Abstract：
OBJECTIVE: The relationship between metabolic dysfunction-associated steatotic liver disease (MASLD) and type 2 diabetes mellitus, insulin resistance and the metabolic syndrome is well established. While zinc finger BED-type containing 3 (ZBED3) has been linked to type 2 diabetes mellitus and the metabolic syndrome, its role in MASLD remains unclear. In this study, we aimed to investigate the function of ZBED3 in the context of MASLD.
METHODS: Expression levels of ZBED3 were assessed in individuals with MASLD, as well as in cellular and animal models of MASLD. In vitro and in vivo analyses were conducted using a cellular model of MASLD induced by NEFA and an animal model of MASLD induced by a high-fat diet (HFD), respectively, to investigate the role of ZBED3 in MASLD. ZBED3 expression was increased by lentiviral infection or tail-vein injection of adeno-associated virus. RNA-seq and bioinformatics analysis were employed to examine the pathways through which ZBED3 modulates lipid accumulation. Findings from these next-generation transcriptome sequencing studies indicated that ZBED3 controls SREBP1c (also known as SREBF1; a gene involved in fatty acid de novo synthesis); thus, co-immunoprecipitation and LC-MS/MS were utilised to investigate the molecular mechanisms by which ZBED3 regulates the sterol regulatory element binding protein 1c (SREBP1c).
RESULTS: In this study, we found that ZBED3 was significantly upregulated in the liver of individuals with MASLD and in MASLD animal models. ZBED3 overexpression promoted NEFA-induced triglyceride accumulation in hepatocytes in vitro. Furthermore, the hepatocyte-specific overexpression of Zbed3 promoted hepatic steatosis. Conversely, the hepatocyte-specific knockout of Zbed3 resulted in resistance of HFD-induced hepatic steatosis. Mechanistically, ZBED3 interacts directly with polypyrimidine tract-binding protein 1 (PTBP1) and affects its binding to the SREBP1c mRNA precursor to regulate SREBP1c mRNA stability and alternative splicing.
CONCLUSIONS: This study indicates that ZBED3 promotes hepatic steatosis and serves as a critical regulator of the progression of MASLD.
METHODS: RNA-seq data have been deposited in the NCBI Gene Expression Omnibus ( www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE231875 ). MS proteomics data have been deposited to the ProteomeXchange Consortium via the iProX partner repository ( https://proteomecentral.proteomexchange.org/cgi/GetDataset?ID=PXD041743 ).
METHODS: Expression levels of ZBED3 were assessed in individuals with MASLD, as well as in cellular and animal models of MASLD. In vitro and in vivo analyses were conducted using a cellular model of MASLD induced by NEFA and an animal model of MASLD induced by a high-fat diet (HFD), respectively, to investigate the role of ZBED3 in MASLD. ZBED3 expression was increased by lentiviral infection or tail-vein injection of adeno-associated virus. RNA-seq and bioinformatics analysis were employed to examine the pathways through which ZBED3 modulates lipid accumulation. Findings from these next-generation transcriptome sequencing studies indicated that ZBED3 controls SREBP1c (also known as SREBF1; a gene involved in fatty acid de novo synthesis); thus, co-immunoprecipitation and LC-MS/MS were utilised to investigate the molecular mechanisms by which ZBED3 regulates the sterol regulatory element binding protein 1c (SREBP1c).
RESULTS: In this study, we found that ZBED3 was significantly upregulated in the liver of individuals with MASLD and in MASLD animal models. ZBED3 overexpression promoted NEFA-induced triglyceride accumulation in hepatocytes in vitro. Furthermore, the hepatocyte-specific overexpression of Zbed3 promoted hepatic steatosis. Conversely, the hepatocyte-specific knockout of Zbed3 resulted in resistance of HFD-induced hepatic steatosis. Mechanistically, ZBED3 interacts directly with polypyrimidine tract-binding protein 1 (PTBP1) and affects its binding to the SREBP1c mRNA precursor to regulate SREBP1c mRNA stability and alternative splicing.
CONCLUSIONS: This study indicates that ZBED3 promotes hepatic steatosis and serves as a critical regulator of the progression of MASLD.
