Innate-Adaptive Immune Interface in Liver Ischemia-Reperfusion Injury

肝脏缺血再灌注损伤中的先天适应性免疫界面

• 批准号: 9975698
• 负责人: Jerzy W Kupiec-Weglinski
• 金额: $ 38.23万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9975698
• 关键词: Adoptive Transfer; Affect; Apoptotic; Autophagocytosis; Biological; Biometry; Bone Marrow; CD4 Positive T Lymphocytes; CEACAM1; Cell Death; Cells; Cessation of life; Coculture Techniques; Communication; Cryopreservation; Cyclin D1; Cytoprotection; Data; Development; Environment; Functional disorder; Galectin 3; Gene Targeting; Genes; HMGB1 gene; Hepatic; Hepatocellular Damage; Hepatocyte; Homeostasis; Human; Hydrogen Peroxide; Immune; Immunity; Immunoglobulins; Immunologics; Impairment; In Vitro; Inflammation; Knockout Mice; Ligands; Link; Liver; Liver diseases; Liver neoplasms; Metabolic; Microsurgery; Mouse Strains; Mus; Nuclear; Organ Donor; Organ Transplantation; Organ failure; PPBP gene; Pathway interactions; Patients; Phenotype; Phosphorylation; Process; Regulation; Reperfusion Injury; Reporting; Research; Resistance; Role; Signal Transduction; Site; Sterility; Stress; System; T cell regulation; T-Cell Activation; T-Lymphocyte; TLR4 gene; Testing; Tissues; Tumor Immunity; Warm Ischemia; beta catenin; carcinoembryonic antigen-related cell adhesion molecules; clinically relevant; direct application; exhaust; exhaustion; experimental study; gain of function; hepatocellular injury; immune activation; immunoregulation; in vivo; insight; liver ischemia; liver transplantation; loss of function; macrophage; migration; mouse model; mutant; novel; null mutation; overexpression; programs; regenerative; response; success; transplant model

PROJECT I - SUMMARY/ABSTRACT Ischemia-reperfusion injury (IRI) remains the primary obstacle limiting the success of orthotopic liver transplantation (OLT) in patients with end-stage liver disease and those with tumors of hepatic origin. Our group has pioneered the concept that hepatic IRI requires activated CD4+ T cells to facilitate liver tissue damage. T cell immunoglobulin-3 (TIM-3; encoded by Havcr2 gene) is the central negative regulator of T cell activation. CEACAM1 (carcinoembryonic antigen-related cell adhesion molecule 1; encoded by CC1 gene) has been identified as a new cellular ligand determining TIM-3 function. First, we found that compared with CEACAM1 proficient (WT) livers, CEACAM1 null-mutation (CC1-/-) exacerbated IRI in OLT. Second, we discovered that the benefit of recipient CD4+TIM-3+ signaling in IR-stressed OLT (WT→TIM-3Tg) was completely lost when CEACAM1 KO mice served as organ donors (CC1-/-→TIM-3Tg). These preliminary data have led us to central hypothesis that 1/ TIM-3 – CEACAM1 negative regulation is essential to control IRI by imposing exhaustion-like dysfunction in OLT-infiltrating CD4+ T cells; and 2/ CEACAM1 in the donor liver promotes hepatoprotection. Project I will test this hypothesis through two interlocked specific aims: Aim 1: Define mechanisms of TIM-3 – CEACAM1 negative T cell regulation in IR-stressed iso-OLT. A panel of mice available to us for Aim 1 experiments include CD4+ T cell mutants, which are: i/ CEACAM1Tg; ii/ double TIM-3Tg and CEACAM1-/-; as well as: iii/ TIM-3Tg and TIM-3-/- mice. Aim 1.1. Hypothesis: CEACAM1 - TIM-3 signaling on host circulating CD4+ T cells promotes exhaustion- type phenotype in IRI–OLT. Aim 1.2. Hypothesis: Under dominant CAECAM1 signaling, TIM-3+CD4+ exhausted T cells inhibit the development and progression of IRI in OLT. Aim 2: Define mechanisms by which hepatocellular CEACAM1 in donor liver regulates IRI in iso-OLT. Gene- targeted strains for Aim 2 studies include: i/ hepatic CEACAM1 inactivation (loss-of- function; L-CC1-/-); or ii/ forced hepatic CEACAM1 overexpression (gain-of-function; L-CC1Tg). Aim 2.1. Hypothesis: Enhancement of hepatocyte-specific CEACAM1 – β-catenin regenerative functions facilitates hepatoprotection. Aim 2.2. Hypothesis: TIM-3 – CECACAM1 signaling enhances hepatocyte autophagy program. Integration with PPG: By providing novel insights into TIM-3 – CEACAM1 checkpoint regulation at the innate – adaptive immune interface in IR-stressed iso-OLT, Project I naturally informs/precedes studies assessing how host rejection regulates innate immune activation/IRI sequel in allo-OLT (Project II). Direct application of approaches blunting inflammation while promoting hepatoprotection in mouse OLT models will accelerate assessments of immune phenotypes in human liver transplants (Project III).

