Investigating the role of CaV1.2 in aortic valve stenosis

研究 CaV1.2 在主动脉瓣狭窄中的作用

基本信息

批准号：
10421276
负责人：
Geoffrey S Pitt
金额：
$ 79.49万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10421276
关键词：
Alternative Therapies Aortic Valve Stenosis Bioinformatics Calcium Channel Blockers Cells Clinical Clinical Data Complementary DNA Coronary Arteriosclerosis Data Development Disease Disease Progression Early treatment Etiology Family suidae Gene Expression Profile Gene set enrichment analysis High Fat Diet Human Hyperlipidemia Inflammatory Response L-Type Calcium Channels Lesion Life Medical Meta-Analysis Motivation Mus Myofibroblast Nodule Pathology Pathway Analysis Pathway interactions Patients Process Quantitative Trait Loci Reporting Risk Factors Role Signal Pathway Signal Transduction Signaling Molecule Signaling Protein Susceptibility Gene System Testing Tissues Transfection Validation advanced disease antagonist aortic valve aortic valve disorder calcification candidate identification effective therapy efficacy testing genetic association genome wide association study in vivo innovation interstitial cell macrophage mouse model normotensive novel novel therapeutics osteogenic overexpression pre-clinical prevent success targeted treatment therapeutic target voltage

项目摘要

Calcific aortic valve disease (CAVD) is a progressive life-threatening disorder characterized by dystrophic and/or osteogenic transformation of valve cells (VICs), both leading to calcification. There are no medical therapies to prevent or delay CAVD. Building on a recent meta-analysis of genome-wide association studies that unexpectedly identified CACNA1C, encoding the pore-forming α subunit of the voltage gated L-type calcium channel CaV1.2, as a candidate CAVD susceptibility gene, we present preliminary data suggesting that Ca2+ influx through CaV1.2 in VICs is causal for CAVD. This raises the tantalizing possibility that clinically used Ca2+ channel antagonists (CCBs) may be an effective therapy. Although CCBs are generally contraindicated in patients with severe CAVD, they would be reasonable therapies earlier in the disease process. We propose to test the overall hypotheses that: 1) increased Ca2+ influx through CaV1.2 in VICs leads to dystrophic and/or osteogenic transformation; and 2) reduction of Ca2+ influx through CaV1.2 with CCBs and/or by targeting Ca2+- dependent signaling molecules downstream of CaV1.2 in VICs delays CAVD progression. We developed novel mouse models and innovative VIC cultures to test those overall hypotheses with the following Aims: Aim 1: Can elevated Ca2+ influx through CaV1.2 activate VICs and/or increase expression of signaling pathway genes leading to dystrophic or osteogenic calcification in the AoV? Using porcine and murine VICs enhanced by transfection of informative cDNAs, we have developed means to dissect the Ca2+-dependent signaling pathways downstream of CaV1.2 that contribute to CAVD, and thereby to discover new potential therapeutic targets. We propose a multi-pronged approach to identify the critical downstream Ca2+-dependent signaling molecules and pathways leading to activation of dystrophic VICs and/or osteogenic transformation of VICs; Aim 2: Can inhibition of CaV1.2 signaling pathways in the aortic valve decrease the valve pathology? With novel mouse models that mimic the increased CaV1.2 expression and signaling in human CAVD, we propose to test if CCBs ameliorate valve lesions and reverse the disease process once calcification is initiated. We further propose to build upon our identification of novel candidate targets from Aim 1 to test if manipulating these targets similarly reduces or slows CAVD progression. Aim 3: Does excess Ca2+ influx through CaV1.2 contribute to CAVD in the context of hyperlipidemia? Hyperlipidemia is a prominent CAVD risk factor. In coronary artery disease, a related disorder, long ignored data point to a synergistic interaction between CaV1.2 and hyperlipidemia. We propose: to test with novel mouse models whether Ca2+ influx through CaV1.2 in VICs or macrophages accelerates AoV lesions in the context of hyperlipidemia and, as a corollary, whether CCBs are effective when the disease process is driven by hyperlipidemia, thereby determining whether targeting the CaV1.2 pathway is broadly applicable in patients.

钙化性主动脉瓣疾病 (CAVD) 是一种进行性危及生命的疾病，其特征为营养不良和/或瓣膜细胞（VIC）的成骨转化，均会导致钙化，目前尚无治疗方法。以最近对全基因组关联研究的荟萃分析为基础，预防或延迟 CAVD。意外鉴定出 CACNA1C，编码电压门控 L 型钙的成孔 α 亚基 CaV1.2通道作为候选CAVD易感基因，我们提供的初步数据表明Ca2+ 通过 CaV1.2 流入 VIC 是导致 CAVD 的原因，这提出了临床使用 Ca2+ 的诱人可能性。通道拮抗剂 (CCB) 可能是一种有效的治疗方法，尽管 CCB 通常是禁忌的。对于患有严重 CAVD 的患者，我们建议在疾病过程的早期进行合理的治疗。检验总体假设：1) VIC 中通过 CaV1.2 增加的 Ca2+ 流入导致营养不良和/或成骨转化；2) 通过 CaV1.2 使用 CCB 和/或靶向 Ca2+- 减少 Ca2+ 流入 VIC 中 CaV1.2 下游的依赖性信号分子可延迟 CAVD 进展。小鼠模型和创新的 VIC 培养物来测试这些总体假设，目标如下：目标 1：可以通过 CaV1.2 增加的 Ca2+ 流入会激活 VIC 和/或增加信号通路基因的表达，从而导致使用转染增强的猪和鼠 VIC 来治疗 AoV 中的营养不良或成骨性钙化？凭借信息丰富的 cDNA，我们开发了剖析 Ca2+ 下游信号通路的方法 CaV1.2 有助于 CAVD，并发现新的潜在治疗靶点。多管齐下的方法来识别关键的下游 Ca2+ 依赖性信号分子和途径导致营养不良的 VIC 激活和/或 VIC 成骨转化；目标 2：可以抑制 CaV1.2；主动脉瓣中的信号通路可以减少瓣膜病理学吗？人 CAVD 中 CaV1.2 表达和信号传导增加，我们建议测试 CCB 是否能改善瓣膜病变一旦钙化开始，我们进一步建议以我们的鉴定为基础来逆转疾病过程。目标 1 中的新候选目标，以测试操纵这些目标是否同样会减少或减缓 CAVD 目标 3：在高脂血症的情况下，通过 CaV1.2 的过量 Ca2+ 流入是否会导致 CAVD？高脂血症是冠状动脉疾病（一种相关疾病）中的一个重要危险因素，但长期被忽视的数据。指出 CaV1.2 和高脂血症之间存在协同相互作用我们建议：用新型小鼠进行测试。模拟 Ca2+ 通过 VIC 或巨噬细胞中的 CaV1.2 流入是否会加速 AoV 病变高脂血症，以及作为推论，当疾病过程由以下因素驱动时，CCB 是否有效：高脂血症，从而确定靶向 CaV1.2 通路是否广泛适用于患者。