Zhang J, Liu C, Tang Y, Gao K, Xu W, Zhang J, Zhao L, Wang G.
Erbin destabilization by O-GlcNAcylation promotes 5-FU resistance via homologous recombination activation in colorectal cancer.
Biochemical pharmacology2026
243(Pt 1)
41072816
Abstract: The emergence of 5-fluorouracil (5FU) resistance critically compromises chemotherapy efficacy in colorectal cancer (CRC). While O-GlcNAcylation-a dynamic post-translational modification linked to tumor progression-has been widely investigated, its functional role in 5FU chemoresistance remains poorly defined. Through mass spectrometry and tissue microarray analysis, we identified aberrantly elevated levels of O-GlcNAcylation and its catalytic enzyme O-GlcNAc transferase (OGT) in CRC tissues. Genetic silencing of OGT significantly reduced global O-GlcNAcylation, suppressing CRC cell proliferation, migration, and invasion while enhancing 5FU sensitivity both in vitro and in vivo. Notably, 5FU-resistant CRC models (HCT8/HCT116 5FUR) exhibited upregulated OGT and O-GlcNAcylation versus parental cells. Strikingly, OGT knockdown in resistant cells reversed chemoresistance, inhibited proliferation, and induced apoptosis. O-GlcNAcylated proteomic profiling identified Erbin, a tumor suppressor, as a hyper-modified protein with diminished expression in resistant cells, validated by co-immunoprecipitation. Remarkably, Erbin overexpression restored 5FU sensitivity in resistant cells, whereas OGT depletion stabilized Erbin by blocking ubiquitination. Site-directed mutagenesis pinpointed Thr1070 as Erbin's critical O-GlcNAcylation site; its mutation reduced ubiquitination, linking O-GlcNAcylation to Erbin degradation. Mechanistically, RNA sequencing (RNA-seq) revealed that Erbin suppresses homologous recombination (HR), causing unresolved DNA damage accumulation and heightened 5FU sensitivity, confirmed in vitro and in vivo experiments. Critically, elevated OGT in 5FU-resistant cells drives hyper-O-GlcNAcylation of Erbin at Thr1070, which facilitates its ubiquitin-dependent degradation, alleviating HR suppression to sustain chemoresistance. This defines the OGT-Erbin-HR axis as a central driver of 5FU resistance, proposing therapeutic targeting of this pathway to overcome CRC chemoresistance.
Moniaux N, Geoffre N, Deshayes A, Dos Santos A, Job S, Lacoste C, Nguyen TS, Darnaud M, Friedel-Arboleas M, Guettier C, Purhonen J, Kallijärvi J, Amouyal G, Amouyal P, Bréchot C, Vivès RR, Buendia MA, Issad T, Faivre J.
Tumor suppressive role of the antimicrobial lectin REG3A targeting the O -GlcNAc glycosylation pathway.
Hepatology (Baltimore, Md.)2025
81(5)
38975812
Abstract: Antimicrobial proteins of the regenerating family member 3 alpha (REG3A) family provide a first line of protection against infections and transformed cells. Their expression is inducible by inflammation, which makes their role in cancer biology less clear since an immune-inflammatory context may preexist or coexist with cancer, as occurs in HCC. The aim of this study is to clarify the role of REG3A in liver carcinogenesis and to determine whether its carbohydrate-binding functions are involved.
Liang Y, Chen L, Huang Z, Li Y, Weng H, Guo L.
O-GlcNAcylation of progranulin promotes hepatocellular carcinoma proliferation.
Biochemical and biophysical research communications2025
742
39657353
Abstract: Progranulin (PGRN) is overexpressed and implicated in hepatocellular carcinoma (HCC) development; however, its post-translational modifications and regulatory mechanisms in HCC remain largely unexplored. Here, the expression levels of PGRN, OGT, and O-GlcNAcylation were found to be elevated in both HCC samples and cell lines. LC-MS/MS analysis and immunoprecipitation revealed that PGRN underwent O-linked N-acetylglucosamine (O-GlcNAc) modification at threonine 272 (Thr272). Co-immunoprecipitation and confocal microscopy confirmed the interaction and colocalization of O-GlcNAc transferase (OGT) with PGRN. Reducing O-GlcNAcylation increased the ubiquitination of PGRN, while increasing O-GlcNAcylation inhibited ubiquitination and elevated PGRN stability, as measured by cycloheximide (CHX) chase experiments. This regulation of PGRN stability was directly linked to its expression levels. Moreover, mutation at the primary O-GlcNAc site Thr272 inhibited the activity of the PI3K/AKT/mTOR signaling pathway and suppressed HCC cell proliferation. Together, our findings indicate that O-GlcNAcylation at Thr272 is essential for PGRN-driven HCC cell proliferation.
