Netrin-1 and Netrin-1 Preconditioned EPCs in Vascular Protection

Netrin-1 和 Netrin-1 预处理 EPC 在血管保护中的作用

• 批准号: 9917420
• 负责人: Hua Linda Cai
• 金额: $ 45.86万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9917420
• 关键词: Address; Angioplasty; Apolipoprotein E; Apoptosis; Atherosclerosis; Attenuated; Blood Vessels; Cell Survival; Cell physiology; Clinical; Cyclic GMP; Dependence; Development; Dyslipidemias; Endothelium; Fatty acid glycerol esters; Homing; Infiltration; Injury; Intervention; Knockout Mice; Lead; Mediating; MicroRNAs; Molecular; Mus; Mutation; NTN1 gene; Oxidants; Oxidative Stress; Pathology; Pathway interactions; Phosphotransferases; Physiological; Procedures; Role; Signal Pathway; Signal Transduction; attenuation; blood vessel occlusion; conditioning; design; endothelial stem cell; feeding; femoral artery; in vivo; innovation; macrophage; migration; monocyte; netrin receptor; novel; novel therapeutics; preconditioning; protective effect; restenosis

ABSTRACT The application aims to identify potential roles and molecular mechanisms of netrin-1 and netrin-1 pre-conditioned endothelial progenitor cells (EPCs) in vascular protection, specifically related to their anti-restenosis and anti- atherosclerosis effects. Four entirely novel hypotheses will be addressed: 1) Netrin-1 pre-conditioning protects EPCs from oxidative stress induced apoptosis through PI3K-dependent p70s6 kinase activation and Bim inhibition; In addition, mechanistic pathways mediating novel regulatory miRNAs-dependent enhancement of EPC survival by netrin-1 will be fully delineated. By targeting these novel mechanisms, EPC functions can be augmented to attenuate restenosis and atherosclerosis, leading to development of novel therapeutics. 2) Netrin-1 inhibits monocyte activation in a UNC5B (repellant class of netrin-1 receptor)-dependent fashion; and that UNC5B is innovatively regulated by a p47phox-dependent mechanism. Therefore, inhibition of p47phox may potentiate the beneficial effects of netrin-1 in limiting restenosis and atherosclerosis via attenuation of UBC5B expression and UNC5B-dependent monocyte activation. Netrin-1 also inhibits VSMC migration and proliferation via a NO/cGMP/PKG/p38MAPK-dependent mechanism. 3) Administration with netrin-1 or netrin-1 preconditioned EPCs attenuates atherosclerosis in high-fat fed apoE null and LDLR deficient mice. These protective effects are at least in part attributed to augmented EPC function and abrogated monocyte activation, as well as attenuated VSMC proliferation and migration. 4) Endogenous netrin-1 signaling is physiologically protective against restenosis and atherosclerosis, loss of which exaggerates vascular pathologies. We hypothesize that femoral artery injury/high-fat feeding into the netrin-1 or netrin-1/apoE double knockout mice will result in exaggerated restenosis/atherosclerosis. The overall hypothesis is that administration of netrin-1 and netrin-1 preconditioned EPCs remarkably attenuate restenosis and atherosclerosis via mechanisms of augmented EPC function (increased survival, proliferation and homing) to lead to rapid re-endothelialization, attenuated monocyte and VSMC activation, as well as diminished dyslipidemia. Endogenous netrin-1 signaling is physiologically vascular protective, amplification of which with exogenous administration of netrin-1 is necessary to result in sufficient protection, whereas deficiency in the endogenous signaling of netrin-1 (i.e. due to genetic defects) is prone to deteriorated vascular pathologies. Four specific aims are designed to fully address these hypotheses, and accomplishments of the aims may establish netrin-1, netrin-1 pre-conditioned EPCs, and modulators of related pathways as novel therapeutic options for vascular pathologies of restenosis and atherosclerosis.

抽象的 该应用旨在确定 netrin-1 和 netrin-1 预处理的潜在作用和分子机制 内皮祖细胞（EPC）在血管保护中的作用，特别是与它们的抗再狭窄和抗- 动脉粥样硬化的影响。将讨论四个全新的假设：1) Netrin-1 预处理保护 氧化应激中的 EPC 通过 PI3K 依赖性 p70s6 激酶激活和 Bim 抑制诱导细胞凋亡； 此外，介导新型调控 miRNA 依赖性增强 EPC 存活的机制途径 netrin-1 将被完全描绘。通过针对这些新机制，EPC 功能可以增强到 减轻再狭窄和动脉粥样硬化，导致新疗法的开发。 2) Netrin-1 抑制 单核细胞以 UNC5B（netrin-1 受体排斥类）依赖性方式激活； UNC5B 是 通过 p47phox 依赖性机制进行创新性调节。因此，抑制 p47phox 可能会增强 netrin-1 通过减弱 UBC5B 表达和限制再狭窄和动脉粥样硬化的有益作用 UNC5B 依赖性单核细胞激活。 Netrin-1 还通过以下途径抑制 VSMC 迁移和增殖： NO/cGMP/PKG/p38MAPK 依赖性机制。 3) 使用 netrin-1 或 netrin-1 预处理的 EPC 进行给药 减轻高脂肪喂养的 apoE 缺失和 LDLR 缺陷小鼠的动脉粥样硬化。这些保护作用至少在 部分归因于 EPC 功能增强、单核细胞活化消除以及 VSMC 减弱 扩散和迁移。 4) 内源性 netrin-1 信号传导对再狭窄具有生理保护作用 动脉粥样硬化，其丧失会加剧血管病变。我们假设股动脉损伤/高脂肪 喂养 netrin-1 或 netrin-1/apoE 双基因敲除小鼠将导致过度的再狭窄/动脉粥样硬化。 总体假设是，给予 netrin-1 和 netrin-1 预处理的 EPC 会显着减弱 通过增强 EPC 功能的机制（增加生存、增殖和 归巢）导​​致快速再内皮化、单核细胞和 VSMC 激活减弱以及 血脂异常。内源性 netrin-1 信号传导具有生理性血管保护作用，其放大作用是 外源性给予 netrin-1 对于产生充分的保护是必要的，而 netrin-1 的缺乏 netrin-1 的内源性信号传导（即由于遗传缺陷）很容易导致血管病理恶化。四 设计具体目标是为了充分解决这些假设，并且目标的实现可能会建立 netrin-1、netrin-1 预处理的 EPC 和相关途径的调节剂作为新的治疗选择 再狭窄和动脉粥样硬化的血管病理学。