Mesenchymal stem cells in the treatment of hindlimb ischemia in diabetic mice

间充质干细胞治疗糖尿病小鼠后肢缺血

• 批准号: 7987780
• 负责人: Louis Michael Messina
• 金额: $ 41.13万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7987780
• 关键词: Acetylcysteine; Activities of Daily Living; Adipocytes; Aftercare; Angiogenic Factor; Antioxidants; Arteries; Autologous Transplantation; Automobile Driving; Bioavailable; Biology; Blood Vessels; Blood capillaries; CXCR4 gene; Cell Lineage; Cell physiology; Cells; Comorbidity; Diabetes Mellitus; Diabetic mouse; Disease; Endothelial Cells; Epidemic; Exhibits; Functional disorder; Genetic Engineering; Goals; Healthcare; Hindlimb; Homing; Hydrogen Peroxide; Infiltration; Insulin; Ischemia; Knowledge; Light; Limb structure; Lower Extremity; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Mus; Muscle; Non-Insulin-Dependent Diabetes Mellitus; Outcome; Oxidants; Palliative Care; Participant; Pathology; Pathway interactions; Patients; Pear; Perfusion; Peripheral arterial disease; Phenotype; Population; Production; Property; Recovery; Resveratrol; Risk Factors; Role; Stem cell transplant; System; Testing; Therapeutic Intervention; Transgenic Organisms; Transplantation; Treatment Efficacy; Vascular Endothelial Growth Factors; Wild Type Mouse; Work; adipocyte differentiation; base; capillary; chemokine receptor; diabetic; diabetic patient; improved; in vivo; injured; neovascularization; novel therapeutics; overexpression; oxidant stress; paracrine; pluripotency; public health relevance; response; rosiglitazone; rosuvastatin; tetrahydrobiopterin

DESCRIPTION (provided by applicant): The goal of this project is to enhance the means to use mesenchymal stem cells (MSC) for the treatment of peripheral artery disease, particularly in the diabetic patient. MSC transplantation hold great promise as a therapeutic intervention for PAD based on their pluripotency, as well as their efficacy in paracrine delivery of proangiogenic factors. However, we have determined that diabetic MSC manifest greater oxidant stress than healthy (WT) MSC. Diabetic MSC display restricted pluripotency, favoring adipocytic over endothelial differentiation; when transplanted into a WT host, diabetic MSC impair post-ischemic neovascularization and generate fatty infiltration in the ischemic hindlimb. The project hypothesis is that oxidant stress in diabetic MSC restricts their pluripotency and hence their neovascularization capacity. Three aims are proposed to test this hypothesis. Specific Aim 1 will demonstrate that oxidant production is the basis for restricted pluripotency in diabetic MSC, using a reductionist approach in cultured MSC. Exp [1] will determine if Nox4-derived H2O2 drives adipocyte differentiation in diabetic MSC. Exp [2] will determine the role of PPAR3-overexpression in diabetic MSC in generating adipocyte differentiation. Exp [3] will determine the role of uncoupled eNOS in generating oxidant stress in diabetic MSC and evaluate strategies for eNOS recoupling. Exp [4] will determine if deficiencies of the VEGF-Akt-eNOS pathway are the basis for impaired endothelial differentiation in diabetic MSC. Specific Aim 2 will use an in vivo MSC transplant paradigm to demonstrate that antioxidant treatment of diabetic MSCs or the diabetic host improves the efficacy of MSC transplant vis-a-vis post-ischemic neovascularization. Exp [1] will determine if ex vivo treatment of diabetic MSC with N-acetylcysteine (NAC) or other agents with direct or indirect antioxidant properties (resveratrol, rosiglitazone, rosuvastatin) improve their function upon subsequent transplant into a WT host in the setting of hindlimb ischemia. Exp [2] will determine if treatment of the db/db recipient mouse with NAC, or the other agents just noted, improve the outcome of MSC transplant in the setting of hindlimb ischemia. Specific Aim 3 will demonstrate that genetic engineering of MSC to enhance their expression of proangiogenic factors improves their participation in post-ischemic neovascularization. Exp [1] will determine if overexpression of wild type eNOS or constitutively active Akt in MSCs prior to transplant increases eNOS activation and eNOS-derived NO production, and in turn improves their efficacy in the treatment post-ischemic neovascularization. Exp [2] will determine if selection MSC expressing the chemokine receptor CXCR4 increases the homing of these cells to the ischemic hindlimb. If homing is improved, then this CXCR4+ MSC will undergo genetic engineering to maximize their in vivo functional capacity. PUBLIC HEALTH RELEVANCE: Current treatments for peripheral artery disease (PAD) are either palliative and treatment options are particularly limited in patients with the common co-morbidity of diabetes mellitus; discovery of novel therapeutic options are an urgent matter of national health care priority. Autologous transplantation with mesenchymal stem cells (MSC), a pluripotent cell that par- ticipates in post-ischemic neovascularization, holds great promise as a means to effect symptomatic relief by reversing disease pathology. The proposed work will elucidate the mechanisms responsible for the loss of pluripotency and therapeutic efficacy of diabetic MSC, as well as greatly increase knowledge regarding the basic and applied vascular biology of MSC in the treatment of lower extremity ischemia.

