Mesenchymal Stem Cell Therapeutics in Hibernating Myocardium

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在急性缺血模型中进行。建立良好的猪冬眠心肌模型，它非常类似于人类冠状动脉疾病的慢性和稳定性，将用于开发和优化基于联合干细胞和基因治疗的治疗策略。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，并能产生多种生长因子和细胞因子。虽然有希望作为衰老和疾病的再生医学，但MSC等待进一步分析其生长，分化，存活，组织归巢和衰老特征的机制。生长因子对间充质干细胞多向分化潜能的调节，衰老对间充质干细胞功能的影响，以及同种异体间充质干细胞的应用将被描述。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。该提案的第一部分依赖于MSC的广泛细胞培养表征，为该提案的第二部分奠定了基础，该部分解决了工程MSC的生理效应。目的1将确定多种VEGF异构体对猪MSCs生长和多向分化潜能的差异作用。目的2分析心肌间充质干细胞归巢过程中趋化因子受体的表达及调控。目的3将表征细胞和动物衰老对MSC生长能力、细胞存活能力、多谱系潜能和趋化迁移潜能的影响。目的4优化心肌内植入MSCs的体内命运追踪和鉴定策略，评价同种异体和老化MSCs的可行性。目的5将确定经工程化以增强存活能力、血管生成潜能或归巢潜能的MSC是否可以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换将导致优化的MSC治疗方法，这些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.