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Inactivation of Rhbdf2–MAP3K7 axis is essential for Trim31 function a Representative immunofluorescence images of Rhbdf2 and p-MAP3K7 co-expression in mice liver sections isolated from Flox, THKO- NTG, and THTG mice that were treated with a HFD for 16 weeks (magnification, ×40; n = 10 images per group for each staining) (**P < 0.01 vs. Flox HFD groups or NTG groups). b Representative immunoblotting bands for expression alterations of total amounts or phosphorylated forms of critical indicators associating with the Rhbdf2-MAP3K7 axis and its downstream events cascades including ADAM17, TNFR1/2, MKK7, p-MKK7, c-Jun, and p-c-Jun (Ser63) in the liver of 16-week HFD-fed Flox, THKO- NTG and THTG mice (n = 6 per experiment) (**P < 0.01 vs. Flox HFD groups or NTG groups). The GAPDH was used as a loading control. c Representative immunoblotting bands for expression changes of total amounts or phosphorylated forms of critical indicators associating with the Rhbdf2–MAP3K7 axis and its downstream events cascades including ADAM17, TNFR1/2, MKK7, p-MKK7, c-Jun, and p-c-Jun (Ser63) in primary hepatocytes isolated from the Flox, THKO- NTG, and THTG mice that were incubated with 400 μM PA for 10 h (n = 6 per experiment) (**P < 0.01 vs. Flox palmitate groups or NTG palmitate groups). The GAPDH was used as a loading control. d Representative immunoblotting bands for expression changes of total amounts or phosphorylated forms of critical indicators associating with the insulin signaling, including IRS1, p-IRS1(Ser307), AKT, p-AKT, GSK3β, p-GSK3β, FOXO1, and p-FOXO1, in the adenovirus-packed full-length Rhbdf2 sequences (AdRhbdf2) or shRNA targeting Rhbdf2 (AdshRhbdf2)-transfected THKO or THTG primary hepatocytes that were treated with 400 μM PA for 10 h (n = 4 per experiment) (**P < 0.01 vs. Palmitate/THKO-AdRhbdf2 or Palmitate/THTG-AdRhbdf2). The corresponding AdGFP was used as controls. The GAPDH was used as a loading control. Data are expressed as mean ± SEM. The relevant experiments presented in this part were performed independently at least three times. Significance determined by one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparisons test (a–c) and Student’s two-tailed t test analysis (d).
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Systemic metabolic syndrome significantly increases the risk of morbidity and mortality in patients with non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH). However, no effective therapeutic strategies are available, practically because our understanding of its complicated pathogenesis is poor. Here we identify the t...
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This work was to study the regulatory mechanism of large intergenic non-coding RNA 0196 (LINC0196), miR-584-5p, miR-34a-5p, and tripartite motif 59 (TRIM59) on neuroblastoma. The interaction among the four was analyzed to provide a research basis for the clinical treatment of neuroblastoma at the molecular level. The human neuroblastoma SK-N-SH cel...
Citations
... The role of TRIM31 in NAFLD has recently been investigated. Surprisingly, hepatically expressed TRIM31 prevented HFD-induced hepatic steatosis, IR, inflammatory response, and associated metabolic syndrome in mice (25,26). Similarly, in human subjects, the investigators found that TRIM31 tended to be reduced in the livers of patients with NASH compared to human livers without steatosis, and TRIM31 expression was negatively correlated with the severity of NASH (25). ...
... This inhibits Rhbdf2-TAK1 signaling and downstream JNK/NF-kB phosphorylation, and thus exerting a protective effect against NAFLD, including attenuating IRS1 phosphorylation-mediated insulin resistance and thereby inhibiting glucose production. which in turn inhibits glucose production, attenuates hepatic fibrosis by reducing CTGFmediated deposition of collagen fibers, and attenuates the accumulation of pro-inflammatory factors, cytokines, and chemokines mediated by c-jun phosphorylation, thereby reducing the inflammatory response (26). Another research team discovered a natural compound 'mulberrin' in mulberry branches that can effectively target activation of TRIM31, reduce oxidative stress and liver inflammation, thereby alleviating NASH (27). ...
Nonalcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases and is closely associated with metabolic abnormalities. The causes of NAFLD are exceedingly complicated, and it is known that a variety of signaling pathways, endoplasmic reticulum stress, and mitochondrial dysfunction play a role in the pathogenesis of NAFLD. Recent studies have shown that ubiquitination and deubiquitination are involved in the regulation of the NAFLD pathophysiology. Protein ubiquitination is a dynamic and diverse post-translational alteration that affects various cellular biological processes. Numerous disorders, including NAFLD, exhibit imbalances in ubiquitination and deubiquitination. To highlight the significance of this post-translational modification in the pathogenesis of NAFLD and to aid in the development of new therapeutic approaches for the disease, we will discuss the role of enzymes involved in the processes of ubiquitination and deubiquitination, specifically E3 ubiquitin ligases and deubiquitinating enzymes that are important in the regulation of NAFLD.
... Trim31 expression is altered in various pathological states. Trim31 levels have been reported to decrease in several diseases, such as colitis [37], gastritis [40], nonalcoholic fatty liver disease [41], and hypertensive nephropathy [11]. Nonetheless, it remains unclear whether Trim31 expression changes in HF. ...
Tripartite motif-containing protein 31 (Trim31) is known to be involved in various pathological conditions, including heart diseases. Nonetheless, its specific involvement in heart failure (HF) has yet to be determined. In this study, we examined the function and mechanism of Trim31 in HF by using mice with cardiac-specific knockout (cKO) of Trim31. The HF mouse model was induced via the subcutaneous injection of isoproterenol (ISO). We observed a decrease in Trim31 expression in the heart tissues of mice with HF. Compared with wild-type (WT) mice, Trim31 cKO mice presented more severe characteristics of HF, including worsened cardiac dysfunction, hypertrophy, and fibrosis. However, these symptoms in Trim31 cKO mice were significantly reversed when they received an intramyocardial injection of recombinant adeno-associated virus (AAV) expressing Trim31. Excessive activation of the NLRP3 inflammasome, manifested by increased levels of NLRP3, ASC, cleaved Caspase-1, cleaved GSDMD, IL-1β, and IL-18, was observed in Trim31 cKO mice with HF. However, Trim31 overexpression effectively reversed the NLRP3 inflammasome activation in Trim31 cKO mice with HF. Selective inhibition of the NLRP3 inflammasome with the NLRP3 inhibitor MCC950 effectively reversed the worsened cardiac dysfunction, hypertrophy, and fibrosis observed in Trim31 cKO mice with HF. Overall, the findings from this study reveal a crucial role of Trim31 in HF. Trim31 deficiency may contribute to the progression of HF by promoting cardiac hypertrophy, fibrosis, and inflammation by facilitating the activation of the NLRP3 inflammasome. Therefore, Trim31 may hold significant potential as a therapeutic target for the treatment of HF.
... Likewise, a more recent study has shown that TRIM16 attenuates hepatocyte steatosis and inflammation in a mouse NASH model by directly interacting with the phosphorylated form of TAK1 and promoting its degradation, leading to suppressed NASH development (16). Two more recent studies have demonstrated that TRIM31 alleviates NAFLD and NASH pathologies by targeted degradation of rhomboid 5 homolog 2 (Rhbdf2) (35) and TAK1 (36) in the liver. TRIM31 is also responsible for the antifibrotic effects of mulberrin (a bioactive phytochemical from the traditional Chinese medicine Ramulus Mori) in CCl 4 -induced liver fibrosis (37). ...
