Research and development prospects of TRIM65

Abstract

Ubiquitination, the prevalent posttranslational modification, plays a crucial role in regulating protein function, localization, and degradation within cellular environments. As an E3 ubiquitin ligase, TRIM65 has been shown in various studies to facilitate the ubiquitination of specific substrates, thereby controlling inflammation, innate immune responses, cell proliferation, apoptosis, and tumor progression. Given the multifaceted and significant role of TRIM65, this review compiles existing research on TRIM65 and lays the groundwork for future studies aimed at uncovering the mechanisms of TRIM65. Further understanding of TRIM65’s interactions with its substrate proteins will offer valuable insights into the molecular underpinnings of certain diseases. Additionally, by identifying small molecules or inhibitors that target TRIM65, we may be able to develop novel drugs that modulate its activity. Such research could lead to more precise and effective treatments for conditions such as chronic inflammation, autoimmune diseases, and cancer. In summary, the study of TRIM65 not only enhances our understanding of fundamental cellular processes but also opens up new perspectives and avenues for the development of innovative therapies.

Introduction

The posttranslational modification of proteins through ubiquitination is a widespread phenomenon among eukaryotic organisms. This process involves the carboxyl terminus of ubiquitin, a small regulatory protein, forming a covalent bond with the ε-amino group of a lysine residue on the target protein via an isopeptide linkage. Ubiquitin can construct polyubiquitin chains by linking additional ubiquitin molecules to itself through any of its seven lysine residues (K6, K11, K27, K29, K33, K48, K63) or an N-terminal methionine (M1) site.The ubiquitination pathway typically requires the coordinated actions of three enzymes: the ubiquitin-activating enzyme (E1), the ubiquitin-conjugating enzyme (E2), and the ubiquitin ligase (E3). During this process, the E1-activating enzyme utilizes energy obtained from the hydrolysis of ATP to form a thioester bond between an active site cysteine sulfhydryl group and the carboxyl terminus of the ubiquitin. Following activation, the ubiquitin is transferred onto the sulfhydryl group of the E2-conjugating enzyme. Subsequently, the E3 ligase identifies the target protein and facilitates the transfer of ubiquitin to the designated substrate(Komander et al., 2012). The E3 ligase is essential for initiating the ubiquitination modification of proteins. In humans, there are over 600 distinct E3 ligases encoded, and their diversity in type and function is closely associated with the specificity of ubiquitination(Borlepawar et al. 2018). Ubiquitination is a critical regulatory mechanism of cellular processes, allowing for the control of target protein stability, activity, localization, and interactions. It is crucial for numerous biological functions, as protein ubiquitination is integral to processes such as protein degradation, signal transduction, cell cycle regulation, and the modulation of gene expression(van der Veen et al., 2012;Oh et al. 2018).

Tripartite motif-containing proteins (TRIMs) constitute a family of proteins with E3 ligase activity, distinguished by their unique structural features, which include the N-terminal RBCC (RING finger, B-box, coiled-coil) domain and a variable C-terminal domain. Recent studies have indicated that the RING domain not only possesses E3 ligase activity but also serves as the binding site for the E2 conjugating enzyme and the target protein. Furthermore, the E2 conjugating enzyme facilitates the transfer of ubiquitin to the target protein, thereby initiating the process of ubiquitination(Cai et al. 2022). One or two highly variable C-terminal domains (PRY, SPRY, COS, FNIII, PHD) are typically appended to the RBCC domain and are likely responsible for differential substrate recognition and enzyme activity regulation. Variations in the C-terminal domain determine the specificity of each TRIM protein and serve as the basis for the classification of TRIM subfamilies(Deng et al. 2024). However, a few TRIM proteins, such as TRIM14, TRIM20, and TRIM16, lack the RING domain. It has been reported that TRIM16 can exert E3 ligase function through its B-box domain(Di Rienzo et al. 2020). The ubiquitination of target proteins mediated by TRIMs family proteins typically results in two outcomes: degradation by the 26 S proteasome, which is facilitated by K48 or K11 ubiquitination, or non-proteolytic pathways, such as the activation of protein kinases due to K63 ubiquitination and the regulation of protein stability through K27 ubiquitination(Azuma et al. 2021). Many important biological processes, including intracellular signal transduction, protein quality control, innate immunity, autophagy, and carcinogenesis, have been shown to involve TRIM family members(Cai et al. 2022; Ahsan et al. 2024).

Current studies on the specific biological mechanisms and functions of TRIM65, a member of the TRIMs family, focus on its biological effects on disease processes through the ubiquitination of target proteins. Initially identified as an SNP gene associated with white matter lesions(Fornage et al. 2011; Freudenberger et al. 2012; Schmidt et al. 2012), TRIM65 was later shown to be a cofactor that regulates microRNA function (Li et al. 2014a, b)by ubiquitinating TNRC6, while also establishing its own E3 ligase activity(Li et al. 2014a, b). As research progresses, TRIM65 is emerging as a promising target for the treatment of related diseases. For instance, TRIM65 has been shown to inhibit inflammatory responses by mediating the ubiquitination (Tang et al. 2021)of NLRP3 and to regulate innate antiviral immune responses by mediating the ubiquitination of the innate immune receptor melanoma differentiation-associated protein 5 (MDA5)(Lang et al. 2017). TRIM65 has been demonstrated to either promote or counteract the stability of specific oncogenic proteins within the context of tumor biology through ubiquitination, thereby influencing tumor cell growth and survival(Yang et al. 2017). Our study has revealed that TRIM65 plays a dual role in atherosclerotic disease. TRIM65 mediates K48-linked ubiquitination and degradation of VCAM-1, suppressing monocyte adhesion to activated endothelial cells and monocyte infiltration in tissues(Li et al. 2020), which leads to the improvement of endothelial dysfunction and atherosclerosis. Interestingly, in smooth muscle cells, TRIM65 can also promote the phenotypic transition of these cells from a contractile to a synthetic state by activating the PI3K/Akt/mTOR signaling pathway, thereby facilitating atherosclerosis(Zhou et al. 2024). Further exploration of the more extensive and deeper biological functions of TRIM65 in atherosclerosis is ongoing.

This review article aims to summarize clinical and basic research related to TRIM65, elucidate its biological functions across various pathophysiological conditions and diseases, provide guidance for future research, and propose new protein targets for the intervention or treatment of certain diseases.

