Mycardial Repair Using Human iPSC Derived Cardiac Muscle Patch

描述(申请人提供)：尽管最近报道了细胞疗法对心肌梗死后左室重构(MI)心脏的有益效果，但在心肌梗死表面应用预制的心肌组织等效物可能还有额外的好处。我们的目标是开发一种“心肌贴片”，将来自人诱导多能干细胞(HiPSC)的人心肌细胞、内皮细胞和平滑肌细胞包裹在3D多孔纤维蛋白生物材料中。“心肌贴片”能有效防止左室梗死后瘢痕膨大，降低BZ壁应力，改善心肌生物能。心血管生理学中最重要的问题之一是如何在活体心脏中调节ATP的产生和利用的速率，以及这一速率的限制如何导致衰竭心脏的收缩功能障碍。准确评价心肌ATP转换率(ADP+PI) 由于活体心脏中的心肌游离无机磷(PI)水平太低，无法直接测量，因此通常不成功。我们的目标是证明一种新的双饱和转移(MRS-MST)方法，使我们能够在不测量PI水平的情况下计算ATP水解率，从而克服了在体内确定ATP周转率的主要障碍。虽然关于高场(&gt；7Tesla，[T])人脑功能成像的报道已经产生了重大的新发现，推动了神经科学领域的发展，但由于缺乏线圈工程，7T下用于人类的心脏MRI和光谱学还不存在。具体目标(SA)是：SA1。目的：制备并鉴定人心肌斑块(HcMP)。我们将人诱导多能干细胞(HiPSC)分化为心肌细胞(CM)、内皮细胞(EC)和平滑肌细胞(SMC)，并利用这些心肌细胞构建hcMP，并表征hcMP在基础状态和起搏反应中的机械特性。SA2.利用免疫抑制的猪心肌梗死后左室重构模型，研究CM-、EC-和SMC-hiPSC用于心肌修复的新方法。我们将研究移植预制的hcMP是否会减少左室瘢痕隆起和室壁应力，并改善心肌生物能量学和收缩功能。我们还将通过使用环路记录器全天候监测8周的EKG，并进行PES研究，来比较接受和不接受C移植的心脏的电稳定性。SA3.在高场磁铁中检测心肌生物能量学，具有以前无法达到的能力、灵敏度和空间定位水平。3A)展示一部小说 核磁共振MRS-MST技术，可以测量在体心脏的心肌ATP周转率。3b)开发用于7T/125 CM大口径磁体的第一个身体线圈，该线圈可用于进行1H和31P心脏核磁共振研究，以测量心肌功能、血流灌注和ATP转换率；该线圈的成功将使这些方法易于在临床环境中应用，并为监测人体心脏的心肌功能、代谢和血流灌注提供一种全新的有价值的方法。

DESCRIPTION (provided by applicant): Although the beneficial effects of cellular therapy in hearts with post myocardial infarction LV remodeling (MI) have been reported recently, there could be additional benefit in applying a prefabricated cardiac muscle tissue equivalent over the surface of myocardial infarct. We aim to develop a "cardiac muscle patch", formed by entrapping human cardiac myocytes, endothelial cells and smooth muscle cells that derived from human induced pluripotent stem cells (hiPSC) in a 3D porous fibrin biomaterial. A "cardiac muscle patch" could effectively prevent LV infarct scar bulging, reduce the BZ wall stress and improve the myocardial bioenergetics. One of the most significant problems in cardiovascular physiology is how the rate of ATP production and utilization is regulated in the in vivo heart and how the limitation of this rate may contribute to contractile dysfunction in failing hearts. The accurate evaluation of myocardial ATP turnover rates (ADP+Pi ATP), has generally not been successful because the level of myocardial free inorganic phosphate (Pi) in the in vivo heart is too low to be measured directly. We aim to demonstrate a novel double saturation transfer (MRS-MST) method enabling us to calculate the ATP hydrolysis rate without measuring Pi levels, which overcomes the primary barrier in determining the ATP turnover rate in vivo. Although reports of human brain function imaging at high field (>7Tesla, [T]), have generated significant novel findings that advance the field of neuroscience, cardiac MRI and spectroscopy for humans at 7T is nonexistent because of the lack of coil engineering. The specific aims (SA) are: SA1. To fabricate and characterize the human cardiac muscle patch (hcMP). We will differentiate human induced pluripotent stem cells (hiPSC) into cardiomyocytes (CM), endothelial cells (EC), and smooth muscle cells (SMC); and use these cardiac cells to fabricate an hcMP and characterize the mechanical properties of the hcMP during baseline and in response to pacing. SA2. To examine novel delivery of CM-, EC- and SMC-hiPSC for myocardial repair using an immuno-suppressed swine model of post infarction LV remodeling. We will examine whether the transplantation of a prefabricated hcMP will result in reductions in LV scar bulging and wall stress, and improvements in myocardial bioenergetics and contractile function. We will also compare the electrical stability of hearts with or without C transplantation by using a Loop Recorder to monitor EKGs 24/7 for 8 weeks, and by conducting a PES study. SA3.To examine myocardial bioenergetics in a high field magnet with previously unattainable levels of capability, sensitivity and spatial localization. 3a) To demonstrate a novel NMR MRS-MST technology that can measure the myocardial ATP turnover rate in the in vivo heart. 3b) To develop the first body coil for a 7T/125 CM large-bore magnet that can be used to perform 1H- and 31P- cardiac NMR studies for measurements of myocardial function, perfusion, and ATP turnover rate; The success of this coil will enable these methods to be readily applied in a clinical setting and provide an entirely new and valuable method for monitoring myocardial function, metabolism, and perfusion in the human heart.