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

基本信息

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
项目类别：
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

项目摘要

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满足的临床和科学需求，以增强供体肝脏供应通过器官“复兴”。我们报道了肝脏 CEACAM1（由 CC1 基因编码；CD66a）决定供体肝脏质量，并防止小鼠和人类的早期 OLT 损伤 CC1 mRNA 发生选择性剪接。 (AS) 生成剪接变体，其特征是包含（CC1-L；长）或排除（CC1-S；短）外显子 7. 假设：肝细胞 CC1-S 充当细胞保护哨兵，而 OLT 浸润受体中的 CC1-L 衍生的中性粒细胞，调节肝脏红外炎症并微调其分辨率。目标 1：描述肝脏 CC1-S 亚型促进 IR- 细胞保护的机制应激小鼠 OLT 假设：反义寡聚体吗啉代 (MO) 强制肝 CC1 的 AS 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（在冷红外应激下），同时促进 GPX4 并靶向铁死亡（在热红外应激下）。在这里，我们将确定 1.1：CC1-AS 是否会加速急性和急性期受损的肝功能恢复。 IR炎症消退阶段1.2：CC1-S通过调节肝细胞死亡途径控制IRI-OLT。 1.3：CC1-S 介导的细胞保护作用在热肝 IR 应激与冷肝 IR 应激下受 HIF-1α 信号传导控制。目标 2：定义中性粒细胞 CC1-L 亚型发挥抗炎和抗炎作用的机制 IR 应激小鼠 OLT 中的细胞保护功能假设：受体 CC1-L 中性粒细胞对抗急性。 IRI-OLT，通过协调 N1/N2 极化刺激炎症消退，从而改善肝脏我们将研究2.1：受体CC1-L缺陷的免疫细胞是否存在稳态/再生重塑。加剧肝脏 IRI-OLT 2.2：CC1-L 极化 N2 中性粒细胞以促进抗炎表型/ 2.3：CC1-L 熟练的中性粒细胞极化 M2 巨噬细胞/促进肝自噬。目标 3：阐明肝脏 CC1-S 亚型使废弃人类肝脏恢复活力的机制进行常温机器保存 (NMP) 假设：NMP，辅以 HIF-1α – CC1-。 S轴调节剂，通过减轻铁死亡同时促进废弃人类肝脏的功能我们将把新兴的 NMP 策略与辅助 CC1 干预结合起来，以评估是否存在自噬。 3.1: MOs 条件下的人源化 CC1 转基因小鼠 (huCC1-Tg) 模拟 CC1-AS 对肝脏 IR 的影响正常小鼠的压力，但可转化为人肝脏 3.2：CC1-S 亚型与 HIF-1α 协同作用。 3.3：CC1-S与HIF-1α协同作用，增强人体肝脏的细胞保护作用。与 PPG 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 的分辨率。项目 1 中的目标 1-2 将通过人类 OLT 活检筛查进行验证评估项目 3 中的临床先天免疫表型。项目 1 依赖于核心 A （行政）、核心 B（小鼠和人类 OLT 手术）和核心 C（计算和生物统计学）。