Defining SMC phenotypes critical in late stage atherosclerosis pathogenesis

定义在晚期动脉粥样硬化发病机制中至关重要的 SMC 表型

• 批准号: 10084307
• 负责人: Gary K Owens
• 金额: $ 74.09万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10084307
• 关键词: Age; Anti-Inflammatory Agents; Aortic Segment; Apolipoprotein A-I; Apolipoprotein E; Arterial Fatty Streak; Atherosclerosis; Biological Assay; CETP gene; Cause of Death; Cell Lineage; Cells; Clinical; Collaborations; Collagen; Coronary artery; Development; Diet; Epigenetic Process; Event; Genes; Genomics; Goals; High Density Lipoproteins; Human; In Situ Hybridization; Inflammatory; Investments; Knock-out; Laboratories; Lead; Lesion; Lesion by Stage; Ligation; Link; Lipids; Mechanics; Medicine; Methods; Modeling; Mus; Myocardial Infarction; Nature; Necrosis; Operative Surgical Procedures; Pathogenesis; Pharmaceutical Preparations; Phenotype; Phenylephrine; Play; Probability; Progressive Disease; Research; Resolution; Risk; Role; Rupture; Smooth Muscle Myocytes; Stem cell pluripotency; Stroke; Testing; Thick; Thinness; Transplantation; Vascular Smooth Muscle; Western World; atheroprotective; base; biomarker panel; cell type; conditional knockout; cytokine; feeding; in vivo; indexing; inhibitor/antagonist; innovation; macrophage; new therapeutic target; novel; novel marker; novel strategies; novel therapeutic intervention; risk variant; stem cells; thrombotic; transcriptome sequencing; virtual; western diet

Atherosclerosis is a progressive disease that is a leading cause of death in the Western world. Remarkably, despite decades of research, there remain major ambiguities regarding the role of smooth muscle cells (SMC) in lesion pathogenesis, as well as mechanisms that control plaque stability and the probability of plaque rupture with possible myocardial infarction (MI) or stroke. The general dogma is that SMC are primarily involved in late not early stage lesions, and that their primary role is atheroprotective by contributing to formation of a fibrous cap. However, recent Nature Medicine studies by our lab involving simultaneous SMC-specific lineage tracing and knockout (KO) of the stem cell pluripotency genes, Oct4 or Klf4 provided compelling evidence that SMC play a much greater role in lesion pathogenesis than has been generally appreciated. Key findings included our showing that: 1) >80% of SMC-derived cells within advanced lesions of ApoE-/- mice lack detectable expression of typical SMC markers; 2) 30-40% of SMC- derived cells within both advanced mouse and human lesions lack detectable SMC markers and have activated markers of MФs; and 3) SMC can have major beneficial or detrimental effects on lesion pathogenesis depending on the nature of their phenotypic transitions. Studies in this proposal will test the hypothesis that SMC phenotypic transitions can exert dominant atheroprotective or atheropromoting effects on late stage lesion pathogenesis and represent a novel therapeutic target for treating advanced atherosclerosis. Aim 1 will test the hypothesis that Klf4-dependent transitions in SMC phenotype and function exacerbate lesion pathogenesis in early and late stages of atherosclerosis, and will include defining distinct atheroprotective SMC phenotypes induced by SMC-specific conditional KO of Klf4 based on complementary flow cytometric and high resolution confocal analyses of unique marker panels derived from our in vivo RNAseq and Klf4/Oct4 ChIPseq genomic analyses of advanced brachiocephalic (BCA) lesions and cross referenced to genes linked to increased human CAD risk. We will also determine if delayed SMC specific conditional KO of Klf4 after establishment of advanced lesions also induces favorable changes in lesion pathogenesis. Aim 2 will test the hypothesis that Oct4-dependent transitions in SMC phenotype and function promote plaque stabilization by enhancing investment of SMC into a protective fibrous cap and reducing their transition to a pro-inflammatory state. We will also test if SMC phenotypes identified in our mouse studies also occur in human lesions and define which phenotypes correlate with stable versus unstable lesions. Aim 3 will determine if SMC phenotypic changes within advanced lesions are reversible, and if treatment SMC lineage tracing mice with advanced lesions with a PCSK9 inhibitor promotes beneficial changes in SMC phenotype and/or overall lesion pathogenesis. Studies may lead to identification of novel therapeutic approaches for reducing late stage clinical complications of atherosclerosis by inducing beneficial (plaque stabilizing) changes in SMC phenotype and function.

