The roles of Cryptochrome-1: the circadian clock as a control point in cancer therapy.

Introduction

The internal circadian clock system aligns physiology and behavioral processes with the external environment’s rhythmic changes over a 24-h day cycle [ 1 ]. This is accomplished through clock gene oscillations, which govern the daily activities of the majority of light-sensitive species, including humans and cyanobacteria [ 2 ]. The core clock genes identified in humans include Cryptochrome-1 (CRY1), which is the focus of our review [ 3 ]. Cryptochrome (CRY) was initially identified as a potential photoreceptor in Arabidopsis thaliana. It was cloned as the gene that compensates for the hy4 photomorphogenic mutant, which exhibits a defect in the blue-light-dependent suppression of hypocotyl elongation [ 4 ]. CRYs was subsequently found to be the blue-light absorbing flavoproteins that share important structure and sequence resembleness with the light-activated DNA-repair enzyme (known as photolyase) [ 5 , 6 ]. In birds and insects, photoexcited cryptochrome has a role in magnetoreception [ 7 ].

Cancer development arises from cell overproliferation and genetic instability [ 8 – 10 ]. Historically, cancer rarely contributed to mortality due to the shorter average life expectancy. However, advancements in medical care over the past century have greatly enhanced life expectancy. As a consequence, the total incidence of cancer has increased, resulting in a significantly higher number of cases. According to the World Health Organization (WHO), one in six deaths globally are due to cancer, making it the second most common cause of death [ 11 ].

Various studies have revealed the connection between CRY1 and tumors, including the environmental disruption of circadian ryhthms, polymorphisms (SNPs) and other related elements, which offers valuable understanding into the possible therapeutic applications of CRY1 in the treatment of cancer [ 12 ]. The first evidence suggesting that environmental circadian disruption affects breast cancer was observed in 1969 [ 13 ]. Subsequently, WHO have demonstrated thata link between a disturbed circadian rhythm and an elevated risk of cancer through multiple humans and animal models [ 12 , 14 – 16 ]. Furthermore, groundbreaking studies in the field of molecular oncology have uncovered a multitude of genetic variations that affect clock genes. The relationship between these genetic differences in circadian genes and the risk of cancer has been the subject of numerous studies [ 17 – 19 ]. They has discovered the alterations in Circadian locomotor output cycles protein kaput (CLOCK), Aryl hydrocarbon receptor nuclear translocator-like (ARNTL), CRY1-2, Casein kinase 1 epsilon (CK1ε), Neuronal PAS domain protein 2 (NPAS2) and Differentiated ambryo-chondrocyte expressed gene 1/2 (DEC1/2) are frequently associated with malignancies of the reproductive and pancreatic tissues [ 20 ]. Therefore, CRY1 may be a prognostic biomarker and control point in cancer therapy.

Overview of CRY1

Genes encoding for CRY1

CRY1 is a protein coding gene. A flavin adenine dinucleotide-binding protein that is essential to the circadian core oscillator complex, which controls the circadian clock, is encoded by this gene. With a length of 2.02 kb and 15 exons, the CRY1 gene is found at 12q23.3 on the short arm of chromosome 12 (Source: CRY1 cryptochrome circadian regulator 1 [Homo sapiens (human)] - Gene - NCBI (nih.gov) ). By interacting with other clock proteins including period 2 (PER2) or brain-muscle Arnt-like protein 1 (BMAL1), CRY1 moved to the nucleus [ 21 , 22 ]. Among normal tissue samples, CRY1 is highly expressed in liver and testis [ 23 ].

Structure of the CRY1

CRYs are part of a superfamily of flavoproteins that are found in all domains of life [ 24 ]. The N-terminal photolyase homology (PHR) domain is present in every member of this superfamily, and binds both a 1 5,10-methenyltetrahydrofolate (MTHF) cofactor and a flavin adenine dinucleotide (FAD) cofactor (CRY1 - Cryptochrome-1 - Homo sapiens (Human)| UniProtKB| UniProt). Additionally, the PHR connects with the PER-ARNT-SIM (PAS) regions of mCLOCK and the C-terminal portions of mPER1 and mPER2 [ 25 ]. The three-dimensional architectures of CRY1—including the DNA bindable structure—are similar to those of photolyases [ 26 ].

Function of the CRY1

Circadian clock and circadian rhythms

The model of the mammalian biological clock that is rather detailed has been made possible by significant advancements (Fig. 1 ). Nearly every cell in humans and mice has a clock, which is produced via an autoregulatory transcription-translation feedback loop (TTFL). Situated in the brain’s suprachiasmatic nucleus (SNC), the central clock uses a range of electrical, endocrine, and metabolic signaling pathways to synchronize the peripheral clocks in different tissues and detect light signals to maintain consistency with the time of the light-dark cycle [ 27 , 28 ]. The circadian clock system’s key components, BMAL1 and CLOCK (or NPAS2), work as transcriptional regulators that stimulate the expression of DEC, PER1-3, and CRY1-2 [ 29 ]. By attaching to the BMAL1-CLOCK or BMAL1-NPAS2 dimers and blocking their interactions with different promoters and enhancers, the heterodimers made up of PER and CRY function as co-repressors, turning off BMAL1 (or NPAS2) -CLOCK-induced gene transcription [ 30 , 31 ]. Consequently, this leads to a decrease in CRY and PER levels, allowing the CLOCK-BMAL1 or BMAL1-NPAS2 complex to regain its activity [ 32 , 33 ]. By competitively binding E-box elements via BMAL1-CLOCK or BMAL1-NPAS2, DEC proteins can also form dimers to decrease the transcription of the PER and DEC genes, creating a negative feedback loop to control circadian rhythms [ 34 ]. The ROR responsive element (RRE) is bound by the cytoplasmic BMAL1-CLOCK or BMAL1-NPAS2 dimers in the promoter regions of the Rorα and Rev-erbα genes to enhance transcription [ 35 ]. The resultant proteins serve as transcription factors that, respectively, stimulate and inhibit BMAL1 and NPAS2 transcription [ 36 ]. Additionally, during the day/night shift, CRY is most expressed in the eye and SCN [ 37 ]. Tumor formation is greatly impacted by metabolic activity and endocrine homeostasis, which are regulated by the circadian clock and gene rhythms linked to these processes [ 38 , 39 ].

Cell cycles

The sequence of events in a cell that results in its division and duplication is known as the cell cycle. The cell cycle is regulated by a complex series of checks and balances known as cell cycle checkpoints, which ensure that each phase occurs in the correct sequence and that the cell is ready to proceed to the next phase [ 40 ]. Many proteins, like as cyclins and cyclin-dependent kinases (CDKs), regulate these checkpoints [ 41 ]. It is widely recognized that tumor is a group of dieases in which cells divide constantly and excessively [ 42 ]. Cancer-related mutations change how the cell cycle is regulated, allowing cells to keep dividing without control [ 42 ].

There is increasing evidence that the circadian clock and the cell cycle are physiological oscillators that are molecularly linked through a number of coupling mechanisms, many of which are dysregulated in tumor cells [ 43 ]. Since clock genes were discovered, circadian rhythms have been seen in a number of processes, including cell cycle mitosis, the expression of cell cycle proteins in the mouse model, and the synthesis of nucleic acids (DNA and RNA) [ 33 , 44 , 45 ]. In 2001, biopsies taken from the skin and oral mucosa of eight healthy, diurnally active male research individuals were used to examine the relative RNA levels of clock genes during a 24-hour period by G A Bjarnason and colleagues [ 46 ]. In detail, the oral mucosa and skin were found to express CRY1, and their circadian profile matched that of the rats’ peripheral tissues and suprachiasmatic nuclei. Because G1 cyclins are highly expressed in CRY1/CRY2-deficient animals, osteoblasts from these mice show a faster G1/S transition that results in increased bone mass [ 47 ]. Similarly, Destici et al. have also shown that primary CRY1−/−CRY2−/− murine fibroblasts proliferate more quickly and move through the cell cycle more quickly than wild-type cells [ 48 ]. Furthermore, the mitochondrial transport signal is located in the N-terminal portion of CRY1 [ 49 ], thus parcitipating in the heart of cell proliferation [ 50 ]. CRY influences tumourigenesis through the cell cycle, since is a essential cell cycle regulator [ 45 ].

