Impact of Aging on Stem Cell Repair in Atherosclerosis

衰老对动脉粥样硬化干细胞修复的影响

• 批准号: 7117906
• 负责人: Pascal J. Goldschmidt-Clermont
• 金额: $ 6.06万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-15 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7117906
• 关键词: aging; angiogenesis; animal old age; atherosclerosis; bone marrow; cardiovascular disorder prevention; cardiovascular disorder therapy; cell biology; cell population study; flow cytometry; gene expression; genetically modified animals; integrins; juvenile animal; laboratory mouse; microarray technology; nonhuman therapy evaluation; pathologic process; phenotype; regeneration; selectins; stem cell transplantation; stem cells; vascular cell adhesion molecule; vascular endothelium

DESCRIPTION (provided by applicant): Atherosclerosis represents an aberrant, continuous reparative inflammatory process in response to repeated injuries to the vessel wall. Indeed, the cardiovascular system is continuously exposed to a multitude of insults whose impact cumulates with the passage of time. Two sources of cells participating in the repair process exist: (1) local, differentiated, vascular endothelial and smooth muscle cells that migrate from adjacent vessel segments; and (2) recruited stern cells/vascular progenitor cells from the bone marrow, via peripheral circulation. We have recently discovered that chronic administration of whole bone marrow (BM) cells significantly reduced atherosclerotic lesion formation in an established mouse model of atherosclerosis --ApoE -/- C57/B6 mice fed high fat cholesterol, Western-type diet--even in the absence of restoration of ApoE gene expression and normalization of plasma cholesterol levels (>1200 mg/dl). Furthermore, BM cells from young, but not old, ApoE-/- mice were capable of vascular rejuvenation and atherosclerosis prevention. This data underscores the importance of stem cells/vascular progenitor cells in vascular healing and atherogenesis and provide support for the use of stern/progenitor cell therapy as a novel preventative and/or treatment strategy for atherosclerosis, particularly for individuals whose bone marrow, and its obsolescence, represents the bottleneck for long-term integrity of the cardiovascular system. In this project, we propose to test the hypothesis that after a lifetime of repairing atherosclerotic arteries, the supply of the specific type(s) of vascular progenitor cells (VPCs) needed to maintain the homeostasis of the cardiovascular system is somehow exhausted or these cells are functionally impaired. If the VPC "spare parts" can be outsourced, the repair process can be boosted at appropriate intervals and atherosclerotic consequences delayed, perhaps indefinitely. Specific Aim 1: To determine the quantitative composition (FACS analysis), progeny functional characteristics, and gene expression phenotype (microarray analysis) of whole bone marrow cells or lineage negative side population (lin-SP) cells obtained from young versus old (wild-type, and ApoE -/-) mice; Specific Aim 2: To establish that competent VPCs are encompassed in the lin-SP fraction in the marrow, which convey the anti-atherosclerotic efficacy, by testing the effects of unfractionated bone marrow cells, enriched linSP cells and bone marrow cells deprived of lin-SP fraction obtained from young versus old (wild-type and ApoE -/-) mice in suppressing elevated plasma levels of chemo-cytokines and growth factors and in preventing atherosclerotic lesion formation in ApoE -/- mice; Specific Aim 3: To determine within the lin-SP fraction of young apoE -/- and wild-type BM the relative efficacy of a) candidate VPCs (CVPC, cells that are either depleted or functionally impaired in aging mice) isolated using markers identified in Aim 1, b) unfractionated lin-SP cells minus CVPCs (USP-CVPC), and c) unfractionated linSP cells (USP) in suppressing elevated plasma levels of IL-6, VEGF and other inflammatory markers and in preventing atherosclerosis in ApoE -/- mice; and Specific Aim 4: To determine the role of a4-integrin, VCAM-1, and E-selectin in mediating the engraftment of vascular progenitor cells to chronically injured vessel wall in apoE-/- mice. As a corollary, the relative contribution of cell engraftment versus a non-cell autonomous mechanism to vascular repair will be characterized.

描述（由申请人提供）：动脉粥样硬化代表了对船只壁反复伤害的反复伤害的异常，连续的修复过程。实际上，心血管系统不断暴露于多种侮辱，这些侮辱会随着时间的流逝而累积。参与维修过程的两个细胞来源存在：（1）局部，分化，血管内皮和平滑肌细胞，这些细胞从相邻的血管段迁移； （2）通过周围循环从骨髓募集了船尾细胞/血管祖细胞。我们最近发现，在既定的高脂胆固醇，西方型饮食的建立小鼠模型中，整个骨髓（BM）的长期给药可显着降低动脉粥样硬化病变的形成 - APOE - / - C57/B6小鼠，在缺乏APOE基因表达和正常水平的情况下（plope cloce of Plaste/normolative of Ploce cholsma choLStrAl cong）（>>>>>>>）。此外，来自年轻但不旧的ApoE - / - 小鼠的BM细胞能够预防血管再生和动脉粥样硬化。该数据强调了干细胞/血管祖细胞在血管愈合和动脉粥样硬化中的重要性，并为使用骨骼/祖细胞疗法作为动脉粥样硬化的新型预防和/或治疗策略提供了支持，尤其是对骨髓及其过时的个体，代表了瓶颈的长期完整性。在该项目中，我们建议测试以下假设：在修复动脉粥样硬化动脉的寿命后，维持心血管系统稳态所需的血管祖细胞（VPC）的特定类型的供应会以某种方式疲惫或这些细胞受到功能受损。如果可以将VPC“备件”外包，则可以在适当的时间间隔内提高维修过程，并且可能会无限期地延迟动脉粥样硬化后果。具体目的1：确定整个骨髓细胞的定量组成（FACS分析），后代功能特征和基因表达表型（微阵列分析）或谱系阴性侧群（LIN-SP）细胞（LIN-SP）细胞，这些细胞（LIN-SP）从Young versus versus versus versus（野生型和APOE- apoE--/ - ）小鼠获得；具体目的2：确定骨髓中的LIN-S-S-S-S-S-Smort中包含有效的VPC，通过测试未分离的骨髓细胞，丰富的linsp细胞和dy型骨骨髓细胞的未分离骨骨髓细胞的影响，从而传达了抗动脉粥样硬化的功效，从而传达了抗动脉粥样硬化的疗效。化学 - 赋形剂和生长因子以及防止APOE - / - 小鼠的动脉粥样硬化病变形成； Specific Aim 3: To determine within the lin-SP fraction of young apoE -/- and wild-type BM the relative efficacy of a) candidate VPCs (CVPC, cells that are either depleted or functionally impaired in aging mice) isolated using markers identified in Aim 1, b) unfractionated lin-SP cells minus CVPCs (USP-CVPC), and c) unfractionated linSP cells (USP)在抑制血浆水平升高的IL-6，VEGF和其他炎症标志物以及防止APOE-/ - 小鼠的动脉粥样硬化的过程中；和特定的目标4：确定A4-整合素，VCAM-1和E-选择蛋白在介导血管祖细胞的植入到APOE-/ - 小鼠中长期受伤的血管壁中的作用。作为推论，将表征细胞植入与非自主机制对血管修复的相对贡献。