbl-b inhibition promotes less differentiated phenotypes of T cells with enhanced cytokine production. Cellular Immunol. 2024;403:104863.

113. Zhou Z, Zheng J, Lu Y, Mai Z, Lin Y, Lin P, et al. Optimizing CD8 (+) T cell-based immunotherapy via metabolic interventions: a comprehensive review of intrinsic and extrinsic modulators. Exp Hematol Oncol. 2024;13(1):103.

114. Van der Vreken A, Vanderkerken K, De Bruyne E, De Veirman K, Breckpot K, Menu E. Fueling CARs: metabolic strategies to enhance CAR T-cell therapy. Exp Hematol Oncol. 2024;13(1):66.

115. Feng B, Li R, Li W, Tang L. Metabolic immunoengineering approaches to enhance CD8+ T cell-based cancer immunotherapy. Cell Syst. 2024;15(12):1225–44.

116. Liu Y, Wang F, Peng D, Zhang D, Liu L, Wei J, et al. Activation and antitumor immunity of CD8 (+) T cells are supported by the glucose transporter GLUT10 and disrupted by lactic acid. Sci Transl Med. 2024;16(762):7399.

117. Peralta RM, Xie B, Lontos K, Nieves-Rosado H, Spahr K, Joshi S, et al. Dysfunction of exhausted T cells is enforced by MCT11-mediated lactate metabolism. Nat Immunol. 2024;25(12):2297–307.

118. Qiu Y, Su Y, Xie E, Cheng H, Du J, Xu Y, et al. Mannose metabolism reshapes T cell differentiation to enhance anti-tumor immunity. Cancer Cell. 2025;43(1):103-21.e8.

119. Zhou W, Qu M, Yue Y, Zhong Z, Nan K, Sun X, et al. Acetylcysteine synergizes PD-1 blockers against colorectal cancer progression by promoting TCF1+PD1+CD8+ T cell differentiation. Cell Commun Signal. 2024;22(1):503.

120. Pallavicini I, Frasconi TM, Catozzi C, Ceccacci E, Tiberti S, Haas D, et al. LSD1 inhibition improves efficacy of adoptive T cell therapy by enhancing CD8+ T cell responsiveness. Nat Commun. 2024;15(1):7366.

121. Ma S, Ong L-T, Jiang Z, Lee WC, Lee PL, Yusuf M, et al. Targeting P4HA1 promotes CD8+ T cell progenitor expansion toward immune memory and systemic anti-tumor immunity. Cancer Cell. 2025;43(2):213-31.e9.

122. Wang Y, Jiang C, Zhou H, Han R. Transforming cancer immunotherapy: integration of distinct immune-based approaches as redefined dual immunotherapy with potential third-sensitizer. Exp Hematol Oncol. 2025;14(1):114.

123. Zheng N, Fang J, Xue G, Wang Z, Li X, Zhou M, et al. Induction of tumor cell autosis by myxoma virus-infected CAR-T and TCR-T cells to overcome primary and acquired resistance. Cancer Cell. 2022;40(9):973-85.e7.

124. Drakes DJ, Abbas AM, Shields J, Steinbuck MP, Jakubowski A, Seenappa LM, et al. Lymph node-targeted vaccine boosting of TCR T-cell therapy enhances antitumor function and eradicates solid tumors. Cancer Immunol Res. 2024;12(2):214–31.

125. Stefanidis E, Semilietof A, Pujol J, Seijo B, Scholten K, Zoete V, et al. Combining SiRPα decoy-coengineered T cells and antibodies augments macrophage-mediated phagocytosis of tumor cells. J Clin Invest. 2024;134(11):e161660.

126. Wang X, Yang X, Wang Y, Chen Y, Yang Y, Shang S, et al. Combination of expanded allogeneic NK cells and T cell-based immunotherapy exert enhanced antitumor effects. Cancers. 2022;15 (1).

127. Kortleve D, Hammerl D, van Brakel M, Wijers R, Roelofs D, Kroese K, et al. TCR-engineered T cells directed against Ropporin-1 constitute a safe and effective treatment for triple-negative breast cancer. Cancer Discov. 2024;14(12):2450–70.

128. van Amerongen RA, Tuit S, Wouters AK, van de Meent M, Siekman SL, Meeuwsen MH, et al. PRAME and CTCFL-reactive TCRs for the treatment of ovarian cancer. Front Immunol. 2023;14:1121973.

129. Jin BY, Campbell TE, Draper LM, Stevanović S, Weissbrich B, Yu Z, et al. Engineered T cells targeting E7 mediate regression of human papillomavirus cancers in a murine model. JCI Insight. 2018;3(8):e99488.

