Improving hiPSC cardiomyocyte engraftment and integration with nanowired human cardiac organoids

项目摘要：仅在美国，每年就有超过73.5万例心肌梗塞(MI)， 这表明迫切需要开发修复受伤心脏的治疗方法。由于再生能力有限 成人心脏、人诱导多能干细胞来源的心肌细胞(hiPSC-CMS)的能力 由于他们表现出肌肉重塑和修复的能力而受到极大的关注 移植到受损心脏后的收缩功能。尽管取得了进展，但目前的方法是有限的。 当作为分离的细胞或工程化的心脏组织输送时，由于细胞保留率低和整合不良 帕奇斯。为了应对这些挑战，我们率先使用了导电硅纳米线(e- SiNWs)促进hiPSC-CMS的自组装，形成纳米线的hiPSC心脏球体。我们的活体研究 显示纳米线球体改善了移植后细胞的保留和植入，推测是由于 到他们的3D微组织结构和e-SiNW增强的电子集成。提高细胞存活率 和植入受伤的心脏，我最近开发了一种有机体制造方案，在那里我们种植 支持细胞(如内皮细胞、心脏成纤维细胞、人类脂肪干细胞)与纳米线连接的HiPSC 心脏球体。我的初步数据显示，在心脏器官中植入了相当大的纳米线 缺血/再灌注损伤大鼠心脏，宿主血管迅速渗入并改善组织 与非纳米线心脏器官相比，收缩结构的发展。的目标是 这项建议是为了确定e-SiNWs和有机化合物的预血管对HiPSC-CM的影响 植入和整合(目标1)并展示纳米线人类心脏的翻译潜力 有机化合物在修复心肌梗死中的作用(目标2)。这项提议的中心假设是1)e-SiNW 可促进移植器官的收缩发育；2)移植后的管腔状血管构筑 有机化合物可以与宿主心肌快速吻合。该建议的创新之处在于，对于 第一次，我们将协同e-SiNW和预血管、可注射的3D心脏微组织来开发一种 可扩展的平台，有效地植入HiPSC-CMS，并改善其与成人心肌的整合。我的 长期目标是为推动基于细胞的治疗方法的发展做出重大贡献 修复心脏损伤。因此，我们将追求以下具体目标：1)确定电子- SiNW和纳米线器官中的预血管形成对收缩发育和血管整合的影响 2)确定纳米线对人心脏的治疗效果。 带有受损老鼠心脏的有机化合物。拟议的研究将首次使我们能够调查 E-SiNW和支持细胞类型对hPSC-CM植入和整合的协同作用 红心。本研究将为利用纳米线人类心脏器官来追求大容量的 动物研究和加速其翻译应用。

PROJECT SUMMARY: In the U.S. alone, there are more than 735,000 myocardial infarctions (MI) each year, suggesting a pressing need to develop treatments for repairing injured hearts. Due to the limited regenerative capacity of adult hearts, human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) have received significant attention due to their demonstrated capacity for remuscularization and restoration of contractile function upon transplantation to injured hearts. Despite the progress, the current approach is limited by low cell retention and poor integration when delivered as dissociated cells or engineered cardiac tissue patches. To address these challenges, we pioneered the use of electrically conductive silicon nanowires (e- SiNWs) to facilitate self-assembly of hiPSC-CMs to form nanowired hiPSC cardiac spheroids. Our in vivo studies showed the nanowired spheroids improve cell retention and engraftment after transplantation, presumably due to their 3D microtissue configuration and the e-SiNW enhanced electrical integration. To improve cell survival and engraftment in injured hearts, I recently developed an organoid fabrication protocol where we seed the supporting cells (e.g., endothelial cells, cardiac fibroblasts, human adipose stem cells) onto nanowired hiPSC cardiac spheroids. My preliminary data showed sizeable engraftments of nanowired cardiac organoids in ischemia/reperfusion (I/R) injured rat hearts, with rapid infiltration of host vasculature and improved organization and development of contractile structures, when compared to non-nanowired cardiac organoids. The goal of this proposal is to determine the effects of e-SiNWs and prevascularization of the organoids on hiPSC-CM engraftment and integration (Aim 1) and demonstrate the translational potential of nanowired human cardiac organoids in repairing infarcted hearts (Aim 2). The central hypotheses of this proposal are 1) the e-SiNWs can improve the contractile development of the transplanted organoids, and 2) the lumen-like vasculature in the organoids can allow for rapid anastomosis with host myocardium. The proposal is innovative in that, for the first time, we will synergize e-SiNWs and pre-vascularized, injectable 3D cardiac microtissues to develop a scalable platform to effectively engraft hiPSC-CMs and improve their integration with adult myocardium. My long-term goal is to make significant contributions towards advancing development of cell-based therapies for repairing cardiac injury. Accordingly, we will pursue the following specific aims: 1) Determine the effects of e- SiNWs and prevascularization in nanowired organoids on contractile development and vascular integration with host myocardium in healthy rat hearts, and 2) Determine therapeutic efficacy of nanowired human cardiac organoids with injured rat hearts. The proposed research would, for the first time, allow us to investigate the synergistic effect of e-SiNWs and supporting cell-types on hiPSC-CM engraftment and integration in injured hearts. This research will provide the foundation to use nanowired human cardiac organoid to pursue large animal studies and accelerate their translational applications.