TNNI3K: A cardiac-specific kinase regulating ischemic injury and fibrotic remodel

TNNI3K：调节缺血性损伤和纤维化重塑的心脏特异性激酶

基本信息

批准号：
8465269
负责人：
Thomas Force
金额：
$ 36.41万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-05-05 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8465269
关键词：
Address Alleles Biological Models Biology Cardiac Cell Death Cells Chest Clinical Clinical Trials Data Extracellular Matrix Fibrosis Gene Expression Gene Expression Profile Gene Targeting Genes Genomics Heart Heart failure Hypertension In Vitro Infarction Injury Ischemia Letters Modeling Molecular Mus Mutate Names National Heart, Lung, and Blood Institute Phosphotransferases Placebos Play Pre-Clinical Model Process Protein Kinase Proteomics Reagent Regulation Regulator Genes Reperfusion Injury Reperfusion Therapy Reverse Transcriptase Polymerase Chain Reaction Role Safety Small Interfering RNA Staging Technology Transgenic Mice Translations Troponin I analog base constriction in vivo inhibitor/antagonist injury and repair novel novel strategies oxidant stress patient population pressure public health relevance response screening small molecule tool

项目摘要

DESCRIPTION (provided by applicant): Heart failure commonly results from ischemic or hypertensive disease, and novel strategies are sorely needed to address the ever-expanding population of patients with heart failure world-wide. Over the past two years we have explored the role of a novel cardiac-specific protein kinase, troponin I type 3 interacting kinase (gene name TNNI3K) in regulating ischemic injury, post-MI remodeling, and pressure overload-induced remodeling. We have employed a number of strategies including 1) transgenic mice expressing either wild-type, constitutively active, or kinase-inactive TNNI3K, 2) conditional cardiac-specific deletion of TNNI3K (CKO mice), and 3) novel small molecule inhibitors of TNNI3K generated by our collaborators at Glaxo-Smith-Kline that are able to be used both in vitro and in vivo. All of our data support two key conclusions: 1) Inhibition of TNNI3K is strongly protective, and activation of TNNI3K is highly detrimental, in the setting of ischemia/reperfusion (I/R) injury, and 2) TNNI3K appears to be a central regulator of fibrotic remodeling in the heart. We now propose to determine the molecular mechanisms by which TNNI3K regulates these processes, thereby expanding our understanding of mechanisms of ischemic injury and repair. In our Specific Aims we propose to identify TNNI3K targets from genomic analyses and have already identified a profound role of TNNI3K in regulating expression of genes implicated in fibrotic remodeling. This is consistent with the significant reduction in fibrosis we observe in mice subjected to thoracic aortic constriction that were treated with one of the small molecule inhibitors. We will also employ proteomic approaches to identify direct substrates of TNNI3K using the analog sensitive kinase allele (ASKA) approach pioneered by Shokat and co-workers. Finally, we will focus on the role of select substrates identified in Aim 2, and will determine ther role in ischemic injury and post-infarct remodeling. Given our successful use of small molecule inhibitors targeting TNNI3K in vivo, and the demonstrated safety of this approach in our pre-clinical models, we believe that the stage could be set for fairly rapid translation to clinical trals if our hypotheses are borne out.

描述（由申请人提供）：心力衰竭通常由缺血性或高血压疾病引起，迫切需要新的策略来解决世界范围内不断扩大的心力衰竭患者群体。在过去的两年里，我们探索了一种新型心脏特异性蛋白激酶，即肌钙蛋白 I 3 型相互作用激酶（基因名称 TNNI3K）在调节缺血性损伤、心肌梗死后重塑和压力超负荷诱导的重塑中的作用。我们采用了多种策略，包括 1) 表达野生型、组成型活性或激酶失活 TNNI3K 的转基因小鼠，2) TNNI3K 的条件性心脏特异性缺失（CKO 小鼠），以及 3) TNNI3K 的新型小分子抑制剂由我们在葛兰素史克公司的合作者产生，可以在体外和体内使用。我们所有的数据都支持两个关键结论：1) TNNI3K 的抑制作用很强 TNNI3K 具有保护作用，并且在缺血/再灌注 (I/R) 损伤的情况下，TNNI3K 的激活非常有害，2) TNNI3K 似乎是心脏纤维化重塑的中心调节因子。我们现在建议确定 TNNI3K 调节这些过程的分子机制，从而扩大我们对缺血性损伤和修复机制的理解。在我们的具体目标中，我们建议从基因组分析中识别 TNNI3K 靶点，并且已经确定了 TNNI3K 在调节与纤维化重塑相关的基因表达中的深远作用。这与我们在接受一种小分子抑制剂治疗的胸主动脉缩窄小鼠中观察到的纤维化显着减少是一致的。我们还将采用 Shokat 及其同事首创的模拟敏感激酶等位基因 (ASKA) 方法，采用蛋白质组学方法来鉴定 TNNI3K 的直接底物。最后，我们将重点关注目标 2 中确定的选定底物的作用，并确定它们在缺血性损伤和梗塞后重塑中的作用。鉴于我们在体内成功使用了针对 TNNI3K 的小分子抑制剂，并且在我们的临床前模型中证明了这种方法的安全性，我们相信，如果我们的假设得到证实，就可以为相当快速地转化为临床试验奠定基础。