Project-002

项目-002

基本信息

批准号：
10254291
负责人：
Hind Lal
金额：
$ 45.8万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-18 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10254291
关键词：
Adenovirus Vector Adult Basic Science Bioinformatics Biology Body Weight Cancer Biology Cardiac Cardiac Myocytes Cardiovascular Diseases Cells Data Development Fibroblasts Functional disorder Gene Delivery Gene Expression Goals Heart Heart Diseases Heart Injuries Heart failure Homeostasis Human Knockout Mice Knowledge Lead Literature Mediating Mission Modeling Mus Myocardial Myocardial Ischemia Myocardial dysfunction Myofibroblast Pathology Pathway interactions Phenotype Phosphotransferases Play Prevalence Protein Kinase Protein-Serine-Threonine Kinases Public Health Reagent Regulation Research Role Route Screening Result Serotyping Signal Transduction Specificity Stress Testing Treatment Efficacy Troponin T United States National Institutes of Health adenoviral-mediated base cardiogenesis cardioprotection coronary fibrosis design gene therapy genetic manipulation heart function homeodomain in vivo insight mouse model novel novel strategies p38 Mitogen Activated Protein Kinase periostin prevent promoter public health relevance repaired restoration screening therapeutic target therapeutically effective transcriptome translational approach translational study vector virtual

项目摘要

Kinases play a critical role in the development of heart failure (HF) and, therefore, represent a potential therapeutic target. While the cardiac kinome is comprised of several hundred kinases, the vast majority of the literature is focused on only a few kinases. Indeed, the function of numerous highly expressed cardiac kinases is unexplored; these may include potential therapeutic targets for cardiovascular diseases. In order to identify novel cardiac kinase(s) potentially involved in HF development, we employed an integrated transcriptome and bioinformatics approach (Expression2Kinases (X2K)) with control and failing hearts. Indeed, in our screening results, most of the identified kinases are well-known for their roles in cardiac biology, validating the applied approach. Intriguingly, we also identified a novel kinase, Homeodomain-Interacting Protein Kinase 2 (HIPK2), as potentially being involved in the regulation of cardiac remodeling and HF. Indeed, there is no literature describing the function of HIPK2 in the heart. The overall goal of this proposal is to define the role of HIPK2 in cardiac pathophysiology. As a first step towards this goal, we have generated mouse models including fibroblast (FB)-specific HIPK2 KOs (periostinMCM and TCF21MCM) and cardiomyocyte (CM)-specific HIPK2 KOs (αMHCCre and αMHCMCM). Three specific aims are designed to critically examine the role of HIPK2 in cardiac biology. Aim 1: to define the role of HIPK2 in cardiac fibrosis and remodeling. We will test the hypothesis that HIPK2 exerts a critical break on fibrotic remodeling by inhibiting profibrotic TGF-β1/SMAD-3 and p38 pathways. In Aim 2, we will test the hypothesis that CM-HIPK2 is essential in maintaining basal cardiac homeostasis and required to protect against cardiac stress. We will utilize inducible CM-specific cre mice to delete HIPK2 only in the fully mature CM, evaluate the mechanism(s) whereby HIPK2 may exert its cardioprotective effect. Aim 3 will assess the therapeutic efficacy of AAV9 mediated HIPK2 gene therapy to mitigate the pathologies induced by cardiac stress. Expression of HIPK2 is significantly reduced in failing human hearts, we hypothesize that restoration of HIPK2 in cardiac cells will reduce and may reverse the cardiac pathologies. AAV9-HIPK2 will be used to deliver the FB-specific (under periostin promoter and TCF21 promoter) and CM-specific (troponin-t promoter) gene expression. These studies will directly test the translational potential of the findings from Aims 1 and 2. Thus, the proposed studies are of high significance, not only by contributing translationally-relevant information but also by providing novel mechanistic insights into cardiac pathophysiology.

激酶在心力衰竭（HF）的发展中起关键作用，因此代表了潜力治疗靶标。心脏动物组完成了数百个激酶，但绝大多数文献仅关注几个激酶。确实，许多高度表达的心脏激酶的功能是出乎意料的；这些可能包括心血管疾病的潜在治疗靶标。为了确定新颖的心脏激酶（S）可能参与HF开发，我们工作了一个集成的转录组和具有控制和失败心脏的生物信息学方法（表达2kinass（x2k））。确实，在我们的放映中结果，大多数确定的激酶以其心脏生物学的作用而闻名，验证了所应用的方法。有趣的是，我们还确定了一种新型的激酶，同源域相互作用蛋白激酶2（HIPK2），可能参与心脏重塑和HF的调节。确实，没有文学描述HIPK2在心脏中的功能。该提议的总体目标是确定Hipk2在心脏病理生理学。作为实现此目标的第一步，我们生成了鼠标模型成纤维细胞（FB） - 特异性HIPK2 KOS（Pererostinmcm和TCF21MCM）和心肌细胞（CM） - 特异性HIPK2 KOS （αMHCCRE和αMHCMCM）。设计三个特定目标旨在批判性地检查HIPK2在心脏中的作用生物学。目标1：定义HIPK2在心脏纤维化和重塑中的作用。我们将检验以下假设 HIPK2通过抑制纤维化的TGF-β1/SMAD-3和p38，在纤维化重塑上施加临界破裂途径。在AIM 2中，我们将检验以下假设：CM-IHIPK2对于维持基本心脏至关重要稳态，需要防止心脏压力。我们将利用可诱导的CM特异性CRE小鼠仅在完全成熟的CM中删除HIPK2，评估HIPK2可以执行其的机制心脏保护作用。 AIM 3将评估AAV9介导的HIPK2基因治疗的治疗效率减轻心脏应激引起的病理。失败的HIPK2的表达显着降低人类的心，我们假设心脏细胞中HIPK2的恢复将减少并可能逆转心脏病理。 AAV9-IHIPK2将用于传递FB特异性启动子）和CM特异性（肌钙蛋白-T启动子）基因表达。这些研究将直接测试 AIMS 1和2的调查结果的转化潜力。这是，所提出的研究具有很高的意义，不仅通过贡献翻译相关的信息，还通过提供新颖的机械见解心脏病理生理学。