Metabolism

Research has established a connection between circadian rhythms and metabolic regulation. Night shift workers and individuals with irregular schedules exhibit circadian disruption [ 51 ]. Rodent models with disturbed rhythms show similar patterns [ 52 ]. These impairments correlate strongly with metabolic dysfunction and reduced lifespan [ 53 ]. Additionally, both glucose and fatty acid homeostasis display circadian variations, and the SCN’s disturbance after surgery has a negative impact on glucose homeostasis [ 54 ].

The roles of CRY1 in metabolism, including glucose metabolism, lipid metabolism, and melatonin metabolism have been discussed in several sdudies. CRY1 inhibits the ligand-bound glucocorticoid receptor (GR) from causing the liver’s glycoheterotrophic enzyme phosphoenolpyruvate carboxykinase 1 (Pck1) to be expressed [ 55 ]. Additionally, autophagy degrades CRY1 to control the liver clock and glucose metabolism [ 56 ]. In this case, a number of light chain 3 (LC3)-interacting region motifs found in CRY1 help cargo proteins connect with the autophagosome marker LC3.

It is widely acknowledged that lipid metabolism needs to be coordinated with other body processes and with daily changes in the environment and provides energy for body’ activity [ 57 , 58 ]. Peroxisome proliferator-activated receptor delta (PPARδ) supports the preferred fuel source of fats during physical activity. By modifying PPARδ activity, CRY1 modifies exercise physiology, which in turn modifies energy storage and substrate selection for energy generation [ 59 ]. CRY1 is also stimulated by insulin-induced SREBP1c, which is an essential transcription factor for lipogenesis, and reduces hepatic gluconeogenesis by facilitating the degradation of Forkhead box O 1 (FOXO1) [ 60 ]. CRY1 could be involved in recalibrating energy balance when faced with fluctuating nutritional conditions. Mice that receive a high-fat diet (HFD) do not develop obesity when CRY1 is ablation-free, indicating that CRY1 could be a viable target for anti-obesity treatment [ 61 ].

In mammals and humans, the pineal gland primarily secretes the indolic substance melatonin during the night [ 62 ]. Melatonin regulates the sleep-wake cycle when used at appropriate therapeutic times as an endogenous sleep-promoting circadian factor [ 63 ]. The “master biological clock” in the suprachiasmatic nucleus of the SCN controls the production and release of melatonin [ 64 ]. Melatonin is a hallmark of circadian rhythms because it can also control central clocks and peripheral oscillators in tissues and organs, as well as arrange target organ systems into suitable homeostatic metabolic rhythms [ 65 – 67 ].

Accumulating evidence illustrates the relationship between melatonin and CRY1. CRY1 mRNA expression reaches its peak at night, which aligns with rising melatonin levels in the blood plasma [ 68 ]. CRY1 expression (with diminished PER1) is directly induced by melatonin in the pars tuberalis (PT) [ 68 ]. According to Johnston et al., melatonin infusion in female Soay sheep increased the level of CRY1 in the PT at six distinct time points during a 24-hour period [ 69 ]. INPAS4 was identified by researchers as a potential instantaneous early response gene in the ovine PT, which is stimulated by melatonin and drives CRY1 [ 70 ]. Moreover, melatonin suppression brought on by nighttime light exposure is linked to a higher risk of prostate, breast, and colon cancer [ 71 ]. Melatonin might be useful as adjuvant in cancer therapy, including anaplastic thyroid carcinoma, breast cancer and brain tumors [ 72 – 74 ].

DNA repair

The DNA molecule, which is the fundamental unit of heredity, is inherently reactive and highly prone to chemical alterations by both endogenous and exogenous agents [ 75 ]. Therefore, systems for repairing damaged DNA are necessary for cell viability [ 76 ], which work closely tied with the control of the cell cycle [ 77 ]. For instance, cells cannot undergo mitosis when DNA damage is present because the G2/M checkpoint of the cell cycle is tightly regulated [ 78 ]. DNA damage can change nucleotide sequences and cause malfunctioning proteins to be expressed if it is not repaired, which can result in cancer and other illnesses [ 77 , 79 ].

Due to the similar structure as photolyases, CRY1 continues to play a part in detecting or reacting to damaged DNA, including DNA repair and the G2/M transition. Timeless (Tim), a cell cycle checkpoint protein and evolutionarily conserved ortholog, is involved in DNA replication, cell cycle progression, embryonic development, and the DNA damage response [ 80 , 81 ]. Certain amino-terminal proteins such as Tim interact with CRY1 [ 82 ]. In detail, Tim armadillo binds a C-terminal Tim helix and repeats in a continuous core, resembling the interactions seen between light-insensitive CRYs and their binding partners in mammals. Photolyases also use this mechanism to identify damaged DNA. Hausp was discovered to be the most highly enriched protein in CRY1-containing complexes by Stephanie J. Papp et al. in 2015 when they used mass spectrometry to find new protein partners of mammalian CRYs [ 26 ]. In particular, Hausp-dependent CRY1 evolved to preserve genomic integrity by regulating the downstream transcriptional response and reacting to DNA damage via posttranslational modification. After researching the molecular process in malignancies, Ayesha A. Shafi et al. came to the conclusion later in 2021 that CRY1 continues to be stabilized by genomic damage, modulates the G2/M transition and DNA repair, and improves cell viability through temporal regulatory transcription [ 71 ]. Interestingly, telomerase activity itself exhibits circadian oscillations in cancers, suggesting potential interplay between DNA repair machinery and clock-regulated genomic stability mechanisms [ 83 , 84 ].

Tumor immunity

Monocytes, natural killer cells, T and B lymphocytes, and other immune cell populations are all regulated by the molecular circadian clock of the mammalian immune system [ 85 – 88 ]. Their defensive responses, including the generation of pro-inflammatory cytokines and other effectors, are influenced by the circadian clock and fluctuate in accordance with a roughly 24-hour cycle [ 19 ]. CRY1 overexpression in endothelial cells in a mouse model reduces monocyte adherence by blocking the nuclear factor-kappa B (NF-κB) pathway, inflammatory cytokines, and adhesion molecules [ 89 ]. In addition, through PKA signals and NF-κB, knockout of CRY1 resulted in constitutively increased pro-inflammatory cytokines [ 90 ]. The immune system and cancer have a complex interaction relationship, and the influence of clock genes on the immune system can also act on cancer-related mechanisms [ 91 – 93 ]. Clock gene mutations affect different cells in the tumor microenvironment (TME), influencing the balance between pro-tumor immunosuppressive cells and anti-tumor immune cells, encouraging immune evasion, and accelerating tumor growth. They also modulate the proliferation, migration, infiltrative capacity of tumor-associated macrophage (TAM) cell subsets, as well as the levels of Th17 and regulatory T cells (Tregs), and CD8 T cell numbers [ 94 ].

The neoplastic formation and progression represent a highly intricate biological process. Beyond inducing profound cell cycle dysregulation, it requires multiple critical steps: (1) evasion of apoptosis triggered by tumor cell contact, (2), initiation of tumor angiogenesis (a pivotal transition from benign to malignant states), and (3) acquisition of migratory and invasive capabilities [ 95 – 97 ]. These processes are coordinately regulated by key genes that function as clock-controlled genes, with CRY1 playing a central role. Specifically, CRY1 influences tumor development through both direct and indirect mechanisms by involving in cell cycle regulation, DNA damage repair, metabolism and tumor immunity [ 48 , 68 , 71 , 90 , 94 ].