Nonalcoholic fatty liver disease (NAFLD) encompasses a disease continuum from simple steatosis to nonalcoholic
steatohepatitis (NASH). However, there are currently no approved pharmacotherapies for NAFLD, although several drugs
are in advanced stages of clinical development. Because of the complex pathophysiology and heterogeneity of NAFLD,
the identification of potential therapeutic targets is clinically important. Here, we demonstrated that tripartite motif
56 (TRIM56) protein abundance was markedly downregulated in the livers of individuals with NAFLD and of mice fed
a high-fat diet. Hepatocyte-specific ablation of TRIM56 exacerbated the progression of NAFLD, while hepatic TRIM56
overexpression suppressed it. Integrative analyses of interactome and transcriptome profiling revealed a pivotal role of
TRIM56 in lipid metabolism and identified the lipogenesis factor fatty acid synthase (FASN) as a direct binding partner
of TRIM56. TRIM56 directly interacted with FASN and triggered its K48-linked ubiquitination–dependent degradation.
Finally, using artificial intelligence–based virtual screening, we discovered an orally bioavailable small-molecule inhibitor
of FASN (named FASstatin) that potentiates TRIM56-mediated FASN ubiquitination. Therapeutic administration of
FASstatin improved NAFLD and NASH pathologies in mice with an optimal safety, tolerability, and pharmacokinetics
profile. Our findings provide proof of concept that targeting the TRIM56/FASN axis in hepatocytes may offer potential
therapeutic avenues to treat NAFLD.
... The pathogenesis of metabolic dysfunction-associated steatotic liver disease (MASLD) and metabolic dysfunction-associated steatohepatitis (MASH) is complex and involves increased inflammation, insulin resistance, and impaired glucose tolerance 30,33,34 . Before evaluating the protective effects of T6BP, we first examined the role of PYK2 in the development of steatohepatitis. ...
... TRIM31 inhibits Rhbdf2-MAP3K7 and its downstream signaling by directly binding alongside polyubiquitination-degrading Rhbdf2 to prevent hepatic steatosis [72]. TRIM67 expression was reported to be induced by PGC-1α, HFD, and obesity, which activates liver inflammation, the accumulation of liver lipids, and the progression of MAFLD. ...
The worldwide impact of liver diseases is increasing steadily, with a consistent upswing evidenced in incidence and mortality rates. Chronic liver diseases (CLDs) refer to the liver function’s progressive deterioration exceeding six months, which includes abnormal clotting factors, detoxification failure, and hepatic cholestasis. The most common etiologies of CLDs are mainly composed of chronic viral hepatitis, MAFLD/MASH, alcoholic liver disease, and genetic factors, which induce inflammation and harm to the liver, ultimately resulting in cirrhosis, the irreversible final stage of CLDs. The latest research has shown that tripartite motif family proteins (TRIMs) function as E3 ligases, which participate in the progression of CLDs by regulating gene and protein expression levels through post-translational modification. In this review, our objective is to clarify the molecular mechanisms and potential therapeutic targets of TRIMs in CLDs and provide insights for therapy guidelines and future research.
... The interplay among inflammation, insulin resistance, and hepatic lipid metabolic disorders is central to NAFLD progression [7]. However, the mechanism of NAFLD is yet not fully elucidated and rare drugs have been approved to cure NAFLD to date [8]. The pathologic progression of simple steatosis to NASH can be suppressed more efficiently with an understanding of the underlying pathogenic mechanism. ...
... Transforming growth factor-β-activated kinase-1(TAK1), which belongs to the MAPK kinase kinase (MAP3K) family, is an essential activator of MAPK and NF-κB signaling pathways [23][24][25], and then promote the signaling cascades of pro-inflammatory cytokines and inhabit insulin signaling pathway, and these pathways contribute to liver inflammation, insulin resistance, and hepatic steatosis, all of which are hallmarks of NAFLD. Previous studies indicated TAK1 is a key element in regulating NAFLD in liver steatosis, insulin resistance, and inflammation [5,8,26], and its phosphorylation levels is influenced by the level of ubiquitination [27,28]. USP4 and USP18, as members of the USP family, have been shown to interact directly with TAK1 and subsequently de-ubiquitinate TAK1 and inhabit its activation to alleviate NAFLD [10,12]. ...
