Endothelial Cell to Mesenchymal Cell Transitions Play a Critical Biological Sex- and Aging-Dependent Role in Formation and Maintenance of the Acta2+ Atherosclerotic Lesion Protective Fibrous Cap

内皮细胞向间充质细胞的转变在 Acta2 动脉粥样硬化病变保护性纤维帽的形成和维持中发挥着关键的生物性别和衰老依赖性作用

• 批准号: 10542427
• 负责人: Gary K Owens
• 金额: $ 79.29万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10542427
• 关键词: Aging; Antibodies; Aorta; Apolipoprotein E; Apoptosis; Arterial Fatty Streak; Arteries; Atherosclerosis; Biological Assay; Bone Marrow Transplantation; Cardiovascular Diseases; Carotid Endarterectomy; Cause of Death; Cd68; Cell Aging; Cell Lineage; Cell Proliferation; Cells; Clinical; Clinical Research; Clonal Expansion; Coronary artery; Coronary heart disease; DNA biosynthesis; Deoxyuridine; Dependence; Development; Diet; Endothelial Cells; Event; Excision; Exhibits; Extracellular Matrix; Failure; Female; Functional disorder; Genes; Goals; Growth; Human; IL1R1 gene; Impairment; Incidence; Individual; Inflammation; Inflammatory; Intervention; Investments; Knock-out; Knockout Mice; Label; Lesion; Lesion by Stage; Life Style Modification; Lipids; Macrophage; Maintenance; Mechanics; Medicine; Mesenchymal; Metabolism; Modeling; Morphology; Mus; Myocardial Infarction; Myofibroblast; Nature; Oligonucleotides; PDGFRB gene; PECAM1 gene; Paper; Pathogenesis; Permeability; Pharmaceutical Preparations; Phenotype; Platelet-Derived Growth Factor beta Receptor; Play; Predisposition; Probability; Process; Proliferating; Property; Relaxation; Research; Resistance; Role; Rupture; Sampling; Signal Transduction; Smooth Muscle Myocytes; Stroke; Testing; Thick; Thinness; Thrombosis; Time; United States; Woman; atherothrombosis; biological sex; cell type; conditional knockout; endothelial dysfunction; feeding; high risk; improved; in vivo; indexing; insight; irradiation; male; men; novel; novel therapeutic intervention; prevent; senescence; sex; thrombotic; 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 (2021 Nature Metabolism) showed that smooth muscle cell (SMC) conditional knockout (KO) of the platelet-derived growth factor receptor-β (PDGFRβ) in ApoE-/- mice was associated with nearly complete failure of SMC to invest into lesions or the fibrous cap. However, despite nearly complete absence of SMC in lesions of SMC PDGFRβ KO mice, 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). Further studies showed that: 1) contrary to dogma that Acta2+ fibrous cap cells are derived almost exclusively from SMC, we found they account for only 60-75% of ACTA2+ cells in advanced ApoE-/- brachiocephalic (BCA) or human coronary artery (CA) lesions with the remainder coming from endothelial cell (EC)-to-mesenchymal-to-myofibroblast transitions (EndoMT-MFT, 15-20%) and macrophage-to-myofibroblast transitions (MMFT, 10-15%); 2) loss of SMC investment into lesions with SMC PDGFR? KO or bone marrow transplantation (BMT) was associated with a 2-3 fold increase 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 