Project-002

项目-002

基本信息

批准号：
10020438
负责人：
Hind Lal
金额：
$ 42.88万
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/10020438
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 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) 的发展中发挥着关键作用，因此代表了潜在的虽然心脏激酶组由数百个激酶组成，但绝大多数激酶。事实上，文献只关注少数激酶，但许多高表达的心肌激酶的功能。尚未探索；这些可能包括心血管疾病的潜在治疗靶点。可能参与心力衰竭发展的新型心肌激酶，我们采用了整合的转录组和事实上，在我们的筛选中，生物信息学方法（Expression2Kinases (X2K)）具有控制和衰竭的心脏。结果，大多数已识别的激酶因其在心脏生物学中的作用而闻名，验证了应用有趣的是，我们还发现了一种新的激酶，同源结构域相互作用蛋白激酶 2 (HIPK2)，事实上，尚无文献报道其可能参与心脏重塑和心力衰竭的调节。描述 HIPK2 在心脏中的功能该提案的总体目标是定义 HIPK2 在心脏中的作用。作为实现这一目标的第一步，我们生成了包括以下在内的小鼠模型。成纤维细胞 (FB) 特异性 HIPK2 KO（periostinMCM 和 TCF21MCM）和心肌细胞 (CM) 特异性 HIPK2 KO （αMHCCre 和 αMHCMCM）旨在严格检查 HIPK2 在心脏中的作用。目标 1：确定 HIPK2 在心脏纤维化和重构中的作用我们将检验以下假设。 HIPK2 通过抑制促纤维化 TGF-β1/SMAD-3 和 p38 对纤维化重塑发挥关键作用在目标 2 中，我们将检验 CM-HIPK2 对于维持基础心脏功能至关重要的假设。我们将利用可诱导的 CM 特异性 cre 小鼠来实现体内平衡和防止心脏应激。仅在完全成熟的 CM 中删除 HIPK2，评估 HIPK2 可能发挥其作用的机制目标 3 将评估 AAV9 介导的 HIPK2 基因疗法对心脏的保护作用。减轻心脏应激引起的病理，HIPK2 的表达在失败时显着降低。人类心脏中，我们发现心肌细胞中 HIPK2 的恢复将减少并可能逆转 AAV9-HIPK2 将用于传递 FB 特异性（在骨膜素启动子和 TCF21 下）。这些研究将直接测试 CM 特异性（肌钙蛋白-t 启动子）基因表达。目标 1 和 2 研究结果的转化潜力。因此，拟议的研究具有很高的意义，不仅通过提供翻译相关信息，还通过提供新颖的机制见解心脏病理生理学。