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：
10545046
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/10545046
关键词：
Ablation 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 Inhibition of Apoptosis Knock-out Link Lysine Mediating Mitochondria Molecular Molecular Target Mus Mutation Myocardial Myocardial Ischemia Necrosis Necrosis Induction Oxidative Stress Induction Pathogenesis Pathologic Pathway interactions Phosphotransferases Physiological Pilot Projects Play Polyubiquitin Polyubiquitination Process Proto-Oncogene Proteins c-akt Regulation Role Signal Transduction Signaling Protein Skin Neoplasms TNF receptor-associated factor 2 Testing Therapeutic Transgenic Mice Transgenic Model Tumor Suppressor Proteins Ubiquitination autosome cardioprotection functional loss gain of function in vivo ischemic injury loss of function mouse model new therapeutic target novel overexpression prevent protein transport receptor small hairpin RNA therapeutic target ubiquitin isopeptidase 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.

项目摘要/摘要心脏细胞死亡和/或坏死是心脏缺血性损伤，病理重塑的标志和最终心力衰竭。与凋亡相反，坏死在心脏病发病机理中的作用新兴证据已经确定了几种形式的“编程坏死”，这样由于死亡受体介导的坏死（称为“坏死”），线粒体介导的坏死和氧化性应力诱导的坏死。如何在心脏中调节编程坏死仍在很大程度上仍然未知，并且预防坏死仍然是一个重要的挑战。此外，目前尚无分子策略类似地靶向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（cylindromatisos）的意外作用，赖氨酸63（K63）特异性去泛素酶，作为关键调节剂心肌细胞中的多个细胞死亡途径，包括凋亡，坏死和氧化应激诱导坏死。有趣的是，在缺血性损伤后心脏中明显更新CYLD表达。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消融减弱，而CYLD的过度表达加剧，心脏缺血性损伤。重要的是，消融 CYLD抑制心肌细胞的凋亡，坏死和坏死，而CYLD的过表达显示了相反的效果。从机械上讲，我们的数据揭示了K63连接的多泛素化（K63-UB）依赖 CYLD控制三个细胞死亡的泛素化状态和活动的细胞死亡信号传导机制调节器：TRAF2，TAK1和AKT/PKB。因此，我们假设去泛素酶CYLD是关键调节剂多个细胞死亡途径以及心脏缺血性损伤和重塑的有前途的治疗靶标。使用遗传损失和功能障碍策略，我们将解决两个具体目标：目标1） CYLD作为心肌细胞死亡，缺血性损伤和使用CYLD重塑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 依赖细胞死亡信号传导网络调节心肌细胞中的细胞凋亡，坏死和坏死。这项目研究了一个新型的CYLD介导的细胞死亡信号网络及其功能相关性在心脏缺血性损伤和重塑中。此外，拟议的研究将定义K63-UB依赖性调节细胞凋亡，坏死和坏死的机制，构成了细胞死亡的新范式规定。这些研究还通过提供新的抗细胞死亡具有重要的翻译意义鉴于我们的初步结果揭示了CYLD是多个细胞死亡过程的分子靶标的策略。