Li J, Shao G, Peng B, Xu X, Dong MQ, Li J.
The Role of Polo-Like Kinase 1 (PLK1) O-GlcNAcylation in Mitosis.
Methods in molecular biology (Clifton, N.J.)2025
2874
39614052
Abstract: Polo-like kinase 1 (PLK1) is a crucial mitotic kinase that is implicated in various aspects of cell cycle. Many post-translational modifications have been identified on PLK1 to regulate its activation, stability, and localization. PLK1 has been shown previously to colocalize with the O-linked β-N-acetylglucosamine (O-GlcNAc) transferase (OGT), and OGT regulates PLK1 stability. In our recent work, we show that PLK1 is O-GlcNAcylated by click chemistry. Using stepped collisional energy/higher energy collision dissociation mass spectrometry, we mapped the PLK1 O-GlcNAc site to be T291. We further utilized fluorescent activated cell sorting and time-lapse microscopy to assess the mitotic defects of PLK1 O-GlcNAc mutants. In vivo studies in mouse xenograft demonstrated that it promoted uterine cancer tumorigenesis. In this chapter, we delineate the methodologies we used in studying PLK1 O-GlcNAcylation, including click chemistry, stepped collisional energy/higher energy collision dissociation mass spectrometry, fluorescent activated cell sorting, time-lapse microscopy, and mouse xenograft assays.
Jeon BC, Kim YJ, Park AK, Song MR, Na KM, Lee J, An D, Park Y, Hwang H, Kim TD, Lim J, Park SK.
Dynamic O-GlcNAcylation governs long-range chromatin interactions in V(D)J recombination during early B-cell development.
Cellular & molecular immunology2025
22(1)
39627609
Abstract: V(D)J recombination secures the production of functional immunoglobulin (Ig) genes and antibody diversity during the early stages of B-cell development through long-distance interactions mediated by cis-regulatory elements and trans-acting factors. O-GlcNAcylation is a dynamic and reversible posttranslational modification of nuclear and cytoplasmic proteins that regulates various protein functions, including DNA-binding affinity and protein-protein interactions. However, the effects of O-GlcNAcylation on proteins involved in V(D)J recombination remain largely unknown. To elucidate this relationship, we downregulated O-GlcNAcylation in a mouse model by administering an O-GlcNAc inhibitor or restricting the consumption of a regular diet. Interestingly, the inhibition of O-GlcNAcylation in mice severely impaired Ig heavy-chain (IgH) gene rearrangement. We identified several factors crucial for V(D)J recombination, including YY1, CTCF, SMC1, and SMC3, as direct targets of O-GlcNAc modification. Importantly, O-GlcNAcylation regulates the physical interaction between SMC1 and SMC3 and the DNA-binding patterns of YY1 at the IgH gene locus. Moreover, O-GlcNAc inhibition downregulated DDX5 protein expression, affecting the functional association of CTCF with its DNA-binding sites at the IgH locus. Our results showed that locus contraction and long-range interactions throughout the IgH locus are disrupted in a manner dependent on the cellular O-GlcNAc level. In this study, we established that V(D)J recombination relies on the O-GlcNAc status of stage-specific proteins during early B-cell development and identified O-GlcNAc-dependent mechanisms as new regulatory components for the development of a diverse antibody repertoire.
Du Y, Gao X, Chen J, Chen X, Liu H, He W, Liu L, Jiang Y, He B, Deng Z, Liang C, Guo F.
OGT mediated HDAC5 O-GlcNAcylation promotes osteogenesis by regulating the homeostasis of epigenetic modifications and proteolysis.
Journal of orthopaedic translation2025
50
39659899
Abstract: O-GlcNAc transferase (OGT) is responsible for attaching O-linked N-acetylglucosamine (O-GlcNAc) to proteins, regulating diverse cellular processes ranging from transcription and translation to signaling and metabolism. This study focuses on the role and mechanisms of OGT in osteogenesis.
Qiu Y, Yu W, Zhang X, Zhang M, Ni Y, Lai S, Wu Q.
