Regulatory Mechanisms Addressing Diabetic Vasculopathy

解决糖尿病血管病变的调节机制

基本信息

批准号：
10718850
负责人：
Kanhaiya Singh
金额：
$ 38.25万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-07 至 2028-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10718850
关键词：
Accounting Address Adjuvant Adult Affect Angioblast Blood Vessels CDH5 gene Cadherins Cardiovascular system Cell Compartmentation Cells Clinical Clinical Research Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Complex CpG Islands Cyclic AMP-Dependent Protein Kinases DNA DNA Methylation Dermal Development Diabetes Mellitus Diglycerides Dinoprostone Down-Regulation Eicosanoids Elements Endothelial Cells Endothelium Environment Epigenetic Process Exhibits Gene Combinations Gene Expression Gene Targeting Generations Genes Granulation Tissue Guide RNA Healthcare Human Hyperglycemia Hypermethylation Infection Injury Inositol Ischemia Knockout Mice Literature Macrophage Mediating Medicare Methylation Molecular Mus Myelogenous Myeloid Cells Non-Insulin-Dependent Diabetes Mellitus Obesity Outcome Outcome Study PIK3CG gene PLCgamma2 Pathway interactions Patients Perfusion Phosphatidic Acid Population Prevalence Process Production Protein Dephosphorylation Proto-Oncogene Proteins c-akt Reactive Oxygen Species Recovery of Function Regimen Regulation Reporting Research Role Signal Pathway Signal Transduction Skin TREM2 gene Testing Therapeutic Therapeutic Intervention Tissues Vascular Diseases Vascular Endothelial Growth Factors Vascularization Work aging population angiogenesis beneficiary blood vessel development cadherin 5 chronic wound combination gene therapy cost demethylation diabetic diabetic ulcer diabetic wound healing epigenetic silencing expectation falls functional outcomes improved in vivo injured lipid mediator lymphatic vessel non-diabetic prevent promoter public health relevance single-cell RNA sequencing therapy outcome tool tripolyphosphate vasculogenesis wound wound healing wound vascularization

项目摘要

ABSTRACT The prevalence of diabetes, obesity, and vascular disease in an aging population is fueling a surge in chronic skin wounds, which affect >8 million Medicare beneficiaries at a cost of >$96 billion per year. An estimated 2.5% of US population is affected by chronic wounds. Diabetic ulcers (DU), complicated by infection/ischemia/high reactive oxygen species, often suffer from poor perfusion. Reliance on VEGF therapy to improve perfusion makes logical sense, yet clinical study outcomes fall far short of expectations. Possible factors limiting VEGF therapy outcomes include insufficient local concentration of VEGF signaling partners and regression of immature vessels. It is critically important to troubleshoot barriers and to establish a therapeutic regimen that delivers desirable functional outcomes. To circumvent the low-efficiency hurdle of VEGF single-gene “monotherapy”, development of “combined gene therapy” was initiated to deliver of two or more angiogenic molecules to substantially enhance efficacy. Current literature presents compelling evidence recognizing a critical role of active lipid mediators such as eicosanoids in enabling VEGF-dependent angiogenesis. However, the implementation of VEGF therapy has not tested the significance of these lipid mediators as necessary adjuvant. Recently we have reported that in diabetic ischemic skin endothelial cells, PLCγ2 levels remain low accounting for diminished efficiency of VEGF therapy. However, the underlying mechanisms of such downregulation remain unknown. Emerging evidence demonstrate that hyperglycemia leads to DNA methylation and epigenetic silencing in the complex interplay between genes and the environment in DU subjects. In addition to endothelium, PLCγ2 enables macrophage function via TREM2 signaling. TREM2+ macrophages participate in tissue angiogenesis post-injury. The central hypothesis is that hyperglycemia dependent loss of vascular PLCγ2– myeloid TREM2 crosstalk compromises VEGF-induced increase in diabetic wound-edge vascularization. Gene targeted DNA demethylation has potential to rescue PLCγ2 gene expression. The proposed line of work is the first to study CRISPR/dCas9-based in vivo targeted epigenetic editing of dermal endothelial cells as a therapeutic intervention for enabling VEGF therapy. Three aims are proposed: Aim 1. Determine the epigenetic mechanisms underlying PLCγ2 regulation of VEGF signaling under hyperglycemic conditions. Aim 2. Elucidate the molecular mechanisms by which PLCγ2 regulates wound tissue vascularization. Aim 3. Evaluate the functional significance of endothelial-targeted PLCγ2 augmentation in vascular-myeloid crosstalk as it relates to diabetic wound vascularization. The proposed research will unveil the critical role of PLCγ2 and the impact of this important element in VEGF signaling to help improve the efficiency and robustness of vasculogenic VEGF therapy in diabetic wound healing.

抽象的老龄化人口中糖尿病、肥胖症和血管疾病的流行正在加剧慢性病的激增皮肤伤口，影响超过 800 万医疗保险受益人，每年造成的损失超过 960 亿美元，估计为 2.5%。的美国人口受到慢性伤口 (DU) 的影响，并发感染/缺血/高。活性氧，经常遭受灌注不良的困扰，依赖 VEGF 治疗来改善灌注。逻辑上有道理，但临床研究结果远远低于预期，可能是限制 VEGF 的因素。治疗结果包括 VEGF 信号传导伴侣的局部浓度不足以及未成熟细胞的消退排除障碍并建立有效的治疗方案至关重要。为了规避 VEGF 单基因“单一疗法”的低效障碍，开始开发“联合基因疗法”，将两种或多种血管生成分子传递到当前的文献提供了令人信服的证据，承认了其关键作用。然而，活性脂质介质如类二十烷酸能够促进 VEGF 依赖性血管生成。 VEGF治疗的实施尚未测试这些脂质介质作为必要佐剂的意义。最近我们报道，在糖尿病缺血性皮肤内皮细胞中，PLCγ2水平仍然较低然而，这种下调的潜在机制仍然存在。新的证据表明高血糖会导致 DNA 甲基化和表观遗传。 DU 受试者中基因和环境之间复杂的相互作用被沉默。 PLCγ2 通过 TREM2 信号传导激活巨噬细胞功能，参与组织。损伤后血管生成的中心假设是高血糖依赖性血管 PLCγ2- 损失。骨髓 TREM2 串扰损害 VEGF 诱导的糖尿病伤口边缘血管化基因的增加。靶向 DNA 去甲基化有可能挽救 PLCγ2 基因表达。第一个研究基于 CRISPR/dCas9 的真皮内皮细胞体内靶向表观遗传编辑作为治疗方法提出了三个目标：目标 1. 确定表观遗传机制。高血糖条件下 PLCγ2 对 VEGF 信号传导的潜在调节目标 2. 阐明分子机制。 PLCγ2 调节伤口组织血管化的机制目标 3. 评估功能意义。血管-骨髓串扰中内皮靶向 PLCγ2 增强与糖尿病伤口相关的研究拟议的研究将揭示 PLCγ2 的关键作用以及这一重要作用的影响。 VEGF 信号传导元件有助于提高血管生成 VEGF 治疗的效率和稳健性糖尿病伤口愈合。