THE RELAXIN RECEPTOR GR/RXFP1 SIGNALING IN LIVER TRANSPLANT ISCHEMIA-REPERFUSION INJURY AND THE INFLAMMATION RESOLUTION

松弛素受体 GR/RXFP1 信号在肝移植缺血再灌注损伤和炎症消退中的作用

• 批准号: 10268216
• 负责人: Jerzy W Kupiec-Weglinski
• 金额: $ 39万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-22 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10268216
• 关键词: Acute; Affect; Agonist; Anti-Inflammatory Agents; Antiinflammatory Effect; Attenuated; Autophagocytosis; Binding; Biological; Cell physiology; Cells; Chronic; Clinical; Epidemic; Formulation; Genes; Glucocorticoid Receptor; Glucocorticoids; Goals; Gold; Heart failure; Hepatic; Hepatic Tissue; Hepatocyte; Hepatology; Homeostasis; Human; Immune; Inflammation; Inflammatory; Intervention; Knock-in Mouse; Link; Liver; Liver Fibrosis; Liver diseases; Liver neoplasms; Macrophage Activation; Molecular; Mus; Organ; Organ Donor; Organ Transplantation; Outcome; Pathology; Pathway interactions; Patients; Perfusion; Pharmacology; Phase; Phase III Clinical Trials; Phenotype; Pilot Projects; Primary carcinoma of the liver cells; Property; Publishing; Receptor Signaling; Recombinants; Regulation; Rejuvenation; Relaxin; Reperfusion Injury; Resistance; Resolution; Role; SIRT1 gene; Severities; Signal Transduction; Solid; Somatomedins; Stress; System; T-Lymphocyte; Testing; Tissues; Transplant Recipients; Transplantation; alternative treatment; chronic liver disease; editorial; experimental study; glucocorticoid-induced orphan receptor; immunoregulation; improved; innovation; insight; liver ischemia; liver transplantation; macrophage; mimetics; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; peptide hormone; preservation; prevent; receptor; regeneration function; relating to nervous system; relaxin receptor; response; side effect; small molecule; standard of care; success; tissue injury; tool; transplant model

PROJECT SUMMARY/ABSTRACT Hepatic ischemia/reperfusion injury (IRI), an innate immune-driven inflammation response, is a major obstacle limiting the success of orthotopic liver transplantation (OLT) in patients with end-stage liver disease and those with tumors of hepatic origin. Although significant progress has been made in better appreciation of the liver inflammatory cascade by IR-stress, much less is known about its resolution, which may affect not only the severity of tissue injury itself but also, more importantly, the long-term outcomes. Recent studies document striking cytoprotective functions of hrRLX (recombinant human relaxin-2) against IR-stress in mouse OLT models via hepatocyte glucocorticoid receptor (GR) signaling; while polarizing macrophage activation via Notch1 promoted IRI-OLT resistance. These experimental findings, supported by a clinical evidence of enhanced GR/Notch1 phenotype needed for IRI resistance in human OLT, prompted to propose that rhRLX may function as a novel GR agonist and glucocorticoid (GC) mimetic in liver transplantation. Pilot studies also point to anti- fibrotic functions of the cognate RXFP1 receptor, a GR-independent RLX-2 binding partner. As the conventional murine OLT model offers a limited translational utility, this project will be dissecting GR – RXFP1 molecular interplay in mice expressing human RXFP1 gene; as well as testing new concepts of RXFP1-driven hepatic rejuvenation of discarded human livers during hypothermic machine preservation. A newly discovered divergent role of GR–RXFP1 signaling axis in the “acute” and “resolution” phase of IRI-OLT inflammation, has prompted to put forth a novel and heretofore untested overall hypothesis, that: 1/ pharmacological rhRLX-induced GR enhancement will rescue OLT from acute IR-insult; while 2/ harnessing GR-independent rhRLX signaling via its cognate receptor, RXFP1, will promote homeostatic/anti-fibrotic functions in the inflammation resolution phase. Two interlocked aims explore this hypothesis: Aim 1: Delineate molecular mechanisms of rhRLX – GR hepatocellular protection in OLT (acute IRI-inflammation phase). Aim 1.1: Test hypothesis that hepatocellular Keap1-dependent Nrf2 signaling is indispensable for rhRLX – GR axis to prevent DAMPs release and innate inflammation in cold-stored donor livers. Aim 1.2: Test hypothesis that SIRT1 enhances GR-induced hepatocyte regenerative functions/autophagy in IR-stressed OLT. Aim 2. Delineate molecular mechanisms of Notch1 / RXFP1 anti-fibrotic functions in OLT (IRI-inflammation “resolution” phase). Aim 2.1: Test hypothesis that Notch1 (macrophage) – RXFP1 (T cell) cross-regulation is essential in the resolution of IRI – OLT inflammation. Aim 2.2: Test hypothesis that the activation of human RLX receptor exerts anti-fibrotic functions in the resolution of IRI-OLT inflammation in humanized RXFP1 “knockin” mouse system. Aim 2.3: Test hypothesis that activation of RXFP1 receptor during hypothermic machine preservation attenuates inflammation and rejuvenates discarded human donor livers.