130. Li X, Wang W, Wang J, Jiang M, He J, Tan S. Identification of HPV-E7 specific TCRs for tumor immunotherapy. Mol Immunol. 2024;171:56–65.

131. Zhang H, Sun M, Wang J, Zeng B, Cao X, Han Y, et al. Identification of NY-ESO-1157–165 specific murine T cell receptors with distinct recognition pattern for tumor immunotherapy. Front Immunol. 2021;12:644520.

132. Xu R, Wang Q, Zhu J, Bei Y, Chu Y, Sun Z, et al. Membrane fusogenic nanoparticle-based HLA-peptide-addressing universal T cell receptor-engineered T (HAUL TCR-T) cell therapy in solid tumor. Bioeng Transl Med. 2023;8(6):e10585.

133. Wang Q, Peng R, Qi H, Xu R, Liu W, Meng F, et al. Liposome-based in situ antigen-modification strategy for “universal” T-cell-receptor engineered T cell in cancer immunotherapy. MedComm. 2024;5(7):e618.

134. Lu D, Chen Y, Jiang M, Wang J, Li Y, Ma K, et al. KRAS G12V neoantigen specific T cell receptor for adoptive T cell therapy against tumors. Nat Commun. 2023;14(1):6389.

135. Long X, Zhang Z, Li Y, Deng K, Gao W, Huang M, et al. ScRNA-seq reveals novel immune-suppressive T cells and investigates CMV-TCR-T cells cytotoxicity against GBM. J Immunother Cancer. 2024;12(4):e008967.

136. Wang J, Zhang T, Li P, Gai J, Chen S, Espinoza G, et al. Engineered TCR T-cell therapy targeting mass spectrometry-identified natural epitope in PDAC. Cancer Lett. 2023;573:216366.

137. Liu Q, Tian Y, Li Y, Zhang W, Cai W, Liu Y, et al. In vivo therapeutic effects of affinity-improved-TCR engineered T-cells on HBV-related hepatocellular carcinoma. J Immunother Cancer. 2020;8(2):e001748.

138. Vercher E, Covo-Vergara Á, Conde E, Hernández-Rueda M, Elizalde E, Mancheño U, et al. Human T cells engineered with an HLA-A2-restricted murine T-cell receptor targeting glypican 3 effectively control human hepatocellular carcinoma in mice. Hepatology. 2024;82(2):326–43.

139. Yao P, Gao M, Hu W, Wang J, Wang Y, Wang Q, et al. Proteogenomic analysis identifies neoantigens and bacterial peptides as immunotherapy targets in colorectal cancer. Pharmacol Res. 2024;204:107209.

140. Dillard P, Köksal H, Maggadottir SM, Winge-Main A, Pollmann S, Menard M, et al. Targeting telomerase with an HLA Class II-restricted TCR for cancer immunotherapy. Mol Ther. 2021;29(3):1199–213.

141. Wang C, Chen J, Li J, Xu Z, Huang L, Zhao Q, et al. An EBV-related CD4 TCR immunotherapy inhibits tumor growth in an HLA-DP5+ nasopharyngeal cancer mouse model. J Clin Invest. 2024;134(8):e172092.

142. Liang Y, Xu Q, Liu S, Li J, Wang F, Li Z, et al. Single-Cell Transcriptomics Reveals Killing Mechanisms of Antitumor Cytotoxic CD4+ TCR-T Cells. Front Immunol. 2022;13:939940.

143. Delfanti G, Cortesi F, Perini A, Antonini G, Azzimonti L, de Lalla C, et al. TCR-engineered iNKT cells induce robust antitumor response by dual tar geting cancer and suppressive myeloid cells. Sci Immunol. 2022;7(74):eabn6563.

144. Li Y-R, Hu J, Li Z, Zhu E, Chen Y, Halladay T, et al. Multimodal targeting of metastatic renal cell carcinoma via CD70-directed allogeneic CAR-NKT cells. Cell Rep Med. 2025;6(9):102321.

145. Li Y-R, Li Z, Zhu Y, Li M, Chen Y, Lee D, et al. Overcoming ovarian cancer resistance and evasion to CAR-T cell therapy by harnessing allogeneic CAR-NKT cells. Med. 2025;6(10):100804.

146. Yu G, Wang W, He X, Xu J, Xu R, Wan T, et al. Synergistic therapeutic effects of low dose decitabine and NY-ESO-1 specific TCR-T cells for the colorectal cancer with microsatellite stability. Front Oncol. 2022;12:895103.

