iProXProteomicsBanglun PanMus Musculushttp://www.iprox.org/page/project.html?id=IPX00116310001335563838@qq.comFujian Medical UniversityiProXLTβR-overexpressing CAR-T cells have demonstrated surprising effectiveness against solid tumors, exhibiting strong anti-exhaustion and proliferation capabilities. However, the role of LTβR in CD4⁺ T cell differentiation and anti-tumor activity remains unclear. In this study, we employed primary or subcutaneous mouse hepatocellular carcinoma (HCC) models and flow cytometry to study the impact of conditional knock-in of Ltbr on CD4⁺ T cell differentiation and response, particularly the Th17/Treg cell ratio, and its influence on HCC progression. Immunoprecipitation, immunoblotting, RT-qPCR, molecular docking, and Chromatin Immunoprecipitation-qPCR were performed to investigate the molecular mechanism of CD4⁺ T cell differentiation. Adeno-associated virus-modified T cells were introduced into patient-derived orthotopic xenograft (PDOX) model to assess the combined impact of LTβR and glycolysis inhibitors on the Th17/Treg cell differentiation. We found that LTβR reduced PELI1 expression, preventing TRAF3 protein degradation in Th17 cells. TRAF3 then competed with RORC for SMURF1 binding, enhancing RORC stability and Th17 cell differentiation. LTβR also blocked PRDM1 expression, delaying Foxp3 transcription and Treg cell infiltration. Additionally, N-glycosylation supported the stability of LTβR by protecting it from ubiquitination. From a therapeutic perspective, glycolysis inhibitors helped LTβR balance the proportion of Th17/Treg cells in PDOX model to inhibit tumor growth. In conclusion, our findings indicated that LTβR N-glycosylation prevented RORC ubiquitination and Foxp3 transcription, raising the Th17/Treg cell ratio and hindering HCC progression.N-glycosylated LTβR increases the Th17/Treg cell ratio in liver cancer by blocking RORC ubiquitination and FOXP3 transcription.Pan Banglun B, Yao Yuxin Y, Wu Hao H, Ye Dongjie D, Zhang Zhu Z, Zhang Xinyu X, Wang Xiaoqian X, Tang Nanhong NNEOPL LIVER, primary tumour of the liver, resectable malignant neoplasm of the liver, Regulatory T-Lymphocyte, Regulatory T-Lymphocytes, Neoplasms, bacterial transcription., Suppressor T-Lymphocytes, RORG, T-Lymphocyte, adult hepatoma, hepatic, Hepatic Neoplasm, hepatocellular carcinoma plus intrahepatic cholangiocarcinoma, Th3, primary liver cancer, liver, NR1F3, FOXP3, Naturally-Occurring Suppressor T-Cells, Regulatory T Cells, TOR, IPEX, XPID, primary, RGD1562112, not specified as primary or secondary, Hepatic Cancer, Ubiquitylation, Liver Cancer, primary malignant neoplasm of Liver, T-Cells, JM2, Cancer of the liver, Thor, Tr1, adult primary hepatocellular carcinoma, Liver Cancers, sf, Treg Cells, Hepatic Neoplasms, regulatory T-cell, malignant neoplasm of liver, hepatic cancer, RZRG, AIID, Tr1 Cells, T Cells, malignant hepato-biliary neoplasm, Nr1f3, suppressor T-lymphocyte, Resectable malignant neoplasm of Liver, Regulatory T Lymphocyte, primary malignant neoplasm of liver, PIDX, Naturally-Occurring, Hepatic, Cancer of the Liver, ubiquitination, hepatic neoplasm, transcription from bacterial-type RNA polymerase promoter, T Lymphocytes, Cancer, liver Cancer, Treg cell, Liver, Suppressor T Cells, Regulatory, Regulatory T-Cell, Hepatic Cancers, Naturally-Occurring Suppressor T-Lymphocytes, primary tumor of the liver, DIETER, Naturally-Occurring Suppressor T-Cell, Cancer of Liver, primary cancer of liver, liver Cancers, Regulatory T, Suppressor T-Cell, Cell, RZR-GAMMA, Regulatory T Cell, T-Cell, T suppressor cell, Liver Neoplasm, Neoplasm, Naturally-Occurring Suppressor T-Lymphocyte, Th3 Cells, Th3 