Upregulation of OGT-mediated EZH2 O-GlcNAcylation Promotes Human Umbilical Vein Endothelial Cell Proliferation, Invasion, Migration, and Tube Formation in Gestational Diabetes Mellitus.
Cell biochemistry and biophysics202539751742
Abstract: O-linked N-acetylglucosamine transferase (OGT)-catalyzed O-linked N-acetylglucosamine glycosylation (O-GlcNAcylation) is closely associated with diabetes progression. This study aims to investigate the mechanism of OGT in regulating endothelial dysfunction in gestational diabetes mellitus (GDM). Expressions of OGT, O-linked N-acetylglucosamine (O-GlcNAc), enhancer of zeste homolog 2 (EZH2), and HEK27me3 in human umbilical vein endothelial cells (HUVECs) and GDM-derived HUVECs (GDM-HUVECs) were assessed by western blot. RT-qPCR and western blot assays were used to test the OGT overexpression and EZH2 silencing levels. CCK-8, EdU, wound healing, and transwell invasion assays were used to analyze the cell proliferative, migratory, and invasive abilities. Tube formation assay was performed to evaluate angiogenesis ability of cells. Western blot assay was performed to estimate vascular endothelial growth factor (VEGF) and p-VEGFR2 levels in cells. The binding of O-GlcNAc and EZH2 after OGT overexpression was measured by Co-IP assay. The results showed that OGT, O-GlcNAc, EZH2, and HEK27me3 expressions were declined in GDM-HUVECs. OGT overexpression induced the proliferation, migration, and invasion of GDM-HUVECs, and also elevated angiogenesis and the expressions of VEGF and p-VEGFR2 in cells. O-GlcNAc, EZH2, and HEK27me3 expressions were upregulated after OGT overexpression. OGT upregulation induced the binding between O-GlcNAc and EZH2. EZH2 silencing attenuated the promotion of OGT overexpression on the proliferative, invasive, migratory, and angiogenic capacities of GDM-HUVECs. To be concluded, OGT overexpression stabilized EZH2 expression by promoting O-GlcNAcylation modification of EZH2, and further enhanced proliferation, migration, and invasion as well as angiogenesis of GDM-HUVECs. While these effects were decayed after EZH2 absenting. Overall, the modulation of OGT on endothelial dysfunction in GDM provides a novel perspective for the clinical treatment of GDM.
Sitosari H, Fukuhara Y, Weng Y, Zheng Y, He Y, Zheng X, Ikegame M, Okamura H.
High Glucose Inhibits O-GlcNAc Transferase Translocation in Early Osteoblast Differentiation by Altering Protein Phosphatase 2A Activity.
Journal of cellular physiology2025
240(1)
39800889
Abstract: Our previous study revealed a link between O-GlcNAc transferase (OGT) localization and protein phosphatase 2A (PP2A) activity in osteoblast. Given the association of PP2A downregulation with osteoblast differentiation, we hypothesized that OGT localization changes during this process. We examined OGT localization in MC3T3-E1 cells undergoing differentiation under normal and high glucose conditions. Changes in PP2A activity were followed by alterations in OGT localization. Organ culture of calvaria revealed similar OGT localization changes in bone-surrounding osteoblasts near the suture area. Furthermore, the levels of O-GlcNAc modification in various proteins including Runt-related transcription factor 2, Osterix, and ATP synthase subunit alpha (ATP5A) were shifted in parallel with OGT translocation. These findings suggest a regulatory role of OGT, under the influence of PP2A, during osteoblast differentiation.
Zhou X, Yang Y, Qiu X, Deng H, Cao H, Liao T, Chen X, Huang C, Lin D, Ni G.
Antioxidant taurine inhibits chondrocyte ferroptosis through upregulation of OGT/Gpx4 signaling in osteoarthritis induced by anterior cruciate ligament transection.
Journal of advanced research202539778769
Abstract: The aim of this study was to investigate the potential molecular mechanisms by which taurine protects against cartilage degeneration.
Yan Z, Li Y, Wang M, Xu K, Liu Y, Wang L, Luo H, Chen Z, Liu X.
O-GlcNAcylation of DJ-1 suppresses ferroptosis in renal cell carcinoma by affecting the transsulfuration pathway.
