Mechanisms of L-type Calcium Channel Regulation in Heart Health and Disease

L 型钙通道在心脏健康和疾病中的调节机制

基本信息

批准号：
10734121
负责人：
Jonathan Satin
金额：
$ 64.37万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-31 至 2027-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10734121
关键词：
Acceleration Address Adrenergic Receptor Agonist Angiotensins Animal Model Attenuated Binding Biophysics Bypass C-terminal Ca(2+)-Calmodulin Dependent Protein Kinase Calcium Channel Cardiac health Cessation of life Clinical Trials Cyclic AMP-Dependent Protein Kinases Data Disease model Diuretics Down-Regulation EFRAC Failure Family Family suidae Goals Health Heart Heart Diseases Heart failure Human Hydrophobicity Inotropism Kinetics Knock-in Knock-in Mouse Knock-out L-Type Calcium Channels Macromolecular Complexes Measures Mediating Modeling Modification Molecular Monomeric GTP-Binding Proteins Mouse Strains Mus Myocardial Contraction Pathologic Patients Phosphatidylinositol 4,5-Diphosphate Phosphorylation Pilot Projects Preparation Receptor Signaling Regulation Risk Risk Reduction Safety Sarcomeres Signal Pathway Signal Transduction Slice Structural Models Structure Sympathomimetics Testing Therapeutic Therapeutic Uses United States Work antagonist early phase clinical trial follow-up gain of function genetic approach improved in vivo loss of function member novel novel therapeutics pharmacologic phosphoric diester hydrolase protein complex response sensor stoichiometry success translational potential voltage

项目摘要

Inotropic support for hearts progressing towards failure represents an unmet need. Direct attempts to improve systolic function by activating -adrenergic receptor (-AR) signaling increases associated risk of heart failure and death. Calcitropes (agents influencing Ca2+ handling) that bypass -AR signaling are not necessarily proarrhythmic. Our working global hypothesis based on our recent and ongoing work studying RAD regulation of the L-type Ca2+ channel (LTCC) is that bypassing -AR signaling to increase Ca2+-induced Ca2+ release (CICR) provides safe, stable gain of function to counter heart failure progression. In this application we focus on the mechanisms of Rad – LTCC interactions as a novel means to instill inotropic support to the heart. The LTCC is a macromolecular hub that integrates multiple signaling pathways including protein kinase A (PKA) and Ca2+- calmodulin kinase II (CaMKII). RAD is a member of the RGK family of monomeric G-proteins. RAD binds to auxiliary CaV2 and CaV1.2, the pore-forming subunit of the LTCC. Deletion, or phosphorylation of RAD causes LTCC current (ICa,L) modulation and facilitation. Modulation of ICa,L is commonly observed after -AR signaling to activate PKA; facilitation is caused by CaMKII activation. In Specific Aim 1 we will dissect how RAD integrates each of these signaling pathways using a combination of pharmacological and genetic approaches. In Specific Aim 2 we will explore RAD – CaV1.2 structure-function using knock-in models of genetically modified mice that allow us to explore RAD – LTCC effects retaining native stoichiometry of the LTCC heteromultimeric protein complex. Specific Aim 3 explores RAD – LTCC interplay as an approach to attenuate progression of heart failure. To achieve these goals, we will integrate findings among Aims using in vivo, ex vivo and cellular/molecular approaches in animal models. Ex vivo human heart slices will be tested to evaluate the translational potential of RAD – LTCC regulation in heart health and disease.

对心脏朝着失败的心脏的肌力支持代表了未满足的需求。直接尝试改进通过激活肾上腺素能受体（-AR）信号传导增加相关心力衰竭的风险和死亡。无需旁路信号传导的钙化（影响Ca2+处理的代理）心律不足。根据我们最近和正在进行的研究RAD监管的工作，我们工作的全球假设 L型Ca2+通道（LTCC）的是绕过-AR信号以增加Ca2+诱导的Ca2+释放（CICR）提供安全，稳定的功能增益，以应对心力衰竭进展。在此应用中，我们专注于 RAD - LTCC相互作用的机制作为一种新颖的方法，用于灌输对心脏的肌力支持。 LTCC是大分子枢纽整合了包括蛋白激酶A（PKA）和Ca2+ - 在内的多个信号通路钙调蛋白激酶II（CAMKII）。 Rad是RGK单体G蛋白家族的成员。 rad结合辅助Cav2和cav1.2，LTCC的孔子亚基。删除或rad原因的磷酸化 LTCC电流（ICA，L）调制和设施。 ICA的调节，通常在-AR信号传导后观察到L 激活PKA；设施是由CAMKII激活引起的。在特定目标1中，我们将剖析RAD集成方式这些信号通路都结合了药物和遗传方法。具体 AIM 2我们将使用一般修饰的小鼠的敲入模型来探索RAD - CAV1.2结构功能允许我们探索RAD - LTCC效应，以保留LTCC异类蛋白的天然化学计量法复杂的。特定的目标3探索了RAD - LTCC相互作用，作为减轻心力衰竭进展的方法。为了实现这些目标，我们将使用体内，离体和细胞/分子的目标整合目标动物模型的方法。将测试过体内的人心脏切片，以评估 RAD - 心脏健康和疾病中的LTCC调节。