Cell, Suppressor T-Cells, Suppressor T Lymphocytes, ca liver - primary, Tr1 Cell, Suppressor T-Lymphocyte, non-resectable primary hepatic malignant neoplasm, Regulatory T Lymphocytes, Treg Cell, Cancers, malignant tumor of liver, Naturally-Occurring Suppressor, Naturally Occurring, suppressor T-cell, neoplasm of liver, RORgamma, Treg, Ca liver - primary, Regulatory T-Cells, Cells, Hepatocellular Cancers, Hepatocellular, LIVER NEOPL, Cancer of liver, T Cell, Ts cell, Hepatocellular Cancer, scurfinTNFR-RP, Immunoelectroblotting, O-linked Glycosylations, Laboratory, FON1, Embden-Meyerhof, Galactosylations, NSHPT, growth and development, adult hepatoma, TRANSPL HETEROL, Tumor, FLORAL ORGAN NUMBER 1, DLEU5, House Mouse, Liver Cell Carcinoma, Th3, Adeno-Associated Viruses, NR1F3, Ly-4, symptoms, MCVADR, CAR4|6, Analysis, IPEX, Cell Sortings, imprinted and ancient gene protein, Molecular Docking Analyses, Th17 T-cell, Molecular, ChIP, T-Cells, Bdr, D12S370, Dependoparvovirus, adult primary hepatocellular carcinoma, T, HLD5, Pathways, EIG8, Sialylation, SNAP-25, Hepatocellular Carcinoma, MCAR, Nr1f3, suppressor T-lymphocyte, Xenotransplantations, PIDX, O-linked, Role Concepts, Naturally-Occurring, Type 17 Helper T Cells, T cell development, xenotransplant, blimp1, Precipitations, transcription from bacterial-type RNA polymerase promoter, Thymus Dependent Lymphocytes, N-Acetylglucosaminylations, T helper 17 cell, Tumors, Cell Sorting, screening, Liver, Treg cell, Pathway, Degradations, Suppressor T Cells, Tnfrsf3, CAR-beta, Naturally-Occurring Suppressor T-Cell, anaerobic glycolysis, Regulatory T, Docking, Embden-Meyerhof-Parnas, Suppressor T-Cell, Embden-Meyerhof-Parnas Pathway, LT-BETA-R, Benign, Role Concept, T-cell differentiation, A930031K15Rik, Role, GENA70, Th3 Cell, Suppressor T Lymphocytes, DmelCG4394, Carcinomas, Reverse Immunoblottings, Xenotransplantation, Hepatoma, Protein Digestions, Swiss Mouse, signs, Benign Neoplasms, Regulatory T Lymphocytes, CD4mut, AI528849, Treg Cell, Fluorescence-Activated Cell, 4930431E10Rik, TH-17, Malignant Neoplasms, dTRAF3, contractile actomyosin ring, glycolysis, TNFR3, antagonists and inhibitors, Reverse Immunoblotting, Cells, T Cell, Mouse, Hepatocellular., HYPOC1, Molecular Docking, other neoplasm, E430016J11Rik, Embden-Meyerhof Pathways, Care2, scurfin, FLORAL DEFECTIVE 10, HYCC1, Immunoblotting, Neoplasms, Regulatory T-Lymphocytes, SPG5C, RORG, SUPERMAN, N-linked Glycosylations, GPRC2A, TNFRSF3, ADOHR, Fluorescence-Activated, N-linked, Fluorescence-Activated Cell Sortings, Microfluorometry, Dot, AW553441, actomyosin ring, FOXP3, Immune Precipitations, Mus musculus domesticus, Adult Liver Cancer, PGN, TOR, CNC, Immune Precipitation, ZNFPR1A1, Cytofluorometry, RGD1562112, HCVADR, RNF77, 3110001L12Rik, AA409794, Liver Cancer, Flow Cytofluorometries, RGD1309707, ligand, Coxsackievirus B-adenovirus receptor, Tr1, AI586297, TH-17 Cell, Neoplasias, Tr1 Cells, Phosphoglycosylation, HHC1, Flow, Traf-related, Flow Microfluorometries, Cytometry, Immunoelectroblottings, Heterologous, N-linked Glycosylation, Sortings, PKR1, Cancer, CD40bp, inhibitors, Traf3, findings, Malignant Neoplasm, Protein Glycosylation, Synaptosomal-associated 25 kDa protein, Flow Cytometries, LEU5, Digestions, Cell, Concept, Regulatory T Cell, XENOTRANSPL, Dot Immunoblottings, Proteolyses, T-cell differentiation antigen L3T4, T suppressor cell, Neoplasm, imprinted and ancient gene protein homolog, IMPACT, GlcNAcylations, TRAF3, Th3 Cells, Flow Microfluorometry, Adeno-Associated Virus, Th17 T cell, BLIMP1, Chromatin, amn, pheromone catabolic process, Tr1 Cell, Q-PCR, Hepatocellular carcinoma, underdeveloped, mKIAA1625, LAP-1 