动脉粥样硬化是一种进行性疾病，是西方世界死亡的主要原因。 值得注意的是，尽管进行了数十年的研究，但关于平滑的作用仍然存在很大的模糊性。 肌细胞（SMC）在病变发病机制中的作用，以及控制斑块稳定性和斑块稳定性的机制 斑块破裂并可能导致心肌梗塞 (MI) 或中风的概率 一般的规律是： SMC 主要参与晚期而非早期病变，其主要作用是动脉粥样硬化 然而，我们实验室最近的《自然医学》研究涉及纤维帽的形成。 同时 SMC 特异性谱系追踪和干细胞多能性基因的敲除 (KO)，Oct4 或 Klf4 提供了令人信服的证据，表明 SMC 在病变发病机制中发挥的作用比 我们的主要发现包括：1) >80% 的 SMC 衍生细胞。 ApoE-/- 小鼠的晚期病变缺乏可检测到的典型 SMC 标志物的表达 2) 30-40% 的 SMC- 晚期小鼠和人类病变中的衍生细胞缺乏可检测的 SMC 标记，并且具有 MФs 的激活标记物；3) SMC 对病变有重大的有益或不良影响 发病机制取决于其表型转变的性质。本提案中的研究将进行测试。 SMC 表型转变可以发挥显着的动脉粥样硬化保护作用或动脉粥样硬化促进作用的假设 对晚期病变发病机制的影响，代表了治疗晚期病变的新治疗靶点 目标 1 将检验 SMC 表型和功能中 Klf4 依赖性转变的假设。 动脉粥样硬化早期和晚期病变发病机制恶化，包括定义不同的 基于互补流的 SMC 特异性条件性敲除 Klf4 诱导的动脉粥样硬化 SMC 表型 对源自我们的体内 RNAseq 的独特标记组进行细胞计数和高分辨率共聚焦分析 对晚期头臂 (BCA) 病变进行 Klf4/Oct4 ChIPseq 基因组分析并交叉引用基因 我们还将确定是否延迟 SMC 对 Klf4 的特定条件 KO。 晚期病变的建立也会引起病变发病机制的有利变化，目标 2 将测试。 假设 SMC 表型和功能中的 Oct4 依赖性转变通过以下方式促进斑块稳定： 增强 SMC 对保护性纤维帽的投资，并减少其向促炎性纤维帽的转变 我们还将测试小鼠研究中发现的 SMC 表型是否也出现在人类病变中并定义。 哪些表型与稳定病变和不稳定病变相关，目标 3 将确定 SMC 表型是否发生变化。 晚期病变内是可逆的，并且如果使用 SMC 谱系追踪具有晚期病变的小鼠 PCSK9 抑制剂可促进 SMC 表型和/或整体病变发病机制的有益变化。 导致确定减少晚期临床并发症的新治疗方法 通过诱导 SMC 表型和功能的有益（斑块稳定）变化来预防动脉粥样硬化。

项目成果

Characterization of protein isoform diversity in human umbilical vein endothelial cells via long-read proteogenomics.

• DOI: 10.1080/15476286.2022.2141938
• 发表时间: 2022-01
• 期刊: RNA BIOLOGY
• 影响因子: 4.1
• 作者: Mehlferber, Madison M.;Jeffery, Erin D.;Saquing, Jamie;Jordan, Ben T.;Sheynkman, Leon;Murali, Mayank;Genet, Gael;Acharya, Bipul R.;Hirschi, Karen K.;Sheynkman, Gloria M.
• 通讯作者: Sheynkman, Gloria M.

Smooth muscle cells-specific loss of OCT4 accelerates neointima formation after acute vascular injury.

• DOI: 10.3389/fcvm.2023.1276945
• 发表时间: 2023
• 期刊: FRONTIERS IN CARDIOVASCULAR MEDICINE
• 影响因子: 3.6
• 作者: Shin, Junchul;Tkachenko, Svyatoslav;Gomez, Delphine;Tripathi, Rupande;Owens, Gary K.;Cherepanova, Olga A.
• 通讯作者: Cherepanova, Olga A.