CEACAM1 Alternative Splicing in Liver Ischemia-Reperfusion Injury

CEACAM1 选择性剪接在肝脏缺血再灌注损伤中的作用

• 批准号: 10622462
• 负责人: Jerzy W Kupiec-Weglinski
• 金额: $ 54.09万
• 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 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10622462
• 关键词: Acceleration; Acute; Address; Advocate; Alternative Splicing; Anti-Inflammatory Agents; Autophagocytosis; Biological Markers; Biometry; Biopsy; Brain Death; Cardiac Death; Cell Culture System; Cell Death; Cells; Clinical; Complement; Complement component C1; Cryopreservation; Cytoplasm; Cytoprotection; Data; Exclusion; Exons; Genes; Hepatic; Hepatic Tissue; Hepatocyte; Human; Immune; Impairment; Infiltration; Inflammation; Injury; Intervention; Ischemia; Life; Liver; Lymphocyte; MAP Kinase Gene; Macrophage; Mediating; Messenger RNA; Mucosal Immunity; Mus; Myelogenous; Natural Immunity; Natural regeneration; Operative Surgical Procedures; Organ; Organ Donor; Organ Preservation; Organ Transplantation; Outcome; Pathway interactions; Patients; Phase; Phenotype; Protein Isoforms; RNA Splicing; Recovery; Regulation; Rejuvenation; Reperfusion Injury; Reporting; Resistance; Resolution; Sentinel; Signal Transduction; Sterility; Stress; Tissues; Transgenic Mice; Transplantation; Transplantation Surgery; Variant; Waiting Lists; Warm Ischemia; carcinoembryonic antigen-related cell adhesion molecules; clinically relevant; end stage liver disease; experimental study; functional improvement; immunoregulation; improved; insight; liver function; liver inflammation; liver ischemia; liver transplantation; mortality; mouse model; neutrophil; novel; novel therapeutics; p38 Mitogen Activated Protein Kinase; preservation; prevent; programs; regenerative; repaired; screening; standard of care; success; synergism; transplant model

PROJECT 1 – SUMMARY/ABSTRACT Project 1 competitive renewal addresses the unmet clinical and scientific needs to enhance donor liver supply through organ “rejuvenation.” We reported that hepatic CEACAM1 (encoded by CC1 gene; CD66a) dictates donor liver quality, and prevents early OLT injury in mice and humans. CC1 mRNA undergoes alternative splicing (AS) to generate splice variants, characterized by the inclusion (CC1-L; long) or exclusion (CC1-S; short) of exon 7. Hypothesis: Hepatocyte CC1-S functions as a cytoprotective sentinel, while CC1-L in OLT-infiltrating recipient- derived neutrophils, regulates liver IR-inflammation and fine-tunes its resolution. Aim 1: Delineate mechanisms by which hepatic CC1-S isoform promotes cellular protection in IR- stressed mouse OLT. Hypothesis: Antisense oligomer morpholinos (MOs)-enforced AS of hepatic CC1 generates CC1-S isoform in the donor mouse liver, which under the control of HIF-1α, stimulates cytoprotection by blocking p-p38 (under cold IR-stress) while promoting GPX4 and targeting ferroptosis (under warm IR-stress). Here, we will ascertain whether 1.1: CC1-AS accelerates otherwise impaired hepatic recovery in the acute and the resolution phase of IR-inflammation. 1.2: CC1-S controls IRI-OLT by regulating hepatic cell death pathways. 1.3: CC1-S-mediated cytoprotection is controlled by HIF-1α signaling under warm vs. cold hepatic IR-stress. Aim 2: Define mechanisms by which neutrophil CC1-L isoform exerts anti-inflammatory and cytoprotective functions in IR-stressed mouse OLT. Hypothesis: Recipient CC1-L neutrophils counteract acute IRI-OLT, and stimulate inflammation resolution by orchestrating N1/N2 polarization, with resultant liver homeostatic/regenerative remodeling. We will study whether 2.1: Recipient CC1-L deficient immune cells exacerbate hepatic IRI-OLT. 2.2: CC1-L polarizes N2 neutrophils to promote an anti-inflammatory phenotype/ improve OLT outcomes. 2.3: CC1-L proficient neutrophils polarize M2 macrophages/promote hepatic autophagy. Aim 3: Elucidate mechanisms by which hepatic CC1-S isoform rejuvenates discarded human livers subjected to normothermic machine preservation (NMP). Hypothesis: NMP, supplemented with HIF-1α – CC1- S axis modifiers, improves the function of discarded human livers by mitigating ferroptosis while promoting autophagy. We will incorporate the emerging NMP strategy with adjunctive CC1-intervention to assess whether 3.1: MOs-conditioned humanized CC1 transgenic mice (huCC1-Tg) mimic the effects of CC1-AS upon liver IR- stress in normal mice but be translatable to the human liver. 3.2: CC1-S isoform synergizes with HIF-1α to improve liver function. 3.3: CC1-S synergizes with HIF-1α to enhance cytoprotection in human livers. Integration with PPG: Project 1 complements studies assessing homeostatic mechanisms in the resolution of IRI-OLT in Project 2. Aim 1-2 in Project 1 will be validated by the screening of human OLT biopsies to accelerate assessments of clinical innate immune phenotypes in Project 3. Project 1 is dependent on Core A (Administrative), Core B (Mouse and Human OLT Surgery), and Core C (Computational and Biostatistics).