项目概要/摘要 肝缺血/再灌注损伤（IRI）是一种先天免疫驱动的炎症反应，是一个主要障碍 限制了终末期肝病患者和那些患者的原位肝移植（OLT）的成功 尽管在更好地了解肝脏方面已经取得了重大进展。 IR应激引起的炎症级联反应，对其解决方案知之甚少，这可能不仅影响 组织损伤本身的严重程度，更重要的是，最近的研究记录了长期结果。 hrRLX（重组人松弛素-2）在小鼠 OLT 模型中对 IR 应激具有显着的细胞保护功能 通过肝细胞糖皮质激素受体 (GR) 信号传导，同时通过 Notch1 极化巨噬细胞激活； 促进了 IRI-OLT 耐药性，这些实验结果得到了增强的临床证据的支持。 人类 OLT 中 IRI 抗性所需的 GR/Notch1 表型提示，表明 rhRLX 可能发挥作用 作为肝移植中的新型 GR 激动剂和糖皮质激素 (GC) 模拟物，初步研究也指出了抗- 同源 RXFP1 受体（一种不依赖 GR 的 RLX-2 结合伴侣）的纤维化功能。 小鼠 OLT 模型提供有限的转化效用，该项目将剖析 GR – RXFP1 分子 表达人类 RXFP1 基因的小鼠中的相互作用以及测试 RXFP1 驱动的肝脏的新概念 低温机器保存过程中废弃人类肝脏的复兴新发现的分歧。 GR–RXFP1 信号轴在 IRI-OLT 炎症“急性”和“消退”阶段的作用，提示 提出一个新颖且迄今为止未经检验的总体假设，即： 1/ 药理学 rhRLX 诱导的 GR 增强将拯救 OLT 免受急性 IR 损伤；同时 2/ 通过其利用 GR 独立的 rhRLX 信号传导 同源受体 RXFP1 将在炎症消退阶段促进稳态/抗纤维化功能。 探索这一假设有两个相互关联的目标： 目标 1：描绘 OLT（急性 IRI 炎症）中 rhRLX – GR 肝细胞保护的分子机制 目标 1.1：检验肝细胞 Keap1 依赖性 Nrf2 信号传导对于 rhRLX 不可或缺的假设 – GR 轴防止冷藏供体肝脏中 DAMP 释放和先天性炎症。目标 1.2：测试。 假设 SIRT1 在 IR 应激的 OLT 中增强 GR 诱导的肝细胞再生功能/自噬。 目标 2. 描绘 OLT 中 Notch1 / RXFP1 抗纤维化功能的分子机制（IRI-炎症） “解决”阶段）。目标 2.1：测试 Notch1（巨噬细胞）-RXFP1（T 细胞）交叉调节的假设。 目标 2.2：测试人类 RLX 激活的假设。 受体在人源化RXFP1“敲入”中在IRI-OLT炎症消退中发挥抗纤维化功能 目标 2.3：测试低温机器期间 RXFP1 受体激活的假设。 保存可减轻炎症并使废弃的人类供体肝脏恢复活力。