Cell Specific Gene Editing to Close Diabetic Wounds

细胞特异性基因编辑闭合糖尿病伤口

• 批准号: 10628884
• 负责人: Chandan K Sen
• 金额: $ 57.93万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10628884
• 关键词: Affect; Amputation; Caring; Cells; Chronic; Complication; Complications of Diabetes Mellitus; DNA Methylation; Data; Dermatologic; Diabetes Mellitus; Diabetic Foot Ulcer; Discipline; Fruit; Gene Expression; Gene Silencing; Gene Targeting; Genes; Genetic study; Genomics; Genotype; Goals; Healthcare; Human; Human Genome Project; Hypermethylation; Impairment; Individual; Knowledge; Longevity; Longitudinal cohort study; Malignant Neoplasms; Morbidity - disease rate; NOTCH1 gene; Nucleotides; Ontology; Operative Surgical Procedures; Patients; Persons; Pharmacotherapy; Positioning Attribute; Prevalence; Provider; Quality of life; Research; Risk; Signal Transduction; Techniques; Testing; Transcription Initiation Site; Trauma; United States National Institutes of Health; Variant; Vision; Work; care burden; chronic wound; cohort; combinatorial; diabetic; diabetic ulcer; diabetic wound healing; epithelial wound; genetic risk factor; genome-wide; healing; lifetime risk; mortality; novel; personalized medicine; skin ulcer; skin wound; transcription factor; wound; wound closure; wound healing

Diabetic skin ulcers (DU) represent a major healthcare burden that is known to lead to amputations. It is estimated that 15–20% of all diabetics develop skin wounds across their lifespan. These include wounds caused by trauma and those caused by planned surgery. Notably, the 5-year mortality rate of DFU is higher than most cancers. Majority of these skin wounds are complicated, become chronic and are cared for by surgical and dermatological providers. It is estimated that only half of all diabetics with ulcerative skin wounds survive more than five years after their initial manifestation. The proposed work is inspired by the observation that single nucleotide variations (SNV) are important genetic factors that predispose an individual to diabetic complications. SNV are likely to engage in crosstalk with sequence-specific transcription factors and influence DNA methylation which could silence gene expression required for wound closure. Diabetic ulcer patient-based preliminary genotyping studies, identified greater than 17,000 SNV. Gene ontology analyses identified 53 SNV-containing genes relevant to wound epithelialization. NOTCH1 gene emerged as a major signaling hub associated with DU. Two NOTCH1 specific SNV, rs10870081 and rs34485221 (upstream of NOTCH1 transcription start site) were enriched in human diabetic wound-edge. The significance of SNV in wound closure remains unknown. The proposed work seeks to systematically study SNV and its association with diabetic wound closure in chronic wound patients. The combinatorial approach of collecting SNV, hypermethylation and healing trajectory data from the same patients from a longitudinal cohort study as proposed is likely to establish a new paradigm in the discipline of diabetic wound healing. Systematic patient-based genomic studies of DU are scanty and the proposed work is aimed at seeding a novel paradigm in wound healing research. The following specific aim is proposed: Aim 1. In patients with diabetic ulcer, identify SNV responsible for impaired wound closure. 1.1 Diabetes-dependent SNVs are associated with impaired wound closure. Loci affected by such SNV will be identified and tested for their significance in wound closure. 1.2 The functional significance of diabetes dependent SNV identified above, in the context of impaired wound closure, is determined by hypermethylation status of the corresponding loci.

糖尿病皮肤溃疡 (DU) 是一项重大的医疗负担，已知会导致截肢。据估计，15-20% 的糖尿病患者在其一生中都会出现皮肤伤口。这些包括外伤造成的伤口和计划手术造成的伤口。值得注意的是，DFU 的 5 年死亡率高于大多数癌症。大多数皮肤伤口都很复杂，会变成慢性伤口，需要由外科和皮肤科医生进行护理。据估计，只有一半患有溃疡性皮肤伤口的糖尿病患者在首次出现症状后存活五年以上。这项工作的灵感来自于这样的观察：单核苷酸变异（SNV）是使个体易患糖尿病并发症的重要遗传因素。 SNV 可能与序列特异性转录因子发生串扰，并影响 DNA 甲基化，从而沉默伤口闭合所需的基因表达。基于糖尿病溃疡患者的初步基因分型研究，确定了超过 17,000 个 SNV。基因本体分析确定了 53 个与伤口上皮化相关的含有 SNV 的基因。 NOTCH1 基因成为与 DU 相关的主要信号中枢。两个NOTCH1特异性SNV，rs10870081和rs34485221（NOTCH1转录起始位点的上游）在人类糖尿病伤口边缘富集。 SNV 在伤口闭合中的重要性仍不清楚。拟议的工作旨在系统地研究 SNV 及其与慢性伤口患者糖尿病伤口闭合的关系。所提出的从纵向队列研究中收集同一患者的 SNV、高甲基化和愈合轨迹数据的组合方法可能会在糖尿病伤口愈合学科中建立一个新的范例。基于患者的 DU 系统性基因组研究很少，拟议的工作旨在为伤口愈合研究播下新的范例。提出以下具体目标： 目标 1. 在糖尿病溃疡患者中，确定导致伤口闭合受损的 SNV。 1.1 糖尿病依赖性 SNV 与伤口闭合受损有关。受此类 SNV 影响的基因座将被识别并测试其在伤口闭合中的重要性。 1.2 在伤口闭合受损的情况下，上述确定的糖尿病依赖性 SNV 的功能意义由相应基因座的高甲基化状态决定。