Based on the disccusion in this article, collectively, the pleiotropic nature of CRY1 establishes it as a master regulator capable of suppressing hallmark cancer phenotypes. Both CRY1 deficiency and overexpression disrupt the temporal coordination of these processes, ultimately driving sustained proliferation, evasion of growth suppressors, genomic instability, metabolic reprogramming, induced angiogenesis and immune evasion (Table 1 ).

Mechanism of related signaling pathways

cAMP/PKA/CREB signaling way

cAMP is responsible for activation of the protein kinase A (PKA) as the intracellular second messenger [ 98 ]. PKA, which consists of 2 regulatory and 2 inactive catalytic subunits, is an essential effector that phosphorylates CREB [ 98 – 100 ]. Numerous clinicopathological characteristics, such as tumor stage, grade, metastasis, elevated recurrence rates, poor prognosis, and decreased survival among cancer patients, have been linked to CREB overexpression [ 98 , 101 , 102 ]. The cAMP/PKA/CREB signaling pathway exhibits a dual function in tumor biology, manifesting both tumor-promoting and tumor-suppressive effects across a variety of human malignancies [ 103 ]. The effects of cAMP/PKA/CREB signaling pathway and circadian rhythm are bidirectional. The cAMP/PKA/CREB signaling pathway has been demonstrated to be instrumental in the modulation of circadian rhythms [ 104 ]. In the early morning, a decrease in melatonin levels can lead to a surge in intracellular cAMP levels, which results in the transcriptional reaction of cAMP-regulated genes [ 104 ]. For the first time, Zhang et al. clarified the non-nuclear function of the circadian clock protein CRY in mice [ 105 ]. Through its possible interaction with the G protein Gs α subunit, CRY controls the circadian fluctuations in hepatic gluconeogenesis. By facilitating the temporal modulation of glucagon signaling, this relationship seems to eventually cause the classical cAMP/PKA pathway to phosphorylate CREB, which in turn inhibits melanogenesis (Fig. 2 ) [ 106 , 107 ].

HIF1α signaling way

Many human and animal cells have the oxygen-sensitive transcriptional activator HIF-1. An alpha subunit (HIF-1α) and a beta subunit (HIF-1β) form a heterodimer that aids in the development of mammals and the advancement of tumors [ 108 ]. Under low oxygen conditions, HIF-1α induces the levels of vascular endptheila growth factor (VEGF), which signals blood vessels to facilitate tumor migration [ 109 ]. Tumor cells, stromal cells (including pericytes and endothelial cells), immune cells and the substances they release, and the extracellular matrix make up the majority of the TME [ 110 ]. By controlling a range of immune cells in the TME, HIF-1α encourages immunity escape from tumors. For example, in 2022, Haixia Wei et al. demonstrated that HIF-1α initiated PD-1 expression in CD4+ T cells to modulate immune response [ 111 ]. Additionally, HIF-1α can induce tumor immune escape by overexpressing PD-L1 [ 112 ].

Particularly necessary for resetting the molecular clock by oxygen cycles, HIF-1α responds to variations in oxygen levels by acting upstream of the clock [ 113 ]. Increased cellular HIF levels and improved proliferation and migration can result from CRY1 deficiency; these effects can be countered by CRISPR/Cas9- or short hairpin RNA-mediated HIF knockout. When the CRY1-HIF crosstalk is disrupted at the cellular level, it becomes evident that HIF-1α and CRY1 have conflicting effects on cell proliferation [ 114 ]. Through its tail region, CRY1 mechanically interacts with the basic-helix-loop-helix domain of HIF-1α, lowering its half-life and facilitating HIF binding to the target gene promoter [ 115 ], which is intimately linked to the onset and spread of hematological cancers, including multiple myeloma, lymphoma, and leukemia [ 116 ]. Additionally, there is strong evidence that the clock components influence the level of angiogenic proteins in cancer cells, including VEGF, ARNT, and HIF1α, which promotes tumor development and metastasis [ 36 , 117 , 118 ].

NF-κB signaling pathway

There are canonical and non-canonical routes in the NF-κB signaling pathway [ 119 ]. A variety of stimuli, such as ligands for different cytokine receptors, pattern recognition receptors (PRRs), T-cell receptors (TCRs), members of the tumor necrosis factor receptor (TNFR) superfamily, and B-cell receptors, can activate the canonical NF-κB pathway [ 119 , 120 ]. By connecting to TRADD, the canonical route triggered by TNFR1 signaling creates an assembly platform for the recruitment of FADD and TRAF2, which then bind to RIP1 to activate IKK [ 121 ]. On the other hand, the non-canonical route produces the p52-RelB complex by partially digesting p100 and relying on IKKα-mediated phosphorylation of RelB-associated p100 [ 121 ]. The formation of several immune cell layers is brought about by the non-canonical route. For example, thymic epithelial cells, which are essential for T cell formation in the thymus, require this pathway to mature and function [ 119 , 122 ].

One nuclear transcription factor that is necessary for preventing the development and spread of cancer is p53. The mutations in TP53 gene undermine the tumor suppressor functions of the p53 protein, making these alterations the most prevalent single-gene mutations found in human cancers [ 123 ]. Cryptochrome mutations make p53 mutant and oncogenically altered cells more susceptible to tumor necrosis factor α (TNFα)-induced apoptosis, as Jin Hyup Lee et al. showed in 2011 [ 124 ]. This effect is accomplished by using the GSK3β kinase to interface with the NF-κB signaling pathway, which reduces prosurvival NF-κB signaling [ 124 ]. Later in 2015, Lei Yang found that the overexpression of CRY1 led to a significant reduction in the expression of toll-like receptors (TLR) 2 and 4, as well as phosphorylated p65 (p-p65), which is a member from NF-κB family [ 121 , 125 ]. The luciferase reporter assay results demonstrated that the overexpression of CRY1 effectively inhibited the stimulation of NF-κB. As was indicated in the section on “tumor immunity,” CRY1 regulates the NF-κB signaling pathway, which is essential for inhibiting the inflammatory response [ 89 , 90 ].

STAT3 signaling pathway

Cell proliferation, differentiation, apoptosis, angiogenesis, inflammation, and immunological actions are among the biological mechanisms that are regulated by the cytoplasmic transcription factor namely signal transducer and activator of transcription 3 (STAT3) [ 126 ]. The transactivation domain, linker domain, DNA binding domain, coiled-coiled domain, N-terminal domain, and Src homology 2 domain make up the majority of STAT3’s structural components [ 127 ]. Numerous receptors, such as those for growth factor receptors, G-protein coupled receptors (GPCRs), interleukin-6 (IL-6) and IL-6 family cytokines (including IL-11 and IL-23), and Toll-like receptors, activate the conventional STAT3 signaling pathway. Their particular ligands, which include angiotensin, sphingosine-1-phosphate, lipopolysaccharides (LPS), cytokines, and hormones, among others, stimulate these receptors [ 128 ]. Persistent activation of STAT3 has been demonstrated to lead to unregulated gene transcription, resulting in the promotion of tumor development and progression by its gene products [ 127 , 129 , 130 ]. Therefore, STAT3 signaling pathway represents potential therapeutic targets for altering metabolic reprogramming in cancer [ 131 ].

