Autogeneic Mitochondria: Surgical Cardioprotection

描述(申请人提供)：基于细胞的心肌修复或再生疗法显示出巨大的潜力；然而，关于特定细胞群的有效性、细胞采集和扩增的后勤以及基于细胞的心肌修复/再生的机制仍有待阐明。最重要的是，细胞分离、免疫耐受、细胞植入和整合方面的困难依然存在。因此，增加细胞递送、细胞功能/存活的策略是通过细胞治疗成功修复/再生心肌的关键。最近，我们已经证明，从患者自身和骨骼组织中分离出的自体线粒体，不受缺血的影响，然后在再灌注早期注入缺血区，可以显著减少心肌坏死(坏死和凋亡)，并显著增强缺血后的功能。移植的线粒体是有活力的，呼吸正常，维持膜电位，在注射后至少21天存在于心肌中，并从心外膜分布到距离注射部位很远的心内膜下。从远端骨骼肌中分离和制备自体线粒体的速度很快，可以在90分钟内完成。-在冠状动脉旁路移植(CABG)和经皮冠状动脉介入治疗ST段抬高心肌梗死(PCI-STEMI)冠状动脉血运重建术(PCI-STEMI)的临床干预范围内的合理时间框架。自体线粒体移植在不使用抗排斥药物治疗的情况下，为实际应用提供了免疫学上的优势，可以作为独家干预或作为后续自体、异体或异种细胞再生干预之前的主要干预措施，以改善心肌坏死和增强心肌功能。我们建议在临床相关的原位冠状动脉搭桥术和冠状动脉介入性心肌梗死模型中优化使用自体线粒体移植来改善心肌坏死和增强心肌功能；并通过生化/免疫组织化学、核磁共振以及综合转录和蛋白质组学分析来确定自体线粒体移植显著增强外科心脏保护的具体机制(S)。 公共卫生相关性：最近，我们已经证明，从患者自己的身体、遥远的未受缺血影响的骨骼组织中分离出自体线粒体，然后在再灌注早期直接注入心肌缺血区，可以显著减少心肌坏死(坏死和凋亡)，并显著促进缺血后功能恢复。在这一应用中，我们建议在临床相关的原位冠状动脉旁路移植(CABG)和经皮冠状动脉介入ST段抬高型心肌梗死(PCISTEMI)模型中优化使用自体线粒体移植来改善心肌坏死和增强心肌功能；并通过生化/免疫组织化学、核磁共振以及综合转录和蛋白质组学分析来确定自体线粒体移植显著增强外科心脏保护的特定机制(S)。

DESCRIPTION (provided by applicant): Cell-based therapies for myocardial repair or regeneration have shown great potential; however, debate as to the efficacy of specific cell populations, the logistics of cell harvesting and expansion, the mechanisms of cell- based myocardial repair/regeneration remain to be elucidated. Most importantly difficulties over cell isolation, immune tolerance, cellular engraftment and integration remain. Therefore strategies to augment cell delivery, cell function/survival are crucial in permitting successful myocardial repair/regeneration through cellular therapy. Recently, we have demonstrated that autogeneic mitochondria isolated from the patient's own body, from skeletal tissue, unaffected by ischemia and then injected into the ischemic zone during early reperfusion significantly decreases myonecrosis (necrosis and apoptosis) and significantly enhance post-ischemic function. The transplanted mitochondria are viable, respiration competent, maintain membrane potential, are present in the myocardium for at least 21 days after injection and are distributed from the epi- to the sub- endocardium at significant distance from the site of injection. The isolation and preparation of autogeneic mitochondria from remote skeletal muscle is rapid and can be performed in < 90 min. - a time frame reasonable within the clinical interventions of both coronary artery bypass grafting (CABG) and percutaneous coronary intervention for coronary revascularization for ST segment elevation myocardial infarction (PCI-STEMI). Autogeneic mitochondrial transplantation provides immunological advantages for practical application without the use of anti-rejection drug therapy and could be used either as an exclusive intervention or as a primary intervention prior to subsequent auto-, allo- or xeno-geneic cellular regenerative interventions to ameliorate myonecrosis and enhance myocardial function. We propose to optimize the use of autogeneic mitochondrial transplantation for the amelioration of myonecrosis and enhancement of myocardial function in the clinically relevant in situ CABG and PCI-STEMI model; and to identify the specific mechanism(s) through which autogenic mitochondrial transplantation significantly enhances surgical cardioprotection using biochemical /immunohistochemical, NMR and integrated transcriptomic and proteomic analysis. PUBLIC HEALTH RELEVANCE: Recently, we have demonstrated that autogeneic mitochondria isolated from the patient's own body, from remote skeletal tissue unaffected by ischemia, and then directly injected into the ischemic zone of the myocardium during early reperfusion, significantly decrease myonecrosis (necrosis and apoptosis) and significantly enhance post-ischemic functional recovery. In this application we propose to optimize the use of autogeneic mitochondrial transplantation for the amelioration of myonecrosis and enhancement of myocardial function in the clinically relevant in situ coronary artery bypass grafting (CABG) and percutaneous coronary intervention for coronary revascularization for ST segment elevation myocardial infarction (PCI-STEMI) model; and to identify the specific mechanism(s) through which autogenic mitochondrial transplantation significantly enhances surgical cardioprotection using biochemical /immunohistochemical, NMR and integrated transcriptomic and proteomic analysis.