International immunopharmacology2025
148
39842141
Abstract: Renal cell carcinoma (RCC) is one of the most common urological malignancies worldwide, and advanced patients often face challenges with chemotherapy resistance and poor prognosis. Ferroptosis, a novel form of cell death, offers potential therapeutic prospects. In this study, we found that DJ-1 was elevated in kidney renal clear cell carcinoma (KIRC), and this abnormal expression pattern was closely associated with clinical pathological characteristics and worse prognosis. Our experiments both in vivo and in vitro revealed that DJ-1 enhanced the malignant characteristics of KIRC, leading to increased tumor growth. Additionally, DJ-1 inhibited ferroptosis through promoting homocysteine (Hcy) synthesis in the transsulfuration pathway in KIRC cells. Mechanistic studies revealed that O-GlcNAc transferase (OGT) mediated O-GlcNAcylation of DJ-1 was crucial for maintaining its homodimeric structure. Importantly, O-GlcNAcylation-deficient mutation of DJ-1 at T19 residue enhanced the interaction between S-adenosyl homocysteine hydrolase (SAHH) and the negative regulatory factor S-adenosyl homocysteine hydrolase-like-1 (AHCYL1), thereby inhibited the activities of SAHH and transsulfuration pathway. In summary, the oncogenic role of DJ-1 in KIRC was closely related to the reduction of ferroptosis, and the O-GlcNAcylation of DJ-1 exerted an antioxidant effect by activating the transsulfuration pathway. Therefore, DJ-1, specifically O-GlcNAcylation of DJ-1 could represent an important target for ferroptosis-based anti-tumor therapy.
Gou J, Bi J, Wang K, Lei L, Feng Y, Tan Z, Gao J, Song Y, Kang E, Guan F, Li X.
O-GlcNAcylated FTO promotes m6A modification of SOX4 to enhance MDS/AML cell proliferation.
Cell communication and signaling : CCS2025
23(1)
39849461
Abstract: Fat mass and obesity-associated protein (FTO) was the first m6A demethylase identified, which is responsible for eliminating m6A modifications in target RNAs. While it is well-established that numerous cytosolic and nuclear proteins undergo O-GlcNAcylation, the possibility of FTO being O-GlcNAcylated and its functional implications remain unclear. This study found that a negative correlation between FTO expression and O-GlcNAcylation in patients with myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). The decreased O-GlcNAcylation on FTO can result in diminished m6A modification of SRY-related high mobility group box 4 (SOX4). This led to the promotion of cell apoptosis and inhibition of cell proliferation in MDS/AML. The O-GlcNAcylation of FTO stabilized SOX4 transcripts in an m6A-dependent manner, resulting in increased AKT and MAPK phosphorylation and decreased cell apoptosis. Inhibiting FTO O-GlcNAcylation significantly slowed AML progression in vitro, a finding supported by clinical data in MDS/AML patients. In conclusion, our study highlights the crucial role of FTO O-GlcNAcylation in RNA m6A methylation and the progression of MDS/AML, thereby providing a potential therapeutic avenue for these formidable diseases.
Rodrigues BDC, Dos Santos Lucena MC, Costa ACR, Oliveira IA, Thaumaturgo M, Paes-Colli Y, Beckman D, Ferreira ST, de Mello FG, De Melo Reis RA, Todeschini AR, Dias WB.
O-GlcNAcylation regulates tyrosine hydroxylase serine 40 phosphorylation and L-DOPA levels.
American journal of physiology. Cell physiology202539870381
Abstract: O-GlcNAcylation is a post-translational modification characterized by the covalent attachment of a single moiety of GlcNAc on serine/threonine residues in proteins. Tyrosine hydroxylase (TH), the rate-limiting step enzyme in the catecholamine synthesis pathway and responsible for production of the dopamine precursor, L-DOPA, has its activity regulated by phosphorylation. Here, we show an inverse feedback mechanism between O-GlcNAcylation and phosphorylation of TH at serine 40 (TH pSer40). First, we showed that, during PC12 cells neuritogenesis, TH O-GlcNAcylation decreases concurrently with the increase of pSer40. In addition, an increase in O-GlcNAcylation induces a decrease in TH pSer40 only in undifferentiated PC12 cells, while the decrease in O-GlcNAcylation leads to an increase in TH pSer40 levels in both undifferentiated and differentiated PC12 cells. We further show that this feedback culminates on the regulation of L-DOPA intracellular levels. Interestingly, it is noteworthy that decreasing O-GlcNAcylation is much more effective on TH pSer40 regulation than increasing its levels. Finally, ex vivo analysis confirmed the upregulation of TH pSer40 when O-GlcNAcylation levels are reduced in dopaminergic neurons from C57Bl/6 mice. Taken together, these findings demonstrate a dynamic control of L-DOPA production by a molecular crosstalk between O-GlcNAcylation and phosphorylation at Ser40 in tyrosine hydroxylase.
