Cell-specific role and therapeutic potential of KCa3.1 in atherosclerosis

KCa3.1 在动脉粥样硬化中的细胞特异性作用和治疗潜力

• 批准号: 10586148
• 负责人: DOUGLAS K BOWLES
• 金额: $ 55.68万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10586148
• 关键词: Angiography; Animal Model; Apolipoprotein E; Area; Arterial Fatty Streak; Atherosclerosis; Benign; Calcium-Activated Potassium Channel; Cause of Death; Cells; Cessation of life; Characteristics; Clinical; Clinical Trials; Coagulation Process; Collagen; Complex; Coronary; Coronary Angiography; Coronary Arteriosclerosis; Coronary heart disease; Development; Disease Progression; Disease model; Drug Targeting; FDA approved; Familial Hypercholesterolemia; Family suidae; Foam Cells; Foundations; Gene Expression; Genes; Genetic; Goals; Heart Diseases; Histology; Hospitals; Human; Immunohistochemistry; In Vitro; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Knockout Mice; Lesion; Link; Macrophage; Mammals; Medial; Mediating; Mediator; Modeling; Mus; Myocardial Infarction; Necrosis; Outcome; Pathogenesis; Pharmaceutical Preparations; Phenotype; Play; Proliferating; Regulator Genes; Reporter; Role; Rupture; Set protein; Signal Transduction; Smooth Muscle; Solid; Stroke; Symptoms; Testing; Therapeutic; Tomatoes; Translating; United States; acute coronary syndrome; cell type; combat; coronary lesion; coronary plaque; cost; drug testing; in vivo; inhibitor; migration; myocardin; next generation; novel; pharmacologic; porcine model; pre-clinical; recruit; therapeutically effective; tool; transcription factor REST; transcriptome sequencing; transdifferentiation; translational potential; ultrasound

Pathogenesis of coronary artery disease is complex, with multiple cell types contributing to lesion size and composition. Acute coronary syndromes are most often associated with rupture of complex, vulnerable plaques that are otherwise clinically benign. The progression to either a relatively benign, stable lesion or a rupture-prone, vulnerable plaque has been linked to key lesion characteristics, i.e. smooth muscle (SM) and collagen content, macrophage infiltration and necrotic core area within the lesion. The objectives of this proposal are to 1) determine the SM-specific role and underlying mechanism(s) by which the intermediate conductance, Ca2+-activated K+ channel, KCa3.1 (encoded by Kcnn4), dictates atherosclerotic lesion formation and composition and 2) determine the translational potential of clinically approved KCa3.1 inhibitors on lesion development in a large mammal model of coronary artery disease (CAD). In support, we provide the first genetic evidence of a causal link between KCa3.1 and lesion size and SM and macrophage recruitment. The overall hypothesis is that KCa3.1 activation increases migration of SM and macrophages into the intima and contributes to lesion formation. Conversely, blocking KCa3.1, both by genetic silencing or pharmacologically, will decrease atherosclerotic lesion size and beneficially alter composition. Aim 1 will determine the contribution of KCa3.1 in smooth muscle to atherosclerotic lesion formation and composition. Specifically, we will use SM-specific, inducible KO mice to examine the role of KCa3.1 in determining plaque size, composition and gene expression. Aim 2 will define both upstream (REST) and downstream (DOCK2) mechanisms determining KCa3.1 effects on SM and atherosclerosis. We will use genetically modified mice to examine the role of REST and DOCK2 in mediating SM effects of KCa3.1 on phenotype, proliferation, migration, plaque size and composition. In addition, we will use RNA sequencing to identify novel mechanisms of atherosclerosis development by KCa3.1. We will use VSM lineage-tracking in Aim 3 will use SM lineage-tracking to determine role of SMC-KCa3.1 in mediating SMC intimal to medial migration and foam cell transdifferentiation during atherosclerotic lesion development. Finally, Aim 4 will determine the effect of the FDA approved KCa3.1 inhibitor, senicapoc, on atherosclerosis development in a swine model of CAD. We longitudinally track coronary artery disease progression using angiography and IVUS in familial hypercholesterolemic (FH) swine to test the ability of KCa3.1 inhibition with senicapoc, to decrease the size and promote a more favorable composition of coronary lesions. The long-term goal is to provide the pre-clinical foundation for translating current therapeutic tools and developing the next generation drugs targeting KCa3.1 and/or downstream signaling to beneficially manipulate atherosclerotic lesion composition.

冠状动脉疾病的发病机理很复杂，多种细胞类型导致病变大小 和构图。急性冠状动脉综合症通常与复合物破裂有关 否则在临床上是良性的脆弱斑块。向相对良性的发展， 稳定的病变或容易破裂的脆弱斑块与关键病变特征有关，即 平滑肌（SM）和胶原蛋白含量，巨噬细胞浸润和坏死核心区域内 病变。该提案的目标是1）确定特定于SM的作用和基础 中间电导Ca2+激活的K+通道KCA3.1的机制（由 KCNN4），决定动脉粥样硬化病变的形成和组成，2）确定翻译 在大型哺乳动物模型中，临床认可的KCA3.1抑制剂对病变发育的潜力 冠状动脉疾病（CAD）。为了支持，我们提供了因果关系的第一个遗传证据 在KCA3.1和病变大小以及SM和巨噬细胞募集之间。总体假设是 KCA3.1激活增加了SM和巨噬细胞的迁移到内膜中，并导致病变 形成。相反，无论是通过遗传沉默还是药理上阻止KCA3.1，都会减少 动脉粥样硬化病变的大小和有益改变组成。 AIM 1将确定 KCA3.1在平滑肌中至动脉粥样硬化病变形成和组成。具体来说，我们将使用 SM特异性的，可诱导的KO小鼠以检查KCA3.1在确定斑块大小的作用，成分中的作用 和基因表达。 AIM 2将定义上游（休息）和下游（DOCK2）机制 确定KCA3.1对SM和动脉粥样硬化的影响。我们将使用转基因的小鼠 检查REST和DOCK2在介导KCA3.1对表型，增殖的SM效应中的作用 迁移，斑块大小和组成。此外，我们将使用RNA测序来识别新颖 KCA3.1动脉粥样硬化发展的机制。我们将在AIM 3中使用VSM谱系跟踪 使用SM谱系跟踪来确定SMC-KCA3.1在介导SMC内膜迁移中的作用 和动脉粥样硬化病变发育过程中的泡沫细胞转分化。最后，AIM 4将 确定FDA批准的KCA3.1抑制剂Senicapoc对动脉粥样硬化发展的影响 在CAD的猪模型中。我们纵向追踪使用冠状动脉疾病进展 家族性高胆固醇（FH）猪的血管造影和IVU测试KCA3.1抑制作用的能力 使用Senicapoc，可以减少大小并促进更有利的冠状动脉病变组成。 长期目标是为翻译当前的治疗工具和 开发靶向KCA3.1和/或下游信号的下一代药物以有益 操纵动脉粥样硬化病变组成。