Several studies have demonstrated a strong association between STAT3 and twelve distinct cancers, including hematologic tumors (lymphomas, acute myeloid leukemia, chronic myeloid leukemia, etc.) and solid tumors (pancreatic adenocarcinoma, esophageal squamous cell carcinoma, non-small cell lung cancer, ovarian carcinoma, breast cancer, cervical cancer, colon cancer, and, etc.) [ 128 ]. The regulating function of CRY1 via the STAT3 signaling pathway in the development of cancer has been documented in the literature. For instance, hydrogen peroxide modulates CRY1 expression via STAT3 signaling pathway, thereby involving in cancer [ 132 ]. Tissue microarray research in 2021 revealed that cervical cancer overexpressed CRY1 and NANOG [ 133 ]. In this instance, CRY1 silencing caused cervical cancer cell lines to undergo cell apoptosis, exhibit decreased NANOG expression, block STAT3 signaling, and activate p53 signaling.

FOXO signaling pathway

FOXO transcription factors, including FOXO1, FOXO3, FOXO4, FOXO6 [ 134 ], serve as critical tumor suppressors and exert their anticancer effects through the following mechanisms: First, in terms of cell cycle regulation, FOXO induces cell cycle arrest by activating target genes such as p27 KIP1 and GADD45A [ 135 ]. Second, FOXO triggers apoptosis by upregulating pro-apoptotic genes like Bim [ 136 ]. Additionally, FOXO suppresses tumor cell proliferation and metastatic potential [ 137 ]. Consistent with the aforementioned metabolic roles of FOXO, these transcription factors actively participate in regulating cancer cell metabolic processes, particularly in modulating glucose and lipid metabolism [ 60 ]. Another prime example is the AMPK-SIRT1-FOXO pathway, which exhibits a dual role in tumor regulation: it suppresses tumor growth by restricting energy supply and inducing apoptosis, while paradoxically supporting cancer cell survival under stress conditions through enhancing DNA repair and metabolic flexibility [ 138 ].

Tomas Duffy and colleagues conducted pioneering research that first elucidated FOXO1-mediated transcriptional regulation of circadian clock genes [ 139 ]. Their genome-wide analysis revealed that among 2985 FOXO1-binding sites identified in chondrocytes, the circadian rhythm pathway showed the most striking enrichment (logP = -16.21), with 30 directly regulated target genes [ 139 ]. Moreover, the circadian regulation of autophagy involves Foxo3a-mediated transcriptional control of multiple autophagy-related factors, particularly Bcl2 interacting protein 3 like (Bnip3l), GABA type A receptor associated protein like 1 (Gabarapl1), Bcl2 interacting protein 3 (Bnip3), and microtubule-associated protein light chain 3B (LC3B) [ 140 ]. This molecular mechanism establishes Foxo3a as an essential regulator that coordinates autophagic activity with circadian rhythms, revealing its pivotal role in the clock-dependent induction of autophagy [ 140 ]. Swip Draijer and colleagues also demonstrated that hepatic FoxO3 serves as a critical modulator of circadian regulation [ 141 ].

The insulin-SREBP1c axis upregulates circadian protein CRY1, which binds FOXO1 and recruits MDM2 E3 ligase to promote FOXO1 ubiquitination and proteasomal degradation [ 60 ]. This CRY1-mediated FOXO1 suppression complements acute AKT-mediated FOXO1 phosphorylation, providing sustained inhibition of gluconeogenic genes (PEPCK/G6Pase) during prolonged insulin signaling [ 60 ]. In diabetic db/db mice, defective CRY1 induction leads to FOXO1 accumulation and hyperglycemia [ 60 ]. The pathway thus couples circadian (CRY1) and metabolic (FOXO1) regulation for postprandial glucose control [ 60 ].

Hedgehog signaling pathway

The Hedgehog (HH) signaling pathway is an evolutionarily conserved developmental regulatory system mediated by three secreted ligands: Sonic hedgehog (SHH), Indian hedgehog (IHH), and Desert hedgehog (DHH) [ 142 , 143 ]. Ligand binding to the 12-pass transmembrane receptor Patched1 (Ptch1) relieves its suppression on the 7-transmembrane protein Smoothened (Smo), inducing Smo conformational change and ciliary translocation [ 144 ]. Activated Smo promotes dissociation of SUFU-Gli complexes, allowing the transcription factor Gli to enter the nucleus and regulate target genes (e.g., Ptch1, Gli1), thereby orchestrating embryogenesis, tissue regeneration, and stem cell self-renewal [ 145 , 146 ].

The HH signaling pathway plays a pivotal role in tumor initiation, progression, and metastasis [ 147 ]. Its aberrant activation enhances tumor proliferation, invasion, and drug resistance. In medulloblastoma, particularly the SHH subtype, mutations in genes such as Ptch1 lead to constitutive HH pathway activation [ 147 ]. In pancreatic ductal adenocarcinoma (PDAC), dysregulated HH signaling induces epithelial-mesenchymal transition (EMT)—a process where epithelial cells lose adhesion molecules (e.g., E-cadherin) and acquire mesenchymal traits (e.g., upregulated N-cadherin, Vimentin), thereby increasing migratory and invasive properties to facilitate metastasis [ 147 , 148 ].

SHH signaling is rhythmically activated by circadian oscillations in primary cilium shape in neuromedin S (NMS)-producing SCN neurons [ 149 ]. By encouraging the nuclear translocation of Gli transcription factors, this activation modifies the production of neuropeptides (VIP/GRP) and core clock components (PER1/CRY1/BMAL1) [ 149 ]. Using a Smoothened knockout (SAC-KO) mouse model, another study found that the diurnal oscillations of hepatic lipid metabolism are influenced by the HH signaling system, which controls the rhythmic expression of clock genes through the Gli transcription factors [ 150 ]. Similarly, BMAL1/CLOCK directly activates IHH transcription via E-box elements in its promoter, driving diurnal expression in growth plate chondrocytes [ 151 ]. The PER/CRY complex (including CRY1) inhibits BMAL1/CLOCK activity, suppressing IHH rhythmicity [ 151 ].

The association with specific cancers in previous studies

The association with specific cancers in previous studiesAn extensive body of literature supports that CRY1 is a double-edge sword in cancer development. On the one hand, according to Shafi AA et al., CRY1 is considered to be an protumorigenic factor [ 71 ]. Another study further highlighted the oncogenic role of CRY1, showing that it enhances the degrading of p53 by binding to the E3 ubiquitin ligase MDM2 in bladder cancer cells. This interaction increases sensitivity to anticancer drugs [ 152 ]. On the other hand, CRY1 has a major function in antitumor efficacy [ 153 ]. For instance, CRY1 is closely related with the homeostasis maintenance and tumor immunity [ 89 ]. Additionally, CRY1 expression is commonly suppressed in various cancers (Table 2 ) [ 154 ], which we will discuss later in this section.

This section of the review focuses on the current comprehension of the intricate role of how CRY1 plays in the development of several solid tumors and how its function serves as a novel therapeutic target that could pave the way for the treatment of cancers.

Cervical cancer

According to global figures from 2022, cervical cancer is the fourth most prevalent type of cancer diagnosed worldwide and the top cause of death for women [ 155 ]. Cervical cancer is typically a result of contamination with the human papillomavirus (HPV), a ubiquitous virus recognized for inducing abnormal cellular transformations in the cervix, which, if not addressed promptly, may ultimately progress into cancer [ 156 ]. Some sexually transmitted illnesses (HIV and Chlamydia trachomatis), smoking, high fertility, and long-term use of oral contraceptives are additional significant cofactors [ 157 ].

Some studies have explored the mechanism of CRY1 related with cervical cancer. For example, Gwan Hee Han et al. proposed that the CRY1-STAT3-NANOG axis, which is a key prognostic biomarker for cervical cancer prognosis, is involved in cervical cancer through p53 [ 133 ]. After data-mining of the oncomine gene expression database using the data from two independent stuies [ 158 , 159 ], and Real-time polymerase chain reaction (RT-PCR) and Western blotting evaluation utilizing samples from frozen tissue specimens, Watt et al. confirmed notable suppression of core circadian clock including CRY1 in cervical cancer as opposed to healthy cervical epithelium [ 160 ]. In this study, CRY1 was also found to be methylated in cell line models, which were part of the mechanism in tumor formation.

