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 - 摘要/摘要 缺血再灌注损伤（IRI）仍然是限制原位肝脏成功的主要障碍 终末期肝病和肝源性肿瘤患者的移植（OLT）。 研究小组率先提出了肝脏 IRI 需要激活的 CD4+ T 细胞来促进肝组织的概念 T 细胞免疫球蛋白-3（TIM-3；由 Havcr2 基因编码）是 T 细胞的中央负调节因子。 CEACAM1（癌胚抗原相关细胞粘附分子1；由CC1基因编码）已被激活。 被确定为决定TIM-3功能的新细胞配体首先，我们发现与此相比。 CEACAM1 精通 (WT) 肝脏、CEACAM1 无效突变 (CC1-/-) 加重了 OLT 中的 IRI。 发现受体 CD4+TIM-3+ 信号传导在 IR 应激的 OLT (WT→TIM-3Tg) 中的益处是 当CEACAM1 KO小鼠作为器官供体时（CC1-/-→TIM-3Tg）完全丢失了这些初步数据。 使我们得出中心假设：1/ TIM-3 – CEACAM1 负调节对于通过以下方式控制 IRI 至关重要： 在供体肝脏中的 OLT 浸润性 CD4+ T 细胞和 2/ CEACAM1 中造成衰竭样功能障碍； 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： 目标 1：定义 IR 应激 iso-OLT 中 TIM-3 – CEACAM1 阴性 T 细胞的调节机制。 我们可用于 Aim 1 实验的小鼠包括 CD4+ T 细胞突变体，它们是：i/ CEACAM1Tg； TIM-3Tg 和 CEACAM1-/-；以及：iii/ TIM-3Tg 和 TIM-3-/- 小鼠。 目标 1.1：宿主循环 CD4+ T 细胞上的 CEACAM1 - TIM-3 信号传导促进耗竭 - IRI-OLT 中的类型表型。 目标 1.2 假设：在 CAECAM1 信号传导下，TIM-3+CD4+ 耗尽的 T 细胞抑制 OLT 中 IRI 的发展和进展。 目标 2：确定供体肝脏中肝细胞 CEACAM1 调节 iso-OLT 基因中 IRI 的机制。 Aim 2 研究的目标菌株包括： i/ 肝脏 CEACAM1 失活（功能丧失；L-CC1-/-）；或 ii/ 强迫肝脏 CEACAM1 过度表达（功能获得；L-CC1Tg）。 目标 2.1. 假设：增强肝细胞特异性 CEACAM1 – β-catenin 再生功能 有利于肝脏保护。 目标 2.2. 假设：TIM-3 – CECACAM1 信号增强肝细胞自噬程序。 与 PPG 集成：通过提供对 TIM-3 – CEACAM1 检查点监管的新颖见解 IR 应激 iso-OLT 中的先天-适应性免疫界面，项目 I 自然地通知/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 后果（项目 II）。 在小鼠 OLT 模型中应用抑制炎症同时促进肝脏保护的方法将 加速人类肝脏移植免疫表型的评估（项目 III）。