项目1--摘要/摘要 项目1竞争性更新解决了未得到满足的临床和科学需求，以增加供体肝脏的供应 通过器官“返老还童”。我们报道了肝脏CEACAM1(由CC1基因编码；CD66a)决定了 供体肝脏质量，并防止小鼠和人类的早期原位肝移植损伤。CC1基因经历选择性剪接 (AS)产生剪接变体，特征是外显子的包含(CC1-L；长)或排除(CC1-S；短) 7.假设：肝细胞CC1-S起细胞保护作用，而CC1-L在原位肝移植浸润性受体中起细胞保护作用。 衍生的中性粒细胞，调节肝脏IR炎症并微调其分辨率。 目的1：探讨肝脏CC1-S亚型促进细胞保护的机制。 应激小鼠原位肝移植。假设：反义寡聚吗啡(MOS)--作为肝脏CC1的增强因子 在供体小鼠肝脏中产生CC1-S亚型，在HIF-1α的控制下，刺激细胞保护 通过阻断p-p38(在冷IR应激下)，同时促进Gpx4和靶向铁下垂(在热IR应激下)。 在这里，我们将确定1.1：CC1-AS是否加速了急性和慢性肝炎患者原本受损的肝脏恢复。 IR炎症消退阶段。CC1-S通过调节肝细胞死亡途径控制IRI-OLT。 CC1-S介导的细胞保护作用受HIF-1α信号调控。 目的2：明确中性粒细胞CC1-L异构体抗炎作用的机制。 IR应激小鼠原位肝移植的细胞保护作用。假设：受体CC1-L中性粒细胞中和急性 IRI-OLT，并通过协调N1/N2极化刺激炎症消退，并产生肝脏 动态平衡/再生重塑。我们将研究2.1：受体CC1-L是否存在免疫细胞缺陷 加重肝脏IRI-OLT。2.2：CC1-L极化中性粒细胞促进抗炎表型/ 改善OLT结果。L熟练的中性粒细胞极化M2巨噬细胞/促进肝脏自噬。 目的3：阐明肝CC1-S亚型使废弃的人肝脏恢复活力的机制 进行常温机器保存(NMP)。假设：NMP，补充HIF-1α-CC1- S轴修饰剂，通过在促进的同时缓解铁下垂来改善废弃人体肝脏的功能 自噬。我们将把新兴的国家管理计划战略与辅助的CC1干预结合起来，以评估 3.1：MOS条件化人源化CC1转基因小鼠(huCC1-TG)模拟CC1-AS对肝脏IR的影响 应激在正常小鼠，但可翻译到人的肝脏。CC1-S亚型与HIF-1α协同作用 改善肝功能。3.3CC1-S与HIF-1α协同作用增强人肝脏细胞保护作用。 与PPG的整合：项目1补充了评估体内平衡机制的研究 项目2中IRI-OLT的解析。项目1中的目标1-2将通过对人类OLT活检的筛选来验证 在项目3中加速临床天然免疫表型的评估。项目1依赖于核心A (行政)、核心B(老鼠和人类原位移植手术)和核心C(计算和生物统计学)。