Tissue reprogramming in diabetic wound healing

糖尿病伤口愈合中的组织重编程

基本信息

批准号：
10224448
负责人：
Sashwati Roy
金额：
$ 48.15万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10224448
关键词：
Affect Antisense Oligonucleotide Therapy Antisense Oligonucleotides Blood Vessels Cell physiology Cells Characteristics Chronic Data Dermal Devices Diabetes Mellitus Diabetic mouse Endothelium FLI1 gene FOXC2 gene Fibroblasts Foundations Future Gene Silencing Healthcare Human Immunologic Surveillance Intervention Investigation Ischemia Longevity Molecular Mus Nanochip Analytical Device Outcome Perfusion Plasmids Point of Care Technology Population Regenerative Medicine Reporting Role Silicon Site Skin Skin wound healing System Technology Testing Tissues Topical application Untranslated RNA Vascular Diseases Vascularization Work base chronic wound diabetic diabetic patient diabetic ulcer diabetic wound healing endothelial dysfunction gene therapy humanized mouse improved in vivo knock-down mouse model nanofabrication novel regenerative response skin ulcer skin wound transcription factor vasculogenesis wound wound closure wound healing wound vascularization

项目摘要

ABSTRACT An estimated 2% of US population is affected by chronic wounds. About 15–20% of all diabetic patients develop skin wounds across their lifespan, the majority evolving toward chronicity. It is estimated that only 56% of diabetics with ulcerative wounds survive more than 5 years after their initial manifestation. This proposal seeks to achieve successful vascularization of diabetic wounds utilizing the novel regenerative principles of non-viral in vivo tissue reprogramming. Tissue nanotransfection technology (TNT), recently reported by the PI, achieves tissue reprogramming, under immune surveillance in vivo, using a topical electrophoretic delivery system enabled by a novel nanofabricated silicon chip hardware. The proposed studies emanate from prior work by the applicant on the role of TNT delivery of specific transcription factors Etv2, Foxc2, and Fli1 (EFF) to achieve vasculogenic reprograming in vivo. EFF was thus discovered as a vasculogenic plasmid cocktail capable of vascularizing ischemic tissue. The proposed work is based on first evidence that instead of the use of plasmid cocktails that have been used by many groups to achieve cell reprogramming, a single anti-sense oligonucleotide (ASO) is highly effective in producing induced vasculogenic cells (iV) from skin fibroblasts. Preliminary data show that under in vivo conditions, ASO can improve perfusion of diabetic wound tissue. Because diabetes is known to cause endothelial dysfunction and vasculopathy, TNT delivery of ASO will be tested to improve wound outcomes. Vasculogenic effects of ASO is achieved by turning on a major vascular switch Fli1. The proposed work will delineate the molecular mechanisms of ASO-induced perfusion of diabetic cutaneous wound perfusion (Aim 1). To enable invasive mechanistic studies, a humanized mouse model has been thus proposed in Aim 2. The following two specific aims are proposed: Aim 1: Elucidate the significance and molecular mechanisms by which ASO induces accelerated wound closure in diabetic mice. 1.1 ASO delivery by TNT rescues healing of cutaneous wounds in murine models of diabetes; 1.2 ASO delivery improves diabetic wound vascularization by rescuing Fli1 from miR-200b dependent gene silencing; 1.3 A subset of wound-site fibroblasts contribute to the pre-existing vasculature by acquiring vasculogenic endothelial-like characteristics post-ASO treatment. Aim 2: Test significance of ASO reprogramming in a humanized diabetic mouse model. 2.1 ASO delivery by TNT accelerates wound healing and improves wound vascularization in a humanized NSG diabetic mouse model. The proposed work lays the foundation stone to future trials attempting to reprogram human skin stroma towards improved perfusion of the wound-site.

抽象的据估计，有 2% 的美国人受到慢性伤口的影响，大约 15-20% 的糖尿病患者在一生中都会出现皮肤伤口，其中大多数会发展成慢性伤口。据估计，只有 56% 的溃疡性伤口患者能够存活超过 10 年。该提案旨在利用非病毒体内组织重编程技术（TNT）的新再生原理成功实现糖尿病伤口的血管化。 PI 最近报道，在体内免疫监视下，使用由新型纳米制造硅芯片硬件实现的局部电泳传递系统实现了组织重编程。拟议的研究源于申请人之前关于 TNT 传递特定转录的作用的工作。因此，EFF 被发现是一种能够使缺血组织血管化的血管生成质粒混合物。拟议的工作基于第一个证据，即单一反义寡核苷酸（ASO）可以非常有效地从皮肤产生诱导血管生成细胞（iV），而不是使用许多团体使用的质粒混合物来实现细胞重编程。初步数据表明，在体内条件下，ASO 可以改善糖尿病伤口的灌注，因为已知糖尿病会导致内皮功能障碍和血管病变，因此 ASO 的 TNT 递送会受到影响。经测试可改善伤口结果。ASO 的血管生成作用是通过打开主要血管开关 Fli1 来实现的。拟议的工作将描述 ASO 诱导的糖尿病皮肤伤口灌注的分子机制（目标 1）。因此，在目标 2 中提出了人源化小鼠模型。提出了以下两个具体目标：目标 1：阐明 ASO 诱导加速伤口闭合的意义和分子机制1.1 TNT 递送 ASO 可挽救糖尿病小鼠模型的皮肤伤口愈合；1.2 ASO 递送可通过挽救 Fli1 免受 miR-200b 依赖性基因沉默的影响而改善糖尿病伤口血管化；1.3 伤口部位成纤维细胞的一个亚群有助于预目标 2：测试 ASO 治疗后的血管生成内皮样特征。 2.1 人源化糖尿病小鼠模型中的 ASO 重编程可加速人源化 NSG 糖尿病小鼠模型中的伤口愈合并改善伤口血管化。这项工作为未来尝试重新编程人类皮肤基质以改善皮肤基质的灌注奠定了基础。伤口部位。