Zuo Y, Wang Q, Tian W, Zheng Z, He W, Zhang R, Zhao Q, Miao Y, Yuan Y, Wang J, Zheng H.
β-hydroxybutyrylation and O-GlcNAc modifications of STAT1 modulate antiviral defense in aging.
Cellular & molecular immunology202539979583
Abstract: Aging changes the protein activity status to affect the body's functions. However, how aging regulates protein posttranslational modifications (PTMs) to modulate the antiviral defense ability of the body remains unclear. Here, we found that aging promotes STAT1 β-hydroxybutyrylation (Kbhb) at Lys592, which inhibits the interaction between STAT1 and type-I interferon (IFN-I) receptor 2 (IFNAR2), thereby attenuating IFN-I-mediated antiviral defense activity. Additionally, we discovered that a small molecule from a plant source, hydroxy camptothecine, can effectively reduce the level of STAT1 Kbhb, thus increasing antiviral defense ability in vivo. Further studies revealed that STAT1 O-GlcNAc modifications at Thr699 block CBP-induced STAT1 Kbhb. Importantly, fructose can improve IFN-I antiviral defense activity by orchestrating STAT1 O-GlcNAc and Kbhb modifications. This study reveals the significance of the switch between STAT1 Kbhb and O-GlcNAc modifications in regulating IFN-I antiviral immunity during aging and provides potential strategies to improve the body's antiviral defense ability in elderly individuals.
Jaiswal R, Liu Y, Petriello M, Zhang X, Yi Z, Fehl C.
A reference dataset of O-GlcNAc proteins in quadriceps skeletal muscle from mice.
Glycobiology2025
35(3)
39927985
Abstract: A key nutrient sensing process in all animal tissues is the dynamic attachment of O-linked N-acetylglucosamine (O-GlcNAc). Determining the targets and roles of O-GlcNAc glycoproteins has the potential to reveal insights into healthy and diseased metabolic states. In cell studies, thousands of proteins are known to be O-GlcNAcylated, but reference datasets for most tissue types in animals are lacking. Here, we apply a chemoenzymatic labeling study to compile a high coverage dataset of quadriceps skeletal muscle O-GlcNAc glycoproteins from mice. Our dataset contains over 550 proteins, and > 80% of the dataset matched known O-GlcNAc proteins. This dataset was further annotated via bioinformatics, revealing the distribution, protein interactions, and gene ontology (GO) functions of these skeletal muscle proteins. We compared these quadriceps glycoproteins with a high-coverage O-GlcNAc enrichment profile from mouse hearts and describe the key overlap and differences between these tissue types. Quadriceps muscles can be used for biopsies, so we envision this dataset to have potential biomedical relevance in detecting aberrant glycoproteins in metabolic diseases and physiological studies. This new knowledge adds to the growing collection of tissues with high-coverage O-GlcNAc profiles, which we anticipate will further the systems biology of O-GlcNAc mechanisms, functions, and roles in disease.
Rucli S, Descostes N, Ermakova Y, Chitnavis U, Couturier J, Boskovic A, Boulard M.
Functional genomic profiling of O-GlcNAc reveals its context-specific interplay with RNA polymerase II.
Genome biology2025
26(1)
40128797
Abstract: How reversible glycosylation of DNA-bound proteins acts on transcription remains scarcely understood. O-linked β-N-acetylglucosamine (O-GlcNAc) is the only known form of glycosylation modifying nuclear proteins, including RNA polymerase II (RNA Pol II) and many transcription factors. Yet, the regulatory function of the O-GlcNAc modification in mammalian chromatin remains unclear.
Liu Z, Yuan J, Din MA, Tian Y, Mao F.
HucMSC-Ex alleviates inflammatory bowel disease by regulating O-GlcNAcylation modification of RACK1 in intestinal epithelial cells.