TRIM65 facilitates NLRP3 ubiquitination to suppress inflammasome activation

The study of the NLRP3 inflammasome has been a hot topic since Agostini L reported its discovery in Immunity in 2004(Agostini et al. 2004). The NLRP3 inflammasome, a multi-protein complex consisting of the “sensor protein” NLRP3, the “adaptor protein” ASC, and pro-caspase-1, assembles in response to cellular perturbation(Lu et al. 2014a, b). Inflammasomes detect pathogen-associated molecular patterns (PAMPs) or host-derived damage-associated molecular patterns (DAMPs) to recruit and activate the pro-inflammatory protease Caspase-1. The activation of Caspase-1 facilitates the cleavage of inactive IL-1β and IL-18 precursors, resulting in the production of mature IL-1β and IL-18. Additionally, activated Caspase-1 cleaves gasdermin D (GSDMD) into its reactive amino (N) and carboxyl (C) termini. The N-terminal domain, which exhibits lipid selectivity, binds to phosphatidylinositol (unique to eukaryotic membranes) and fructophospholipids (unique to prokaryotic membranes, including those containing phosphatidylmuscle and cardiolipin liposomes) and polymerizes to form hollow, ring-shaped oligomers with diameters ranging from 10 to 20 nanometers upon membrane formation. The microlumen creates an imbalance between the interior and exterior of the cell, resulting in cell lysis and death. Simultaneously, pores facilitate the secretion of small molecules, such as IL-1β and IL-18, to the exterior of the cell. Additionally, an increased number of immune cells are recruited, triggering an inflammatory response that ultimately leads to cellular pyroptosis (Mariathasan et al. 2006)(Fig. 1a). The NLRP3 inflammasome, a central component of innate immunity, plays a crucial role in regulating inflammatory responses through communication with other cellular compartments(Dupaul-Chicoine et al. 2015). The secretion of proinflammatory cytokines, mediated by the NLRP3 inflammasome, is detrimental in the pathogenesis of chronic inflammatory and metabolic diseases, including diabetes(Zeng et al. 2024), obesity(Javaid et al. 2023), atherosclerosis(Guo et al. 2024), and tumors(Sorrentino et al. 2015). However, some studies have reported that these proinflammatory cytokines play a beneficial role in certain infectious diseases and tumors(Christgen et al. 2021; Sharma et al., 2021). It has been discovered that the NLRP3 inflammasome can function as a negative regulator of tumorigenesis in colitis-associated colorectal cancer(Allen et al. 2010). In conclusion, regulating the activity of the NLRP3 inflammasome is crucial for maintaining sustained normal cellular and health homeostasis.

Fig. 1.

a Upon detection of an activation stimulus, NLRP3 undergoes a transformation from inactive homomultimers to active multimeric inflammasomes. This process facilitates the assembly of NLRP3 inflammasomes with their ASC and pro-caspase-1, leading to the activation of caspase-1. Subsequently, activated caspase-1 promotes the proteolytic cleavage and activation of protease family cytokines of the IL-1 family and gasdermin D, triggering terminal lytic cell death, termed pyroptosis. b TRIM65 facilitates the ubiquitination of NLRP3 and inhibits the interaction between NLRP3 and NEK7, thereby diminishing the assembly and activation of the NLRP3 inflammasome. UroB enhances the competitive binding of TRIM65 to NLRP3, impeding the binding of TXNIP to NLRP3. Berberine promotes the TRIM65-mediated ubiquitination of NLRP3

Many E3 ligases can mediate the ubiquitination of NLRP3, thereby regulating the activity of the NLRP3 inflammasome(Lopez-Castejon 2019; Deng et al. 2022, 2024). Yang Lu’s team observed that berberine (BBR), a compound known for its anti-inflammatory effects(Zhu et al. 2022), can promote the binding of TRIM65 to NLRP3, effectively reducing inflammatory damage to hippocampal neuron function(Yang et al. 2023). Mechanistically, TRIM65 binds to the NACHT domain of NLRP3, promotes lys48- and lys63-linked ubiquitination of NLRP3, and inhibits the interaction between NLRP3 and Nima-associated protein kinase-7 (NEK7)(Tang et al. 2021). This is a crucial step that precedes NLRP3 oligomerization, assembly, and caspase-1 activation (Shi et al. 2015; He et al. 2016; Schmid-Burgk et al. 2016; Zhao et al. 2020)Fig. 1b). It has been widely confirmed that lysine 48-linked ubiquitination regulates protein degradation, whereas lysine 63-linked ubiquitination facilitates signal transduction(Lopez‐Castejon 2020). The team led by Kawashima discovered that the E3 ligase Ariadne homolog 2 can also catalyze the mixed lysine 48- and lysine 63-linked ubiquitination of NLRP3, yet it does not influence the expression levels of NLRP3(Kawashima et al. 2017). This is consistent with the phenomenon observed in the study by Tiantian Tang’s team(Tang et al. 2021), indicating that TRIM65-mediated mixed lys48- and lys63-linked ubiquitination of NLRP3 inhibits the oligomerization of the “sensor protein” NLRP3 without leading to its proteasomal degradation.

Thioredoxin interacting protein (TXNIP) is recognized as an endogenous regulator of cellular oxidative stress and inflammation. When exposed to oxidative stress conditions, TXNIP dissociates from thioredoxin (TRX) and binds to NLRP3, thereby activating the NLRP3 inflammasome(Zhou et al. 2011; Tsubaki et al. 2020). Downregulation of TXNIP expression has been shown to inhibit NLRP3 inflammasome activity in various diseases, including vascular dementia(Du et al. 2018), non-alcoholic fatty liver disease(NAFLDHe et al. 2017a, b; He et al. 2017a, b), and neonatal hypoxic-ischemic brain damage(Chen et al. 2018). Urolithin B (UroB), a metabolite resulting from the metabolism of ellagitannins by intestinal microbiota, has demonstrated therapeutic potential in various neurological disorders(Eidizade et al. 2023; Rahimi-Kalateh Shah Mohammad et al. 2023). In our study, we observed that treatment with UroB significantly upregulated TRIM65 expression, which had previously been downregulated in the brain tissue of rats subjected to middle cerebral artery occlusion (MCAO), while simultaneously downregulating TXNIP expression. These findings suggest that UroB promotes the competitive binding of TRIM65 to NLRP3, thereby inhibiting the binding rate of TXNIP to NLRP3 (Luo et al. 2024)(Fig. 1b). This mechanism effectively suppresses neuroinflammation and mitigates cerebral ischemia/reperfusion injury.

Finally, we demonstrate that TRIM65 is indispensable for NLRP3 ubiquitination and the regulation of inflammation. TRIM65 acts as a negative regulator of NLRP3 inflammasome activation, thereby providing new avenues and therapeutic approaches for dealing with inflammatory diseases mediated by the NLRP3 inflammasome.

