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.

抽象的 衰老是 2 型患者出现慢性、不愈合足部溃疡的一个重要危险因素 糖尿病（T2DM）。患有 T2DM 的老年人的糖尿病足溃疡具有毁灭性的后果，导致 截肢和全因死亡的风险增加。开发治疗方法的需求迫切但尚未得到满足 治疗该人群中这些无法愈合的伤口；然而，合适的动物模型可用于 开发对人类疾病具有高度可转化性的生物疗法和干细胞疗法 不存在。因此，本提案的总体目标是开发老年T2DM人源化小鼠 模型来研究衰老和 T2DM 的综合影响使人类失调的机制 T2DM 患者在伤口愈合过程中的免疫系统，并根据这些新见解测试治疗方法。 为了实现这一目标，提出了两个具体目标。具体目标 1 的目的是开发人性化的 移植了来自老年 T2DM 人类捐赠者的 CD34+ 造血干细胞 (HSC) 的小鼠模型用于研究 伤口愈合。该假设是，人源化小鼠被移植了来自患有老年痴呆症的老年人的 CD34+ HSCs。 糖尿病忠实地再现了不愈合的伤口表型和伤口的偏向极化 已在人类 T2DM 患者中记录到浸润性巨噬细胞。这个模型是基于我们的 已发表和未发表的研究结果表明，T2DM 和衰老会通过氧化应激损害伤口愈合 依赖HSC自主机制。因此，衰老-T2DM 相关的伤口愈合损伤将 由老年 T2DM 患者的供体 HSC 赋予。具体目标 2 的目的是确定 ATLAS 疗法对移植老年 T2DM HSC 的人源化小鼠伤口愈合的影响。这 假设是 ATLAS 对来自老年 T2DM 人类供体的 HSC 进行处理，然后将其植入 NSG-SGM3 小鼠减少 HSC 氧化应激并增加一氧化氮生物利用度，从而恢复正常 伤口中的巨噬细胞数量和极化。我们的结果表明，在 T2DM 小鼠伤口模型中 愈合 ATLAS 是 L-精氨酸、四氢生物蝶呤和 L-抗坏血酸的组合，可降低 HSC 氧化 压力并增加 HSC NO 生物利用度，恢复正常伤口愈合。的发展 提出的人源化小鼠模型弥补了小鼠伤口愈合模型与人类伤口愈合模型之间的主要差距 患有 T2DM 的老年人的不愈合足部溃疡；因此，创建一个高度可翻译的工具来开发 治疗患有 T2DM 的老年人不愈合伤口的新型生物疗法满足了一个重大的未满足需求 对于这些患者。由于该项目的成果，我们希望大大减少以下疾病的痛苦： 这一毁灭性问题对全世界老年人造成了不成比例的影响。