A K63-Ubiquitination Dependent Necroptosis Signaling Network in the Heart

心脏中 K63 泛素化依赖性坏死性凋亡信号网络

基本信息

批准号：
10543111
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：
2021
资助国家：
美国
起止时间：
2021-01-08 至 2025-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10543111
关键词：
Ablation Acute Adult Apoptosis Attenuated Biological Cardiac Cardiac Myocytes Caspase Cell Death Signaling Process Cell membrane Cessation of life Complex Data Dilated Cardiomyopathy Disease Enzymes Gene Transfer Genetic Goals Heart Heart Injuries Heart failure Inflammation Investigation Ischemia Link MAP3K7 gene Mediating Mitochondria Modeling Molecular Mus Myocardial Myocardial Ischemia Necrosis Organelles Pathologic Pathway interactions Phosphorylation Physiology Pilot Projects Play Polyubiquitination Process RIPK1 gene RIPK3 gene Regulation Reperfusion Injury Role Signal Transduction Swelling TNF receptor-associated factor 2 TNFRSF1A gene Testing Therapeutic Ubiquitination cardioprotection experimental study gain of function in vivo insight ischemic injury loss of function myocardial injury novel overexpression prevent receptor therapeutic target ubiquitin isopeptidase ubiquitin ligase ubiquitin-protein ligase vector

项目摘要

Project Summary/Abstract Loss of cardiomyocytes by apoptosis and/or necrosis is a hallmark of cardiac ischemic injury, pathological remodeling, and end-stage heart failure. Although necrosis was traditionally regarded as a passive and unregulated process, emerging evidence has shifted this paradigm and identified several forms of “programmed necrosis”, including death receptor-mediated necrosis (termed “necroptosis”), mitochondria-mediated necrosis, and other regulated necrotic processes. Despite recent progress, how necroptosis is regulated in the heart remain largely unknown and preventing necroptotic cardiomyocyte death is still an important challenge. Our pilot studies identified a new K63-linked polyubiquitination (K63-Ub) dependent necroptosis signaling network that critically regulates cardiac ischemic injury and pathological remodeling. Intriguingly, the K63-deubiquitinase CYLD (cylindromatosis) was markedly up-regulated, whereas the E3 ubiquitin ligase TRAF2 (TNF receptor associated factor-2) was down-regulated, in the heart after ischemia-reperfusion injury. Our preliminary results further identified CYLD and TRAF2 as an activator and a suppressor of myocardial necroptosis, respectively. Importantly, cardiomyocyte-specific ablation of CYLD attenuated ischemic injury and adverse remodeling by inhibiting necroptosis. Acute deletion of TRAF2 in the adult heart caused dilated cardiomyopathy by promoting necroptosis, whereas TRAF2 gene transfer elicited cardioprotection. Mechanistically, we propose a new K63- Ub dependent necroptotic regulatory mechanism whereby CYLD deubiquitinates TRAF2 and TAK1, disrupts TAK1-RIP1 interaction, and promotes the necroptotic complex. As an opposing mechanism, TRAF2 acts as an E3 ligase for TAK1 to inhibit necroptosis signaling. Thus, the reversible K63-Ub (mainly non-proteolytic) constitutes a new paradigm of necroptosis signaling with important biological implications. We hypothesize that the deubiquitinase CYLD, in conjunction with the E3 ligase TRAF2, critically regulate myocardial necroptosis, ischemic injury, and remodeling, thus representing promising therapeutic targets. Aim 1 will investigate the opposing roles of K63-Ub modifying enzymes CYLD and TRAF2 in regulating myocardial necroptosis and ischemic injury in vivo. Aim 2 will define a CYLD/TRAF2 mediated, K63-Ub dependent necroptosis signaling network in cardiomyocytes. Using genetic and molecular strategies, we will investigate the opposing roles of CYLD and TRAF2 in regulating myocardial necroptosis under basal conditions and in the setting of ischemic injury. The proposed studies will provide new insights into the molecular regulation and functional significance of necroptosis in the heart, which has important translational implications, especially given our preliminary results revealing CYLD/TRAF2 as key necroptotic regulators and promising targets for ischemic heart disease.

项目概要/摘要心肌细胞因凋亡和/或坏死而损失是心脏缺血性损伤、病理性损伤的标志。尽管坏死传统上被认为是一种被动的、慢性的疾病。不受监管的过程，新出现的证据已经改变了这种范式，并确定了几种形式的“程序化” 坏死”，包括死亡受体介导的坏死（称为“坏死性凋亡”）、线粒体介导的坏死，尽管最近取得了进展，但坏死性凋亡在心脏中是如何调节的。仍然很大程度上未知，预防坏死性心肌细胞死亡仍然是我们试点的一个重要挑战。研究发现了一个新的 K63 连接的多聚泛素化 (K63-Ub) 依赖性坏死性凋亡信号网络有趣的是，K63-去泛素酶对心脏缺血性损伤和病理重塑具有重要调节作用。 CYLD（圆柱瘤病）显着上调，而 E3 泛素连接酶 TRAF2（TNF 受体）我们的初步结果是，缺血再灌注损伤后心脏中相关因子2）下调。进一步确定 CYLD 和 TRAF2 分别是心肌坏死性凋亡的激活剂和抑制剂。重要的是，CYLD 的心肌细胞特异性消融减轻了缺血性损伤和不良重塑抑制成人心脏中 TRAF2 的急性缺失，通过促进扩张性心肌病。坏死性凋亡，而 TRAF2 基因转移引起心脏保护作用，我们提出了一种新的 K63-。 Ub 依赖性坏死性凋亡调节机制，CYLD 使 TRAF2 和 TAK1 去泛素化，破坏 TAK1-RIP1 相互作用，并促进坏死性凋亡复合物作为相反机制，TRAF2 充当一种拮抗机制。 TAK1 的 E3 连接酶抑制坏死性凋亡信号传导，因此，K63-Ub 可逆（主要是非蛋白水解）。构成了具有重要生物学意义的坏死性凋亡信号传导的新范例。去泛素酶 CYLD 与 E3 连接酶 TRAF2 结合，关键地调节心肌坏死性凋亡，缺血性损伤和重塑，因此代表了有希望的治疗目标，目标 1 将研究这一点。 K63-Ub 修饰酶 CYLD 和 TRAF2 在调节心肌坏死性凋亡中的相反作用目标 2 将定义 CYLD/TRAF2 介导的 K63-Ub 依赖性坏死性凋亡信号传导。使用遗传和分子策略，我们将研究心肌细胞中的相反作用。 CYLD 和 TRAF2 在基础条件下和缺血环境下调节心肌坏死性凋亡拟议的研究将为分子调控和功能意义提供新的见解。心脏坏死性凋亡，这具有重要的转化意义，特别是考虑到我们的初步结果揭示 CYLD/TRAF2 作为关键的坏死性凋亡调节因子和缺血性心脏病的有希望的靶点。