qualitative differences in the ability of fibrous cap cells to sustain lesion stability depending on their origin; 4) the contribution of EC to the ACTA2+ fibrous cap increased with lesion regression by feeding mice 18 weeks of WD followed by 12 weeks of chow diet; and 5) female but not male mice exhibit IL1R1-dependent EndoMT. Studies herein will test the hypothesis that the contribution of EC to the ACTA2+ fibrous cap increases with lethal irradiation-BMT, SMC PDGFRβ KO, aging, female sex, and/or plaque regression resulting in: 1) increased EC proliferation and replicative senescence; 2) loss of EC integrity and/or reduced anti- thrombotic properties; and 3) increased susceptibility to plaque erosion or rupture. Aim 1 will determine if the marked increase in the contribution of EndoMT-MFT to the ACTA2+ fibrous cap following lethal irradiation-BMT, SMC PDGFR? KO, or with plaque regression is associated with increased proliferation, clonal expansion, replicative senescence, and/or dysfunction of EC. Aim 2 will determine if treatment of our SMC- or EC-lineage tracing ApoE-/- or our novel Myh11-CreERT2/Rosa-eYFP/SR-BIΔCT/ΔCT/LDLR- (ΔCT) mice with advanced BCA and CA lesions with senolytic drugs is associated with beneficial changes in indices of plaque stability, improved survival, reduced MI or stroke, reduced dependence on EndoMT-MFT for maintenance of the ACTA2+ fibrous cap, and/or improved EC integrity and/or function. All aims include human validation studies on stable versus high risk carotid endarterectomy samples, and assessment of sex-dependent determinants of late-stage lesion pathogenesis. The proposed studies will provide novel insights regarding mechanisms that regulate the cellular and extracellular matrix (ECM) composition of the fibrous cap and may lead to development of novel therapeutic 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（2021年自然代谢）表明平滑肌细胞（SMC） APOE-/ - 小鼠中血小板衍生的生长因子受体-β（PDGFRβ）的条件敲除（KO）是相关的 SMC几乎完全未能投资于病变或纤维帽。但是，多皮几乎完成 在SMCPDGFRβKO小鼠病变中没有SMC，令人惊讶的是，我们观察到病变大小或指标没有变化 在西方饮食（WD）18周之后，包括Acta2+纤维帽的厚度，包括斑块稳定性。 进一步的研究表明：1）与教条相反，Acta2+纤维帽细胞几乎完全来自 SMC，我们发现它们仅占晚期APOE - / - 腕骨（BCA）或人类中ACTA2+细胞的60-75％ 冠状动脉（CA）病变，其余的来自内皮细胞（EC） - 至间质至肌纤维细胞 过渡（内摩特-MFT，15-20％）和巨噬细胞到肌纤维细胞转变（MMFT，10-15％）； 2）SMC的损失 用SMC PDGFR投资病变？ KO或骨髓移植（BMT）与2-3倍有关 内分鼠ft和mmft的增加； 3）增加的endomt-mft，MMFT并不能保持稳定指标 WD喂养延长至26周，表明纤维帽细胞能力的质量差异 病变稳定性取决于其起源； 4）EC对ACTA2+纤维帽的贡献随病变增加而增加 通过喂养小鼠18周的WD，然后进行12周的Chow饮食来消退； 5）女性但不是雄性老鼠 展示IL1R1依赖性恩赐。这里的研究将检验以下假设：EC对ACTA2+的贡献 纤维帽随着致命的辐照-BMT，SMCPDGFRβKO，衰老，女性和/或斑块回归而增加 导致：1）EC增殖和复制感应增加； 2）EC完整性丧失和/或抗抗 血栓性特性； 3）增加对斑块侵蚀或破裂的敏感性。 AIM 1将确定是否标记 致命辐照BMT后，内摩特-MFT对ACTA2+纤维帽的贡献增加了SMC PDGFR？ KO，或带有斑块回归与增殖，克隆膨胀，复制性的增加有关 EC的感应和/或功能障碍。 AIM 2将确定是否处理我们的SMC或EC-Linege跟踪APOE - / - 是 或我们的新型MyH11-Creert2/rosa-eyfp/sr-biΔct/Δct/ldlr-（Δct）小鼠，带有晚期BCA和CA病变 鼻溶剂与牙菌斑稳定指数的有益变化有关，生存率提高，MI降低或 中风，降低对维护ACTA2+纤维帽的依赖性的依赖性和/或改进的EC 完整性和/或功能。所有目的包括有关稳定与高风险颈动脉内部切除术的人类验证研究 样品以及晚期病变发病机理的性依赖性决定剂的评估。提出的研究 将提供有关调节细胞和细胞外基质（ECM）组成的机制的新见解 纤维帽的含量，并可能导致新型热方法的发展，以增强斑块稳定性。