描述（由申请人提供）：本项目的目标是加强使用间充质干细胞（MSC）治疗外周动脉疾病的方法，特别是在糖尿病患者中。MSC移植作为基于其多能性的PAD的治疗干预，以及其在促血管生成因子的旁分泌递送中的功效，具有很大的前景。然而，我们已经确定，糖尿病MSC表现出比健康（WT）MSC更大的氧化应激。糖尿病MSC显示出有限的多能性，有利于脂肪细胞而不是内皮细胞分化;当移植到WT宿主中时，糖尿病MSC损害缺血后新血管形成并在缺血后肢中产生脂肪浸润。该项目的假设是，糖尿病MSC中的氧化应激限制了它们的多能性，因此限制了它们的新血管形成能力。提出了三个目标来检验这一假设。具体目标1将证明氧化剂的产生是糖尿病MSC中限制性多能性的基础，在培养的MSC中使用还原方法。Exp [1]将确定Nox 4衍生的H2 O2是否驱动糖尿病MSC中的脂肪细胞分化。实验[2]将确定糖尿病MSC中PPAR 3过表达在产生脂肪细胞分化中的作用。Exp [3]将确定未偶联的eNOS在糖尿病MSC中产生氧化应激的作用，并评估eNOS重新偶联的策略。实验[4]将确定VEGF-Akt-eNOS通路的缺陷是否是糖尿病MSC中内皮分化受损的基础。具体目标2将使用体内MSC移植范例来证明糖尿病MSC或糖尿病宿主的抗氧化剂治疗 改善MSC移植相对于缺血后新生血管形成的功效。Exp [1]将确定在后肢缺血的情况下，用N-乙酰半胱氨酸（NAC）或其他具有直接或间接抗氧化特性的药物（白藜芦醇、罗格列酮、瑞舒伐他汀）离体治疗糖尿病MSC是否能改善其随后移植到WT宿主中的功能。实验[2]将确定用NAC或刚刚提到的其他药剂治疗db/db受体小鼠是否改善了后肢缺血情况下MSC移植的结果。具体目标3将证明，基因工程的MSC，以提高其表达的促血管生成因子，改善其参与缺血后新血管形成。Exp [1]将确定移植前MSC中野生型eNOS或组成型活性Akt的过表达是否增加eNOS活化和eNOS衍生的NO产生，并进而提高其在治疗缺血后新血管形成中的功效。实验[2]将确定表达趋化因子受体CXCR 4的选择MSC是否增加这些细胞向缺血后肢的归巢。如果归巢得到改善，那么这种CXCR 4 + MSC将经历基因工程，以最大限度地提高其体内功能能力。 公共卫生关系：目前外周动脉疾病（PAD）的治疗要么是姑息性的，治疗选择特别有限的患者与常见的合并症的糖尿病;发现新的治疗选择是一个紧迫的问题，国家卫生保健的优先事项。间充质干细胞（MSC）是一种参与缺血后新生血管形成的多能细胞，其自体移植作为通过逆转疾病病理来实现症状缓解的手段具有很大的前景。拟议的工作将阐明负责糖尿病MSC的多能性和治疗效果的损失的机制，以及大大增加有关MSC在下肢缺血治疗中的基础和应用血管生物学的知识。