147. Wada A, Hirohashi Y, Kutomi G, Murata K, Iwabuchi S, Mizue Y, et al. Eribulin is an immune potentiator in breast cancer that upregulates human leukocyte antigen class I expression via the induction of NOD-like receptor family CARD domain-containing 5. Cancer Sci. 2023;114(12):4511–20.

148. Chen A, Yu Z, Ma N, Lu X, Zhang Y, Xu W, et al. Atovaquone enhances antitumor efficacy of TCR-T therapy by augmentation of ROS-induced ferroptosis in hepatocellular carcinoma. Cancer Immunol Immunother. 2024;73(3):49.

149. Wang L, Liang Z, Guo Y, Habimana JDD, Ren Y, Amissah OB, et al. STING agonist diABZI enhances the cytotoxicity of T cell towards cancer cells. Cell Death Disease. 2024;15(4):265.

150. Cieniewicz B, Bhatta A, Torabi D, Baichoo P, Saxton M, Arballo A, et al. Chimeric TIM-4 receptor-modified T cells targeting phosphatidylserine mediates both cytotoxic anti-tumor responses and phagocytic uptake of tumor-associated antigen for T cell cross-presentation. Mol Ther. 2023;31(7):2132–53.

151. Hafezi M, Lin M, Chia A, Chua A, Ho ZZ, Fam R, et al. Immunosuppressive drug-resistant armored T-cell receptor T cells for immune therapy of HCC in liver transplant patients. Hepatology. 2021;74(1):200–13.

152. Tan AT, Meng F, Jin J, Zhang JY, Wang SY, Shi L, et al. Immunological alterations after immunotherapy with short lived HBV-TCR T cells associates with long-term treatment response in HBV-HCC. Hepatol Commun. 2021;6(4):841–54.

153. Lin M, Bhakdi SC, Tan D, Lee JJX, Tai DWM, Pavesi A, et al. Lytic efficiency of immunosuppressive drug-resistant armoured T cells against circulating HBV-related HCC in whole blood. Immunother Adv. 2023;3(1):015.

154. Degirmencay A, Thomas S, Mohammed F, Willcox BE, Stauss HJ. Modifications outside CDR1, 2 and 3 of the TCR variable β domain increase TCR expression and antigen-specific function. Front Immunol. 2023;14:1148890.

155. Orlando D, Miele E, De Angelis B, Guercio M, Boffa I, Sinibaldi M, et al. Adoptive immunotherapy using prame-specific T cells in medulloblastoma. Can Res. 2018;78(12):3337–49.

156. Yarza R, Bover M, Herrera-Juarez M, Rey-Cardenas M, Paz-Ares L, Lopez-Martin JA, et al. Efficacy of T-cell receptor-based adoptive cell therapy in cutaneous melanoma: a meta-analysis. Oncologist. 2023;28(6):e406–15.

157. Hong DS, Butler MO, Pachynski RK, Sullivan R, Kebriaei P, Boross-Harmer S, et al. Phase 1 clinical trial evaluating the safety and anti-tumor activity of ADP-A2M10 SPEAR T-cells in patients with MAGE-A10+ head and neck, melanoma, or urothelial tumors. Front Oncol. 2022;12:818679.

158. Hong DS, Van Tine BA, Biswas S, McAlpine C, Johnson ML, Olszanski AJ, et al. Autologous T cell therapy for MAGE-A4+ solid cancers in HLA-A*02+ patients: a phase 1 trial. Nat Med. 2023;29(1):104–14.

159. Pan Q, Weng D, Liu J, Han Z, Ou Y, Xu B, et al. Phase 1 clinical trial to assess safety and efficacy of NY-ESO-1-specific TCR T cells in HLA-A∗02:01 patients with advanced soft tissue sarcoma. Cell Rep Med. 2023;4(8):101133.

160. D’Angelo SP, Furness AJS, Thistlethwaite F, Burgess MA, Riedel RF, Haanen J, et al. Lete-cel in patients with synovial sarcoma or myxoid/round cell liposarcoma: planned interim analysis of the pivotal IGNYTE-ESO trial. J Clin Oncol. 2024;42(16_suppl):2500.38828957

161. Meng F, Zhao J, Tan AT, Hu W, Wang S-Y, Jin J, et al. Immunotherapy of HBV-related advanced hepatocellular carcinoma with short-term HBV-specific TCR expressed T cells: results of dose escalation, phase I trial. Hep Intl. 2021;15(6):1402–12.