Breast cancer

The most prevalent kind of cancer worldwide is breast cancer [ 157 ]. Three primary tumor subtypes of breast cancer are distinguished by the presence of the estrogen or progesterone receptor and the amplification of the ERBB2 gene [ 161 ]. Early-stage breast cancer is thought to be treatable, with systemic therapy administered either before or after surgery if required, and locoregional therapy (surgery and radiation) as the cornerstone [ 162 ].

Hongji Dai et al. discovered that variations in Chinese communities’ risk of breast cancer are caused by genetic diversity in genes related to the circadian rhythm control pathway [ 163 ]. SNPs in the CLOCK and CRY1 genes were linked to an increased risk of breast cancer, according to this study’s univariate logistic regression analysis. Nevertheless, the initial study to conclusively investigate the connection between breast cancer and circadian rhythm gene variants across several ethnic groups found no correlations between clock gene variants and breast cancer [ 164 ]. Although two SNPs (rs11113179 in CRY1 and rs3816360 in ARNTL) were substantially linked to breast cancer, they lost their significance after the correction for the rate of erroneous discoveries [ 164 ].

It has been demonstrated that the Hippo pathway is crucial for carcinogenesis [ 165 ]. The Hippo pathway’s central element, Yes-associated protein/transcriptional co-activators with PDZ-binding motifs (YAP/TAZ), encourages tumor development, metastasis, and treatment-resistant behavior [ 166 ]. CRY1 inhibits the transcription of the Hippo pathway effectors TAZ and YAP in the MDA-MB-231 and MCF7 breast cancer cell lines [ 167 ]. To elaborate, YAP had a significantly greater impact on CRY1 transcriptional regulation than TAZ.

Ovarian cancer

In 2020, ovarian cancer is expected to account for 3.7% of cases and 4.7% of cancer-related deaths worldwide, making it the eighth most frequent disease in women [ 157 ]. There are five main clinically and genetically different histotypes of ovarian cancer, including the most prevalent kind, epithelial ovarian cancer. Ovarian cancer is a very diverse illness [ 168 ].

When ovarian cancer was compared to normal ovaries, the expression of CRY2, CLOCK, PER1, and PER2 was much reduced [ 169 ]. The highest expression, on the other hand, was CRY1, followed by PER3 and BMAL1, indicating that ovarian cancer may still exhibit inverted expression of CRY1 and BMAL1 [ 169 ]. Ovarian cancers have clock gene dysregulation, which may be connected to the tumors’ vulnerability to chemotherapy [ 170 ]. The treatment of high-grade, advanced epithelial ovarian cancer (EOC), particularly that which lacks homologous recombination (HR), has been revolutionized by poly(ADP-ribose) polymerase (PARP) inhibitors [ 171 ]. According to a recent study, anti-angiogenic drugs can effectively block the VEGF/VEGFR/PI3K signaling pathway, which suppresses the increase of CRY1 expression and lowers HR [ 172 ]. Furthermore, CRY1 inhibition made EOC cells more sensitive to olaparib [ 172 ]. These results show that the combination of antiangiogenic medications and CRY1 inhibitors holds great potential as a therapeutic approach to the care of HR-deficient epithelial EOC alongside PARP inhibitors [ 172 ].

Gastric cancer

Gastric cancer is the fifth most common type of cancer worldwide and ranks third in terms of cancer-related mortality [ 173 ]. It often classified into two distinct subsites, cardia and noncardia, exhibits variations in risk factors, oncogenic effects, and epidemiological patterns [ 157 ]. Noncardia stomach cancer is thought to be primarily caused by a chronic Helicobacter pylori infection, and this bacterium is responsible for nearly all instances [ 157 ]. Due to Helicobacter pylori prevention, noncardia stomach cancer rates of occurrence and death have been continuously decreasing in the majority of populations over the past 50 years [ 174 – 176 ]. According to epidemiology, eating a lot of fat is positively linked to a number of malignancies, such as colorectal, breast, prostate, and stomach cancers [ 177 , 178 ]. Numerous research address the effects of dietary modifications and content on the intestinal epithelium and the milieu around it [ 179 ]. Consumption of salted pickles, red meat, cooked meats, and salty dishes is commonly believed to increase the risk of gastric cancer [ 180 , 181 ], whereas intake of antioxidant vitamins, fresh fruits, and veggies might lower the chance of stomach cancer [ 182 – 184 ].

Some research studied the interrelationship of CRY1 and obesity. The harmful effects of type 2 diabetes and obesity is increased when the circadian rhythm is disturbed [ 185 , 186 ]. As previously stated, CRY1 ablation shields mice from obesity brought on by a high-fat diet [ 61 ]. At the conclusion of the 16-week trial, CRY1(-/-) mice on the HFD acquired significantly less body weight (18%) and had less fat formation (-61%), although having identical calorie intake, indicating an increase in energy expenditure [ 61 ]. Mechanically, CRY1-induced Cystathionine β-synthase activation affects fatty acid, amino acid and one-carbon metabolic pathways, suggesting that CRY1 regulates cellular metabolism through non-transcriptional post-transcriptional modulation of CBS enzyme activity [ 187 ].

Using RT-PCR to detect the circadian clock genes expression from 29 gastric cancer patients, Ming Luen Hu et al. determined the CRY1 was up-regulated strongly connected with phases III and IV, which are more sophisticated [ 188 ]. Stomach adenocarcinoma (STAD) ranks third in terms of cancer-related mortality among all malignancies and fifth in incidence, making it among the most prevalent cancerous growths worldwide [ 189 , 190 ]. Researchers investigated the expression and prognostic significance of key circadian clock genes in STAD in 2021 using bioinformatics techniques [ 191 ]. According to the results, there was an upregulation of the CRY1, NR1D, and CLOCK mRNA levels.

Colorectal cancer

Colorectal cancer is the second highest mortality (9.3%) of all cancers reported by the global statistics 2022 [ 155 ]. More than 90% of deaths from colorectal cancer can be avoided with early detection, but despite the wide range of tests available, more than one-third of those eligible for screening are not screened in a timely manner [ 192 ].

Growing evidence highlights the critical role of clock genes in regulating intestinal physiological functions. For instance, colonic epithelial cells exhibit circadian rhythmicity through the expression of core clock components, including CRY1, REV-ERBα, BMAL1, and CLOCK [ 193 ]. Disruption of the circadian timing system not only perturbs this rhythm but also accelerates cancer progression [ 194 ]. Furthermore, experimental studies in rats demonstrate that forced circadian rhythm alterations can dysregulate the gut microbiota, promoting intestinal low-grade inflammation—a potential contributor to carcinogenesis [ 195 ].

