Role of Metabolic Reprogramming in Formation and Maintenance of the Acta2+ Atherosclerotic Lesion Protective Fibrous Cap

代谢重编程在 Acta2 动脉粥样硬化病变保护性纤维帽形成和维持中的作用

• 批准号: 10441555
• 负责人: Gary K Owens
• 金额: $ 67.28万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10441555
• 关键词: Accounting; Apolipoprotein E; Arterial Fatty Streak; Atherosclerosis; Carotid Endarterectomy; Cause of Death; Cell Lineage; Cells; Cessation of life; Clinical; Clinical Research; Collagen; Coronary artery; Coronary heart disease; Cytoskeleton; Data; Databases; Development; Diet; Endothelial Cells; Endothelium; Energy Metabolism; Energy Metabolism Pathway; Event; Exhibits; Extracellular Matrix; Failure; Flow Cytometry; Genetic; Glucose; Glutamine; Goals; Histologic; Human; In Vitro; Individual; Inflammation; Investigation; Investments; Knock-out; Lactate Dehydrogenase; Lead; Lesion; Lesion by Stage; Life Style Modification; Lipids; Maintenance; Manuscripts; Mechanics; Medicine; Mesenchymal; Metabolic; Microscopic; Modeling; Mus; Myocardial Infarction; Myofibroblast; Nature; Paper; Pathogenesis; Pathway interactions; Pharmacology; Phenotype; Platelet-Derived Growth Factor; Platelet-Derived Growth Factor beta Receptor; Play; Probability; Property; Research; Role; Rupture; Sampling; Smooth Muscle Myocytes; Stroke; Testing; Thick; Transforming Growth Factor beta; United States; aerobic glycolysis; antagonist; aortic arch; atherothrombosis; cell type; chronic inflammatory disease; conditional knockout; dietary; dietary approach; feeding; high risk; in vivo; indexing; inhibitor; insight; knock-down; macrophage; metabolic profile; multiplexed imaging; novel; novel strategies; prevent; pyruvate dehydrogenase; response; sex; small hairpin RNA; thrombotic; transcriptome sequencing; transcriptomics; validation studies; western diet

Atherothrombosis, resulting from rupture or erosion of unstable atherosclerotic plaques, is the leading cause of death worldwide. However, the mechanisms that regulate the stability of late stage atherosclerotic lesions remain poorly understood. Recent studies from our lab showed that smooth muscle cell (SMC) conditional knockout of the platelet-derived growth factor receptor-β (PdgfrbSMC-Δ/Δ) in ApoE-/- mice was associated with nearly complete failure of SMC to invest into lesions or the fibrous cap. However, surprisingly we observed no changes in lesion size or indices of plaque stability, including the thickness of the Acta2+ fibrous cap, following 18 weeks of Western diet (WD) feeding. Further investigation provided novel insights regarding the mechanisms underlying these changes. Key findings included: 1) contrary to long-standing dogma that Acta2+ fibrous cap cells are derived almost exclusively from SMC, we showed that they account for only 60-70% of Acta2+ cells in advanced ApoE-/- brachiocephalic (BCA) or human coronary artery lesions with the remainder coming from endothelial cell-to-mesenchymal-to-myofibroblast transitions (EndoMT-MFT, 20-25%) and macrophage-to-myofibroblast transitions (MMFT), 10-15%) respectively; 2) loss of SMC investment into lesions with SMC PDGFRB KO was associated with large adaptive increases in EndoMT-MFT and MMFT; 3) increased EndoMT-MFT, and MMFT did not sustain indices of stability when WD feeding was extended to 26 weeks suggesting there may be qualitative differences in Acta2+ fibrous cap cells depending on their origin; 4) RNA-seq analysis on the BCA of 18-week WD-fed PdgfrbSMC-Δ/Δ ApoE-/- mice versus control littermate mice showed that energy metabolism pathways were the top ten upregulated pathways suggesting that metabolic reprogramming may be required for SMC-MF, EndoMT-MF, and MMF transitions; and 5) inhibition of aerobic glycolysis in cultured SMC prevented their transition to a MF state following treatment with PDGF and TGFβ. Studies in this proposal will test the hypothesis that SMC, EC, and macrophage adaptive responses for maintaining plaque stability require major shifts in energy metabolism and that the metabolic state of these cells can be manipulated to stimulate beneficial changes in the phenotype of lesion cells and overall increases in plaque stability. Aim 1 will determine if genetic knockout or pharmacologic inhibition of aerobic glycolysis in SMC and EC lineage tracing ApoE-/- mice with advanced atherosclerosis is associated with evidence of reduced plaque stability. Aim 2 will determine if genetic or pharmacologic promotion of aerobic glycolysis in SMC and EC lineage tracing ApoE-/- lineage tracing mice with advanced atherosclerosis is associated with increased plaque stability. Studies for both aims will include human validation studies using scRNAseq and histological data from stable asymptomatic versus unstable symptomatic carotid endarterectomy samples. All studies will assess sex-dependent determinants of late-stage lesion pathogenesis. Taken together, the proposed studies will provide novel insights regarding mechanisms by which metabolic reprogramming of cells contribute to the cellular and extracellular matrix (ECM) composition of the fibrous cap and may lead to development of novel pharmacological and dietary approaches for enhancing plaque stability.

