Signaling Mechanisms of FGF2-induced Cardioprotection

该方案的总体目标是研究成纤维细胞生长因子2介导的分子靶点(S) 心脏保护。最初提案的发现表明，一氧化氮(NO)、蛋白激酶C (PKC)和丝裂原活化蛋白激酶(MAPK)信号转导是FGF2诱导所必需的 心脏保护。然而，这些信号通路介导FGF2的下游靶点- 诱导的抗心肌功能障碍和心肌梗死的心脏保护作用仍有待阐明。 因此，我们将进行药理学、电生理学、亚蛋白质组学和 基于生理学的多学科方法来识别已知或新的信号底物 与成纤维细胞生长因子2诱导的心脏保护相关的通路。该提案将对参与程度进行评估 已知的与FGF2活性或与FGF2活性相关的激酶下游底物 心肌保护及FGF2亚组诱导心肌保护的新靶点 分析。该研究计划将蛋白质和细胞水平的基本信息与 整个器官/动物层面的信息。这些研究还将使我们能够直接联系 心肌梗死后变化与缺血后心功能恢复的生物学关系 FGF2的活性与其蛋白激酶通路的特定下游靶点有关。以确定 ATP敏感钾(KATP)通道参与FGF2诱导的心肌保护，我们将 使用药理学、电生理学和分子方法。将使用类似的技术 在细胞水平上确定FGF2在调节钙稳态中的重要性 Sarco(Endo)质网(SR)蛋白和收缩装置，最终影响后 缺血性心功能。初步数据显示线粒体、肌浆网和收缩 作为FGF2诱导的心肌保护靶点的装置及其候选和新的底物 细胞器将通过磷酸蛋白质组学(免疫印迹和2-D Gel/MS)技术进行询问。 这项提案的结果将为FGF2诱导的心脏保护提供新的见解，并可能 最终导致FGF2作为抗缺血治疗的药理或遗传学发展 心脏病。

The overall goal of this proposal is to investigate the molecular target(s) underlying FGF2-mediated cardioprotection. Findings from the original proposal indicate that nitric oxide (NO), protein kinase C (PKC), and mitogen-activated protein kinase (MAPK) signaling are necessary for FGF2-induced cardioprotection. Yet, the downstream targets by which these signaling pathways mediate FGF2- induced cardioprotection against myocardial dysfunction and infarction remain to be elucidated. Therefore, we will undertake a pharmacological-, electrophysiological-, subproteomic- and integrative physiological-based multidisciplinary approach to identify known or novel substrates of the signaling pathways associated with FGF2-induced cardioprotection. The proposal will evaluate the involvement of known downstream substrates of kinases that have been affiliated with FGF2 activity or with cardioprotection, and the identify novel targets of FGF2-induced cardioprotection by subproteomic analysis. The research plan will integrate basic information at the protein and cellular level with information at the whole organ/animal level. These studies will also enable us to directly relate changes in myocardial infarction and post-ischemic recovery of ventricular function to the biological activity of FGF2 and to specific downstream targets of its protein kinase pathways. To ascertain the involvement of ATP-sensitive potassium (KATP) channels in FGF2-induced cardioprotection, we will employ pharmacologic, electrophysiological, and molecular methods. Similar techniques will be used to determine the importance of FGF2 in regulating calcium homeostasis at the level of sarco(endo)plasmic reticulum (SR) proteins and contractile apparatus, ultimately influencing post- ischemic cardiac function. With preliminary data implicating the mitochondria, SR, and contractile apparatus as targets of FGF2-induced cardioprotection, candidate and novel substrates of these organelles will be interrogated via phosphoproteomic (immunoblotting and 2-D gel/MS) techniques. Results of this proposal will provide new insights into FGF2-induced cardioprotection and may eventually lead to the pharmacologic or genetic development of FGF2 as a therapy against ischemic heart disease.