There is currently a wealth of concrete evidence that suggests CRY1 has a role in colorectal cancer development. A multicenter study revealed CRY1 expression patterns in colorectal cancer [ 196 ]. In detail, low CRY1 correlated with older age and female patients [ 196 ]. Additionally, high CRY1 was associated with distal colon tumors [ 196 ]. In 2013, Hongyan Yu and colleages investigated the function of CRY1 in colorectal cancer related with human [ 197 ], demonstrating it’s prognostic significance in the treatment of cancer. Later in 2016, Gianluigi Mazzoccoli et al. firstly proved that cryptochrome gene (CRY1 and CRY2) expression is altered in colorectal cancer [ 198 ]. Based on that, K Hasakova et al. detected a significant correlation between tumour situation and CRY1 protein levels in neighbouring tissues, and the combined results suggest that the effect of tumors on neighbouring tissues is dependent on tumour location [ 199 ]. They came to the conclusion that certain patient subgroups with colorectal cancer should be taken into account due to altered clock gene expression. Kristina Hasakova et al. evaluated clock-regulated (VEGF-a, early growth response protein 1, and oestrogen receptor β) and clock-controlled (CRY1, CRY2, and PER2) gene expression in colorectal cancer and surrounding tissues in 2018 [ 200 ]. Ultimately, they discovered sex-independent relationships between patient survival and the expression of the aforementioned genes. Furthermore, a 2021 study found that individuals with colorectal cancer who had high expression of CRY1 had a worse OS and a lower disease-free survival at five years [ 201 ]. Derived from preceding research, CRY1 may aggravate colorectal cancer progression [ 202 ]. Furthermore, this conclusion has already been applied in other fields, such as the interaction between emerging environmental factors and miRNA-mediated regulation [ 203 ].

Prostate cancer

In 2020, China has the age-standardized rate (ASR) of 4.6 deaths per 100,000 population [ 157 ]. Because of its enormous population, China accounts for 8.2% of new instances of prostate cancer and 13.6% of deaths from the disease worldwide, despite having low incidence and mortality rates. This highlights the urgent need for increased attention to prostate cancer in China [ 157 ]. Prostate biopsy and analysis, magnetic resonance imaging (MRI), rectal examinations, PSA testing, or health screenings are used to diagnose and predict the outcome of prostate cancer [ 204 , 205 ]. However, current clinical and pathological features do not reliably and accurately predict the prognosis of individual patients.

Prior research has demonstrated that CRY1 expression is linked to poor outcomes in prostate cancer and can block nuclear receptors implicated in endocrine malignancies, including the androgen receptor, indicating a possible treatment target for the condition [ 71 ]. According to one hypothesis-driven analysis, five SNPs—the genes for ARVCF, IL4, CRY1, RNASEL, and LEPR—were confirmed to be substantially linked to prostate cancer [ 206 ]. This represents the inaugural population-based investigation demonstrating that germline genetic variants offer prognostic insights regarding survival specific to prostate cancer. In 2015, the gene-based analysis conducted showed that a variation in the CRY1 gene was ostensibly linked to prostate cancer death [ 207 ]. Two SNPs in CRY1 were shown to have nominally significant correlations with the risk of fatal disease in HPFS and AGES-Reykjavik within the respective cohorts.

Viewed from the perspective of circadian rhythm, emerging evidence suggests that prostate cancer risk may be modulated by chronodisruption — a hypothesis investigated less comprehensively than in breast cancer, and with currently conflicting epidemiological support [ 208 ]. A large-scale meta-analysis by Dapang Rao et al. found night-shift workers had significantly increased prostate cancer incidence (meta-relative risk: 1.24, 95% CI: 1.05–1.46; P = 0.011) [ 209 ]. This association persists across multiple professions with night-shift exposure, including healthcare workers, firefighters, and police officers [ 208 , 210 ]. The pathogenic mechanism likely involves artificial light-at-night-induced melatonin suppression and subsequent CRY1 dysregulation, as demonstrated in circadian biology studies [ 71 , 211 ]. Contrary to the established link mentioned before, the PROtEuS case-control study ( n = 3,869) found null associations between night-shift work and prostate cancer risk across all exposure metrics [ 212 ].

Pancreatic cancer

According to data from 2020, pancreatic cancer is the seventh greatest cause of cancer-related fatalities in both sexes, accounting for almost as many deaths (466,000) as new diagnoses (496,000) [ 157 ]. It is anticipated that pancreatic ductal adenocarcinoma will be the second most frequent cause of cancer-related fatalities by 2030 [ 213 ]. Patients with pancreatic cancer generally face a grim prognosis once metastasis occurs, as the chances of a cure diminish significantly [ 214 ].

As mentioned in the section about the circadian clock, there is evidence from experimental studies showing that the SCN acts as a tumor suppressor by regulating circadian rhythms. According to one study by Filipski E et al., mice with SCN lesions had quicker growth rates for pancreatic tumors [ 215 ]. In particular, malignant tumor growth is accelerated when mouse circadian rhythms are disrupted, indicating that the host biological clock may be crucial for the endogenous regulation of tumor progression. Moreover, epidemiological studies have connected circadian rhythm disruption to heightened vulnerability to cancer development, such as pancreatic cancer, in all key human organ systems [ 216 ].

Pancreatic cancer is primarily characterized by its most common form, known as PDAC, which has an average 5-year survival rate of fewer than 10% [ 217 ]. PDAC is an aggressive cancer most often detected at an advanced stage, limiting treatment options to systemic chemotherapy with a modest clinical response [ 218 ]. Quantitative RT-PCR was used to examine the circadian clock genes, including CRY1, in PDAC and surrounding and benign tissue [ 219 ]. For the first time, the data showed that all circadian genes had lower expression levels than the corresponding adjacent tissue. Later in 2014, Francesca Tavano et al. demonstrated that ARNLT, CRY1-2, PER1-3, and SIRT1 were all significantly down-regulated in pancreatic cancer samples [ 220 ]. According to this study, dysregulated SIRT1 and clock gene expression must be weighed against opposing changes in pro- and anti-tumor genes since it may be environmentally dependent and implicated in oncogenic or tumor suppressor signaling [ 220 ].

Glioma

According to the WHO classification of central nervous system (CNS) malignancies, gliomas are categorized into CNS WHO grades 1–4, ranging from low grade to high grade malignancy, according to their chemical properties and microscopic appearance [ 221 ]. Gliomas are primary CNS tumors that generated from glial progenitor cells [ 222 ], whose incidence and mortality rates are expected to rise in the coming years, particularly in developing countries [ 223 ].

In 2012, Yong Luo et al. observed significantly reduced levels of CRYs in glioma cells in contrast with the surrounding non-glioma cells [ 224 ], which leads to hypothesize that alterations in the expression of CRYs may disrupt the regulation of the circadian rhythm, potentially providing survival benefit to glioma cells. Melissa H. Madden and colleages evaluated the relationship between glioma risk and genetic variations in the circadian gene pathway [ 225 ]. Using TCGA data, this clinic-based investigation demonstrated how CRY1 expression differs in glioblastoma multiform compared to normal brain. In addition, Madden et al. 2014 [ 225 ] (with a 10-fold larger speciment than Yong Luo et al. 2012) and Wang et al. 2021 [ 226 ] (using information from three distinct databases) analysed the level of circadian genes in glioma tissues compared to healthy tissues, and concluded that glioma tissues exhibit overexpression of CRY1.

Emerging innovations have explored the mechanisms behind the glioma. One of the most prevalent oncogenes in human malignancies, including gliomas, is c-myc [ 227 – 229 ]. In 2022, Patrica Jarabo et al. found that the PI3K pathway regulates the expression of cryptochrome in glioma cells and, in turn, cryptochrome is essential and adequate to encourage MYC accumulation in gliomas [ 230 ]. According to computational biology studies, CRY1 is linked to gliomas’ increased mutation rate and mutation hotspots in particular genes, like EGFR and PTEN, which may encourage tumor growth by creating genomic instability [ 231 ]. Together with REV-ERB proteins, CRY1 negatively regulates important glycolytic enzymes (like HK2 and LDHA) and genes linked to the tricarboxylic acid cycle [ 231 ]. This may suppress the Warburg effect, which limits the amount of energy that tumor cells can use [ 231 , 232 ]. Furthermore, by affecting fatty acid synthases (such as SCD and FAS), CRY1 may indirectly alter lipid metabolism; nevertheless, its precise function in glioblastoma is still debatable [ 231 ].

