YAP-IRGM1 REGULATES HEPATIC INFLAMMATION IN ORTHOTOPIC LIVER TRANSPLANTATION

YAP-IRGM1 调节原位肝移植中的肝脏炎症

• 批准号: 10605200
• 负责人: Haofeng Ji
• 金额: $ 23.4万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-07 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10605200
• 关键词: Adoptive Transfer; Antigens; Area; Attenuated; Autophagocytosis; Autophagosome; Biological; Cell Hypoxia; Cells; Cellular Stress; Cessation of life; Chronic; Circulation; Clinical; Clinical Data; Coupling; Cryopreservation; Cyclic GMP; Cytoprotection; Data; Deterioration; Development; Dissection; Equilibrium; Event; Excision; Family; GTP-Binding Proteins; Gene Family; Genes; Genetic Transcription; Guanosine Triphosphate Phosphohydrolases; Hepatic; Hepatic Tissue; Hepatocyte; Homeostasis; Human; IRF3 gene; ITGAM gene; Immune; Immunity; Inflammation; Inflammatory; Inflammatory Response; Injury; Innate Immune Response; Interferon Activation; Interferon Type II; Ischemia; Knock-out; Macrophage; Mediating; Metabolism; Modeling; Molecular; Morbidity - disease rate; Mouse Strains; Mus; Myelogenous; Natural Immunity; Natural regeneration; Operative Surgical Procedures; Organ Donor; Organ Retrievals; Organ Size; Oxidative Stress; Pathogenicity; Pathway interactions; Phase; Population; Prognostic Marker; Property; Protein Deficiency; Proteins; Publications; Publishing; Reactive Oxygen Species; Regulation; Reperfusion Injury; Reporting; Risk Factors; Role; Shock; Signal Pathway; Signal Transduction; Signaling Protein; Stress; TLR4 gene; Therapeutic; Tissues; Transplant Recipients; Transplantation; Warm Ischemia; clinically relevant; deprivation; graft dysfunction; improved; interest; ischemic injury; liver function; liver inflammation; liver injury; liver ischemia; liver transplantation; member; mortality; mouse model; novel; novel therapeutic intervention; organ transplant recipient; pathogen; prevent; prognostic; protein activation; protein distribution; protein expression; repaired; response; self-renewal; therapeutic target; tissue regeneration

PROJECT SUMMARY/ABSTRACT Hepatic ischemia and reperfusion injury (IRI) remains a common problem in liver transplant, hepatic resection, and shock. As it often leads to primary graft dysfunction (PGD), late chronic rejection, and contributes to the shortage of donor organs available for transplantation, liver IRI represents one of the most challenging yet understudied complications in clinical orthotopic liver transplantation (OLT). As intrinsic YAP expression negatively correlated with liver function and tissue damage in human OLT, treatment with YAP activator ameliorated liver IRI by depressing macrophage function and improving hepatocyte protection in a non-transplant model of hepatic “warm” IRI. Here, the function of distinct YAP signaling pathways is proposed to study in a clinically relevant mouse model of prolonged hepatic cold ischemia followed by OLT. Hypothesis: downregulated peri-transplant YAP expression is an important driver of graft injury and development of PGD, and that such reduced YAP expression in liver graft may be of prognostic and therapeutic significance. Cellular specific YAP distribution, has prompted to put forth a dissection that YAP signaling at the macrophage (innate immune) regulates inflammation (pathogenic), autophagic reprogramming (homeostatic) and cytoprotection (anti-oxidative) during IR-stress in cold-stored murine OLTs. The following three specific aims are proposed: Aim 1: Determine whether downregulated macrophage YAP expression is a risk factor for developing PGD in cold-stored murine OLTs? Based on preliminary data, genetically modified mice strain (YAPM-KO) will be used to evaluate pre-OLT and post-OLT YAP expression on residential and circulation macrophage in relation to PGD events in a clinically relevant mouse model of extended hepatic cold storage followed by OLT. Aim 2: Define regulatory mechanisms by which YAP signaling controls immunity-related GTPase family M member 1 (IRGM1)-dependent macrophage inflammation in IR-stressed OLTs. Hypothesis: YAP activation modulates macrophage IRGM1 in liver IRI by coupling cyclic GMP-AMP synthase (cGAS) and interferon regulatory factor 3 (IRF3), which in turn mitigates liver IRI by diminishing innate pro-inflammatory responses. YAP-mediated IRGM1 deprivation will be screened in IRI-OLTs. Then, a new model of liver IRI in CD11b-DTR mice in which adoptive transfer of distinctive macrophage populations (after conditional depletion of native CD11b+ cells) will be utilized to dissect YAP–IRGM1 cross-regulation in hepatic IR-inflammation. Aim 3: Define molecular mechanisms by which YAP controls IRGM1-dependent macrophage autophagy in IR-stressed OLTs. Hypothesis: YAP deficiency augments IRGM1-mediated macrophage autophagy and leads into hepatocellular deterioration in IRI-OLT. YAP–IRGM1 mediated major autophagosomal formation in governing macrophage M1/M2 balance, and the biological significance and mechanisms of IRGM1-derived macrophage polarization will be assessed in IR-stressed YAPnull OLTs.

