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）是一个主要问题，严重损害了患者的生活质量，导致住院时间延长，可能需要重大截肢，这表明迫切需要为这些和其他慢性伤口开发下一代治疗。为了填补患者护理中的这一关键空白，需要立即需要更具修复能力的自体细胞来源。我们的长期目标是基于诱导的多能干细胞（IPSC）技术开发一种治疗方法，该技术将通过直接在患者的伤口中处理这些细胞，而无需去除和培养它们，从而将DFU成纤维细胞从非污染转向愈合表型。 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 tr​​igger wound repair. 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”的关键调节剂的出现，DFU成纤维细胞可能受到表观遗传机制（miRNA，组蛋白修饰和DNA甲基化）的控制。结果，我们希望IPSC重新编程将重置表观基因组和反向“代谢记忆”，以改善IPSC分化后的维修。总体目的是使用IPSC技术来开发新的自体维修能力成纤维细胞来源，这些技术将显着改善DFU疗法。我们的中心假设是，当重新编​​程为IPSC时，DFU成纤维细胞将获得维修，然后通过获取将通过表观遗传控制介导的修复促进功能来区分成纤维细胞谱系。我们的研究的基本原理是基于令人兴奋的新初步数据，该数据已经确定了IPSC衍生的成纤维细胞（IPDK）的增强愈合潜力和我们实验室中开发的糖尿病伤口修复的体内模型，这些模型将确定这种修复潜力。为了检验这一假设，我们通过确定IPSC衍生的成纤维细胞比其父母的成纤维细胞更广泛的成纤维细胞来开发技能和数据，以支持这种方法的可行性。我们计划通过追求以下特定目的来测试我们的中心假设：目标1-识别细胞功能，当维修不足时，将dfu成纤维细胞重新编程为IPSC时触发修复的函数，并区分到成纤维细胞，AIL与2- reveal相连，如果切换到不同的维修功能与不同的DNA甲基化拟合功能相关，则具有不同的DNA甲基化拟合功能，或者是模拟的DNA甲基化拟合功能，其历史偶发性型号，历史偶然的型号，历史偶然的DNA甲基化拟合功能，历史偶然甲基化的型号，其历史偶发性的型号，历史偶然的DNA甲基化功能，其历史偶发性的含量，历史偶然的型号，或者是iRNA的含量。 IPDK成纤维细胞，AIM 3-建立IPSC衍生的成纤维细胞在两种糖尿病的动物模型中的修复功效，并优化其递送以治愈糖尿病伤口。最终，这种知识有可能改变患有DFU的患者的护理，并将广泛适用于许多其他类型的非愈合伤口，以及牙周疾病和与衰老有关的疾病。 公共卫生相关性：拟议的研究与公共卫生有关，因为发现可以将糖尿病足溃疡的非治疗细胞逆转到可以治愈这些伤口的人，这从根本上将伤口护理领域转移到了直接在患者伤口中的原位治疗阈值中。它也将显着提高 局部，FDA批准的疗法目前在大多数糖尿病足溃疡患者中无效。因此，拟议的研究与NIH使命的一部分有关 开发基本知识，有助于减轻糖尿病和伤口护理的负担，并改善广泛患者（例如老年和免疫受损患者）中其他慢性伤口的治疗。