Adenoma

Adenomas can form in several sections of the body (encompassing but not restricted to the breast, thyroid, adrenal glands, and intestine) and originate mainly from glandular tissue. Although every adenoma has the capacity to evolve malignantly, only a minority of adenomas actually develop into invasive cancers (progressive adenomas) [ 233 ]. In this part, we describe the expression of CRY1 in aldosterone-producing adenoma (APAs), cortisol secreting adenomas, and parathyroid adenomas.

Pituitary tumors are classified by the WHO into three main classes: normal adenomas, atypical adenomas, and pituitary carcinomas [ 234 ]. Pituitary adenomas are tumors that originate in the anterior pituitary gland [ 235 ]. In adults, they rank third in frequency of CNS malignancies [ 235 ]. In the human anatomical pituitary gland, protein levels of CRY1 do not fluctuate with the time of day [ 236 ].

In the research conducted by Martina Tetti et al., it revealed that CRY1 and CRY2 are regulated by AngII and may play a vital part in the function of adrenal cells, particularly in the context of APAs [ 237 ]. The study uses human adrenocortical cells HAC15 as an in vitro model and comprises 20 paired adrenal samples and 46 sporadic APAs. In contrast to the nearby adrenal cortex, APAs showed differential expression of both CRY1 and CRY2. Additionally, cortisol-secreting adenomas showed much lower expression of PER1, CRY1, and Rev-ERB as compared to normal tissue [ 238 ]. When APA was compared to normal tissue, every gene linked to the clock was overexpressed, though not substantially [ 238 ]. In parathyroid adenomas, mRNA expression of CRY1 was also downregulated [ 170 ].

Hepatocellular carcinoma

In 2020, hepatocellular carcinoma (HCC)—which is developing in the context of chronic liver disease and cirrhosis—accounted for 906,000 new cases and 830,000 deaths worldwide, making it the third most prevalent cause of cancer-related deaths and the sixth most often diagnosed malignancy [ 157 ]. The majority of HCC instances occur in developing nations, with Asia and sub-Saharan Africa accounting for nearly 80% of all cases [ 239 ].

While some studies report significantly decreased expression of core clock genes (CRY1-2, CLOCK, BMAL1, and PER1-2) in HCC [ 240 ], other investigations found no statistically significant difference in the expression of CRY1-2, PER1-2, NPAS2, CLOCK, and BMAL1 between tumor and adjacent normal tissues ( p > 0.05) [ 241 ].

HCC progression involves complex interactions between cellular processes and circadian regulation. Current research indicates potential involvement of the circadian protein CRY1 in certain HCC pathogenic pathways. CRY1 may potentially influence EMT in HCC through its known regulatory effects on HIF-1α stabilization [ 115 ] and NF-κB signaling [ 89 , 90 ], both of which are established mediators of metastatic progression in liver cancer. Furthermore, CRY1 regulates hepatocyte survival through modulation of FOXO1 activity [ 60 ], which specifically implicated in promoting Th17 cell-dependent HCC recurrence following hepatic ischemia-reperfusion injury [ 242 ]. When autophagy is lost or inhibited, preclinical research utilizing mice models does show decreased metastases [ 243 ]. An in vivo model of hepatocellular carcinoma, for instance, showed that autophagy facilitated the spread of metastases and anoikis resistance [ 243 ]. Notably, these relationships are reciprocal - HCC-associated hypoxia and inflammation can disrupt CRY1 expression [ 244 ], creating a vicious cycle that accelerates disease progression. CRY1 potentially ameliorates HCC-associated cachexia by blocking hepatic gluconeogenesis through FOXO1 degradation [ 60 , 245 ]. This intricate crosstalk positions CRY1 as a central node connecting circadian biology to multiple hallmarks of HCC, with important implications for chronotherapeutic strategies.

Future directions and innovations

Target CRY1 with small molecules

Emerging studies have shown promising results in finding small molecules that directly target CRY1 for therapeutic effects (Table 3 ). Researchers discovered KL001, a tiny chemical that selectively interacts with CRY to lengthen the circadian period, in 2012 [ 21 ]. Additionally, in primary hepatocytes, glucagon-induced gluconeogenesis was reduced by KL001-mediated CRY stability, therefore promising for the therapy of metabolic disorders. In vitro and in vivo, KL001 and the derivative SHP656 were shown to selectively kill glioblastoma stem cells [ 246 ].

Benzophenone derivatives were discovered by Dušan Kolarski et al. in 2021 to bind specifically with the CRY1 photolyase homology area, activating CRY1 and lengthening the circadian period [ 247 ]. In particular, the benzophenone component altered the “lid loop”—an area of CRY1 that is not inside the compound-binding pocket—and created a special interaction in the lid loop with Phe409. In order to change this crucial interaction, the benzophenone was carefully swapped out for a switchable azobenzene molecule that can go through light-controlled cis-trans isomerization.

CLOCK-BMAL1-evoked E-box-mediated transcription was activated in 2014 when researchers discovered the first new chemical CRY1/2 inhibitors, substance 15 (KS15), a derivative of 2-ethoxyprognaoic acid [ 248 ]. Sung Kook Chun et al. discovered later in 2015 that KS15 had no effect on MCF-10 A cells but lowered cell growth rate and enhanced MCF-7 cells’ chemosensitivity to tamoxifen and adriamycin [ 249 ]. These results imply that in some forms of breast cancer, KS15’s pharmacological suppression of CRY has an antiproliferative effect and raises susceptibility to antitumor drugs. In summary, the use of a novel photopharmacological technique makes it possible to manipulate circadian rhythms in space and time.

CRY1 as diagnostic and therapeutic tools

Innovative research in molecular cancer epidemiology reveals how clock gene variants manipulate cancer and multiple diseases. Leena Kovanen et al. provided evidence in 2014 that CRY1 is linked to arterial hypertension and high blood pressure, which can impact the development of tumors [ 250 ]. Sandra P. Smieszek et al. discovered that people with the CRY1Δ11 variation may be more likely to develop Delayed Sleep-Wake Phase Disorder (DSWPD), and that early detection and management may be helpful for comorbidities such mood disorders, depression, and ADHD [ 251 ]. A model of diabetes and obesity with elevated gluconeogenesis is the db/db mouse [ 252 ]. When adenovirus causes the liver of db/db mice to overexpress CRY1, the mice show decreased levels of blood glucose under fasting conditions [ 253 ]. This indicates that acute manipulation of clock components could be a method to control blood glucose levels and interfere with gluconeogenesis. As highlighted in the “Metabolism” section, CRY1 may be a viable target for anti-obesity therapy [ 61 ].

Gene regulation of CRY1 can also be a therapeutic targets in cancer therapy. In 2021, researchers came to the conclusion that CRY1 over-expression may be a new therapeutic target and is a powerful predictive biomarker for prognosis and responsiveness to chemoradiation in patients with cervical cancer [ 133 ]. A recent study by Yasushi lida et al. suggested that combining antiangiogenic drugs with CRY1 inhibitors may enhance the effectiveness of PARP inhibitors when treating ovarian cancer that is HR-proficient [ 172 ]. Furthermore, identifying specific changes in physiological rhythms may help identify people who are more susceptible to disease and those who stand to gain from circadian-based treatments [ 254 ]. For example, Qihang Sun et al. suggested a innovative lung adenocarcinoma diagnosis model based on genes linked to the circadian clock [ 255 ]. Notably, liquid biopsy—a technique analyzing circulating tumor components in blood—has emerged as a promising tool for real-time monitoring of molecular dynamics in cancer [ 9 , 85 , 256 ]. When integrated with circadian gene signatures (e.g., CRY1 expression patterns), this approach could potentially refine early detection and personalize chronotherapy strategies [ 9 , 85 , 256 ]. Additionally, repurposing existing drugs combined with vitamin supplementation (e.g., vitamin D as a prophylactic agent with immunomodulatory effects) may synergize with CRY1-targeted therapies, offering a sustainable approach to enhance anti-cancer efficacy [ 257 – 259 ].

