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.

背景及意义。外周动脉疾病（PAD）是动脉粥样硬化的一种表现， 导致供应腿部的动脉逐渐变窄和闭塞。 PAD通常表现为 跛行（腿部疼痛和严重行走受限），但有些患者进展为肢体威胁性缺血 和截肢。跛行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方法，同时减少昂贵的护理和手术的需要。 前期工作。我们开发并验证了猪后肢缺血模型（髂股动脉 动脉切除术）概括了人类 PAD/跛行病理生理学的关键方面，并使用 它用于测试脂肪源性间充质干细胞（ADMSC）的功效。我们的前期工作表明 将 ADMSC 注射到结扎/切除的髂动脉床中会产生 (i) 动脉生成增加，(ii) 与相比，ADMSC 处理的猪的肌肉灌注增强，并且 (iii) 增加了在跑步机上行走的能力 未经处理的猪。 假设，具体目标。我们的中心假设是自体 ADMSC 的动脉外递送 或衍生的外泌体将改善缺血后肢的血流动力学、组织学和功能终点 与 PAD 猪模型中的相关动脉生成有关。我们将用猪来探索我们的假设 高胆固醇血症和高血压背景下的后肢缺血模型（用高胆固醇诱导） 脂肪/高果糖/高盐饮食），模仿 PAD。中心假设将通过三个方面进行探讨 具体目标： 目标 1. 确定 ADMSC 或 ADMSC 衍生的外泌体的动脉外给药是否会影响 刺激动脉生成，改善血流动力学/灌注终点，并调节局部炎症 猪 PAD 模型中的环境。 目标 2：确定动脉外注射 ADMSC 或外泌体是否会改善缺血 PAD 猪模型的肌病和跑步机表现。 目标 3：确定 ADMSC 和单核细胞联合应用是否会诱导动脉生成 在灌注的猪动脉系统中。 创新。我们将使用临床相关的 PAD 大型动物模型和新技术 由我们的研究小组开发，用于研究缺血肢体的生理学和组织病理学。我们也会 利用猪 ADMSC 的衍生物（即外泌体），如果有效的话，它比 常规细胞疗法。此外，我们将描述基于细胞的治疗对局部的影响。 缺血肢体的炎症环境。