Colloids and surfaces. B, Biointerfaces2025
251
40068238
Abstract: Inflammatory Bowel Disease (IBD) is a chronic autoimmune disorder that severely affects the gastrointestinal tract and is difficult to cure. This study explored the mechanism by which human umbilical cord mesenchymal stem cell-derived exosomes (HucMSC-Ex) alleviate IBD through O-GlcNAc glycosylation modification and the expression of related proteins. The study analyzed the effects of HucMSC-Ex on the inhibition of pro-inflammatory factors and promotion of intestinal epithelial cells regeneration in vitro and in vivo, with a focus on the role of the O-GlcNAc glycosylation of the RACK1 protein. The findings indicated that HucMSC-Ex reverses epithelial-mesenchymal transition (EMT) by upregulating O-GlcNAc glycosylation levels and effectively alleviates IBD symptoms and inflammatory responses in mouse intestinal epithelial cells. By modulating O-GlcNAc glycosylation, HucMSC-Ex exhibits significant therapeutic potential in immune regulation and gut microbiota remodeling, offering new perspectives for IBD treatment.
Hao Y, Li Z, Du X, Xie Q, Li D, Lei S, Guo Y.
Characterization and chemoproteomic profiling of protein O-GlcNAcylation in SOD1-G93A mouse model.
Molecular medicine (Cambridge, Mass.)2025
31(1)
40021952
Abstract: Amyotrophic lateral sclerosis (ALS) is a devastating motor neuron disease. Protein O-linked β-N-acetylglucosamine (O-GlcNAc) modification has been found to affect the processing of several important proteins implicated in ALS. However, the overall level and cellular localization of O-GlcNAc during ALS progression are incompletely understood, and large-scale profiling of O-GlcNAcylation sites in this context remains unexplored.
Jia L, Zheng H, Feng J, Ding Y, Sun X, Yu Y, Hao X, Wang J, Zhang X, Tian Y, Chen F, Cui J.
Upregulation of protein O-GlcNAcylation levels promotes zebrafish fin regeneration.
Molecular & cellular proteomics : MCP202540044042
Abstract: As one of the most important post-translational modifications, glycosylation participates in various cellular activities in organisms and is closely associated with many pathogeneses. It has been reported that glycosylation affects liver, spinal cord, and heart tissue regeneration. The zebrafish fin has become a valuable model due to its high regenerative capacity. The molecular mechanism of regeneration has been a hot research topic in the field for a long time. However, studies on the influence of glycosylation during limb regeneration in zebrafish are relatively scarce. We discovered that O-GlcNAc expression, identified by WGA, was elevated during the regeneration of the injured fin in zebrafish using lectin microarray. This phenomenon is due to the upregulation of the expression of OGT enzymes and elevated O-GlcNAcylation levels. To investigate the effects on the fin regeneration when O-GlcNAcylation changes, we used OSMI-1 or Alloxan unilateral microinjection to decrease O-GlcNAcylation and observed that it prevented the fin regeneration. Conversely, the O-GlcNAcylation was impressed by a unilateral microinjection of Thiamet-G or Glucose into the fin, leading to a stimulation of the fin regeneration. To further understand the role of O-GlcNAcylation in fin regeneration, LC-MS/MS was performed to identify O-GlcNAc-glycoproteins. The results demonstrated that the O-GlcNAc glycoproteins, such as THBS4 and HSPG, were involved in the regulation of zebrafish fin regeneration process and were closely associated with certain biological processes, such as stem cell differentiation, ECM-receptor interaction pathway, tissue remodeling, etc. We demonstrated that O-GlcNAc glycoproteins are crucial for zebrafish fin regeneration, during which OGT promotes the process by upregulating the O-GlcNAcylation levels in the zebrafish fin.
Jin H, He H, Li J, Liu X, Cai Q, Shi J, Hao Z, He J.
Mannose Inhibits NSCLC Growth and Inflammatory Microenvironment by Regulating Gut Microbiota and Targeting OGT/hnRNP R/JUN/IL-8 Axis.
International journal of biological sciences2025
21(4)
39990658
Abstract: Recent studies have reported direct antitumor effects of mannose, a natural six-carbon monosaccharide, in the treatment of cancer. Herein, we utilized cancer cell lines, animal models, organoids and experimental techniques such as multi-omics and cellular experiments to investigate the regulatory effects of mannose on NSCLC growth and the inflammatory microenvironment. We demonstrated that mannose can inhibit cancer cell growth, inflammatory cell infiltration and inflammatory cytokine expression in NSCLC tissue, and enhance the antitumor efficacy of immune checkpoint inhibitor both in vitro and in vivo. Orally administered mannose increased the proportion of probiotics in the gut microbiota, the abundance of anti-inflammatory and antitumor metabolites in the blood and feces of NSCLC-bearing mice. In NSCLC cells, mannose reduced JUN mRNA stability and subsequent IL-8 transcription of NSCLC cells by directly targeting OGT to suppress the O-GlcNAc glycosylation of hnRNP R, which bound and stabilized JUN mRNA in an O-GlcNAc glycosylation dependent manner. Taken together, our study demonstrated that mannose can suppress NSCLC by inhibiting tumor growth and the inflammatory microenvironment, and serve as a promising adjunct medication.
