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，是西方世界中死亡的主要原因。 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主要的歧义 病变发病机理中的肌肉细胞（SMC），以及控制斑块稳定性和 牙菌斑破裂的可能性，可能是心肌梗塞（MI）或中风。一般的教条是 SMC主要参与后期而不是早期病变，其主要作用是动脉保护 通过形成纤维帽。但是，我们的实验室最近的自然医学研究涉及 干细胞多能基因的简单SMC特异性谱系跟踪和敲除（KO），OCT4 或KLF4提供了令人信服的证据，表明SMC在病变发病机理中的作用要比 普遍赞赏。主要发现包括我们的表明：1）> 80％的SMC衍生单元 APOE - / - 小鼠的晚期病变缺乏典型SMC标记的可检测表达； 2）SMC的30-40％ 高级小鼠和人病变中的细胞都缺乏可检测的SMC标记，并且具有 MSS的激活标记； 3）SMC可能对病变产生重大有益或有害影响 发病机理取决于其表型过渡的性质。该提案中的研究将测试 SMC表型过渡可以发挥主导性动力保护或动脉保护作用的假设 对晚期病变发病机理的影响，代表了治疗晚期的新型热靶 动脉粥样硬化。 AIM 1将检验以下假设：SMC表型和功能中KLF4依赖性过渡 在动脉粥样硬化的早期和晚期，病变发病机理加剧，将包括定义不同的 基于完成流程，由SMC特异性有条件KO诱导的动脉保护SMC表型基于完整的流量 从我们的体内RNASEQ和 高级臂头（BCA）病变的KLF4/OCT4 CHIPSEQ基因组分析和引用基因的交叉 与人类CAD风险增加有关。我们还将确定在 晚期病变的建立还会影响病变发病机理的有利变化。 AIM 2将测试 假设SMC表型和功能中的OCT4依赖性转变促进了通过 将SMC的投资提高到受保护的纤维帽上，并减少其过渡到促炎症 状态。我们还将测试小鼠研究中发现的SMC表型是否也发生在人体病变中并定义 哪种表型与稳定与不稳定病变相关。 AIM 3将确定SMC表型是否发生变化 在晚期病变中是可逆的，如果治疗SMC谱系用高级病变跟踪小鼠 PCSK9抑制剂促进了SMC表型和/或整体病变发病机理的有益变化。研究可以 导致鉴定出新的治疗方法，以减少晚期临床并发症 动脉粥样硬化通过在SMC表型和功能中引起有益（斑块稳定）的变化来引起动脉粥样硬化。

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.