TRIM65 mediates MDA5 ubiquitination for enhanced immune response

Melanoma differentiation-associated protein 5 (MDA5), an innate immune receptor, is pivotal in the innate immune response against RNA viruses(Gitlin et al. 2006; Kato et al. 2006). Upon recognizing viral RNA, both retinoic acid-inducible gene I (RIG-I) and MDA5 activate their common adaptor protein, mitochondrial antiviral signaling protein (MAVS). This activation triggers type I interferon (IFN-I) signaling and subsequent antiviral responses, while also promoting apoptosis in virus-infected cells(Dixit et al. 2010; Wu et al., 2014;Rehwinkel et al., 2020). Rongbin Zhou’s team observed that TRIM65 interacts with MDA5, enhancing MDA5’s ability to stimulate IFN-I signaling. Mechanistically, TRIM65 mediates K63-linked polyubiquitination at lysine 743 of MDA5(Lang et al. 2017). However, the unanchored K63-linked polyubiquitin chains of MDA5 are essential for its oligomerization and activation(Jiang et al. 2012; Song et al. 2021). Kazuki Kato further discovered that TRIM65 utilizes Spry bivalently to selectively recognize MDA5 filaments, with TRIM65 Spry binding to the α1/α3 helix of MDA5’s Hel2 domain (Kato et al. 2021)(Fig. 2a).

Fig. 2.

a The TRIM65 Spry domain interacts with the Hel2 domain of MDA5, facilitating K63-linked polyubiquitination at lysine 743 of MDA5. This process promotes MDA5 oligomerization and activation. The arrestin-like N domain of ARRDC4 binds to MDA5, further recruiting TRIM65 to mediate K63-linked polyubiquitination of MDA5. Consequently, downstream signaling pathways are activated, leading to the transcription of pro-inflammatory cytokines. b TRIM65’s coiled-coil (CC) and SPRY domains interact with the N-terminal nuclear localization signal (NLS) and HMG box region of TOX4 to mediate the K48-linked ubiquitination and subsequent degradation of TOX4, resulting in increased Bcl2 expression, decreased Bax expression, and inhibition of intestinal cell apoptosis

ARRDC4, a member of the arrestin family, exhibited significantly increased expression in patients with hand, foot, and mouth disease (HFMD) infected with enterovirus 71 (EV71). This elevated expression positively correlated with serum concentrations of IL-6, TNF-α, and CC motif chemokine 3 (CCL3). ARRDC4 functions as an adapter that mechanistically links MDA5 with TRIM65. The ARRDC4 N domain interacts with MDA5, and upon additional engagement of TRIM65, it induces K63 polyubiquitin chains on MDA5(Meng et al. 2012) (Fig. 2a), signaling downstream activation and transcription of proinflammatory cytokines.

TRIM65 also undergoes ubiquitination by other E3 ligases. Two studies on the proinflammatory Salmonella effector SopA, an E3 ligase similar to HECT, suggest diametrically opposing modes of action for SopA in mediating TRIM65 ubiquitination. The results from Jana Kamanova’s team suggest that SopA-catalyzed TRIM65 ubiquitination is nondegradable and can promote TRIM65-mediated MDA5 ubiquitination, enhancing its ability to stimulate interferon-β signaling(Kamanova et al. 2016). Conversely, Evgenij Fiskin and colleagues did not detect stimulation of TRIM65-mediated type I interferon expression by SopA in their study. Instead, they observed that SopA activity led to a decrease in interferon-β transcription. Under standard Salmonella infection conditions, endogenous TRIM65 protein levels gradually decrease, which is considered one of the strategies Salmonella employs to evade the host immune response(Fiskin et al. 2017). The discrepancy between the results of the two studies may be explained by the relative expression levels of secreted SopA, TRIM65 in vivo, and host E3 ligase activity, which determine the different outcomes of host-pathogen targeting events.

Finally, we conclude that TRIM65 serves as a regulator of the innate immune response by functioning in MDA5 ubiquitination, MDA5 oligomerization, and activation. Clearly identifying the interactions among TRIM65, MDA5, ARRDC4, and SopA can greatly deepen our understanding of host-pathogen interactions and provide clues in the development of novel antiviral strategies.

TRIM65’s role in apoptosis regulation

TRIM65 plays a complex and significant role in the regulation of apoptosis. It has been demonstrated that administering isoproterenol increases the expression of TRIM65 in murine myocardial tissues. Conversely, treating TRIM65 knockout mice with isoproterenol resulted in reduced cardiac function and, in some instances, heart failure. The overexpression of TRIM65 significantly downregulates the levels of Jak1, Stat1, cytochrome c, and caspase3, and promotes the upregulation of Bcl2, thereby preventing cardiomyocyte apoptosis in vitro. This was observed in experiments with isoproterenol-treated cardiomyocytes(Liu et al. 2023a, b). Jak1-activated Stat1 promotes the release of mitochondrial cytochrome c, thereby activating caspase3 to induce apoptosis. Additionally, Stat1 downregulates Bcl2 and upregulates the proapoptotic protein Bcl2-associated X protein (BAX), further facilitating apoptosis(Cao et al. 2015; Berlanga-Acosta et al. 2024). TRIM65 is hypothesized to inhibit the Jak1/Stat1 signaling pathway by mediating Jak1 ubiquitination and subsequent degradation, thereby mitigating cardiomyocyte mitochondria-dependent apoptosis(Liu et al. 2023). Nonetheless, the exact molecular mechanisms underlying this process necessitate further elucidation.

Intestinal epithelial cell apoptosis serves as a primary factor contributing to structural and functional damage in intestinal tissue(Günther et al. 2013). Research conducted by Yingjie Huang’s team observed a significant decrease in TRIM65 expression and a concurrent increase in thymocyte selection-associated high mobility group box factor 4 (TOX4) expression in hypoxia-reoxygenation (H/R)-induced intestinal epithelial cells. Previous studies have confirmed TOX4’s role in promoting apoptosis(Liang et al. 2021; Sun et al. 2021). The overexpression of TRIM65 was found to significantly increase Bcl2 levels, decrease Bax levels, and alleviate intestinal cell apoptosis. Mechanistically, the CC and SPRY domains of TRIM65 interact with the nuclear localization sequence (NLS) and the HMG cassette region of TOX4’s N-terminus. This interaction mediates K48-linked ubiquitination and degradation of TOX4(Huang et al. 2024)(Fig. 2b), thereby inhibiting H/R-induced apoptosis in intestinal epithelial cells. Interestingly, in studies on hepatocellular carcinoma (HCC), small interfering RNA (siRNA) targeting TRIM65 was transfected into HCC cells. The results indicated that silencing TRIM65 expression induced apoptosis in these cells, significantly inhibited cell proliferation, attenuated cell growth and colony formation ability, and arrested the cell cycle at the G0-G1 phase(Bian et al. 2024). The biological mechanisms of TRIM65 in HCC will be elaborated upon in the subsequent section, which will discuss TRIM65’s biological functions in HCC.

