Project-002

激酶在心力衰竭(HF)的发展中起着关键作用，因此代表着一种潜在的 治疗靶点。虽然心脏动态组由数百个激酶组成，但绝大多数 文学作品只关注少数几个蛋白。事实上，许多高表达的心脏激酶的功能 尚未探索；这些可能包括心血管疾病的潜在治疗靶点。为了辨认 新的心肌激酶(S)可能参与心力衰竭的发展，我们使用了一个整合的转录组和 生物信息学方法(Expression2Kinase(X2K))与对照和衰竭的心脏。的确，在我们的放映中 结果，大多数已鉴定的激酶因其在心脏生物学中的作用而广为人知，验证了其应用价值 接近。有趣的是，我们还发现了一种新的激酶，同源域相互作用蛋白激酶2(HIPK2)， 可能参与心脏重构和心衰的调节。事实上，没有文学作品 描述HIPK2在心脏中的功能。该提案的总体目标是定义HIPK2在 心脏病理生理学。作为实现这一目标的第一步，我们已经生成了包括 成纤维细胞(FB)特异性HIPK2 KO(骨膜蛋白MCM和TCF21MCM)和心肌细胞(CM)特异性HIPK2 KO (αMHCCre和αMHCMCM)。三个特定的目标被设计来批判性地研究HIPK2在心脏中的作用。 生物学。目的1：明确HIPK2在心肌纤维化和重构中的作用。我们将检验这一假设 HIPK2通过抑制促纤维化转化生长因子-β-1/sMAD-3和p38在纤维化重塑中发挥关键作用 小路。在目标2中，我们将检验CM-HIPK2在维持基础心脏方面是必不可少的假设 动态平衡，并需要防止心脏应激。我们将利用可诱导的CM特异性cre小鼠来 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(S) 心脏保护作用。目的3评价AAV9介导的HIPK2基因治疗HIPK2的疗效。 减轻心脏应激引起的病理改变。HIPK2在衰竭时的表达显著降低 我们假设HIPK2在心肌细胞中的恢复将减少并可能逆转 心脏病理学。AAV9-HIPK2将用于传递FB特异性的(在Periostin启动子和TCF21下) 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 目标1和目标2的结论的翻译潜力。因此，拟议的研究具有很高的意义， 不仅通过提供与翻译相关的信息，还通过提供对 心脏病理生理学。

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.