Background
The molecular mechanisms underlying nonalcoholic fatty liver disease (NAFLD) remain to be fully elucidated. Ubiquitin specific protease 13 (USP13) is a critical participant in inflammation-related signaling pathways, which are linked to NAFLD. Herein, the roles of USP13 in NAFLD and the underlying mechanisms were investigated.
Methods
L02 cells and mouse primary hepatocytes were subjected to free fatty acid (FFA) to establish an in vitro model reflective of NAFLD. To prepare in vivo model of NAFLD, mice fed a high-fat diet (HFD) for 16 weeks and leptin-deficient (ob/ob) mice were used. USP13 overexpression and knockout (KO) strategies were employed to study the function of USP13 in NAFLD in mice.
Results
The expression of USP13 was markedly decreased in both in vitro and in vivo models of NAFLD. USP13 overexpression evidently inhibited lipid accumulation and inflammation in FFA-treated L02 cells in vitro. Consistently, the in vivo experiments showed that USP13 overexpression ameliorated hepatic steatosis and metabolic disorders in HFD-fed mice, while its deficiency led to contrary outcomes. Additionally, inflammation was similarly attenuated by USP13 overexpression and aggravated by its deficiency in HFD-fed mice. Notably, overexpressing of USP13 also markedly alleviated hepatic steatosis and inflammation in ob/ob mice. Mechanistically, USP13 bound to transforming growth factor β-activated kinase 1 (TAK1) and inhibited K63 ubiquitination and phosphorylation of TAK1, thereby dampening downstream inflammatory pathways and promoting insulin signaling pathways. Inhibition of TAK1 activation reversed the exacerbation of NAFLD caused by USP13 deficiency in mice.
Conclusions
Our findings indicate the protective role of USP13 in NAFLD progression through its interaction with TAK1 and inhibition the ubiquitination and phosphorylation of TAK1. Targeting the USP13-TAK1 axis emerges as a promising therapeutic strategy for NAFLD treatment.
... The ready-made system for knockout cell lines construction in the indicated targeted gene-deficiency was founded as described previously [44,45]. In brief, the stomach cancer NCI-N87 cell line with ZDHHC1-ZDHHC24 knockout was prepared by clustered regulatory interspaced short palindromic repeats-CRISPR-associated protein 9 (CRISPR-Cas9) system. ...
... The in vivo or in vitro interaction ubiquitination assay were respectively performed in accordance with the previously described methods [44,45]. Briefly, the in vivo ubiquitination assay was performed as follows: cells were lysed in SDS lysis buffer (20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1 mM EDTA and 1 % SDS) containing Protease Inhibitor Cocktail Tablets (Roche), and denatured by heating for 5 min. ...
The significant regulatory role of palmitoylation modification in cancer-related targets has been demonstrated previously. However, the biological functions of Nrf2 in stomach cancer and whether the presence of Nrf2 palmitoylation affects gastric cancer (GC) progression and its treatment have not been reported. Several public datasets were used to look into the possible link between the amount of palmitoylated Nrf2 and the progression and its outcome of GC in patients. The palmitoylated Nrf2 levels in tumoral and peritumoral tissues from GC patients were also evaluated. Both loss-of-function and gain-of-function via transgenic experiments were performed to study the effects of palmitoylated Nrf2 on carcinogenesis and the pharmacological function of 2-bromopalmitate (2-BP) on the suppression of GC progression in vitro and in vitro. We discovered that Nrf2 was palmitoylated in the cytoplasmic domain, and this lipid posttranslational modification causes Nrf2 stabilization by inhibiting ubiquitination, delaying Nrf2 destruction via the proteasome and boosting nuclear translocation. Importantly, we also identify palmitoyltransferase zinc finger DHHC-type palmitoyltransferase 2 (DHHC2) as the primary acetyltransferase required for the palmitoylated Nrf2 and indicate that the suppression of Nrf2 palmitoylation via 2-bromopalmitate (2-BP), or the knockdown of DHHC2, promotes anti-cancer immunity in vitro and in mice model-bearing xenografts. Of note, based on the antineoplastic mechanism of 2-BP, a novel anti-tumor drug delivery system ground 2-BP and oxaliplatin (OXA) dual-loading gold nanorods (GNRs) with tumor cell membrane coating biomimetic nanoparticles (CM@GNRs-BO) was established. In situ photothermal therapy is done using near-infrared (NIR) laser irradiation to help release high-temperature-triggered drugs from the CM@GNRs-BO reservoir when needed. This is done to achieve photothermal/chemical synergistic therapy. Our findings show the influence and linkage of palmitoylated Nrf2 with tumoral and peritumoral tissues in GC patients, the underlying mechanism of palmitoylated Nrf2 in GC progression, and novel possible techniques for addressing Nrf2-associated immune evasion in cancer growth. Furthermore, the bionic nanomedicine developed by us has the characteristics of dual drugs delivery, homologous tumor targeting, and photothermal and chemical synergistic therapy, and is expected to become a potential platform for cancer treatment.