These findings collectively underscore the diverse and sometimes contradictory biological regulatory functions of TRIM65 across various cellular microenvironments. This complexity suggests the need for further investigation into the molecular mechanisms by which TRIM65 operates within these contexts.

TRIM65: a novel target for tumor treatment

In recent years, there has been considerable interest in studying potential biomarkers for the clinical management of human tumors. Various cancers implicated in TRIM65 include hepatocellular carcinoma (HCC)(Yang et al. 2017; Jiang et al. 2024), non-small cell lung cancer (NSCLC)(Wang et al. 2011; Li et al. 2016; Guo et al. 2022), bladder urothelial carcinoma (UCB)(Wei et al. 2018), renal cell carcinoma (RCC)(Zhang et al. 2024), and rectal cancer(Chen et al. 2019). These findings suggest a close association between TRIM65 and the development and progression of tumors. Numerous studies have indicated that TRIM65 may serve as a potential target for tumor diagnosis and treatment.

TRIM65 overexpression promotes HCC progression

O-GlcNAc-TRIM65-NF2/YAP1 axis: a cascade amplifier driving metabolic reprogramming and immune escape in HCC

HCC continues to be one of the leading causes of cancer-related mortality globally(Llovet et al. 2021). We discovered that TRIM65 is significantly overexpressed in HCC, a process mediated by O-GlcNAcylation via O-GlcNAcylation transferase (OGT), which promotes the progression of HCC, as reported in Advanced Science. In contrast, mice with a conditional TRIM65 knockout exhibited markedly improved liver function and decreased levels of plasma AFP, ALT, and AST. TRIM65 facilitates the ubiquitination of the K44 residue on neurofibromatosis type 2 (NF2), thereby accelerating its degradation(Bian et al. 2024). NF2-Yes associated protein 1 (YAP1) signaling plays a crucial role in the development of HCC, as evidenced by several studies(Hyun et al. 2021; Qi et al. 2023). TRIM65-mediated degradation of NF2 results in the nuclear accumulation of YAP1, which promotes the transcription of downstream targets, including uridine monophosphate synthetase (UMPS) and fatty acid synthase (FASN). UMPS enhances uracil metabolism, subsequently upregulating the O-GlcNAcylation substrate UDP-GlcNAc, which in turn increases TRIM65 expression. This process establishes a positive feedback loop that exacerbates HCC progression. Additionally, FASN increases levels of free fatty acids, particularly palmitic acid, which enhances M2 macrophage polarization and inhibits CD8 + T cell infiltration. These effects foster HCC progression by establishing an immunosuppressive tumor microenvironment (Bian et al. 2024)(Fig. 3a). In conclusion, TRIM65 facilitates HCC progression by acting on the NF2-YAP1 axis.

Fig. 3.

a Role of TRIM65 in HCC: TRIM65 facilitates the ubiquitination and subsequent degradation of NF2, resulting in the nuclear accumulation of YAP1 and the activation of the UMPS (uracil metabolism/UDP-GlcNAc feedback loop) and FASN (which induces M2 macrophage polarization and CD8 + T cell depletion). Additionally, TRIM65 mediates the ubiquitination and degradation of Axin1, suppressing the β-catenin degradation complex, thereby allowing β-catenin/TCF/LEF to enter the nucleus and activate c-myc, cyclin D1, and Bcl-w. b The Role of TRIM65 in NSCLC: TRIM65 facilitates the ubiquitination and degradation of TNRC6A, thereby alleviating the suppression of miR-138-5p on ATG7 expression and fostering the progression of NSCLC. HAR1A enhances TRIM65’s ubiquitination and degradation of ANXA2, which in turn impedes the proliferation, EMT, and metastasis of NSCLC. c TRIM65 facilitates the ubiquitination and degradation of BTG3, resulting in increased Cyclin D1 protein levels and thereby advancing RCC progression. TRIM65 mediates the ubiquitination and degradation of DUSP6, which reduces DUSP6’s negative regulation of the ERK1/2 signaling pathway, thus promoting the progression of endometriosis

TRIM65 mediates the degradation of Axin1: A molecular switch for β-catenin oncogenic activation

β-catenin, a multifunctional protein, undergoes phosphorylation in the cytosol by the Axis Inhibition Protein (Axin) / Colonic adenomatous polyposis protein (APC) / Glycogen synthesis kinase 3 protein (GSK-3) degradation complex and subsequently degraded through the ubiquitin proteasome pathway(Li et al. 2012a, b; Qin et al. 2023). Upon stimulation of Wnt signaling, the Axin/APC/GSK-3 degradation complex is dissociated, and the stability of cytoplasmic β-catenin is enhanced. The stabilized β-catenin then translocates to the nucleus, where it associates with transcription factors, including T-cell factor/lymphoid enhancer factor (TCF/LEF). The activation of β-catenin/TCF/LEF complexes induces the expression of c-myc, cyclin D1, and Bcl-w genes, promotes cell cycle progression, triggers epithelial-mesenchymal transition (EMT), alters cell adhesion, and regulates tissue morphogenesis and tumor development(Nusse et al., 2017;Lecarpentier et al. 2019). Research conducted by YuFeng Yang’s team has demonstrated that TRIM65, which is highly expressed in HCC, directly binds to Axin1 and mediates its ubiquitination. This process accelerates Axin1 degradation through the proteasome, resulting in elevated β-catenin levels(Yang et al. 2017). The increased β-catenin subsequently translocates to the nucleus, exerting oncogenic activity (Fig. 3a).