Le Minh G, Merzy J, Esquea EM, Ahmed NN, Young RG, Sharp RJ, Dhameliya TT, Agana B, Lee MH, Bethard JR, Comte-Walters S, Ball LE, Reginato MJ.
GATAD2B O-GlcNAcylation Regulates Breast Cancer Stem-like Potential and Drug Resistance.
Cells2025
14(6)
40136647
Abstract: The growth of breast tumors is driven and controlled by a subpopulation of cancer cells resembling adult stem cells, which are called cancer stem-like cells (CSCs). In breast cancer, the function and maintenance of CSCs are influenced by protein O-GlcNAcylation and the enzyme responsible for this post-translational modification, O-GlcNAc transferase (OGT). However, the mechanism of CSCs regulation by OGT and O-GlcNAc cycling in breast cancer is still unclear. Analysis of the proteome and O-GlcNAcome, revealed GATAD2B, a component of the Nucleosome Remodeling and Deacetylase (NuRD) complex, as a substrate regulated by OGT. Reducing GATAD2B genetically impairs mammosphere formation, decreases expression of self-renewal factors and CSCs population. O-GlcNAcylation of GATAD2B at the C-terminus protects GATAD2B from ubiquitination and proteasomal degradation in breast cancer cells. We identify ITCH as a novel E3 ligase for GATAD2B and show that targeting ITCH genetically increases GATAD2B levels and increases CSCs phenotypes. Lastly, we show that overexpression of wild-type GATAD2B, but not the mutant lacking C-terminal O-GlcNAc sites, promotes mammosphere formation, expression of CSCs factors and drug resistance. Together, we identify a key role of GATAD2B and ITCH in regulating CSCs in breast cancer and GATAD2B O-GlcNAcylation as a mechanism regulating breast cancer stem-like populations and promoting chemoresistance.
Zeng Z, Liao X, Zhao X.
O-GlcNAc transferase mediates O-GlcNAcylation of NLRP3 regulates pyroptosis in spinal cord injury.
Brain research bulletin2025
222
39892583
Abstract: Spinal cord injury (SCI) represents a severe disorder of the nervous system, imposing significant physical, psychological, and socioeconomic burdens on affected individuals and society.
Alhadidy MM, Stemmer PM, Kanaan NM.
O-GlcNAc modification differentially regulates microtubule binding and pathological conformations of tau isoforms in vitro.
The Journal of biological chemistry2025
301(3)
39909381
Abstract: Tau proteins undergo several posttranslational modifications in physiological and disease conditions. In Alzheimer's disease, O-GlcNAcylation modification of serine/threonine (S/T) residues in tau is reduced. In mouse models of tauopathy, O-GlcNAcase inhibitors lead to increased O-GlcNAcylation and decreased filamentous aggregates of tau. However, various nonfilamentous tau conformations, linked to toxicity and neurodegeneration in tauopathies, involve processes like oligomerization, misfolding, and greater exposure of the phosphatase-activating domain in the amino terminus of tau. Additionally, it is becoming clearer that posttranslational modifications may differently regulate tau pathobiology in an isoform-dependent manner. Therefore, it is crucial to investigate the effects of O-GlcNAcylation on nonfilamentous conformations of both the four-repeat (4R, e.g., hT40) and three-repeat (3R, e.g., hT39) tau isoforms. In this study, we assessed how O-GlcNAcylation impacts pathological tau conformations of the longest 4R and 3R tau isoforms (hT40 and hT39, respectively) using recombinant proteins. Mass spectrometry showed that tau is modified with O-GlcNAc at multiple S/T residues, primarily in the proline-rich domain and the C-terminal region. O-GlcNAcylation of hT40 and hT39 does not affect microtubule polymerization but has opposite effects on hT40 (increases) and hT39 (decreases) binding to preformed microtubules. Although O-GlcNAcylation interferes with forming filamentous hT40 aggregates, it does not alter the formation of pathological nonfilamentous tau conformations. On the other hand, O-GlcNAcylation increases the formation of pathological nonfilamentous hT39 conformations. These findings suggest that O-GlcNAcylation differentially modulates microtubule binding and the adoption of pathological tau conformations in the longest 4R and 3R tau isoforms.