项目摘要/摘要 肝缺血再灌注损伤(IRI)仍然是肝移植、肝切除、肝移植、肝切除、肝移植、肝移植、肝切除、肝移植、肝移植、肝切除、肝移植等的常见问题。 和震惊。因为它经常导致原发性移植物功能障碍(PGD)，晚期慢性排斥反应，并有助于 可供移植的供体器官短缺，肝脏IRI是迄今最具挑战性的问题之一 临床原位肝移植(OLT)并发症研究不足。AS固有的YAP表达式 YAP激活剂治疗的人原位肝移植与肝功能和组织损伤呈负相关 抑制巨噬细胞功能和提高肝细胞保护作用改善小鼠肝脏IRI 非移植肝“温热”IRI模型。在这里，不同的YAP信号通路的功能被提出 肝移植后肝冷缺血时间延长的临床相关小鼠模型的研究。假设： 移植物围手术期YAP表达下调是移植物损伤和PGD发生的重要驱动因素。 YAP在移植肝中的表达降低可能具有预后和治疗意义。蜂窝 YAP的特殊分布，促使人们提出了YAP在巨噬细胞(先天)发出信号的解剖 免疫)调节炎症(致病)、自噬重编程(稳态)和细胞保护 (抗氧化)在IR应激的冷藏小鼠肝移植。提出了以下三个具体目标： 目的1：确定巨噬细胞YAP表达下调是否为发病的危险因素 冷藏小鼠肝移植中的前列腺素D？根据初步数据，转基因小鼠品系(YAPM-KO)将 用来评估移植前和移植后居民和循环巨噬细胞YAP表达的关系 在临床相关的延长肝脏冷藏继之原位肝移植的小鼠模型中发生PGD事件。 目标2：确定YAP信号控制免疫相关GTP酶家族的调控机制 IR应激性OLTS中M成员1(IRGM1)依赖的巨噬细胞炎症。假设：YAP 活化通过偶联cGMP-AMP合成酶(CGAS)调节肝脏IRI中巨噬细胞IRGM1 干扰素调节因子3(IRF3)，它通过减少固有的促炎作用而减轻肝脏IRI 回应。YAP介导的IRGM1缺失将在IRI-OLTS中进行筛查。然后，建立了一种新的肝脏IRI模型 CD11b-DTR小鼠过继转移特殊的巨噬细胞群(条件衰竭后 自然CD11b+细胞)将被用来剖析YAP-IRGM1在肝脏IR炎症中的交叉调节。 目的3：确定YAP控制依赖IRGM1的巨噬细胞自噬的分子机制 在IR应激的OLTS中。假设：YAP缺乏增强IRGM1介导的巨噬细胞自噬和 导致IRI-OLT中的肝细胞恶化。YAP-IRGM1介导的主要自噬小体形成 调节巨噬细胞M1/M2平衡及IRGM1来源的生物学意义和机制 巨噬细胞极化将在IR应激的YAP零OLTS中进行评估。