TRIM65 - JAK1/STAT1 axis: Macrophage polarization switch and regulatory hub for the immunosuppressive environment of HCC

Bioinformatics analyses have revealed a strong correlation between high levels of TRIM65 and several adverse factors in HCC patients, including higher tumor grade, advanced TNM stage, vascular invasion, poor prognosis, and increased macrophage immune invasion levels (M0, M1, M2)(Jiang et al. 2024). M1 macrophages are associated with anticancer properties due to their ability to eliminate tumor cells. In contrast, M2 macrophages promote tumorigenicity by secreting cytokines, growth factors, and proteases that are tumorigenic(Quail et al., 2013;Caux et al. 2016). The activation of STAT1, the central transcription factor responsible for regulating macrophage polarization, induces the M1 phenotype(Caux et al. 2016; Zhu et al. 2020). Experimental observations have shown that downregulating TRIM65 expression significantly inhibits tumor growth and promotes M1 macrophage polarization while inhibiting M2 macrophage polarization. Mechanistically, the knockout of TRIM65 enhances the polarization of peritoneal macrophages and tumor tissue macrophages stimulated by conditioned media towards the M1 phenotype by activating the JAK1/STAT1 signaling pathway, thereby suppressing tumor growth(Jiang et al. 2024). The observed inhibition of the JAK1/STAT1 signaling pathway by TRIM65 in this study is consistent with previous findings in cardiomyocytes.

Immunotherapy and targeted therapy are currently two promising approaches for managing advanced hepatocellular carcinoma (HCC)(Flynn et al. 2019). In light of these findings, TRIM65 exhibits potential as a target for both immunomodulatory and targeted therapies in the management of HCC.

TRIM65 regulates non-small cell lung cancer progression and drug resistance

TRIM65-P53: Influences tumor progression through autophagy and ferroptosis

The downregulation of tumor suppressor gene p53 is associated with the proliferation of tumor cells and the enhanced resistance of tumor cells to chemotherapeutic drugs(Bieging et al. 2014). Studies have demonstrated that p53 exerts an inhibitory effect on autophagy, which is recognized as a major mechanism of chemoresistance in tumor cells(Green et al., 2009;Wu et al. 2011; Sui et al. 2013). Autophagy involves various selective degradation processes within cells and plays a dual role in tumor development(Suzuki 2013). On the one hand, autophagy provides energy and biosynthetic raw materials for the survival (Yang et al. 2011)of tumor cells. Conversely, the timely autophagic removal of damaged proteins and organelles can prevent tumor formation(Liu et al. 2022a, b). TRIM65 has been identified as a mediator of p53 polyubiquitination and proteasomal degradation, thereby promoting the progression of NSCLC(Li et al. 2016) and cervical cancer(Wang et al. 2022)(Fig. 3b). Given p53’s inhibitory effect on autophagy(Green and Kroemer 2009), TRIM65 has also been recognized as a ubiquitin ligase involved in the positive regulation of autophagy in tumor cells(Zhang et al. 2022). In 2015, Gu Wei’s team first revealed that p53 can inhibit tumor development by promoting ferroptosis in tumor cells(Jiang et al. 2015). p53 can enhance ferroptosis induced by glutathione peroxidase 4 (GPX4) inhibitors, thereby exacerbating lipid peroxidation reactions in tumor cells(Rodencal et al. 2023). Then, it is worth considering whether TRIM65 participates in regulating ferroptosis in tumor cells by mediating the degradation of p53 during the progression of many tumors?

TRIM65/miR-138-5p/ATG7 axis: a new target for overcoming NSCLC drug resistance by regulating autophagy

MicroRNAs (miRNAs) have been identified as significant mediators in the regulation of drug resistance and autophagy within tumor cells(Xia et al. 2008; Liang et al. 2010; Pink et al. 2015; Jin et al. 2017). Studies have demonstrated that the expression of miR-138-5p is significantly down-regulated in cisplatin-resistant NSCLC cell lines (A549/DDP)(Wang et al. 2011), while that of TRIM65 is notably up-regulated(Pan et al. 2019). TRIM65 functions mechanistically as a cofactor for microRNA(Li et al. 2014). Furthermore, it has been reported that TRIM65 mediates the ubiquitination and degradation of TNRC6A, thereby negatively regulating microRNA activity(Li et al. 2014). Consequently, the expression of miR-138-5p is inhibited. However, miR-138-5p directly binds to the 3’UTR of the autophagy-specific marker ATG7, inhibiting the expression of ATG7 mRNA(Pan, Chen et al. 2019)(Fig. 3b). TRIM65 activates autophagy and reduces apoptosis in NSCLC cells by upregulating ATG7 levels. Targeting TRIM65 to regulate autophagy in tumor cells may present a promising strategy to overcome chemoresistance in the treatment of NSCLC and related tumors.

TRIM65/HAR1A-ANXA2-Rac1 axis: a bidirectional switch for EMT and metastasis in multiple cancers

Annexin A2 (ANXA2), a calcium-dependent phospholipid-binding protein, plays a complex role in biological regulation. ANXA2 is involved in regulating cell membrane dynamics and migration(Gerke et al. 2005; Grieve et al. 2012), enhancing the RhoA/ROCK pathway while inhibiting the Rac1/WAVE2 pathway(Burridge et al., 2004). This inhibition of actin filament growth(Hayes et al. 2006), lamellipodia formation, and cell migration occurs on one hand(Zhao et al. 2011). On the other hand, ANXA2 promotes the activation of β-catenin(Sarkar et al. 2011), extracellular regulated protein kinases(Li et al. 2023; Liu et al. 2023a, b), and NF-κB(Liu et al. 2022a, b), thereby facilitating tumor progression. ANXA2 overexpression has been reported in hepatocellular carcinoma(Herrera-López et al. 2023), breast cancer(Yuan et al. 2017), and NSCLC(Luo et al. 2013; Ling et al. 2024). Interestingly, the Wen-Su Wei group observed downregulation of ANXA2 expression in UCB, with patients exhibiting low ANXA2 expression having the worst prognosis. However, high expression of TRIM65 in UCB patients positively correlates with advanced clinical stages and poor outcomes. Mechanistically, TRIM65 mediates the ubiquitination and degradation of ANXA2, thereby relieving ANXA2’s inhibition of Rac1. This process facilitates morphological changes in UCB cancer cells and induces epithelial-mesenchymal transition (EMT), migration, and invasion(Wei et al. 2018). Previous studies have identified the long non-coding RNA Highly Accelerated Region 1 A (HAR1A) as a tumor suppressor in NSCLC by regulating the STAT3 signaling pathway(Ma et al. 2022). The team led by Xiaodong Ling observed that overexpression of HAR1A increased the ubiquitination levels of ANXA2 and accelerated its degradation through the proteasome pathway in NSCLC. Mechanistically, HAR1A acts as a scaffold protein, facilitating TRIM65-mediated ubiquitination of ANXA2, thereby suppressing ANXA2-mediated cell proliferation, EMT, migration, and invasion in NSCLC(Ling et al. 2024) (Fig. 3b). This observation seems to contradict the findings of numerous other studies indicating that TRIM65 promotes NSCLC progression. It is conceivable that TRIM65, akin to ANXA2, may have a dual role in the progression of NSCLC or other cancers. It is important to highlight that many studies have reported associations between TRIM65 or ANXA2 and various tumor-associated diseases, positioning them as potential therapeutic targets. Therefore, a comprehensive understanding of the biological roles of both TRIM65 and ANXA2 is crucial.