METHODS: RNA-seq data have been deposited in the NCBI Gene Expression Omnibus ( www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE231875 ). MS proteomics data have been deposited to the ProteomeXchange Consortium via the iProX partner repository ( https://proteomecentral.proteomexchange.org/cgi/GetDataset?ID=PXD041743 ).
摘要：
目的：代谢功能障碍相关脂肪变性肝病（MASLD）与2型糖尿病的关系，胰岛素抵抗和代谢综合征是公认的。虽然锌指BED型含3（ZBED3）与2型糖尿病和代谢综合征有关，其在MASLD中的作用尚不清楚。在这项研究中,我们旨在研究ZBED3在MASLD背景下的功能。
方法：在患有MASLD的个体中评估ZBED3的表达水平，以及MASLD的细胞和动物模型。使用NEFA诱导的MASLD细胞模型和高脂饮食（HFD）诱导的MASLD动物模型进行体外和体内分析。分别,探讨ZBED3在MASLD中的作用。ZBED3表达通过慢病毒感染或腺相关病毒的尾静脉注射而增加。采用RNA-seq和生物信息学分析来检查ZBED3调节脂质积累的途径。这些下一代转录组测序研究的发现表明，ZBED3控制SREBP1c（也称为SREBF1；参与脂肪酸从头合成的基因）；因此，免疫共沉淀和LC-MS/MS用于研究ZBED3调节固醇调节元件结合蛋白1c（SREBP1c）的分子机制。
结果：在这项研究中，我们发现ZBED3在MASLD患者和MASLD动物模型的肝脏中显著上调.ZBED3过表达促进NEFA诱导的甘油三酯在体外肝细胞中的积累。此外，肝细胞特异性Zbed3过表达促进肝脏脂肪变性.相反，肝细胞特异性敲除Zbed3导致HFD诱导的肝脂肪变性耐药.机械上,ZBED3直接与聚嘧啶束结合蛋白1（PTBP1）相互作用，并影响其与SREBP1cmRNA前体的结合，以调节SREBP1cmRNA的稳定性和可变剪接。
结论：本研究表明ZBED3促进肝脂肪变性，并作为MASLD进展的关键调节因子。
方法：RNA-seq数据已保存在NCBI基因表达Omnibus中（www。ncbi.nlm.nih.gov/geo/query/acc。cgi?acc=GSE231875)。MS蛋白质组学数据已通过iProX合作伙伴存储库（https://proteomecentral。proteomexchange.org/cgi/GetDataset?ID=PXD041743)。
方法：在患有MASLD的个体中评估ZBED3的表达水平，以及MASLD的细胞和动物模型。使用NEFA诱导的MASLD细胞模型和高脂饮食（HFD）诱导的MASLD动物模型进行体外和体内分析。分别,探讨ZBED3在MASLD中的作用。ZBED3表达通过慢病毒感染或腺相关病毒的尾静脉注射而增加。采用RNA-seq和生物信息学分析来检查ZBED3调节脂质积累的途径。这些下一代转录组测序研究的发现表明，ZBED3控制SREBP1c（也称为SREBF1；参与脂肪酸从头合成的基因）；因此，免疫共沉淀和LC-MS/MS用于研究ZBED3调节固醇调节元件结合蛋白1c（SREBP1c）的分子机制。
结果：在这项研究中，我们发现ZBED3在MASLD患者和MASLD动物模型的肝脏中显著上调.ZBED3过表达促进NEFA诱导的甘油三酯在体外肝细胞中的积累。此外，肝细胞特异性Zbed3过表达促进肝脏脂肪变性.相反，肝细胞特异性敲除Zbed3导致HFD诱导的肝脂肪变性耐药.机械上,ZBED3直接与聚嘧啶束结合蛋白1（PTBP1）相互作用，并影响其与SREBP1cmRNA前体的结合，以调节SREBP1cmRNA的稳定性和可变剪接。
结论：本研究表明ZBED3促进肝脂肪变性，并作为MASLD进展的关键调节因子。
方法：RNA-seq数据已保存在NCBI基因表达Omnibus中（www。ncbi.nlm.nih.gov/geo/query/acc。cgi?acc=GSE231875)。MS蛋白质组学数据已通过iProX合作伙伴存储库（https://proteomecentral。proteomexchange.org/cgi/GetDataset?ID=PXD041743)。