Promote drug efficacy through CRY1 modulation

Key findings show that optimizing treatment effectiveness may be achieved by coordinating medication distribution with endogenous physiological cycles [ 260 ]. Several chemotherapeutic drugs exhibited elevated lethal effects, particularly during a particular cell-cycle phase, indicating that better treatment outcomes may result from circadian chrono-modulated drug delivery systems [ 261 ]. Furthermore, the actual pharmacological targets may peak at various times [ 254 ]. For instance, in 2014, Ahmed Hamed Salem et al. found that evening doses were associated with 27% lower linifanib bioavailability than morning doses [ 262 ]. Chronomodulated 5-fluorouracil (5-FU) infusion studies identified diurnal pharmacokinetics [ 263 ]. Peak plasma concentrations consistently occurred at 4:00 a.m., which corresponds to maximal chemotherapy tolerance [ 264 ]. PER2 and CRY1 transcription regulate fluctuating VEGF levels in xenograft tumors in a circadian cycle [ 265 ]. Given that VEGF is pivotal in tumor-promoted blood vessel formation, the effectiveness of antitumor efficacy might be improved by timing the drug administration to coincide with periods of elevated VEGF production. Furthermore, the chrono-modulated infusion from 2:00 a.m. to 8:00 a.m. decreased acute and chronic diarrhea in 31 patients receiving irinotecan, an antineoplastic enzyme inhibitor predominantly used to treat colorectal cancer, improving the patients’ quality of life [ 266 ].

Conclusions and clinical perspectives

In this review, a substantial body of evidence demonstrates that CRY1 serves as a double-edged sword in cancer development, with its functional impact being determined by tissue-specific regulatory networks and temporal dynamics. Transcriptomic studies reveal cancer-type-specific mRNA dysregulation patterns: CRY1 is upregulated in ovarian [ 169 ], gastric cancer [ 188 ] and prostate cancers [ 71 ], but downregulated in cervical cancer [ 160 ], pancreatic cacner [ 219 , 220 ] glioma [ 224 ] and parathyroid adenomas [ 170 ]. Notably, changes in circadian clock gene expression, whether up- or down-regulated, throw off the equilibrium of cell division and promote the growth of tumor cells. Therefore, disturbance of the circadian rhythm may be a cause of cancer as well as one of its effects.

At the protein level, CRY1’s functional duality becomes even more apparent through its dynamic interactions with core cellular pathways. On the protumorigenic side, CRY1 promotes c-myc accumulation via PI3K pathway in glioblastoma [ 230 ], sustains STAT3-NANOG signaling to associate with chemoresistance in cervical tumors [ 133 ], and facilitates p53 degradation through MDM2 interaction in bladder cancer [ 152 ]. Conversely, its tumor-suppressive roles include inhibiting hepatic gluconeogenesis through FOXO1 degradation to potentially counteract cancer cachexia [ 60 , 245 ], and mediating DNA repair via HAUSP interaction in response to genomic damage [ 26 ].

Clinical evidence also demonstrates that CRY1 dysregulation impacts cancer progression and treatment outcomes. Elevated CRY1 expression correlates with adverse prognosis in hormone-sensitive malignancies [ 267 ], while epidemiological studies link circadian disruption to shift work-related tumorigenesis [ 164 ]. Therapeutically, emerging research highlights the intersection between CRY1-modulated metabolic pathways and cancer therapy, particularly in obesity-related malignancies—a rapidly evolving research focus. Our analysis further outlines CRY1-targeted antitumor strategies through three key avenues: (1) small-molecule targeting (e.g., KL001 derivatives for glioblastoma stem cells; KS15 for breast cancer chemosensitization); (2) diagnostic repurposing (e.g., CRY1 expression signatures as prognostic biomarkers in cancers); and (3) chronotherapeutic optimization (e.g., timed administration of 5-FU/irinotecan based on CRY1 oscillation phases).

As for the limitations, the current understanding of CRY1 presents several critical research gaps that need to be addressed. First, although numerous CRY1 SNPs have been identified as significant contributors to individual variations in circadian regulation, their specific mechanistic impacts on biological rhythms and associated pathologies remain poorly characterized. Second, while existing genetic association studies and animal models have established connections between CRY1 variants and various disorders - including sleep disturbances, metabolic syndromes, and cardiovascular conditions - these findings predominantly remain at the preclinical stage without clear pathways for clinical application. To bridge these knowledge gaps, future investigations should focus on comprehensive spatiotemporal proteomic analyses to reconcile observed discrepancies between mRNA expression and protein functionality, while particularly emphasizing the characterization of isoform-specific behaviors and the validation of potentially druggable protein-protein interaction interfaces revealed through recent structural biology advances. This multidimensional approach will be essential for translating basic CRY1 research into clinically relevant therapeutic strategies.

Acknowledgements

The authors acknowledge Yijia Liu and Yanzhe Huang for their technical support in this work. We are grateful to everyone who took part in this research.

Abbreviations

Cryptochrome-1

World Health Organization

Polymorphisms

Circadian locomotor output cycles protein kaput

Aryl hydrocarbon receptor nuclear translocator-like

Cryptochrome-1 and − 2

Casein kinase 1 epsilon

Neuronal PAS domain protein 2

Differentiated embryo-chondrocyte expressed gene 1/2

Period 2

Brain-muscle Arnt-like protein 1

Photolyase homology

Methenyltetrahydrofolate

PER-ARNT-SIM

Transcription-translation feedback loop

Suprachiasmatic nucleus

Cyclin-dependent kinases

ROR responsive element

Glucocorticoid receptor

Phosphoenolpyruvate carboxykinase 1

Light chain 3

Peroxisome proliferator-activated receptor delta

High-fat diet

Pars tuberalis

Timeless

Nuclear factor-kappa B

Tumor-associated macrophage

Regulatory T cells

Protein kinase A

Vascular endothelial growth factor

Tumor microenvironment

Pattern recognition receptors

T-cell receptors

Tumor necrosis factor receptor

Tumor necrosis factorα

Toll-like receptors

Phosphorylated p65

Signal transducer and activator of transcription 3

G-protein coupled receptors

Interleukin-6

Lipopolysaccharides

human papillomavirus

Real-time polymerase chain reaction

Yes-associated protein/transcriptional co-activators with PDZ-binding motifs

epithelial ovarian cancer

homologous recombination

poly(ADP-ribose) polymerase

stomach adenocarcinoma

Age-standardized rate

magnetic resonance imaging

Pancreatic ductal adenocarcinoma

Aldosterone-producing adenoma

Central nervous system

Delayed Sleep-Wake Phase Disorder

5-fluorouracil

Hedgehog

Sonic hedgehog

Indian hedgehog

Desert hedgehog

Epithelial-mesenchymal transition

Neuromedin S

Hepatocellular carcinoma

Bcl2 interacting protein 3 like

GABA type A receptor associated protein like 1

Bcl2 interacting protein 3

Microtubule-associated protein light chain 3B

Author contributions

YL: Writing– original draft. YH: Writing– original draft. HL: Writing– review & editing. XL: review & editing. All authors read and approved the final manuscript.

Funding

The author(s) declare that no financial support was received for the research, authorship, and/or publication of this article.

Data availability

Not applicable.

Declarations

Ethics approval and consent to participate

Not applicable.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Footnotes

Publisher’s note

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

Contributor Information

Hongliang Luo, Email: ndefy11008@ncu.edu.cn.

Xueqin Lin, Email: aa13870055778@163.com.

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Associated Data

Data Availability Statement

Not applicable.