Identifying new regulators of cardiac fibrosis and inflammation using zebrafish

使用斑马鱼识别心脏纤维化和炎症的新调节因子

基本信息

批准号：
10892436
负责人：
Juan Manuel Gonzalez-Rosa
金额：
$ 39.13万
依托单位：
BOSTON COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-22 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10892436
关键词：
Acute Adult Adverse effects Affect Alleles Animals Attenuated Cardiac Cardiac Myocytes Cicatrix Clinical Collection Competence Deposition Development Disease Disease Progression Exhibits Extracellular Matrix Feedback Fibrosis Gene Expression Profiling Gene Modified Generations Genes Genetic Models Goals Heart Heart Diseases Heart Injuries Heart failure Hypertrophy Immune Immunocompromised Host Impairment Individual Inflammasome Inflammation Inflammatory Inflammatory Response Injury Ischemia Knowledge Laboratories Link Lymphocytic Infiltrate Macrophage Mediator Modeling Molecular Mus Mutation Myocarditis NF-kappa B Natural regeneration Outcome Pathologic Pathology Persons Phase Phenotype Phosphotransferases Play Population Positioning Attribute Process Production Progressive Disease Proliferating Public Health Reperfusion Injury Reporting Research Resistance Role Serine Proteinase Inhibitors Siblings Signal Transduction Skin Syndrome System T cell infiltration T-Lymphocyte Testing Therapeutic Tissues Transgenic Organisms Translating Trypsin Inhibitors Zebrafish cardiac regeneration chemokine comparison control coronary fibrosis cost design effective therapy experimental study functional decline gain of function genetic variant inter-alpha-inhibitor loss of function mortality mutant new therapeutic target overexpression prevent recruit response tool trait transcriptomic profiling

项目摘要

Project Summary/Abstract Several cardiac diseases involve the progressive or acute loss of cardiomyocytes, which are replaced by fibrotic tissue. Cardiac fibrosis is one of the leading factors in the pathology of heart failure, a progressive disease that affects millions of people every year. Despite its recognized importance, there are no effective therapies to prevent the progression of cardiac fibrosis. Recent findings suggest that naturally occurring genetic variants protect certain individuals against the adverse changes triggered after cardiac injury, which translates into re- duced cardiac fibrosis. One of the genes that modifies the progression of disease is the cardiomyocyte-specific kinase Tnni3k. Mutations that abolish Tnni3k confer resistance to injury, and high Tnni3k levels are associated with rapid functional decline and pathological remodeling. Tnni3k is an understudied kinase, and its downstream targets and specific mechanisms by which it defines injury outcome are unknown. Our goal in this project is to identify the mechanisms linking Tnni3k levels to cardiac fibrosis. In preliminary experiments using the zebrafish as a model, we found that high levels of Tnni3k induce fibrosis deposition shortly after cardiac injury. In contrast to their wild-type siblings, animals overexpressing Tnni3k showed impaired fibrotic regression, while a newly generated tnni3k mutant exhibits minimal fibrosis after injury. Transcriptional profiling revealed that high levels of Tnni3k correlate with an exacerbated inflammatory response, markers of T-cell infiltration, and activation of all the components of the inflammasome, which are all landmarks of cardiomyocyte-induced inflammation. We also identified a potential downstream target, itih5, which has been previously reported to play a role in extracel- lular matrix stabilization in the skin. Based on our preliminary results, our central hypothesis is that Tnni3k plays a previously unappreciated role in cardiac fibrosis via modulation of inflammation, and that Itih5 is a central mediator of these effects. We will test our hypothesis in three integrative aims. In Aim 1, we will determine the role of inflammation in the development of fibrosis in response to high levels of Tnni3k. We will exploit a collection of immunocompromised zebrafish to test the contribution of specific immune populations to this excessive fibrotic response. Additionally, we will use a new cardiomyocyte specific Cas9 line to identify genes required for trigger- ing inflammation in response to high Tnni3k levels. In Aim 2, we will determine the effects of modulating Tnni3k levels pre- and post-injury on cardiac fibrosis and inflammation. We will use a new Tnni3kSWITCH line generated by our lab that will allow us to “turn off” the overexpression of Tnni3k at different times. In Aim 3, we will identify molecular regulators of scar remodeling downstream of Tnni3k. We will start by analyzing how Itih5 regulates fibrosis regression using new gain and loss of function models generated by our laboratory for this application. Collectively, these studies will establish Tnni3k and Itih5 as regulators of cardiac inflammation and fibrosis in zebrafish. We anticipate that this basic knowledge will serve as the springboard for rapid discovery of new targets to treat cardiac fibrosis in the injured heart.

项目摘要/摘要多种心脏疾病涉及心肌细胞的进行性或急性丧失，取而代之的是纤维化组织。心脏纤维化是心力衰竭病理学的主要因素之一，一种进行性疾病每年都会影响数百万人。尽管它的重要性很重要，但没有有效的疗法防止心脏纤维化的进展。最近的发现表明自然存在的遗传变异保护某些人免受心脏损伤后触发的不良变化的影响，这转化为诱导心脏纤维化。改变疾病进展的基因之一是心肌细胞特异性激酶TNNI3K。废除TNNI3K赋予损伤和高tnni3k水平的突变是相关的随着功能迅速下降和病理重塑。 tnni3k是一种知识的激酶，它的下游定义损伤结果的目标和特定机制尚不清楚。我们在这个项目中的目标是确定将TNNI3K水平与心脏纤维化联系起来的机制。在使用斑马鱼的初步实验中作为一个模型，我们发现高水平的TNNI3K会在心脏损伤后不久影响纤维化沉积。相比之下对于它们的野生型兄弟姐妹，表现出过表达TNNI3K的动物纤维化回归受损，而新近的动物损伤后产生的TNNI3K突变体表现出最小的纤维化。转录分析表明高水平 TNNI3K与加重的炎症反应，T细胞浸润的标记以及激活炎性体的所有成分，它们都是心肌细胞引起的炎症的地标。我们还确定了一个潜在的下游目标ITIH5，以前据报道，该目标在外部效果中发挥作用皮肤中的lular基质稳定。基于我们的初步结果，我们的中心假设是TNNI3K 通过调节炎症，在心脏纤维化中扮演以前没有批准的作用，而ITIH5是中心这些影响的中介。我们将在三个综合目的中检验我们的假设。在AIM 1中，我们将确定炎症在对高水平TNNI3K响应纤维化发展中的作用。我们将利用一个集合免疫功能低下的斑马鱼，以测试特异性免疫种群对这种过量纤维化的贡献回复。此外，我们将使用新的心肌细胞特异性CAS9系列来识别触发所需的基因响应高TNNI3K水平的炎症。在AIM 2中，我们将确定调制TNNI3K的影响在心脏纤维化和炎症后伤害后水平。我们将使用生成的新TNNI3KSWWITCH系列通过我们的实验室，这将使我们可以在不同时间“关闭” TNNI3K的过表达。在AIM 3中，我们将确定 TNNI3K下游的疤痕重塑的分子调节剂。我们将首先分析ITIH5如何调节使用我们实验室为此应用产生的新增益和功能模型丧失的纤维化回归。总的来说，这些研究将建立TNNI3K和ITIH5作为心脏感染的调节剂和纤维化的调节剂斑马鱼。我们预计这种基本知识将成为快速发现新目标的跳板治疗受伤的心脏中的心脏纤维化。