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 肝缺血和再灌注损伤（IRI）仍然是肝移植，肝切除， 和震惊。由于它通常会导致原发性移植功能障碍（PGD），慢性拒绝晚期，并有助于 缺乏可用于移植的供体器官，肝脏IRI是迄今为止最挑战的之一 临床原位肝移植（OLT）的并发症正在研究。作为固有的YAP表达 与人类OLT中的肝功能和组织损伤负相关，用YAP激活剂处理 通过降低巨噬细胞功能并改善肝细胞保护来改善肝脏IRI 肝“温暖” IRI的非移植模型。在这里，提出了不同的YAP信号通路的功能 在临床相关的长时间肝冷缺血的临床相关小鼠模型中进行的研究，然后是OLT。假设： 下调移植的YAP表达是移植损伤和PGD发展的重要驱动力， 肝移植中的YAP表达降低可能具有预后和治疗意义。细胞 特定的YAP分布已提示提出解剖，该解剖是在巨噬细胞处的yap信号传导（先天性 免疫调节感染（致病），自噬重编程（稳态）和细胞保护作用 （抗氧化）在冷藏的鼠OLT中IR应力期间。提出了以下三个特定目标： 目标1：确定下调巨噬细胞YAP表达是否是发展的危险因素 PGD​​在冷藏的鼠OLT中？基于初步数据，一般修饰的小鼠应变（YAPM-KO）将 用于评估居民对居民和循环巨噬细胞的表达前和施加后的表达 在扩展肝冷藏的临床相关小鼠模型中，pGD事件随后是OLT。 AIM 2：定义YAP信号传导控制与免疫史的GTPase家族的调节机制 M成员1（IRGM1）依赖性巨噬细胞注射在IR胁迫的OLT中。假设：yap 激活通过耦合循环GMP-AMP合酶（CGA）和 干扰素调节因子3（IRF3），反过来通过减少先天促炎性来减轻肝脏IRI 回答。 YAP介导的IRGM1剥夺将以Iri-olts进行筛选。然后，一种新的肝脏IRI模型 CD11B-DTR小鼠在其中自适应转移了独特的巨噬细胞种群（有条件地耗尽了 天然CD11b+细胞）将用于剖析肝IR炎症中的YAP-IRGM1交叉调节。 AIM 3：定义YAP控制IRGM1依赖性巨噬细胞自噬的分子机制 在IR胁迫的OLT中。假设：YAP缺乏增加IRGM1介导的巨噬细胞自噬和 导致Iri-olt中的肝细胞定义。 YAP – IRGM1介导的主要自噬体形成 管理巨噬细胞M1/M2平衡以及IRGM1衍生的生物识别意义和机制 巨噬细胞极化将在IR胁迫的Yapnull OLT中进行评估。