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”。我们报道说肝癌CeCAM1（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OLT损伤。 CC1 mRNA经历替代剪接 （AS）生成剪接变体，其特征是外显子的包含（CC1-L; Long）或排除（CC1-S; Short） 7。假设：肝细胞CC1-S充当细胞保护前哨，而CC1-L在OLT渗透受体中 - 衍生的中性粒细胞，调节肝脏IR炎症并微调其分辨率。 AIM 1：描述肝CC1-S同工型促进IR-的细胞保护的机制 压力小鼠OLT。假设：反义低聚物形态（MOS） - 肝CC1 在HIF-1α的控制下，在供体小鼠肝脏中生成CC1-S同工型，刺激细胞保护 通过阻止P-p38（在冷ir应力下），同时促进GPX4并靶向铁铁作用（在温暖的IR压力下）。 在这里，我们将确定1.1：cc1- as是否会加速急性中的肝恢复受损 IR炎症的分辨率阶段。 1.2：CC1-S通过控制肝细胞死亡途径来控制iri-olt。 1.3：CC1-S介导的细胞保护受到HIF-1α信号传导的控制，在温暖的肝脏IR应力下。 AIM 2：定义中性粒细胞CC1-L同工型施加抗炎和的机制 IR压力小鼠OLT中的细胞保护功能。假设：接受者CC1-L中性粒细胞抵消急性 iri-olt，并通过策划N1/N2极化来刺激注射分辨率，从而导致肝脏 稳态/再生重塑。我们将研究2.1：受体CC1-L缺乏免疫细胞 加剧肝iri-olt。 2.2：CC1-L对N2中性粒细胞两极化，以促进抗炎表型/ 改善OLT结果。 2.3：CC1-L熟练的中性粒细胞极化M2巨噬细胞/促进肝自噬。 AIM 3：阐明肝CC1-S同工型恢复原状的机制 受到常规机器保存（NMP）的约束。假设：NMP，补充了HIF-1α - CC1- S轴修饰符，通过减轻促进性铁的作用来改善被丢弃的人类生命的功能 自噬。我们将将新兴的NMP策略与辅助CC1干预结合在一起，以评估是否是否 3.1：MOS结构的人源化CC1转基因小鼠（HUCC1-TG）模仿CC1-AS对肝脏IR-的影响 正常小鼠的压力，但可以翻译成人肝。 3.2：CC1-S同工型与HIF-1α协同作用 改善肝功能。 3.3：CC1-S与HIF-1α协同作用，以增强人类生命中的细胞保护作用。 与PPG集成：项目1完成研究评估稳态机制 项目2中的iri-olt的决议。项目1中的目标1-2将通过筛选人类OLT活检来验证 在项目3中加速评估临床先天免疫表型。项目1取决于核心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）。