TRIM65 in renal cell carcinoma, colorectal cancer, gastric cancer, and endometriosis

TRIM65 mediates the ubiquitination and degradation of BTG3: Cyclin D1 promotes the progression of RCC

In clear cell renal cell carcinoma (RCC), TRIM65 expression is significantly elevated and positively correlated with the degree of lymph node metastasis. Patients with high TRIM65 expression in clear cell RCC exhibit a poorer prognosis and reduced survival time(Zhang et al. 2024). The impact of TRIM65 on RCC cell proliferation is associated with the BTG anti-proliferation factor 3 (BTG3). BTG3 has been identified as a transcriptional target of p53 (Ou et al. 2007)and functions as a tumor suppressor in the development and progression of numerous malignancies(Mao et al. 2016; An et al. 2017). Suppressing BTG3 in cancer cells results in a hastened departure from DNA damage-induced G2/M cell cycle arrest(Ou et al. 2007). Mechanistically, TRIM65 promotes K48-linked ubiquitination of BTG3 at lysine 41, which alleviates the G2/M phase cell cycle arrest and increases the protein levels of downstream CyclinD1(Zhang et al. 2024) (Fig. 3c). CyclinD1, a crucial positive regulator of the cell cycle and a significant proto-oncogene, has been demonstrated to promote tumor cell proliferation, DNA damage repair, invasion, and metastasis when overexpressed(Yu et al. 2013; Wang et al. 2018a, b). Consequently, in renal cell carcinoma (RCC), the overexpression of TRIM65 enhances the proliferation and anchorage-independent growth capabilities of tumor cells by modulating the BTG3-CyclinD1 axis.

TRIM65-RhoA/cytoplasmic skeleton remodeling axis: the transfer regulatory hub of the lncrna-mirna sponge network

RhoA, a protein commonly overexpressed in cancer, acts as a critical GTPase necessary for the reorganization of the actin cytoskeleton, which is essential for cell migration in both temporal and spatial dimensions(Sahai et al., 2003;Wang et al. 2003). Concurrently, Rho GTPase activating protein 35 (ARHGAP35) functions as a GTPase activating protein with a well-established role in the inactivation of RhoA(Settleman et al. 1992; Wang et al. 2003; Bartolomé et al. 2008). TRIM65 has been observed to be upregulated in colorectal cancer, correlating with poor patient survival. Mechanistically, TRIM65 mediates the ubiquitination and degradation of ARHGAP35, thereby increasing Rho GTPase activity at the molecular level. (Chen et al. 2019)This, in turn, enhances cancer cell migration through the remodeling of the actin and microtubule cytoskeleton.

Luciferase reporter assays have revealed a direct interaction between miR-1281 and TRIM65, with miR-1281 demonstrating the ability to reduce TRIM65 wild-type reporter gene activity(Hu et al. 2019). Notably, elevated expression of the long non-coding RNA LINC00963 has been detected in colorectal cancer tissues and cells. When highly expressed, LINC00963 functions as a sponge, adsorbing miR-1281, inhibiting its expression in colorectal cancer cells, and leading to TRIM65 upregulation(Lv et al. 2021). In glioma, LINC01857 has been confirmed to act as a sponge adsorbent for miR-1281, resulting in increased TRIM65 expression and enhanced glioma cell migration and invasion(Hu et al. 2019). These findings suggest that targeting miRNAs regulating TRIM65 transcription in tumor cells with high TRIM65 expression may present an effective strategy for indirectly modulating TRIM65 expression levels.

TRIM65-PPM1A-TBK1 axis: inactivation of the phosphorylation brake and inflammation-tumor transformation switch in gastric cancer

The Phosphatase Mg²⁺/Mn²⁺ dependent 1 A (PPM1A) protein is widely recognized as a tumor suppressor gene(Mazumdar et al. 2019). TCGA data indicate that PPM1A expression is decreased in numerous cancer types, including gastric cancer, correlating with reduced patient survival. Yarbrough, W. G.‘s team has demonstrated that low levels of PPM1A can promote tumor development(Lu et al. 2014a, b). Previous studies have indicated that PPM1A can be degraded through ubiquitination mediated by TRIM proteins. For example, TRIM52 is upregulated in HCC and promotes tumor proliferation, migration, and invasion by facilitating the ubiquitination and degradation of PPM1A(Zhang et al. 2018). Likewise, TRIM59 enhances the invasion of ectopic endometrial stromal cells through the same mechanism(Wang et al. 2020). TRIM65 has been found to be highly expressed in gastric cancer tissues, with significantly higher expression in gastric cancer cell lines compared to gastric epithelial cells (GES-1). Inhibition of TRIM65 expression has been shown to suppress gastric cancer cell proliferation and invasion. Chang Liu’s team has demonstrated that downregulating TRIM65 expression in gastric cancer cells inhibits PPM1A ubiquitination and degradation, resulting in increased PPM1A levels and decreased p-TBK1 levels(Liu et al. 2022a, b). The member of the IKK kinase family, TBK1, is also essential for innate immunity and inflammatory responses(Sharma et al. 2003). Studies have demonstrated that an increase in TBK1 phosphorylation occurs under conditions of low PPM1 expression, which in turn activates proliferation and invasion in various cancer cells(Li et al. 2015; Cheng et al. 2018). These findings suggest that targeted regulation of the TRIM65-PPM1A-TBK1 axis may inhibit the proliferation and invasion of gastric cancer cells.

