PPARgamma-regulated mechanisms in hepatocytes that promote NAFLD

肝细胞中 PPARgamma 调节机制促进 NAFLD

• 批准号: 10557828
• 负责人: Jose Cordoba-Chacon
• 金额: $ 41.6万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10557828
• 关键词: Adipocytes; Adult; Anti-Inflammatory Agents; Caring; Cells; Cholesterol; Data; Dependovirus; Development; Diet; Disease; Dose; Enzymes; Extrahepatic; FDA approved; Fatty acid glycerol esters; Fibrosis; Fructose; Gene Expression; General Population; Health; Healthcare Systems; Hepatic; Hepatic Stellate Cell; Hepatic Tissue; Hepatocyte; Homocysteine; Human; In Vitro; Inflammation; Insulin Resistance; Knock-out; Label; Lecithin; Lipids; Liver; Macrophage; Mediating; Messenger RNA; Methionine; Methionine Metabolism Pathway; Methylation; Mus; Non-Insulin-Dependent Diabetes Mellitus; Outcome; PPAR gamma; Patients; Perfusion; Pharmaceutical Preparations; Pharmacological Treatment; Phosphatidylethanolamine N-Methyltransferase; Prevalence; Regulation; Testing; Therapeutic; Therapeutic Effect; Thiazolidinediones; Very low density lipoprotein; betaine-homocysteine methyltransferase; chronic liver disease; clinical care; clinically relevant; defined contribution; diabetogenic; expectation; experimental study; fibrogenesis; improved; in vivo; innovation; liver injury; metabolomics; methyl group; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; restoration

PROJECT SUMMARY / ABSTRACT Non-alcoholic fatty liver disease (NAFLD) is the leading cause of chronic liver disease with a prevalence of 25% in the general population. NAFLD is strongly associated with type 2 diabetes, where it shows a prevalence up to 80%, and a strong progression to non-alcoholic steatohepatitis (NASH, and advance stage of NAFLD). To date, there are no FDA-approved pharmacological treatments for NAFLD. However, thiazolidinediones (TZD), which are effective anti-diabetogenic drugs, may be used to treat NAFLD/NASH. Specifically, TZD activate peroxisome proliferator-activated receptor gamma (PPARγ) in adipocytes, macrophages, and hepatic stellate cells (HSC), and they should reduce insulin resistance, inflammation, and fibrogenesis, respectively. Despite having some positive effects, TZD are not currently used to treat NASH, and it is possible that their true potential as anti- NASH drugs are indeed reduced by direct negative actions on hepatocyte function. In fact, our preliminary studies show that hepatocyte PPARγ is a relevant factor in the regulation of hepatic gene expression that contributes to the progression of NASH, and reduces the therapeutic effects of TZD in the liver of mice with NASH. In this proposal, we hypothesize that hepatocyte PPARγ is a negative regulator of phosphatidylethanolamine methyltransferase (PEMT) and betaine-homocysteine methyltransferase (BHMT), disrupts methionine metabolism, and promotes NASH. In our Aim 1, we will define the contribution of hepatocyte PEMT and BHMT in the TZD-mediated reversal of NASH. Specifically, we will restore the expression of PEMT or BHMT with adeno-associated viruses in PPARγ-intact mice after the development of NASH and treat these mice with TZD to reverse NASH. Also, hepatocyte PEMT and BHMT will be restored in mice with hepatocyte- specific loss of PPARγ expression without a TZD treatment. This aim will show how PEMT and BHMT improve liver health, and reduce steatosis, inflammation and fibrosis to enhance the therapeutic actions of TZD in the reversal of NASH. In aim 2, we will determine if PPARγ directly disrupts the metabolism of methionine in hepatocytes. Briefly, we will use targeted metabolomics in mouse and human primary hepatocytes or in perfused livers of controls and mice with hepatocyte-specific loss of PPARγ expression that are treated with TZD. These experiments will identify how TZD alters the use of methionine in hepatocytes, and contributes, in a hepatocyte- specific PPARγ-dependent manner, to sustain steatosis, inflammation and fibrosis despite the positive actions of TZD on adipocytes, macrophages, and HSC. Overall, in this proposal we will describe how hepatocyte-specific PPARγ negatively regulates methionine metabolism to promote NAFLD, and to limit the potential of TZD as a therapy for NASH. The outcomes of this project will lead us to develop therapeutic strategies that enhance the use of TZD, and to develop new treatments for NAFLD and the care of NASH patients.

项目摘要/摘要 非酒精性脂肪性肝病(NAFLD)是慢性肝病的主要原因，患病率为25% 在普通人群中。非酒精性脂肪肝与2型糖尿病密切相关，在2型糖尿病中，其患病率高达 80%，并强烈进展为非酒精性脂肪性肝炎(NASH，和NAFLD进展期)。到目前为止， 目前还没有FDA批准的治疗NAFLD的药物。然而，噻唑烷二酮(TZD)，它是 是有效的抗糖尿病药物，可用于治疗NAFLD/NASH。具体地说，TZD激活了过氧化酶体 脂肪细胞、巨噬细胞和肝星状细胞中的增殖因子激活受体γ(PPARγ)； 它们应该分别减少胰岛素抵抗、炎症和纤维化。尽管有一些 积极作用，TZD目前还没有用于治疗NASH，有可能其真正的潜在抗炎作用 NASH药物确实是通过对肝细胞功能的直接负面作用而减少的。事实上，我们的初步调查 研究表明，肝细胞PPARγ是调节肝脏基因表达的相关因子 促进NASH的进展，并降低TZD对小鼠肝脏的治疗作用 纳什。在这个方案中，我们假设肝细胞PPARγ是一种负调控因子 磷脂酰乙醇胺甲基转移酶(PEMT)和甜菜碱-同型半胱氨酸甲基转移酶(BHMT)， 扰乱蛋氨酸代谢，促进NASH。在我们的目标1中，我们将定义肝细胞的贡献 PEMT和BHMT在TZD介导的NASH逆转中的作用具体来说，我们将恢复PEMT的表达 PPARγ基因缺失小鼠NASH后与腺相关病毒或BHMT的关系及治疗 TZD逆转NASH的小鼠。此外，肝细胞移植小鼠的肝细胞PEMT和BHMT也将恢复。 未经TZD处理的PPARγ表达特异性缺失。该目标将展示PEMT和BHMT如何改进 保健肝脏，减少脂肪变性、炎症和纤维化，以增强TZD对糖尿病的治疗作用 纳什的逆转。在目标2中，我们将确定PPARγ是否直接干扰蛋氨酸在体内的代谢。 肝细胞。简而言之，我们将在小鼠和人的原代肝细胞或灌流的肝细胞中使用靶向代谢组学。 对照组和肝细胞特异性PPARγ表达缺失的小鼠的肝脏。这些 实验将确定TZD如何改变肝细胞中蛋氨酸的使用，并在肝细胞中贡献- 特异性PPARγ依赖方式，维持脂肪变性、炎症和纤维化，尽管有积极作用 TZD对脂肪细胞、巨噬细胞和HSC的作用。总体而言，在这份提案中，我们将描述肝细胞特异性 PPARγ负性调节蛋氨酸代谢促进非酒精性脂肪肝的发生 纳什的治疗。这个项目的结果将引导我们开发治疗策略，以增强 TZD的使用，以及开发NAFLD的新疗法和NASH患者的护理。