Mesenchymal Stem Cell Therapeutics in Hibernating

描述(申请人提供)：在美国，慢性心肌缺血导致心力衰竭是发病率和死亡率的主要原因。实验性心肌干细胞治疗主要是在急性缺血模型中进行的。已建立的猪冬眠心肌模型与人类冠状动脉疾病的慢性化和稳定性非常相似，将用于开发和优化基于干细胞和基因联合治疗的治疗策略。体外扩增的猪骨髓间充质干细胞(MSCs)具有强大的自我更新和多向分化潜能，能够产生多种生长因子和细胞因子。尽管MSCs在衰老和疾病中作为再生医学很有前途，但关于其生长、分化、存活、组织归巢和衰老特性的机制仍有待进一步分析。内容包括生长因子对骨髓间充质干细胞多向分化潜能的调节、衰老对骨髓间充质干细胞功能的影响以及同种异体间充质干细胞的使用。这些努力的核心是使用表达细胞保护、血管生成和MSC归巢相关基因的重组腺病毒。提案的第一部分依赖于对MSCs的广泛细胞培养特征，为提案的第二部分奠定了基础，后者解决了工程MSCs的生理效应。目的1确定多种血管内皮生长因子亚型对猪骨髓间充质干细胞生长和多系分化潜能的不同影响。目的2分析MSC趋化因子受体在心肌MSC归巢中的表达和调控。目的3描述细胞和动物衰老对骨髓间充质干细胞生长能力、细胞存活能力、多系分化潜能和趋化迁移能力的影响。目的4将优化跟踪和识别植入心肌的MSCs的体内命运的策略，并评估使用同种异体和老年MSCs的可行性。目的5将确定为增强存活能力、血管生成能力或归巢能力而设计的MSCs是否能更好地改善慢性冬眠心肌的血流和功能。从长远来看，基于猪冬眠心肌的MSC疗法与人类慢性冠状动脉疾病的再生药物之间的转换将导致优化的MSC疗法，这可能在管理衰老和治疗疾病方面具有临床价值。

DESCRIPTION (provided by applicant): Chronic myocardial ischemia leading to heart failure is a leading cause of morbidity and mortality in the United States. Experimental myocardial stem cell therapeutics have been performed largely in the acute ischemia model. The well established porcine hibernating myocardium model, which closely resembles the chronicity and stability of human coronary artery disease, will be used to develop and optimize therapeutic strategies based on combined stem cell and gene therapy. Bone marrow-derived porcine mesenchymal stem cells (MSCs) expanded in culture possess robust self-renewal and multilineage differentiation potentials, and are capable of producing many growth factors and cytokines. Although promising as regenerative medicine in aging and disease, MSCs await further analysis regarding the mechanisms governing their growth, differentiation, survival, tissue homing, and aging characteristics. Growth factor modulation of MSC multilineage potential, the influence of aging on the function of MSCs, and the use of allogeneic MSCs will be characterized. Central to these efforts is the use of recombinant adenovirus expressing genes involved in cytoprotection, angiogenesis, and MSC homing. The first part of the proposal relies on extensive cell culture characterizations of MSCs, building the foundation for the second part of the proposal that addresses the physiological effect of engineered MSCs. Aim 1 will determine the differential effects of multiple VEGF isoforms on the growth and multilineage potentials of porcine MSCs. Aim 2 will analyze the expression and regulation of MSC chemokine receptors involved in myocardial MSC homing. Aim 3 will characterize the influence of cellular and animal aging on MSC growth capability, cell survival capacity, multilineage potential, and chemotactic migratory potency. Aim 4 will optimize strategies for tracking and identifying the in vivo fate of implanted MSCs in the myocardium and evaluate the feasibility of using allogeneic and aged MSCs. Aim 5 will determine whether MSCs engineered for enhanced survival capacity, angiogenic potential, or homing potency can better improve flow and function in chronic hibernating myocardium. Long term, the translation between the MSC-based therapy in the porcine hibernating myocardium and regenerative medicine for humans with chronic coronary artery disease will lead to optimized MSC therapeutics that can be of clinical value in managing aging and curing disease.