The TRIM65-DUSP6-ERK1/2-C-myc positive feedback loop promotes endometrial invasion

Extracellular signal-regulated kinases (ERK1/2) translocate from the cytoplasm to the nucleus following phosphorylation, mediating the transcriptional activation of various factors such as Elk-1, ATF, AP-1, c-Fos, and c-Jun. These factors are intricately involved in the regulation of a wide array of biological processes, including cell proliferation, differentiation, maintenance of morphology, cytoskeleton construction, apoptosis, and carcinogenesis(Lake et al. 2016). Research conducted to date has supported the notion that the upregulation of Integrin β1(Chen et al. 2015; Yang et al. 2015), MMP2(Li et al. 2012a, b)d myc (Zhu et al., 2019)through the ERK1/2 pathway is crucial for the proliferation and invasion of endometrial stromal cells. Notably, Jin Wang’s team demonstrated that the downregulation of TRIM65 in malignant lymphoma cell lines, Jurkat (T lymphocytes) and Raji (B lymphocytes), exhibited a tumor suppressor effect remarkably similar to that observed when blocking the ERK1/2 signaling pathway(Wang et al. 2018a, b). In a study conducted by Ying-Ting Wu et al., it was found that TRIM65 expression positively correlates with the levels of p-ERK1/2, C-myc, MMP-2, and integrin β1 in ectopic endometrial tissues from both patients and mouse models. Mechanistically, TRIM65 mediates the ubiquitination and subsequent degradation of DUSP(Wu et al. 2019)6, a known negative regulator of the ERK1/2 signaling pathway(Beaudry et al. 2019)(Fig. 3c). Intriguingly, further investigations revealed that the transcription factor C-myc can induce TRIM65 promoter activity and enhance TRIM65 expression(Wu et al. 2019). In conclusion, TRIM65 promotes endometrial stromal cell invasion by mediating a feedback regulatory loop involving the DUSP6-ERK1/2-C-myc signaling pathway.

Discussion

Extensive research on TRIM65 has revealed its critical role in various pathophysiological processes, including inflammatory responses, immune regulation, and tumor progression. TRIM65 exerts its biological functions by targeting specific proteins (see Table 1).

Table 1.

Summary of potential mechanisms of TRIM65 in diseases

Proteins/RNA	Disease	Effect	Outcome	Ref.
NLRP3	Inflammation	TRIM65 facilitates lys48- and lys63-linked ubiquitination of NLRP3	Suppression of NLRP3 activation, inhibition of inflammasome assembly	(Tang et al. 2021)
MDA5	RNA virus infection	TRIM65 catalyzes K63-linked ubiquitination of MDA5 at lysine 743	Activation of innate antiviral immunity	(Lang et al. 2017)
ARRDC4	EV71 infection	ARRDC4 functions as an adaptor protein, mediating the interaction between TRIM65 and MAVS	Enhancement of anti-EV71 immune response	(Meng et al. 2012)
TOX4	Intestinal II/R injury	TRIM65 orchestrates the K48-linked ubiquitination and subsequent degradation of TOX4	Inhibition of II/R-induced apoptosis in intestinal epithelial cells	(Huang et al. 2024)
NF2	HCC	TRIM65 catalyzes the ubiquitination of NF2 at K44, accelerating its degradation	Promotion of HCC progression	(Bian et al. 2024)
Axin1	HCC	TRIM65 mediates the ubiquitination and degradation of Axin1, enabling β-catenin nuclear translocation and oncogenic activity	Acceleration of HCC progression	(Yang et al. 2017)
JAK1/STAT1	HCC	TRIM65 suppresses the JAK1/STAT1 signaling pathway and promotes M2 polarization of macrophages within tumor microenvironments	Facilitation of HCC progression	(Jiang et al. 2024)
P53	NSCLC/ Cervical cancer	TRIM65 facilitates the ubiquitination and degradation of p53	Enhancement of tumor cell proliferation and chemotherapy drug tolerance	(Li et al. 2016)/ (Wang et al. 2022)
TNRC6A	NSCLC	TRIM65 inhibits miR-138-5p expression by mediating TNRC6A ubiquitination and degradation, subsequently upregulating ATG7 mRNA transcription	Activation of autophagy and suppression of apoptosis in NSCLC cells	(Pan, Chen et al. 2019)
ANXA2	UCB	TRIM65 catalyzes the ubiquitination and degradation of ANXA2, alleviating ANXA2’s inhibitory effect on Rac1	Promotion of migration and invasion in UCB cancer cells	(Wei et al. 2018)
HAR1A	NSCLC	HAR1A serves as a scaffold protein, facilitating the interaction between TRIM65 and ANXA2	Inhibition of AnxA2-mediated cell proliferation, EMT, migration, and invasion	(Ling et al. 2024)
BTG3	RCC	TRIM65 promotes K48-linked ubiquitination of BTG3 at lysine 41, mitigating G2/M phase cell cycle arrest and elevating downstream CyclinD1 protein levels	Promotion of renal cell carcinoma progression	(Zhang et al. 2024)
ARHGAP35	colorectal cancer	TRIM65 orchestrates ARHGAP35 ubiquitination and degradation	Enhancement of Rho GTPase activity and cell migration	(Chen et al. 2019)
LINC01857	Glioma	LINC01857 functions as a miR-1281 sponge, suppressing miR-1281 expression	Upregulation of TRIM65 expression in glioma cells, promoting tumor migration and invasion	(Hu et al. 2019)
LINC00963	colorectal cancer	LINC00963 acts as a miR-1281 sponge, inhibiting miR-1281 expression	Elevation of TRIM65 expression levels in colorectal cancer	(Lv et al. 2021)
PPM1A	Gastric cancer	TRIM65 facilitates ubiquitin-mediated degradation of PPM1A	Enhancement of TBK1 phosphorylation, promoting proliferation and invasion of gastric cancer cells	(Liu et al. 2022)
DUSP6	Endometriosis	TRIM65 mediates ubiquitination and degradation of DUSP6, activating the ERK1/2/C-myc pathway	Promotion of cell proliferation and invasion in EMS cells	(Wu et al. 2019)

Inflammation, a critical characteristic of tumor progression, is intimately linked with proliferation, invasion, angiogenesis, and metastasis(Karki et al. 2016; Karki et al., 2019). The NLRP3 Inflammasome plays a dual role in tumors: it can inhibit colitis-related colorectal cancer(Allen et al. 2010; Zaki et al. 2010), and also promote NSCLC(Sorrentino et al. 2015), gastric cancer(Castaño-Rodríguez et al. 2014), pancreatic cancer(Miskiewicz et al. 2015), and melanoma(Verma et al. 2012). Its activation can trigger pyroptosis(Mariathasan et al. 2006), which is dual in its effects on tumors: it can cause tumor regression or promote the formation of a tumor microenvironment. Tumor cells may inhibit or promote pyroptosis, depending on the specific environment(Loveless et al. 2021). Tang Tian-tian’s team discovered that TRIM65 can mediate the ubiquitination of NLRP3 and inhibit the activation of the NLRP3 inflammasome in THP-1 cells(Tang et al. 2021). This raises a crucial question: Does TRIM65 also mediate the ubiquitination of NLRP3 and inhibit its activation in NSCLC, colorectal cancer, and gastric cancer? Additionally, is the upregulation of TRIM65 in these cancers a compensatory mechanism to inhibit the abnormal activation of NLRP3?