... MAF = 5%). TRIM31 has been identified as a "Janus-faced" regulator of innate immune responses, facilitating signal transduction through target substrate degradation or ubiquitin modification (Xu et al., 2022). Furthermore, in some studies, functional impairment (knockout) of TRIM31 has been shown to significantly increase body weight, fasting blood glucose levels, and fasting insulin levels induced by a high-fat diet (HFD), suggesting that reduced expression of TRIM31 can contribute to weight gain (Xu et al., 2022). ...
... TRIM31 has been identified as a "Janus-faced" regulator of innate immune responses, facilitating signal transduction through target substrate degradation or ubiquitin modification (Xu et al., 2022). Furthermore, in some studies, functional impairment (knockout) of TRIM31 has been shown to significantly increase body weight, fasting blood glucose levels, and fasting insulin levels induced by a high-fat diet (HFD), suggesting that reduced expression of TRIM31 can contribute to weight gain (Xu et al., 2022). Due to the lack of literature supporting the involvement of TDRD10 in weight regulation, the SNP (chr23:29119138) within . ...
... Although the p-value did not reach statistical significance, visualization of the raw body weight differences based on the genotypes at chr23: 29119138 showed a trend where individuals carrying allele A exhibited a lower weight distribution (Supplementary Figure S3). These research findings, while not statistically significant, support the directionality of weight regulation associated with TRIM31 expression previously reported in the literature (Luo et al., 2022;Xu et al., 2022). This suggests that a reduction in TRIM31 expression may contribute to an increase in body weight, and the variant at chr23:29119138 could potentially regulate the expression of TRIM31. ...
While genetic markers related to meat production traits have been identified in many other cattle breeds, research on weight in Hanwoo cattle (Korean native cattle) is still insufficient. In this study, we performed expression quantitative trait loci (eQTL) analysis and differential gene expression analysis to detect candidate genes influencing the weight characteristics of 32 castrated Hanwoo cattle across 22 tissues and, we identified variants that affect gene expression levels. In total, we identified a total of 3,298 differentially expressed genes, among which we discovered key genes such as UBD, RGS2, FASN, and SCD that have functions related to adipogenesis, body weight, obesity, and lipid metabolism. Gene-set enrichment analysis revealed that candidate genes in adipose tissue are involved in metabolic pathways linked to obesity-related traits, adipose metabolism, and lipid metabolism. Additionally, we found that decreased expression of TRIM31 contributes to weight gain which can be explained by the associated candidate cis-eQTL genotypes for TRIM31 and their effect on differential gene expression between the lower and higher weight groups. Our findings revealed candidate genes associated with the weight of Hanwoo cattle and perhaps can provide comprehensive insights into the association of weight with various tissues beyond adipose tissue and muscle, indicating the potential for expanding the focus of livestock trait research.
... However, these degradation pathways have only been studied in mitotic cells of cancer tissues and cell lines, and their role in livers exposed to metabolic stress is unclear. Furthermore, recent in vivo studies of members of TRIM family of E3-ubiquitin ligases have indicated their major role in the control of metabolic homeostasis and development of chronic liver disease (38)(39)(40). ...
