Combined effects of aging and type 2 diabetes on wound healing in a humanized mouse model

衰老和 2 型糖尿病对人源化小鼠模型伤口愈合的综合影响

• 批准号: 10266842
• 负责人: Louis Michael Messina
• 金额: $ 20.94万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10266842
• 关键词: Address; Affect; Age; Aging; Amputation; Animal Model; Aorta; Arginine; Arterial Fatty Streak; Atherosclerosis; Basic Science; Becaplermin; Biological Availability; Biological Response Modifier Therapy; CD34 gene; Cardiovascular Diseases; Cells; Cessation of life; Chronic; Clinical Research; Development; Diabetes Mellitus; Diabetic Foot Ulcer; Disease; Elderly; Engraftment; FDA approved; Failure; Foot Ulcer; Gastrocnemius Muscle; Gene Expression; Goals; Hematopoietic stem cells; Human; Immune; Immune system; Impaired wound healing; Impairment; Inflammatory; Malignant Neoplasms; Modeling; Mus; Natural Immunity; Nitric Oxide; Non-Insulin-Dependent Diabetes Mellitus; Oral; Oxidative Stress; Pathogenesis; Patients; Persons; Phase; Phenotype; Population; Publishing; Reporting; Research; Research Personnel; Risk; Risk Factors; Rodent; Scientist; Supplementation; Testing; Translational Research; Type 2 diabetic; Work; Wound models; age effect; aged; ascorbate; base; cardiovascular risk factor; cell age; chronic ulcer; chronic wound; clinically relevant; diabetic; diabetic patient; diabetic ulcer; hematopoietic stem cell differentiation; human disease; humanized mouse; hypercholesterolemia; immune function; improved; insight; lifetime risk; macrophage; monocyte; mortality; mouse model; non-healing wounds; novel; novel therapeutic intervention; oxidant stress; research and development; risk sharing; stem cell therapy; targeted treatment; tetrahydrobiopterin; therapeutic evaluation; therapy development; tool; wound; wound healing; wound treatment

Abstract Aging is a powerful risk factor for the development of chronic, non-healing foot ulcers in people with type 2 diabetes (T2DM). Diabetic foot ulcers in older adults with T2DM have devastating consequences, leading to increased risks of amputation and all-cause mortality. There is a compelling, unmet need to develop therapies to treat these non-healing wounds in this population; however, an appropriate animal model for use in the development of biotherapeutics and stem cell therapies with a high degree of translatability to human disease does not exist. Therefore, the overall goal of this proposal is to develop an aged-T2DM humanized mouse model to study the mechanisms by which the combined effects of aging and T2DM dysregulate the human immune system of T2DM patients during wound healing and to test therapeutics based on these new insights. To achieve this goal, two Specific Aims are proposed. The purpose of Specific Aim 1 is to develop a humanized mouse model engrafted with CD34+ hematopoietic stem cells (HSCs) from aged-T2DM human donors to study wound healing. The hypothesis is that humanized mice engrafted with CD34+ HSCs from older adults with diabetes faithfully recapitulate the non-healing wound phenotype and skewed polarization of wound- infiltrating macrophages that has been documented in human T2DM patients. This model is based on our published and unpublished findings that T2DM and aging impair wound healing by an oxidant stress- dependent HSC autonomous mechanism. Thus, the aging-T2DM-associated impairment in wound healing will be conferred by the donor HSCs from aged-T2DM patients. The purpose of Specific Aim 2 is to determine the effect of ATLAS therapy on wound healing in humanized mice engrafted with aged-T2DM HSCs. The hypothesis is that ATLAS treatment of HSCs derived from aged T2DM human donors prior to engraftment into NSG-SGM3 mice reduces HSC oxidant stress and increases nitric oxide bioavailability that restores normal macrophage number and polarization in wounds. Our results show that in a T2DM murine model of wound healing ATLAS, a combination of L-arginine, tetrahydrobiopterin, and L-ascorbate, decreases HSC oxidative stress and increases HSC NO bioavailability that restores normal wound healing. The development of the proposed humanized mouse model bridges a major gap between murine wound healing models and human non-healing foot ulcers in older adults with T2DM; thus, the creation of a highly translatable tool to develop novel biological therapies to treat non-healing wounds in older adults with T2DM fulfills a major unmet need for these patients. As a consequence of the results of this project, we hope to greatly reduce the suffering from this devasting problem that disproportionately affects older adults worldwide.

摘要 衰老是2型患者发生慢性、无法愈合足部溃疡的重要危险因素 糖尿病(T2 DM)。患有T2 DM的老年人的糖尿病足部溃疡具有毁灭性的后果，导致 截肢和全因死亡的风险增加。有一种迫切的、尚未得到满足的需求，即开发治疗方法 来治疗这些无法愈合的伤口；然而，一个合适的动物模型用于 对人类疾病具有高度可转译性的生物疗法和干细胞疗法的发展 并不存在。因此，这项建议的总体目标是开发一种老年T2 DM人源化小鼠 研究衰老和2型糖尿病共同作用的人体失调机制的模型 研究创面愈合过程中T2 DM患者的免疫系统，并基于这些新的见解测试治疗方法。 为了实现这一目标，提出了两个具体目标。具体目标1的目的是开发一个人性化的 老年2型糖尿病供者CD34+造血干细胞移植小鼠模型的研究 伤口愈合。假设是人源化小鼠移植了来自老年人的CD34+HSCs， 糖尿病忠实地概括了伤口无法愈合的表型和伤口偏向的极化- 已记录的人类T2 DM患者的巨噬细胞渗入。这个模型是基于我们的 已发表和未发表的研究结果表明，T2 DM和衰老通过氧化应激损害伤口愈合- 依赖的HSC自主机制。因此，与衰老相关的T2 DM对伤口愈合的损害将 由来自老年T2 DM患者的供者HSCs授予。具体目标2的目的是确定 ATLAS治疗对人源化小鼠移植老年T2 DM HSCs伤口愈合的影响这个 假设ATLAS对老年T2 DM供者的HSCs进行移植前的治疗 NSG-SGM3小鼠减少HSC氧化应激，提高一氧化氮生物利用度，恢复正常 创面巨噬细胞的数量和极化。我们的结果表明，在T2 DM小鼠创伤模型中 L-精氨酸、四氢生物蝶呤和L-抗坏血酸的组合物治愈ATLAS减少造血干细胞氧化 压力和增加HSC没有恢复正常伤口愈合的生物利用度。国际贸易组织的发展 提出的人源化小鼠模型弥合了小鼠伤口愈合模型与人类之间的主要差距 患有T2 DM的老年患者的足部溃疡无法愈合；因此，创建了一种高度可翻译的工具来开发 治疗老年T2 DM患者无法愈合的伤口的新生物疗法满足了一个主要的未得到满足的需求 对这些病人来说。由于这一项目的成果，我们希望大大减少 这一毁灭性的问题对世界各地的老年人造成了不成比例的影响。