What we currently understand is that the function of TRIM65 is highly dependent on its substrate selectivity, the type of ubiquitination, the cell type, and the microenvironment. This indeed reveals the significant disease/tissue environment-dependent expression (tissue-specific expression pattern) of TRIM65. Exploring the regulatory mechanisms underlying this specific expression, particularly epigenetic mechanisms like promoter methylation and histone modifications, is crucial for understanding the role of TRIM65 in various pathological states. Utilizing methods such as methylation editing tools, including CRISPR-dCas9-TET1/dnmt, or histone modification inhibitors/agonists for functional intervention to confirm the direct causal regulatory relationship between these epigenetic modifications and TRIM65 expression will be a significant focus of future research. Therefore, the expression level of TRIM65 should not be altered without careful consideration. Particularly during the transition from inflammation to cancer, such as from hepatitis to liver cancer, it is essential to meticulously evaluate the alterations in TRIM65’s substrate specificity to prevent intervention strategies from inadvertently accelerating the malignant progression.

Targeted protein degradation (TPD) refers to the utilization of the body’s own degradation mechanisms to modify the activity and concentration of specific proteins within an organism(Cromm et al., 2017). In the field of TPD, Proteolysis Targeting Chimeras (PROTACs) represent one of the most promising advancements(Luh et al. 2020). PROTACs assist in the formation of a ternary complex that includes the target protein, the PROTAC, and an E3 ligase, thereby facilitating the ubiquitination and proteasomal degradation of the target protein (Burslem et al., 2020). Nonetheless, as of now, out of the roughly 600 E3 ligases encoded in the human genome, only about 1% have been utilized in research for TPD. Currently, the primary protein degradation agents in clinical stages are VHL and CRBN(Luh et al. 2020). However, these enzymes have inherent limitations. For example, PROTACs based on VHL may not achieve the desired effect in tumors with low VHL expression. Given that TRIM65 can regulate substrate degradation and possesses tissue specificity, it may be worth considering its application in PROTACs technology. However, the primary challenges persist due to the scarcity of high-affinity ligands for TRIM65 and its dual biological functions. Consequently, future efforts should concentrate on advancing ligand screening technologies, such as DEL + AI, and on creating tissue-specific degradation agents to minimize systemic risks. Should these endeavors prove successful, TRIM65 could emerge as the next-generation differentiated TPD engine, following CRBN/VHL.

In 2017, researchers published a pioneering protein degradation technology named “TRIM Away” in the journal Cell(Clift et al. 2017). This technology is centered on TRIM21, which recognizes and marks the “antibody-virus” complex, thereby mediating its degradation by the proteasome(Mallery et al. 2010). Inspired by the “TRIM Away” technology, Ye Haifeng’s team developed a new targeted protein degradation method using a truncated variant of TRIM21 (ΔTRIM21) as the core component, named △TRIM-TPD. Its most significant advantage is that it offers highly precise controllability and a modular structure, thereby enhancing the scalability and flexibility of the TPD technology application(Ma et al. 2024). Based on this research foundation, it is worth considering the integration of TRIM65 into the “TRIM Away” technology framework. Further research is necessary to clarify the upstream regulatory factors and downstream effectors of TRIM65 and to construct its regulatory network.

However, several challenges and key issues persist within the TPD technology, including enhancing the accuracy of tissue localization, improving the stability of drug delivery, and increasing the specificity of protein degraders for target proteins to minimize side effects. By employing tissue-specific E3 ligases in TPD technology, we can reduce the unnecessary ubiquitination and degradation of non-target proteins, thereby elevating the selectivity and safety of TPD. According to data from the BioGRID database, TRIM65 can interact with over 20 types of proteins. Therefore, we must consider the challenges and risk management issues associated with treating TRIM65, taking into account the precision medicine strategy. The potential of using TRIM65 as a therapeutic target should be evaluated in conjunction with the precision medicine strategy. For instance, we should comprehensively assess which groups are likely to benefit from the treatment and which are not suitable. Furthermore, we should dynamically monitor inflammatory markers and off-target effects throughout the treatment process and develop intervention strategies tailored to different tissues.

Overall, TRIM65, as a multifunctional E3 ubiquitin ligase, plays a significant role in various biological processes. Future research will delve deeper into its biological mechanisms across different diseases, potentially uncovering new protein targets and strategies for treating these conditions.

Abbreviations

TRIMs

Tripartite motif-containing proteins

MDA5

melanoma differentiation-associated protein 5

PAMPs

pathogen-associated molecular patterns

DAMPs

damage associated molecular patterns

BBR

Berberine

NEK7

Nima-associated protein kinase-7

TXNIP

Thioredoxin interacting protein

UroB

Urolithin B

IFN-I

I interferon

HAR1A

Highly Accelerated Region 1 A

BAX

Bcl2 associated X protein

TOX4

thymocyte selection-associated high mobility group box factor 4

HCC

hepatocellular carcinoma

NSCLC

non-small cell lung cancer

UCB

urothelial carcinoma of the bladder

RCC

renal cell carcinoma

OGT

O-GlcNAcylation transferase

NF2

neurofibromatosis type 2

YAP1

Yes-associated protein 1

UMPS

uridine monophosphate synthetase

FASN

Fatty acid synthase

Axin

Axis Inhibition Protein

APC

Colonic adenomatous polyposis protein

GSK-3

Glycogen synthesis kinase 3 protein

EMT

epithelial-mesenchymal transition

ARHGAP35

Rho GTPase activating protein 35

PPM1A

Phosphatase Mg²⁺/Mn²⁺ dependent 1 A

ERK

Extracellular regulated protein kinases

TPD

Targeted protein degradation

PROTACs

Proteolysis-targeted chimeras

Author contributions

NH.D. and JH.C. wrote the main manuscript text and Zhen Tian prepared Figs. 1, 2 and 3. SB.Q. Wrote-review & editing.

Funding

This work was supported by the Special Project for Clinical and Basic Sci &Tech Innovation of Guangdong Medical University (GDMULCJC2024122 to SBQ) and the Clinical Research Special Fund of Guangdong Medical Association (2024HY-B4018 to NHD).

Data availability

No datasets were generated or analysed during the current study.

Declarations

Ethics approval and consent to participate

Not applicable.

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.