Zou Y, Zhuo M, Chen W, Song W, Jiang Y, Xu J, Wang J.
Multiomics analysis of O-GlcNAcylation in podocytes of diabetic kidney disease.
Diabetes, obesity & metabolism202539950214
Abstract: To investigate the role of O-GlcNAc transferase (OGT)-mediated protein O-GlcNAcylation in podocyte injury during the progression of diabetic kidney disease (DKD).
Uno Y, Hayakawa K.
O-GlcNAcylation on serine 40 of histone H2A promotes proliferation and invasion in triple-negative breast cancer.
Scientific reports2025
15(1)
40128346
Abstract: Triple-negative breast cancer (TNBC) is characterized by resistance to conventional treatment and a poor prognosis. The O-linked β-N-acetylglucosamine (O-GlcNAc) modification of proteins has been reported to affect cancer progression. However, the key O-GlcNAc proteins involved in TNBC phenotypes remain unclear. Our previous study demonstrated that serine 40 of histone H2A was modified by O-GlcNAcylation (H2AS40Gc). Since S40 is located inside the globular domain of H2A, H2AS40Gc may be involved in the regulation of gene expression by altering chromatin conformation and could serve as the molecular basis for TNBC. The present study showed that H2AS40Gc levels were significantly higher in TNBC than in the other breast cancer subtypes. Using TNBC cells in which H2AS40Gc levels were depleted, we found that H2AS40Gc is required to promote cell proliferation and migration. The underlying mechanism of this promotion involves the accumulation of H2AS40Gc in the promoter region of KDM5B, a demethylase for lysine 4 of histone H3 (H3K4) that represses the expression of KDM5B, resulting in increased H3K4 trimethylation and elevated expression of genes related to proliferation and migration. Our findings clearly indicate that H2AS40Gc functions to promote proliferation and migration through KDM5B suppression and provide new insights into potential therapeutic approaches for TNBC.
Li X, Zhang Z, Zhang M, Cao Y, Zhou W, Kou L, Guo W, Zhang B, Li S, Xu B.
Mechanism of O-GlcNAcylation regulating liver lipid synthesis in mice through FASN.
FASEB journal : official publication of the Federation of American Societies for Experimental Biology2025
39(4)
39950311
Abstract: Nonalcoholic fatty liver disease (NAFLD) has become one of the most common chronic liver diseases. O-Linked attachment of beta-N-acetylglucosamine (O-GlcNAc) are ubiquitous post-translational modifications of proteins as "nutrient sensors" and "stress receptors" in the body that are involved in maintaining normal cellular physiological functions. Increased levels of O-GlcNAcylation have been found in the liver samples of patients with NAFLD and nonalcoholic steatohepatitis. However, the role of O-GlcNAcylation in the development and pathogenesis of NAFLD remains unclear. Here, we sought to determine the specific role of O-GlcNAcylation in NAFLD. In this study, the results demonstrated that inhibition of O-GlcNAc transferase (OGT) led to decreased expression of liver lipid synthesis genes and proteins in vitro. In addition, we showed that fatty acid synthase (FASN) expression was positively correlated with O-GlcNAcylation levels. Immunoprecipitation and pulldown assays confirmed the interaction between FASN and OGT at the serine 1483 of FASN, to inhibit K48-linked ubiquitination and degradation of FASN, thereby promoting hepatic lipid accumulation and the development of NAFLD. Administration of the OGT inhibitor OSMI-1 to ob/ob mice led to decreased liver lipid accumulation, further confirming our in vitro experimental results. Finally, we used liver-specific Ogt gene knockout mice fed a high-fat diet to elucidate the specific mechanism of O-GlcNAcylation on NAFLD and found that knockdown of the Ogt gene led to decreased liver lipid accumulation. In conclusion, our findings show that inhibiting the O-GlcNAcylation of FASN at the S1483 site promotes the K48-linked ubiquitination and degradation of FASN and leads to inhibition of lipid accumulation in the liver. Treatment with the OGT inhibitor OSMI-1 leads to decreased lipid accumulation in the liver, suggesting that targeting O-GlcNAcylation sites could be a potential therapeutic strategy for alleviating NAFLD.