项目I -概要/摘要 缺血再灌注损伤（IRI）是限制原位肝移植成功的主要障碍 在终末期肝病和肝源性肿瘤患者中进行原位肝移植（奥尔特）。我们 一个小组率先提出了肝脏IRI需要活化的CD 4 + T细胞来促进肝脏组织 损害T细胞免疫球蛋白-3（TIM-3，由Havcr 2基因编码）是T细胞免疫的中枢负性调节因子， activation. CEACAM 1（癌胚抗原相关细胞粘附分子1;由CC 1基因编码）具有 被鉴定为决定TIM-3功能的新的细胞配体。首先，我们发现， CEACAM 1基因突变（WT）肝细胞中，CEACAM 1基因缺失突变（CC 1-/-）加重了奥尔特中的IRI。二是 发现受体CD 4 +TIM-3+信号传导在IR应激奥尔特（WT→TIM-3 Tg）中的益处是 当CEACAM 1 KO小鼠用作器官供体时（CC 1-/-→TIM-3 Tg）完全丢失。这些初步数据 已经引导我们提出了一个中心假设，即1/ TIM-3 -CEACAM 1负调节对于通过以下途径控制IRI是必不可少的： 在OLT浸润的CD 4 + T细胞中施加衰竭样功能障碍;以及在供体肝脏中2/CEACAM 1 促进肝脏保护。项目I将通过两个相互关联的具体目标来检验这一假设： 目的1：明确IR应激iso-OLT中TIM-3 -CEACAM 1负性T细胞调节的机制。一组 可供我们用于目标1实验的小鼠包括CD 4 + T细胞突变体，它们是：i/CEACAM 1 Tg; ii/双 TIM-3 Tg和CEACAM 1-/-;以及：iii/ TIM-3 Tg和TIM-3-/-小鼠。 目标1.1。假设：宿主循环CD 4 + T细胞上的CEACAM 1- TIM-3信号传导促进耗竭- IRI-OLT的表型。 目标1.2。假设：在显性CAECAM 1信号传导下，TIM-3+ CD 4+耗竭的T细胞抑制CAECAM 1的表达。 IRI在奥尔特中的发展和进展。 目的2：明确供体肝细胞CEACAM 1调控同种异体原位肝移植IRI的机制。吉恩 目标2研究的靶向菌株包括：i/肝CEACAM 1失活（功能丧失; L-CC 1-/-）;或ii/ 强迫性肝CEACAM 1过表达（功能获得性; L-CC 1 Tg）。 目标2.1。假设：增强肝细胞特异性CEACAM 1- β-catenin再生功能 促进肝脏保护。 目标2.2。假设：TIM-3 -CECACAM 1信号转导增强肝细胞自噬程序。 与PPG的整合：通过提供对TIM-3 -CEACAM 1检查点调控的新见解， IR应激iso-OLT中的先天-适应性免疫界面，项目I自然通知/先于研究 评估宿主排斥如何调节allo-OLT中的先天免疫激活/IRI后遗症（项目II）。直接 在小鼠奥尔特模型中应用减轻炎症同时促进肝脏保护的方法， 加速评估人类肝脏移植中的免疫表型（项目III）。