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: 10292012
• 负责人: Gary K Owens
• 金额: $ 74.76万
• 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/10292012
• 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; aortic arch; atherothrombosis; cell type; chronic inflammatory disease; conditional knockout; dietary; dietary approach; feeding; high risk; in vivo; indexing; inhibitor/antagonist; 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.

动脉粥样硬化是由于不稳定的动脉粥样硬化斑块破裂或侵蚀引起的，是导致的主要原因 全球死亡。但是，调节晚期动脉粥样硬化病变稳定性的机制仍然存在 理解不佳。我们实验室的最新研究表明，平滑肌细胞（SMC）条件敲除 APOE-/ - 小鼠中血小板来源的生长因子受体-β（PDGFRBSMC-δ/δ）与几乎完全失败有关 SMC投资于病变或纤维帽。但是，令人惊讶的是，我们观察到病变大小或 在西方饮食（WD）18周之后 进食。进一步研究提供了有关这些变化基础的机制的新见解。钥匙 调查结果包括：1）与长期存在的教条相反，acta2+纤维帽细胞几乎完全得出 从SMC中，我们证明它们仅占晚期APOE - / - 臂头（BCA）中ACTA2+细胞的60-70％ 或人类冠状动脉病变，其余的来自内皮细胞到间质至肌纤维细胞 分别过渡（内摩特-MFT，20-25％）和巨噬细胞到肌纤维细胞转变（MMFT），10-15％）； 2） 使用SMC PDGFRB KO损失SMC投资对病变的投资与自适应增加有关 endomt-mft和mmft; 3）增加的endomt-mft，并且MMFT在WD时无法维持稳定的指标 喂养延长至26周，表明ACTA2+纤维帽细胞可能存在定性差异 取决于它们的起源； 4）关于18周WD-FED PDGFRBSMC-δ/δAPOE-/ - 小鼠BCA的RNA-seq分析 控制同窝仔的小鼠表明能量代谢途径是提示的前十个更新的途径 SMC-MF，Endomt-MF和MMF过渡可能需要该代谢重编程； 5）抑制 有氧糖酵解在培养的SMC中阻止了用PDGF和TGFβ处理后其向MF状态的过渡。 该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用于维持 斑块稳定性需要能量代谢的重大变化，这些细胞的代谢状态可以是 操纵以刺激病变细胞表型的有益变化以及牙菌斑稳定性的总体增加。 AIM 1将确定在SMC和EC谱系中对有氧糖酵解的遗传基因敲除或药物抑制 追踪患有晚期动脉粥样硬化的APOE - / - 小鼠与斑块稳定性降低的证据有关。 AIM 2意志 确定有氧糖酵解在SMC和EC谱系跟踪APOE - / - 谱系中的遗传或药理促进是 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。两个目标的研究将 包括使用SCRNASEQ的人类验证研究以及稳定不对称与不稳定的组织学数据 有症状的颈动脉内膜切除术样品。所有研究将评估性别依赖性确定晚期病变 发病。综上所述，拟议的研究将提供有关机制的新颖见解 细胞的代谢重编程有助于纤维的细胞和细胞外基质（ECM）组成 CAP并可能导致开发新型的药理和饮食方法，以增强牙菌斑的稳定性。