Protein, postnatal growth, Chromatin Immunoprecipitation, TNF Receptor Associated Factor 3, House Mice, Cancers, Naturally Occurring, RORgamma, CNC1, Blimp-1, CD40 Receptor-Associated Factor 1, Regulatory T-Cells, D11Ertd676e, General activity, Neoplasia, Molecular Docking Analysis, Co-Immunoprecipitations, CMAR, Reverse, T-cell surface antigen T4/Leu-3, CVB3-binding protein, Activity, CRAF1, Heterograft, Constitutive androstane receptor, Galactosylation, Mus domesticus, postnatal development, kinetic polymerase chain reaction, Suppressor T-Lymphocytes, T-Lymphocyte, Constitutive active response, Hepatomas, Embden Meyerhof Parnas Pathway, Co Immunoprecipitation, L3T4, Th17 CD4+ T cell, Electroimmunoblotting, T Lymphocyte, Roles, SUP, Virus, ESTM32, Concepts, Naturally-Occurring Suppressor T-Cells, Regulatory T Cells, Embden-Meyerhof Pathway, Flow Microfluorimetry, LAP1, TH-17 Cells, XPID, Docking Analysis, CD40bp Protein, T-BAM, b2b1765Clo, AU016810, DRCTNNB1A, T helper cells type 17, Ubiquitylation, TNFRrp, Th17 T lymphocyte, T Helper 17 Cells, Fluorescence Activated Cell Sorting, TNF-R-III, ATP-dependent proteolysis, Immunoblottings, Type 17 Helper T Cell, HCAR, Co-Immunoprecipitation, Precipitation, hypoplasia, sf, N-Acetylglucosaminylation, Swiss Mice, CD18, regulatory T-cell, Super protein, Adult, RZRG, Protein Degradation, sp, T Cells, Immune, peli1, CG4394, xpellino2, PPNAD1, xpellino1, Therapies, Malignancies, constriction ring, ubiquitination, house mouse, Adeno-Associated, hCAR, Flow Cytofluorometry, Therapy, Carcinoma, Fluorescence-Activated Cell Sorting, Dependoparvoviruses, T-cell surface antigen T4|Leu-3, Tnfbr, mouse, Glycosylations, Liver Cell, qPCR, Fucosylations, LTbetaR, CAP1, Microfluorimetry, RZR-GAMMA, TH 17 Cells, HCC, Th17 T-lymphocyte, T-Cell, Phosphoglycosylations, Digestion, T lymphocyte differentiation, Adult Liver, Suppressor T-Cells, Heterologous Transplantations, TNFCR, Mus musculus, Adeno Associated Viruses, Xenograft, Cytofluorometries, Transplantation, growth pattern, mice, non-developmental growth, T-cell surface glycoprotein CD4, Adeno Associated, N linked Glycosylation, Thymus-Dependent, Adeno Associated Virus, Sialylations, Xenografts, TH 17 Cell, Treatments, Embden-Meyerhof pathway, Thymus-Dependent Lymphocyte, domesticus, CAR1, CAR4/6, Patient, Dependoviruses, N Acetylglucosaminylation, bacterial transcription, xblimp1, Molecular Docking Simulations, Hepatocellular Carcinomas, IL-17-producing CD4+ T helper, pellino2, pellino1, quantitative real time polymerase chain reaction, Thymus-Dependent Lymphocytes, GlcNAcylation, Regulatory T-Lymphocyte, Degradation, Benign Neoplasm, Dot Immunoblotting, Cell Carcinoma, CAP-1, Transplantations, Malignant, Ximpact, heterologous transplantation, Cars, Automobile, Cell Carcinomas, O-linked Glycosylation, reduced, glycosylation, House, Liver Cell Carcinomas, Glycosylation, modified Embden-Meyerhof pathway, RFP2, Rfp2, MB67, tiny, Mice, T-lymphocyte differentiation, HHC, study, AI256028, Immunoprecipitation, CRAF1 Protein, pheromone catabolism, Viruses, Malignancy, Swiss, Protein Glycosylations, Microfluorometries, JM2, Thor, Liver Cancers, Heterografts, inhibiteur, Treg Cells, IIAE5, 2810468L03Rik, Protein Degradations, AIID, helper T cell type 17, Mif1, Regulatory T Lymphocyte, inhibidor, TNFR2-RP, TSE1, Clients, Gprc2a, ACRDYS1, cytokinetic ring, Fucosylation, T Lymphocytes, Molecular Dockings, Lymphocyte, Cytometries, small, Regulatory, Embden Meyerhof Pathway, Regulatory T-Cell, 2610206D03Rik, Naturally-Occurring Suppressor T-Lymphocytes, Blimp1, DIETER, Orphan nuclear receptor MB67, T(H)-17 cell, Docking Simulation, inhibitor, CaR, Th17 