Mesenchymal Stem Cell Therapeutics in Hibernating

描述（由申请人提供）：慢性心肌缺血导致心力衰竭是美国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。建立的猪冬眠心肌模型与人类冠状动脉疾病的慢性和稳定性非常相似，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。培养扩增的猪骨髓间充质干细胞（MSCs）具有强大的自我更新和多系分化潜力，能够产生多种生长因子和细胞因子。尽管MSCs作为治疗衰老和疾病的再生药物前景看好，但其生长、分化、存活、组织归巢和衰老特征的调控机制有待进一步分析。生长因子对间充质干细胞多系潜能的调节，衰老对间充质干细胞功能的影响，以及异体间充质干细胞的使用将被描述。这些努力的核心是使用重组腺病毒表达参与细胞保护、血管生成和MSC归巢的基因。提案的第一部分依赖于MSCs的广泛细胞培养特征，为解决工程MSCs的生理效应的提案的第二部分奠定基础。目的1将确定多种VEGF亚型对猪间充质干细胞生长和多系潜能的不同影响。目的2将分析心肌MSC归巢过程中MSC趋化因子受体的表达和调控。目的3将描述细胞和动物衰老对间充质干细胞生长能力、细胞存活能力、多谱系潜力和趋化迁移能力的影响。目的4将优化跟踪和识别植入骨髓间充质干细胞在心肌中的体内命运的策略，并评估使用同种异体和老化骨髓间充质干细胞的可行性。目的5将确定用于增强存活能力、血管生成潜力或归巢效力的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，猪冬眠心肌中基于MSC的治疗与慢性冠状动脉疾病患者的再生医学之间的转化将导致优化的MSC治疗方法，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.