Effect of cell-based therapies on functional, hemodynamic, and histologic outcomes in a porcine model of peripheral arterial disease

细胞疗法对猪外周动脉疾病模型功能、血流动力学和组织学结果的影响

• 批准号: 10609836
• 负责人: MARK A CARLSON
• 金额: $ 58.27万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10609836
• 关键词: Adipose tissue; Amputation; Anatomy; Animal Model; Aorta; Arteries; Atherogenic Diet; Atherosclerosis; Autologous; Autopsy; Beds; Biological Assay; Biopsy; Bioreactors; Caring; Cell Communication; Cell Therapy; Cells; Diameter; Dimensions; Disease; Dose; Euthanasia; Excision; Exercise Therapy; Family suidae; Fatty acid glycerol esters; Fructose; Functional disorder; Gangrene; Gastrocnemius Muscle; Growth; Hindlimb; Histologic; Histopathology; Human; Hypertension; Inflammation; Injections; Ischemia; Isolated limb perfusion; Label; Leg; Ligation; Limb structure; Measurement; Measures; Mesenchymal Stem Cells; Methods; Mitochondria; Modeling; Muscle; Myopathy; Outcome; Oxidative Stress; Pain; Pain in lower limb; Patient Care; Patients; Performance; Perfusion; Peripheral; Peripheral arterial disease; Pharmaceutical Preparations; Physiology; Procedures; Protocols documentation; Research; Residual state; Specimen; System; Techniques; Testing; Thick; Time; Tissues; Vascularization; Walking; Work; artery occlusion; cell type; claudication; clinically relevant; comorbidity; efficacy testing; exosome; hemodynamics; high salt diet; hypercholesterolemia; iliac artery; improved; indexing; inflammatory milieu; innovation; limb ischemia; monocyte; nanovesicle; novel; novel therapeutic intervention; porcine model; residence; response; stem cell exosomes; stem cell therapy; stem cells; therapy development; treadmill

Background & Significance. Peripheral artery disease (PAD) is a manifestation of atherosclerosis that produces progressive narrowing and occlusion of the arteries supplying the legs. PAD usually presents as claudication (leg pain and severe walking limitation), but some patients progress to limb threatening ischemia and amputation. Standard therapies for claudication have limitations, so there remains a need to develop treatments that improve limb function while decreasing the need for expensive care and procedures. Preliminary Work. We have developed and validated a porcine model of hindlimb ischemia (iliofemoral artery excision) which recapitulates key aspects of the pathophysiology of human PAD/claudication, and used it to test the efficacy of adipose-derived mesenchymal stem cells (ADMSCs). Our preliminary work shows that injection of ADMSCs into the bed of the ligated/excised iliac artery produces (i) increased arteriogenesis, (ii) enhanced muscle perfusion, and (iii) increased treadmill walking capacity in ADMSC-treated pigs compared to untreated pigs. Hypothesis, Specific Aims. Our central hypothesis is that extra-arterial delivery of autologous ADMSCs or derived exosomes will improve hemodynamic, histologic, and functional endpoints of the ischemic hindlimb with associated arteriogenesis in a porcine model of PAD. We will explore our hypothesis using a porcine model of hindlimb ischemia on a background of hypercholesterolemia and hypertension (induced with a high fat/ high fructose/ high salt diet), which mimics PAD. The central hypothesis will be explored with three Specific Aims: Aim 1. To determine whether extra-arterial administration of ADMSCs or ADMSC-derived exosomes will stimulate arteriogenesis, improve hemodynamic/ perfusion endpoints, and modulate the local inflammatory environment in a porcine model of PAD. Aim 2: To determine whether extra-arterial injection of ADMSCs or exosomes will improve ischemic myopathy and treadmill performance in a porcine model of PAD. Aim 3: To determine whether combined application of ADMSCs and monocytes will induce arteriogenesis in a perfused porcine artery system. Innovation. We will use a clinically-relevant large-animal model of PAD and novel techniques that were developed by our research group to study the physiology and histopathology of the ischemic limb. We will also utilize a derivative of porcine ADMSCs (i.e., the exosomes) which, if effective, offer numerous advantages over conventional cellular therapy. In addition, we will characterize the effect of cell-based treatments on the local inflammatory environment of the ischemic limb.

背景和意义。外周动脉疾病（PAD）是动脉粥样硬化的表现， 产生逐渐缩小和阻塞腿部的动脉。垫通常显示为 lautain（腿部疼痛和严重的步行限制），但有些患者发展到肢体威胁性缺血 和截肢。 lau不平的标准疗法有局限性，因此仍然需要开发 改善肢体功能的治疗，同时减少对昂贵的护理和程序的需求。 初步工作。我们已经开发并验证了后肢缺血的猪模型（iliofemoral 动脉切除），概括了人类垫/laudation病理生理学的关键方面，并使用了 它可以测试脂肪衍生的间充质干细胞的功效（ADMSC）。我们的初步工作表明 将ADMSC注射到结扎/切除的乳杆菌动脉的床中会产生（i）动脉生成增加，（ii） 与ADMSC处理的猪相比 未经处理的猪。 假设，具体目的。我们的中心假设是自体ADMSC的动脉外提供 或衍生的外泌体将改善缺血性后肢的血液动力学，组织学和功能终点 与猪垫模型中的相关动脉生成。我们将使用猪探索我们的假设 在高胆固醇血症和高血压的背景下，后肢缺血的模型（以高诱导 脂肪/高果糖/高盐饮食），模仿垫。中央假设将通过三个 具体目的： 目标1。确定动脉外ADMSC或ADMSC衍生的外泌体是否会 刺激动脉生成，改善血液动力学/灌注终点并调节局部炎症 棒球模型中的环境。 目的2：确定动脉外注射ADMSC或外泌体是否会改善缺血性 棒球模型的肌病和跑步机的性能。 目标3：确定ADMSC和单核细胞的联合应用是否会诱导动脉生成 在灌注猪动脉系统中。 创新。我们将使用与临床上相关的大动物垫和新技术的大动物模型 由我们的研究小组开发，以研究缺血性肢体的生理病理学和组织病理学。我们也会 利用猪ADMSC的衍生物（即外泌体），如果有效，则可以提供许多优势 常规的细胞疗法。此外，我们将表征基于细胞的治疗对局部的影响 缺血性肢体的炎症环境。