Reprogramming Macrophages to Improve Vascular Healing in Diabetes

重编程巨噬细胞以改善糖尿病血管愈合

• 批准号: 10674353
• 负责人: Alan Ross Morrison
• 金额: --
• 依托单位: PROVIDENCE VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10674353
• 关键词: Acute; Aging; American; Amputation; Animal Model; Area; Binding; Biology; Blood Glucose; Blood Vessels; Bone Marrow; Bone Marrow Involvement; Bypass; Caring; Cells; Chronic Disease; Cladribine; Complex; Data; Debridement; Diabetes Mellitus; Diabetic mouse; Disease; Effectiveness; Experimental Diabetes Mellitus; Financial Hardship; Genetic Transcription; Goals; Health; Healthcare Systems; Hispanic Americans; IL1R1 gene; Immune; Impaired healing; Impaired wound healing; Impairment; Infection Control; Inflammation; Inflammatory; Injury; Interleukin-1 beta; Laboratories; Limb Salvage; Macrophage; Mechanics; Mediating; Medical; Mexican Americans; Modeling; Molecular; Morbidity - disease rate; Mus; Myelogenous; Nuclear; Parents; Pathway interactions; Patients; Peripheral arterial disease; Process; Production; Promoter Regions; Protein Isoforms; Research; Research Personnel; Role; STAT3 gene; Scientist; Signal Transduction; Source; Stents; Therapeutic; Tissues; Training; Ulcer; Up-Regulation; VEGFA gene; Vascular Diseases; Vascularization; Veterans; Work; angiogenesis; autocrine; career; constitutive expression; design; diabetic; healing; improved; limb loss; military veteran; mortality; necrotic tissue; new growth; novel therapeutics; parent grant; patient subsets; preclinical study; prevent; programs; promoter; response; revascularization surgery; skin regeneration; skin wound; therapeutic angiogenesis; wound healing

Impaired wound healing in US veterans with diabetes mellitus is a major source of morbidity and mortality as well as a large financial strain on the VA health care system. Current treatment paradigms, including debridement of necrotic tissue, infection control, local ulcer care, mechanical off-loading, and management of blood glucose levels, are modestly effective at best. Despite much research in this area, the critical molecular mechanisms regulating angiogenesis-directed wound healing remain minimally defined. Recently, our group identified an important role for early inflammatory macrophage VEGF-A production in promoting consequent angiogenesis/ arteriogenesis required for adequate wound healing. We determined macrophage proangiogenic VEGF-A isoform transcription to be dependent on autocrine IL-1β-IL-1R signaling through activation of nuclear NF-κB and binding of NF-κB to the VEGF-A promoter region. Analogous to macrophage IL-1β-deleted mice, mice with experimental diabetes demonstrated impaired tissue proangiogenic VEGF-A expression and consequent reductions in wound healing and arteriogenesis. Under inflammatory conditions, bone marrow derived macrophages (BMDMs) from diabetic mice expressed reduced VEGF-A consistent with reduced signaling response to IL-1β, and interestingly, there is reduced expression of several key IL-1R signaling complex components, suggesting an impaired responsiveness to IL-1β. Our working model is that DM uncouples IL-1R signaling from VEGF-A transcription, causing impaired proangiogenic VEGF-A expression and consequent decreased angiogenesis-based healing. In this VA Research Supplement to Promote Diversity proposal, we seek to reactivate macrophage VEGF-A expression by effectively bypassing IL-1R signaling and upregulating NF-κB activity directly. The primary hypothesis is that rescue of DM-mediated impairments in VEGF-A-dependent angiogenesis via direct upregulation of NF-κB activity is a viable therapeutic angiogenesis strategy for wound healing. We aim to 1) demonstrate that myeloid expression of constitutively active IKK-2 can rescue proangiogenic VEGF-A expression and VEGF-A-dependent angiogenesis in the context of macrophage IL-1R-deletion; and 2) demonstrate that BMDMs programed to express constitutively active IKK-2 are sufficient to increase wound healing angiogenesis and arteriogenesis using mice with experimental DM. By manipulating mechanistic pathways downstream of IL-1R signaling-dependent VEGF-A expression to therapeutically reactivate diabetic macrophage angiogenic and wound healing responses, we will validate a therapeutic angiogenesis strategy that has tremendous potential to impact the lives of US veterans and all Americans. Moreover, this proposal serves as a robust training platform for Dr. Roberto Mendez, a Mexican American and US veteran, who is committed to developing a VA-based research career focused on veteran health. These studies will provide him with important preliminary data to support his own VA CDA-1 application proposal within the next two years.

美国退伍军人糖尿病患者伤口愈合受损是发病率和死亡率的主要来源， 以及退伍军人管理局医疗保健系统面临的巨大财务压力。目前的治疗模式，包括 坏死组织的清创、感染控制、局部溃疡护理、机械卸载和 血糖水平，充其量是适度有效的。尽管在这一领域进行了大量研究，但关键的分子 调节血管生成导向的伤口愈合的机制仍然最低限度地被定义。最近，我们集团 确定了早期炎症巨噬细胞VEGF-A的产生在促进随后的炎症反应中的重要作用。 充分伤口愈合所需的血管生成/动脉生成。我们确定巨噬细胞促血管生成 VEGF-A亚型的转录依赖于通过激活核因子自分泌IL-1β-IL-1 R信号转导 NF-κB和NF-κB与VEGF-A启动子区的结合。与巨噬细胞IL-1β缺失小鼠类似， 患有实验性糖尿病的小鼠表现出受损的组织促血管生成VEGF-A表达， 从而减少伤口愈合和动脉生成。在炎症条件下，骨髓 来自糖尿病小鼠的衍生的巨噬细胞（BMDM）表达减少的VEGF-A，与减少的VEGF-A表达一致。 IL-1β信号应答，有趣的是，几个关键的IL-1 R信号转导的表达减少， 复合组分，表明对IL-1β的反应性受损。我们的工作模型是DM 将IL-1 R信号与VEGF-A转录解偶联，导致促血管生成VEGF-A表达受损 以及随之而来的基于血管生成的愈合减少。在这个VA研究补充，以促进多样性 我们试图通过有效地绕过IL-1 R信号传导， 直接上调NF-κB活性。主要的假设是，挽救DM介导的损伤， 通过直接上调NF-κB活性的VEGF-A依赖性血管生成是一种可行的治疗性血管生成 伤口愈合的策略。我们的目的是：1）证明组成型活性IKK-2的髓样表达 可以挽救促血管生成VEGF-A表达和VEGF-A依赖性血管生成， 巨噬细胞IL-1 R缺失;和2）证明BMDM编程表达组成型活性IKK-2 足以增加伤口愈合血管生成和动脉生成。通过 操纵IL-1 R信号传导依赖性VEGF-A表达下游的机制途径， 治疗性地重新激活糖尿病巨噬细胞血管生成和伤口愈合反应，我们将验证 治疗性血管生成策略，具有巨大的潜力，影响美国退伍军人和所有 美国人此外，该提案还为墨西哥人罗伯托·门德斯博士提供了一个强大的培训平台。 美国和美国退伍军人，致力于发展以退伍军人为重点的VA研究事业 健康这些研究将为他提供重要的初步数据，以支持他自己的VA CDA-1申请 建议在未来两年内。