162. Yang F, Zheng X, Koh S, Lu J, Cheng J, Li P, et al. Messenger RNA electroporated hepatitis B virus (HBV) antigen-specific T cell receptor (TCR) redirected T cell therapy is well-tolerated in patients with recurrent HBV-related hepatocellular carcinoma post-liver transplantation: results from a phase I trial. Hepatol Int. 2023;17(4):850–9.

163. Wang T, Navenot J-M, Rafail S, Kurtis C, Carroll M, Van Kerckhoven M, et al. Identifying MAGE-A4-positive tumors for TCR T cell therapies in HLA-A∗02-eligible patients. Mol Ther Methods Clin Develop. 2024;32(2):101265.

164. Leidner R, Sanjuan Silva N, Huang H, Sprott D, Zheng C, Shih Y-P, et al. Neoantigen T-cell receptor gene therapy in pancreatic cancer. N Engl J Med. 2022;386(22):2112–9.

165. Kim SP, Vale NR, Zacharakis N, Krishna S, Yu Z, Gasmi B, et al. Adoptive cellular therapy with autologous tumor-infiltrating lymphocytes and T-cell receptor-engineered T cells targeting common p53 neoantigens in human solid tumors. Cancer Immunol Res. 2022;10(8):932–46.

166. Owen K, Ghaly R, Shohdy KS, Thistlethwaite F. Lymphodepleting chemotherapy practices and effect on safety and efficacy outcomes in patients with solid tumours undergoing T cell receptor-engineered T cell (TCR-T) therapy: a systematic review and meta-analysis. Cancer Immunol Immunother. 2022;72(4):805–14.

167. Gyurdieva A, Zajic S, Chang Y-F, Houseman EA, Zhong S, Kim J, et al. Biomarker correlates with response to NY-ESO-1 TCR T cells in patients with synovial sarcoma. Nat Commun. 2022;13(1):5296.

168. Ramachandran I, Lowther DE, Dryer-Minnerly R, Wang R, Fayngerts S, Nunez D, et al. Systemic and local immunity following adoptive transfer of NY-ESO-1 SPEAR T cells in synovial sarcoma. J Immunother Cancer. 2019;7(1):276.

169. Nowicki TS, Berent-Maoz B, Cheung-Lau G, Huang RR, Wang X, Tsoi J, et al. A pilot trial of the combination of transgenic NY-ESO-1-reactive adoptive cellular therapy with dendritic cell vaccination with or without Ipilimumab. Clin Cancer Res. 2019;25(7):2096–108.

170. Ishihara M, Nishida Y, Kitano S, Kawai A, Muraoka D, Momose F, et al. A phase 1 trial of NY‐ESO‐1‐specific TCR‐engineered T‐cell therapy combined with a lymph node‐targeting nanoparticulate peptide vaccine for the treatment of advanced soft tissue sarcoma. Int J Cancer. 2023;152(12):2554–66.

171. He K, Hong DS, Ke D, Kebriaei P, Wang T, Danesi H, et al. Durable control of metastases in an HLA-A2+ patient with refractory melanoma after low-dose radiotherapy in combination with MAGE-A4 T cell therapy: a case report. Melanoma Res. 2023;33(4):332–7.

172. Ding R, Liu S, Wang S, Chen H, Wang F, Xu Q, et al. Single-cell transcriptome analysis of the heterogeneous effects of differential expression of tumor PD-L1 on responding TCR-T cells. Theranostics. 2021;11(10):4957–74.

173. Wang L-X, Shu S, Disis ML, Plautz GE. Adoptive transfer of tumor-primed, in vitro–activated, CD4+ T effector cells (TEs) combined with CD8+ TEs provides intratumoral TE proliferation and synergistic antitumor response. Blood. 2007;109(11):4865–76.

174. Mercier-Letondal P, Kumar A, Marton C, Bonnefoy F, Fredon M, Boullerot L, et al. Characterization of atypical T cells generated during ex vivo expansion process for T cell-based adoptive immunotherapy. Front Immunol. 2024;15:1202017.

175. Peng Q, Xu Y, Yao X. scRNA+ TCR-seq revealed dual TCR T cells antitumor response in the TME of NSCLC. J Immunother Cancer. 2024;12(9):e009376.

176. Borgers JSW, Lenkala D, Kohler V, Jackson EK, Linssen MD, Hymson S, et al. Personalized, autologous neoantigen-specific T cell therapy in metastatic melanoma: a phase 1 trial. Nat Med. 2025;31(3):881–93.

Associated Data

Data Availability Statement

No datasets were generated or analyzed during the current study.