iPSC-derived Repair-Responsive Fibroblasts to Heal Diabetic Foot Ulcers

iPSC 衍生的修复反应性成纤维细胞可治愈糖尿病足溃疡

• 批准号: 8373639
• 负责人: JONATHAN A GARLICK
• 金额: $ 67.88万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8373639
• 关键词: Address; Age; Amputation; Animal Model; Autologous; Biopsy; Caring; Cell physiology; Cells; Chronic; DNA Methylation; Data; Defect; Diabetes Mellitus; Diabetic Foot; Diabetic Foot Ulcer; Diabetic wound; Disease; Elderly; Embryo; Epigenetic Process; Extracellular Matrix; FDA approved; Fibroblasts; Functional disorder; Future; Generations; Genes; Goals; Growth Factor; Harvest; Healed; Hospitalization; Hyperglycemia; Immune; In Situ; Knowledge; Light; Link; Mediating; Memory; Metabolic; MicroRNAs; Mission; Modification; Patient Care; Patients; Pattern; Periodontal Diseases; Phenotype; Process; Production; Public Health; Publishing; Quality of life; Refractory; Research; Site; Skin; Source; Testing; Therapeutic; Tissue Microarray; Ulcer; Wound Healing; age related; angiogenesis; base; cell growth; cell motility; cell type; diabetic; diabetic wound healing; glycemic control; healing; histone modification; improved; in vivo Model; induced pluripotent stem cell; insight; new technology; next generation; novel; novel strategies; promoter; repaired; response; skills; stem cell differentiation; stem cell technology; translational approach; wound

DESCRIPTION (provided by applicant): Diabetic foot ulceration (DFU) is a major problem that significantly impairs quality of life of the patient, leads to prolonged hospitalizations and may require major amputation, demonstrating the urgent need to develop next generation treatments for these and other chronic wounds. To fill this critical gap in patient care, novel sources of autologous cells that are more repair-competent are immediately needed. Our long term goal is to develop a therapeutic approach, based on induced pluripotent stem cell (iPSC) technologies, that will reverse DFU fibroblasts from a non-healing to a healing phenotype by treating these cells in situ, directly in the patient's wound without needing to remove and culture them. To lay the groundwork for this, the immediate goal of our project is to greatly improve the wound repair potency of DFU fibroblasts following their reprogramming to iPSC, that will reveal how "epigenetic memory" (patterns of DNA methylation retained from the cell type reprogrammed) can best be exploited to acquire a spectrum of pleiotrophic effects that will trigger wound repair. In light of the emergence of epigenetics as a critical regulator of the "metabolic memory" linked to diabetic cell dysfunction, DFU fibroblasts are likely to be controlled by epigenetic mechanisms (miRNA, histone modification and DNA methylation). As a result, we expect iPSC reprogramming to reset the epigenome and reverse "metabolic memory" to improve repair after iPSC differentiation. The overall objective is to develop new sources of autologous, repair-competent fibroblasts using iPSC technologies that will dramatically improve DFU therapy. Our central hypothesis is that DFU fibroblasts will become repair- competent when reprogrammed to iPSC and subsequently differentiated to a fibroblast lineage by acquiring repair-promoting functions that will be mediated by epigenetic controls. The rationale for our research is based on exciting new preliminary data that has established the augmented healing potential of iPSC-derived fibroblasts (iPDK) and in vivo models of diabetic wound repair developed in our labs that will determine this repair potential. To test this hypothesis, we have developed skills and data that support the feasibility of this approach by establishing that iPSC-derived fibroblasts are more versatile than their parental fibroblasts. We plan to test our central hypothesis by pursuing the following specific AIMS: AIM 1-Identify cellular functions that trigger repair-competency when repair-deficient, DFU fibroblasts are reprogrammed to iPSC and differentiated to fibroblasts, AIM 2-Reveal if the switch to repair-competency is linked to distinct DNA methylation profiles, histone modifications or miRNA signatures that regulate repair functions in iPDK fibroblasts, and AIM 3- Establish repair efficacy of iPSC-derived fibroblasts in two, well-established animal models of diabetes and optimize their delivery to heal diabetic wounds. Ultimately, this knowledge has the potential to transform the care of patients suffering from DFUs and will be widely applicable to many other types of non- healing wounds, as well as to periodontal disease and aging-related diseases. PUBLIC HEALTH RELEVANCE: The proposed research is relevant to public health because the discovery that non-healing cells from diabetic foot ulcers can be reversed to those that can heal these wounds will fundamentally shift the field of wound care to the threshold of novel in situ treatments, directly in the patient's wound. It will also dramatically improve the efficacy of topical, FDA-approved therapies that are currently not effective in most diabetic foot ulcer patients. Thus, the proposed research is relevant to the part of the NIH's mission that pertains to developing fundamental knowledge that will help reduce the burden of diabetes and wound care, and improve treatment of other chronic wounds in a broad range of patients such as the elderly and immune-compromised patients.

