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The Deubiquitinase CYLD Controls Multiple Cell Death Pathways in the Heart

去泛素酶 CYLD 控制心脏中的多种细胞死亡途径

基本信息

批准号：
10332100
负责人：
Qinghang Liu
金额：
$ 38.88万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10332100
关键词：
Ablation Address Antioxidants Apoptosis Apoptotic Attenuated Benign Cardiac Cardiac Myocytes Cell Death Cell Death Process Cell Death Signaling Process Cell physiology Cessation of life Complex Data Disease Genetic Goals Heart Heart Diseases Heart failure Knock-out Knockout Mice Link Lysine MAP3K7 gene Mediating Mitochondria Molecular Molecular Target Mus Mutation Myocardial Myocardial Ischemia Necrosis Oxidative Stress Pathogenesis Pathologic Pathway interactions Phosphotransferases Physiological Pilot Projects Play Polyubiquitin Polyubiquitination Proto-Oncogene Proteins c-akt Regulation Role Signal Transduction Signaling Protein Skin TNF receptor-associated factor 2 TRAF2 gene Testing Therapeutic Transgenic Mice Transgenic Model Tumor Suppressor Proteins Ubiquitination cardioprotection functional loss gain of function in vivo ischemic injury mouse model new therapeutic target novel overexpression prevent protein transport receptor small hairpin RNA therapeutic target vector

项目摘要

Project Summary/Abstract Cardiac cell death by apoptosis and/or necrosis is a hallmark of cardiac ischemic injury, pathological remodeling, and end-stage heart failure. In contrast to apoptosis, the role of necrosis in the pathogenesis of heart disease has been largely understudied. Emerging evidence has identified several forms of “programmed necrosis”, such as death receptor-mediated necrosis (termed “necroptosis”), mitochondria-mediated necrosis, and oxidative stress-induced necrosis. How programmed necrosis is regulated in the heart remains largely unknown, and preventing necrosis is still an important challenge. Moreover, currently no molecular strategies are available to simultaneously target multiple cell death processes in heart disease. Here, our preliminary studies identified an unexpected role for CYLD (cylindromatosis), a lysine 63 (K63)-specific deubiquitinase, as a key regulator of multiple cell death pathways in cardiomyocytes, including apoptosis, necroptosis, and oxidative stress-induced necrosis. Intriguingly, CYLD expression was markedly upregulated in the heart following ischemic injury. Using Cyld knockout and transgenic mouse models, our preliminary data further show that ablation of CYLD attenuated, whereas overexpression of CYLD exacerbated, cardiac ischemic injury. Importantly, ablation of CYLD inhibited apoptosis, necroptosis, and necrosis in cardiomyocytes, whereas overexpression of CYLD showed the opposite effect. Mechanistically, our data reveal a K63-linked polyubiquitination (K63-Ub) dependent cell death signaling mechanism whereby CYLD controls the ubiquitination status and activity of three cell death regulators: TRAF2, TAK1 and AKT/PKB. Therefore, we hypothesize the deubiquitinase CYLD is a key regulator of multiple cell death pathways and a promising therapeutic target for cardiac ischemic injury and remodeling. Using genetic loss- and gain-of-function strategies, we will address two specific aims: Aim 1) To investigate the novel role of CYLD as a key regulator of myocardial cell death, ischemic injury, and remodeling using Cyld knockout and transgenic models and AAV9-shCYLD vectors. Aim 2) To define a CYLD-mediated, K63-Ub dependent cell death signaling network regulating apoptosis, necroptosis, and necrosis in cardiomyocytes. This project investigates a novel CYLD-mediated cell death signaling network in the heart and its functional relevance in cardiac ischemic injury and remodeling. Moreover, the proposed studies will define a K63-Ub dependent mechanism regulating apoptosis, necroptosis, and necrosis, which constitutes a new paradigm of cell death regulation. These studies also have important translational implications by providing new anti-cell death strategies, given our preliminary results revealing CYLD as a molecular target for multiple cell death processes.

项目概要/摘要心肌细胞因凋亡和/或坏死而死亡是心脏缺血性损伤、病理重塑、和终末期心力衰竭。与细胞凋亡相反，坏死在心脏病发病机制中的作用已被广泛研究。新出现的证据已经确定了几种形式的“程序性坏死”，例如如死亡受体介导的坏死（称为“坏死性凋亡”）、线粒体介导的坏死和氧化应激引起的坏死。心脏中程序性坏死是如何调节的仍然很大程度上未知，并且防止坏死仍然是一个重要的挑战。此外，目前尚无分子策略可用于同时针对心脏病中的多个细胞死亡过程。在这里，我们的初步研究确定了 CYLD（圆柱瘤病）是一种赖氨酸 63 (K63) 特异性去泛素化酶，作为心肌细胞的多种细胞死亡途径，包括细胞凋亡、坏死性凋亡和氧化应激诱导坏死。有趣的是，缺血性损伤后心脏中 CYLD 的表达显着上调。使用 Cyld 敲除和转基因小鼠模型，我们的初步数据进一步表明 CYLD 的消除 CYLD 的过度表达会减轻心脏缺血性损伤，而 CYLD 的过度表达会加剧心脏缺血性损伤。重要的是，消融 CYLD 抑制心肌细胞凋亡、坏死性凋亡和坏死，而 CYLD 过表达表现出相反的效果。从机制上讲，我们的数据揭示了 K63 连接的多泛素化 (K63-Ub) 依赖性细胞死亡信号机制CYLD控制三种细胞死亡的泛素化状态和活性调节因子：TRAF2、TAK1 和 AKT/PKB。因此，我们假设去泛素酶 CYLD 是一个关键的调节因子多种细胞死亡途径的研究和心脏缺血性损伤和重塑的有前途的治疗靶点。使用遗传功能丧失和功能获得策略，我们将实现两个具体目标：目标 1) 研究 CYLD 作为心肌细胞死亡、缺血性损伤和使用 Cyld 进行重构的关键调节剂的新作用敲除和转基因模型以及 AAV9-shCYLD 载体。目标 2) 定义 CYLD 介导的 K63-Ub 依赖的细胞死亡信号网络调节心肌细胞的凋亡、坏死性凋亡和坏死。这该项目研究了心脏中新型 CYLD 介导的细胞死亡信号网络及其功能相关性心脏缺血性损伤和重构。此外，拟议的研究将定义 K63-Ub 依赖性调控细胞凋亡、坏死性凋亡和坏死的机制，构成细胞死亡的新范式规定。这些研究通过提供新的抗细胞死亡也具有重要的转化意义鉴于我们的初步结果表明 CYLD 是多种细胞死亡过程的分子靶点，因此我们采取了一些策略。