动脉粥样硬化血栓形成是由不稳定的动脉粥样硬化斑块破裂或侵蚀引起的，是导致动脉粥样硬化的主要原因。 然而，调节晚期动脉粥样硬化病变稳定性的机制仍然存在。 我们实验室最近的研究表明，平滑肌细胞（SMC）有条件敲除。 ApoE-/- 小鼠中的血小板衍生生长因子受体-β (PdgfrbSMC-Δ/Δ) 与几乎完全失败相关 然而，令人惊讶的是，我们观察到病变大小或纤维帽没有变化。 西式饮食 (WD) 18 周后的斑块稳定性指数，包括 Acta2+ 纤维帽的厚度 进一步的研究提供了关于这些变化背后的机制的新见解。 研究结果包括：1) 与 Acta2+ 纤维帽细胞几乎完全源自这一长期教条相反 从 SMC 中，我们发现它们仅占晚期 ApoE-/- 头臂 (BCA) 中 Acta2+ 细胞的 60-70% 或人冠状动脉病变，其余部分来自内皮细胞到间充质到肌成纤维细胞 分别为转变（EndoMT-MFT，20-25%）和巨噬细胞至肌成纤维细胞转变（MMFT），10-15%）； SMC PDGFRB KO 导致 SMC 对病变的投入损失与适应性的大幅增加相关 EndoMT-MFT 和 MMFT；3) WD 时 EndoMT-MFT 增加，而 MMFT 没有维持稳定性指数 喂养时间延长至 26 周，表明 Acta2+ 纤维帽细胞可能存在质的差异 4) 对 18 周 WD 喂养的 PdgfrbSMC-Δ/Δ ApoE-/- 小鼠的 BCA 进行 RNA-seq 分析 对照同窝小鼠显示，能量代谢途径是前十大上调途径，表明 SMC-MF、EndoMT-MF 和 MMF 转变可能需要代谢重编程；5) 抑制 在用 PDGF 和 TGFβ 处理后，培养的 SMC 中的有氧糖酵解阻止了它们向 MF 状态的转变。 本提案中的研究将检验以下假设：SMC、EC 和巨噬细胞适应性反应以维持 斑块需要能量代谢稳定性发生重大转变，这些细胞的代谢状态可以 进行操作以刺激病变细胞表型的有益变化以及斑块稳定性的总体增加。 目标 1 将确定是否对 SMC 和 EC 谱系中的有氧糖酵解进行基因敲除或药物抑制 追踪患有晚期动脉粥样硬化的 ApoE-/- 小鼠与 Aim 2 斑块稳定性降低的证据相关。 确定 SMC 和 EC 谱系中的有氧糖酵解是否通过遗传或药理学促进，追踪 ApoE-/- 谱系 追踪患有晚期动脉粥样硬化的小鼠与斑块稳定性的增加有关，这两个目的的研究将进行。 包括使用 scRNAseq 的人体验证研究以及稳定无症状与不稳定的组织学数据 有症状的颈动脉内膜切除术样本所有研究都将评估晚期病变的性别依赖性决定因素。 综上所述，拟议的研究将提供有关发病机制的新见解。 纤维细胞和细胞外基质 (ECM) 成分贡献细胞的代谢重编程 上限并可能导致开发新的药理学和饮食方法来增强斑块稳定性。