描述（由申请人提供）：糖尿病足溃疡（DFU）是一个严重问题，会严重损害患者的生活质量，导致住院时间延长，并可能需要大截肢，这表明迫切需要开发针对这些和其他疾病的下一代治疗方法慢性伤口。为了填补患者护理方面的这一关键空白，迫切需要具有更强修复能力的新自体细胞来源。我们的长期目标是开发一种基于诱导多能干细胞 (iPSC) 技术的治疗方法，通过直接在患者伤口中原位治疗这些细胞，将 DFU 成纤维细胞从不愈合表型逆转为愈合表型，而无需去除并培养它们。为了为此奠定基础，我们项目的直接目标是大大提高 DFU 成纤维细胞重编程为 iPSC 后的伤口修复能力，这将揭示“表观遗传记忆”（从重编程的细胞类型中保留的 DNA 甲基化模式）是如何实现的可以最好地利用它来获得一系列多效作用，从而触发伤口修复。 鉴于表观遗传学作为与糖尿病细胞功能障碍相关的“代谢记忆”的关键调节因子，DFU 成纤维细胞很可能受到表观遗传机制（miRNA、组蛋白修饰和 DNA 甲基化）的控制。因此，我们期望 iPSC 重编程能够重置表观基因组并逆转“代谢记忆”，从而改善 iPSC 分化后的修复。总体目标是利用 iPSC 技术开发具有修复能力的自体成纤维细胞的新来源，从而显着改善 DFU 治疗。我们的中心假设是，DFU 成纤维细胞在重编程为 iPSC 时将变得具有修复能力，并随后通过获得由表观遗传控制介导的修复促进功能而分化为成纤维细胞谱系。我们研究的基本原理是基于令人兴奋的新初步数据，这些数据确定了 iPSC 衍生的成纤维细胞 (iPDK) 的增强愈合潜力，以及我们实验室开发的糖尿病伤口修复体内模型，该模型将确定这种修复潜力。为了检验这一假设，我们开发了支持这种方法可行性的技能和数据，确定 iPSC 衍生的成纤维细胞比其亲代成纤维细胞更通用。我们计划通过追求以下具体目标来测试我们的中心假设：AIM 1-识别修复缺陷时触发修复能力的细胞功能，DFU 成纤维细胞被重新编程为 iPSC 并分化为成纤维细胞，AIM 2-揭示是否切换到修复-能力与调节 iPDK 成纤维细胞修复功能的不同 DNA 甲基化谱、组蛋白修饰或 miRNA 特征相关，以及 AIM 3- 建立修复iPSC 衍生的成纤维细胞在两种成熟的糖尿病动物模型中的功效，并优化其递送以治愈糖尿病伤口。最终，这些知识有可能改变 DFU 患者的护理，并将广泛适用于许多其他类型的不愈合伤口，以及牙周病和衰老相关疾病。 公共健康相关性：拟议的研究与公共健康相关，因为糖尿病足溃疡中的不愈合细胞可以逆转为可以治愈这些伤口的细胞这一发现将从根本上将伤口护理领域转向新型原位治疗的门槛，直接在病人的伤口上。也将大幅提升功效 FDA 批准的局部疗法目前对大多数糖尿病足溃疡患者无效。因此，拟议的研究与 NIH 的使命部分相关： 发展基础知识将有助于减轻糖尿病和伤口护理的负担，并改善老年人和免疫功能低下患者等广大患者的其他慢性伤口的治疗。