Cell, Protein Digestion, Client, impact-a, Th17, O linked Glycosylation, development, Constitutive activator of retinoid response, Mus, Protein, Naturally-Occurring Suppressor T-Lymphocyte, CAR, Car, LAP 1 Protein, antagonists, Electroimmunoblottings, Th17 cell, FHH, Suppressor T-Lymphocyte, Adult Liver Cancers, PCAR1, AA209988, Lymphocytes, HETEROL TRANSPL, FLO10, patient, CD40 Receptor Associated Factor 1, HERP, Naturally-Occurring Suppressor, Embden-Meyerhof-Parnas pathway, PRKAR1, Laboratory Mice, suppressor T-cell, FIH, Treg, PRDI-BF1, Therapeutic, SNAP, Sorting, Hepatocellular, TRAF-3, CD4, Treatment, Simulation, T Helper 17 Cell, AI551208, Ts cell, growth, Laboratory Mouse, RWDD5, LtarCD40bp, T-BAM, amn, CRAF1 Protein, Traf3, AA409794, CRAF1, Protein., protein complex, ligand, LAP-1 Protein, Proteins, Gene, TNF Receptor Associated Factor 3, AI528849, CD40 Receptor Associated Factor 1, AI586297, IIAE5, protein, 2810468L03Rik, CAP-1, protein-containing complex, Protein Gene Products, CAP1, Gene Proteins, peli1, dTRAF3, native protein, CG4394, CD40 Receptor-Associated Factor 1, Protein, A930031K15Rik, xpellino2, xpellino1, Traf-related, Gene Products, TRAF-3, LAP1, TRAF3, protein aggregate, D11Ertd676e, pellino2, pellino1, DmelCG4394, LAP 1 Protein, CD40bp ProteinNEOPL LIVER, primary tumour of the liver, resectable malignant neoplasm of the liver, Regulatory T-Lymphocyte, Regulatory T-Lymphocytes, Neoplasms, bacterial transcription., Suppressor T-Lymphocytes, RORG, T-Lymphocyte, adult hepatoma, hepatic, Hepatic Neoplasm, hepatocellular carcinoma plus intrahepatic cholangiocarcinoma, Th3, primary liver cancer, liver, NR1F3, FOXP3, Naturally-Occurring Suppressor T-Cells, Regulatory T Cells, TOR, IPEX, XPID, primary, RGD1562112, not specified as primary or secondary, Hepatic Cancer, Ubiquitylation, Liver Cancer, primary malignant neoplasm of Liver, T-Cells, JM2, Cancer of the liver, Thor, Tr1, adult primary hepatocellular carcinoma, Liver Cancers, sf, Treg Cells, Hepatic Neoplasms, regulatory T-cell, malignant neoplasm of liver, hepatic cancer, RZRG, AIID, Tr1 Cells, T Cells, malignant hepato-biliary neoplasm, Nr1f3, suppressor T-lymphocyte, Resectable malignant neoplasm of Liver, Regulatory T Lymphocyte, primary malignant neoplasm of liver, PIDX, Naturally-Occurring, Hepatic, Cancer of the Liver, ubiquitination, hepatic neoplasm, transcription from bacterial-type RNA polymerase promoter, T Lymphocytes, Cancer, liver Cancer, Treg cell, Liver, Suppressor T Cells, Regulatory, Regulatory T-Cell, Hepatic Cancers, Naturally-Occurring Suppressor T-Lymphocytes, primary tumor of the liver, DIETER, Naturally-Occurring Suppressor T-Cell, Cancer of Liver, primary cancer of liver, liver Cancers, Regulatory T, Suppressor T-Cell, Cell, RZR-GAMMA, Regulatory T Cell, T-Cell, T suppressor cell, Liver Neoplasm, Neoplasm, Naturally-Occurring Suppressor T-Lymphocyte, Th3 Cells, Th3 Cell, Suppressor T-Cells, Suppressor T Lymphocytes, ca liver - primary, Tr1 Cell, Suppressor T-Lymphocyte, non-resectable primary hepatic malignant neoplasm, Regulatory T Lymphocytes, Treg Cell, Cancers, malignant tumor of liver, Naturally-Occurring Suppressor, Naturally Occurring, suppressor T-cell, neoplasm of liver, RORgamma, Treg, Ca liver - primary, Regulatory T-Cells, Cells, Hepatocellular Cancers, Hepatocellular, LIVER NEOPL, Cancer of liver, T Cell, Ts cell, Hepatocellular Cancer, scurfinfalseN-glycosylated LTβR increases the Th17/Treg cell ratio in liver cancer by blocking RORC ubiquitination and FOXP3 transcriptionProteins binding to PELI1 and TRAF3 were identified by protein profilingTue Apr 08 00:00:00 BST 2025PXD0627601009040436857