Epigenetic regulation of keratinocyte function in normal and pathologic skin repair

正常和病理性皮肤修复中角质形成细胞功能的表观遗传调控

基本信息

批准号：
10677308
负责人：
Jadie Moon
金额：
$ 4.17万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
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：
2023-08-01 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10677308
关键词：
Amputation Animals Blood specimen Cell physiology Cells Chromatin Complex Data Defect Development Diabetic mouse Enzymes Epigenetic Process Epithelium Failure Functional disorder Gene Expression Genes Genetic Genetic Transcription Healthcare Systems Histone H3 Human IL17 gene Immune Immunologist Impaired wound healing Impairment Inflammation Inflammatory Interferon-beta Janus kinase Link Lysine Macrophage Mediating Mediator Molecular Morbidity - disease rate Mus NF-kappa B Non-Insulin-Dependent Diabetes Mellitus Pathologic Pathway interactions Patients Phenotype Play Process Production Regulation Repression Research Design Resolution Role STAT protein STAT1 gene Signal Transduction Skin Skin repair Stat3 protein TNF gene Techniques Testing Therapeutic Agents Tissue Sample Treatment Efficacy United States Up-Regulation Wound models cost cytokine diabetic diabetic ulcer diabetic wound healing diet-induced obesity dietary epigenetic regulation experimental study healing histone methylation histone methyltransferase human RNA sequencing improved in vivo keratinocyte migration mortality mouse model new therapeutic target non-healing wounds novel pharmacologic prevent single-cell RNA sequencing skills success targeted treatment therapeutic target tissue repair translational approach translational scientist wound wound care wound healing

项目摘要

PROJECT SUMMARY/ABSTRACT Non-healing wounds in patients with Type 2 Diabetes (T2D) are a major cause of morbidity and mortality and are increasing at an alarming rate. Failure of wound healing in T2D patients represents the most common cause of amputation in the US with a 5-year mortality rate of nearly 50%. Thus, a critical need exists for understanding the wound healing defects in T2D in order to develop targeted therapies. We will utilize both genetic (db/db) and dietary (diet-induced obese) murine models of T2D as well as human wound tissue and blood samples collected from T2D patients to explore mechanisms of impaired wound healing. We present data using human single cell RNA sequencing and murine diabetic wound models, that the repressive histone methyltransferase, SETDB2, is decreased in diabetic wound keratinocytes resulting in increased production of inflammatory genes, namely TNFα. Further, we have identified that interferon-beta (IFNꞵ), via a janus kinase (JAK1) / signal transducer and activator of transcription (STAT) mechanism induces SETDB2 in wound keratinocytes. Our preliminary data also identified that IL-17A is increased in diabetic wounds and may, in addition to reduced levels of IFNꞵ in diabetic wounds, suppress SETDB2 in diabetic wound keratinocytes. These results have led to our hypothesis that induction of SETDB2 in wound keratinocytes represses expression of NFkB-mediated genes (i.e., TNFα) that maintain wound inflammation and directly impair keratinocyte migration, thereby promoting tissue repair. We postulate that in diabetic wounds, it is the failure to induce SETDB2 and repress NFκB-mediated inflammatory (Tnfα) genes in keratinocytes that prevents resolution of inflammation and impairs keratinocyte migration and results in poor wound healing. We will test our hypotheses through three specific aims: Aim 1: To define the in vivo regulation of NFκB-mediated gene expression by SETDB2 in normal and diabetic wound keratinocytes. Aim 2: To determine the JAK/STAT-mediated mechanism(s) that regulate keratinocyte-specific SETDB2 expression in normal and diabetic wound tissue. Aim 3: To examine the therapeutic efficacy and timing of SETDB2-regulated TNFα inhibition on keratinocyte migration and inflammatory wound Mφ phenotype. In this translational approach, our data will pave the way for the development of promising therapeutic agents aimed at the targeting of epigenetic pathways that mediate diabetic wound keratinocyte function and thereby promote diabetic wound repair.

项目概要/摘要 2 型糖尿病 (T2D) 患者的伤口不愈合是发病和死亡的主要原因 T2D 患者伤口愈合失败是最常见的原因。美国的截肢率高达 50%，因此，迫切需要了解这一情况。我们将利用遗传 (db/db) 和 T2D 的伤口愈合缺陷来开发靶向治疗。饮食（饮食诱导肥胖）T2D 小鼠模型以及收集的人类伤口组织和血液样本我们利用人类单细胞从 T2D 患者身上探索伤口愈合受损的机制。 RNA 测序和小鼠糖尿病伤口模型表明，抑制性组蛋白甲基转移酶 SETDB2 糖尿病伤口角质形成细胞减少，导致炎症基因产生增加，即此外，我们还通过 janus 激酶 (JAK1)/信号转导器鉴定了干扰素-β (IFNꞵ)，并且我们的初步数据还表明，转录激活剂 (STAT) 机制会在伤口角质形成细胞中诱导 SETDB2。发现糖尿病伤口中的 IL-17A 增加，并且除了糖尿病患者中 IFNꞵ 水平降低之外，还可能伤口，抑制糖尿病伤口角质形成细胞中的 SETDB2。这些结果导致我们的假设：在伤口角质形成细胞中诱导 SETDB2 会抑制 NFkB 介导的基因（即 TNFα）的表达，维持伤口炎症并直接损害角质细胞迁移，从而促进组织修复。假设在糖尿病伤口中，未能诱导 SETDB2 并抑制 NFκB 介导的炎症角质形成细胞中的 (Tnfα) 基因可防止炎症消退并损害角质形成细胞迁移和我们将通过三个具体目标来检验我们的假设：目标 1：定义 in。 SETDB2 在正常和糖尿病伤口角质形成细胞中体内调节 NFκB 介导的基因表达。 2：确定 JAK/STAT 介导的调节角质形成细胞特异性 SETDB2 表达的机制目标 3：检查 SETDB2 调节的治疗效果和时机。 TNFα 对角质形成细胞迁移和炎症伤口 Mφ 表型的抑制在这种转化方法中，我们的数据将为开发有前景的治疗药物铺平道路，这些治疗药物旨在靶向介导糖尿病伤口角质形成细胞功能并由此促进糖尿病伤口的表观遗传途径维修。