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: 10436073
• 负责人: DOUGLAS K BOWLES
• 金额: $ 55.27万
• 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/10436073
• 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; 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; Mammals; Medial; Mediating; Mediator of activation protein; Modeling; Mus; Myocardial Infarction; Necrosis; Outcome; Pathogenesis; Pharmaceutical Preparations; Pharmacology; Phenotype; Play; 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; macrophage; migration; myocardin; next generation; novel; 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制很复杂，多种细胞类型导致病变大小 和组成。急性冠状动脉综合征通常与复合体破裂有关， 临床上良性的易损斑块。进展到相对良性， 稳定的病变或易于破裂的易损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 病变。该提案的目标是 1) 确定 SM 特定的角色和底层 中间电导、Ca2+ 激活的 K+ 通道、KCa3.1（编码为 Kcnn4)，指示动脉粥样硬化病变的形成和组成，2) 确定平移 临床批准的 KCa3.1 抑制剂对大型哺乳动物模型病变发展的潜力 冠状动脉疾病（CAD）。作为支持，我们提供了因果关系的第一个遗传证据 KCa3.1 与病变大小以及 SM 和巨噬细胞募集之间的关系。总体假设是 KCa3.1 激活增加 SM 和巨噬细胞迁移至内膜并导致病变 形成。相反，通过基因沉默或药理学阻断 KCa3.1 会减少 动脉粥样硬化病变的大小并有益地改变成分。目标 1 将确定的贡献 KCa3.1在平滑肌中参与动脉粥样硬化病变的形成和成分。具体来说，我们将使用 SM 特异性、可诱导 KO 小鼠，用于检查 KCa3.1 在确定斑块大小、组成中的作用 和基因表达。目标 2 将定义上游 (REST) 和下游 (DOCK2) 机制 确定 KCa3.1 对 SM 和动脉粥样硬化的影响。我们将使用转基因小鼠 检查 REST 和 DOCK2 在介导 KCa3.1 对表型、增殖、SM 影响中的作用 迁移、斑块大小和成分。此外，我们将使用RNA测序来鉴定新的 KCa3.1 动脉粥样硬化发展的机制。我们将在 Aim 3 中使用 VSM 谱系追踪 使用 SM 谱系追踪确定 SMC-KCa3.1 在介导 SMC 内膜向内侧迁移中的作用 和动脉粥样硬化病变发展过程中的泡沫细胞转分化。最后，目标 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 猪模型中。我们使用纵向跟踪冠状动脉疾病进展 对家族性高胆固醇血症 (FH) 猪进行血管造影和 IVUS，以测试 KCa3.1 抑制能力 与 senicapoc 一起使用，可减小冠状动脉病变的大小并促进其更有利的组成。 长期目标是为转化当前的治疗工具和方法提供临床前基础。 开发针对 KCa3.1 和/或下游信号传导的下一代药物，以有益于 操纵动脉粥样硬化病变的成分。