... These in vitro-based findings suggest a tumor-suppressor role of TRIM21 by inhibiting metabolic pathways that fuel nucleic acid synthesis, cell growth, and proliferation. Investigations of other members of the TRIM family have suggested that they contribute to the control of inflammation and metabolic homeostasis of chronic liver diseases (38)(39)(40)60). ...
Nonalcoholic steatohepatitis (NASH) is a leading cause for chronic liver diseases. Current therapeutic options are limited due to an incomplete mechanistic understanding of how steatosis transitions to NASH. Here we show that the TRIM21 E3 ubiquitin ligase is induced by the synergistic actions of proinflammatory TNF-α and fatty acids in livers of humans and mice with NASH. TRIM21 ubiquitinates and degrades ChREBP, SREBP1, ACC1, and FASN, key regulators of de novo lipogenesis, and A1CF, an alternative splicing regulator of the high-activity ketohexokinase-C (KHK-C) isoform and rate-limiting enzyme of fructose metabolism. TRIM21-mediated degradation of these lipogenic activators improved steatosis and hyperglycemia as well as fructose and glucose tolerance. Our study identifies TRIM21 as a negative regulator of liver steatosis in NASH and provides mechanistic insights into an immunometabolic crosstalk that limits fatty acid synthesis and fructose metabolism during metabolic stress. Thus, enhancing this natural counteracting force of steatosis through inhibition of key lipogenic activators via TRIM21-mediated ubiquitination may provide a therapeutic opportunity to treat NASH.
... The top 4 indicators, i.e., TRIM26, TRIM31, RNF5, and TRIM8, were distinguished from the different indicated groups. Also, our previous report indicated that TRIM31 confers protection against NASH by suppressing iRhom2/MAP3K7 axis-related inflammation 23 . TRIM26 displays inhibitory effects on NASH progression by regulating CEBPD-HIF1A-related inflammation and NOS2 signalling. ...
... A total of 16 non-steatosis samples, 17 simple steatosis samples, and 16 NASH phenotypic liver samples were procured and included in the present investigation. The human liver samples used in the present manuscript are the same as those used in our previous paper 23 . It is important to mention that liver samples exhibiting steatosis from individuals with the following conditions were excluded from the study: (i) excessive alcohol consumption (defined as consuming more than 70 g of alcohol per week for females or more than 140 g of alcohol per week for males), (ii) viral infection or drug abuse . ...
... A murine model exhibiting NASH phenotype was established by subjecting the mice to a HFHC diet consisting of 42% saturated fat, 14% protein, 44% carbs, and 0.2% cholesterol by weight for a duration of 16 weeks 17,23 . The mice used in the experiment were administered a conventional normal chow diet (NCD) from Research Diets, USA (Cat: D12450J) for a continuous period of 16 weeks. ...
Currently potential preclinical drugs for the treatment of nonalcoholic steatohepatitis (NASH) and NASH-related pathopoiesis have failed to achieve expected therapeutic efficacy due to the complexity of the pathogenic mechanisms. Here we show Tripartite motif containing 26 (TRIM26) as a critical endogenous suppressor of CCAAT/enhancer binding protein delta (C/EBPδ), and we also confirm that TRIM26 is an C/EBPδ-interacting partner protein that catalyses the ubiquitination degradation of C/EBPδ in hepatocytes. Hepatocyte-specific loss of Trim26 disrupts liver metabolic homeostasis, followed by glucose metabolic disorder, lipid accumulation, increased hepatic inflammation, and fibrosis, and dramatically facilitates NASH-related phenotype progression. Inversely, transgenic Trim26 overexpression attenuates the NASH-associated phenotype in a rodent or rabbit model. We provide mechanistic evidence that, in response to metabolic insults, TRIM26 directly interacts with C/EBPδ and promotes its ubiquitin proteasome degradation. Taken together, our present findings identify TRIM26 as a key suppressor over the course of NASH development.
