Improving hiPSC cardiomyocyte engraftment and integration with nanowired human cardiac organoids

改善 hiPSC 心肌细胞的植入以及与纳米线人类心脏类器官的整合

• 批准号: 10058763
• 负责人: Robert Coyle
• 金额: $ 3.68万
• 依托单位: CLEMSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-01 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10058763
• 关键词: 3-Dimensional; Address; Adipose tissue; Adult; Advanced Development; Anastomosis - action; Animals; Anoikis; Attention; Blood Vessels; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cause of Death; Cell Communication; Cell Survival; Cell Therapy; Cells; Cicatrix; Data; Dependence; Development; Diffusion; Echocardiography; Effectiveness; Electrocardiogram; Endothelial Cells; Engraftment; Ensure; Fibroblasts; Foundations; Goals; Heart; Heart Injuries; Histologic; Human; Infiltration; Injectable; Ischemia; Modeling; Myocardial Infarction; Myocardium; Nutrient; Organoids; Protocols documentation; Rattus; Recovery of Function; Reperfusion Injury; Reperfusion Therapy; Research; Seeds; Sepharose; Silicon; Structure; Supporting Cell; Time; Tissues; Transplantation; Treatment Efficacy; Umbilical vein; Ventricular; adult stem cell; cardiac repair; cardiac tissue engineering; cell type; experience; improved; in utero; in vivo; induced pluripotent stem cell; injured; injury and repair; innovation; nanomaterials; nanowire; post-transplant; public health relevance; regenerative; repaired; restoration; self assembly; stem cells

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.

项目摘要：仅在美国，每年就有超过735,000个心肌梗死（MI） 暗示需要开发修复受伤心脏的治疗方法。由于再生有限 成人心脏，人类诱导的多能干细胞衍生的心肌细胞（HIPSC-CMS）的能力 由于其证明了回弹的能力和恢复的能力，因此受到了极大的关注 收缩的功能在移植到受伤的心脏时。尽管取得了进展，但当前的方法是有限的 通过低细胞保留率和较差的整合作为解离细胞或工程性心脏组织 补丁。为了应对这些挑战，我们率先使用了导电硅纳米线的使用（e- sinws）促进hipsc-CM的自组装形成纳米hipsc心脏球体。我们的体内研究 显示纳米球体改善了移植后的细胞保留和植入，大概是由于 到他们的3D微动物配置和E-SINW增强的电气集成。改善细胞存活 并在受伤的心脏中植入，我最近制定了一种类器官制造方案，我们在其中播种 支持细胞（例如内皮细胞，心脏成纤维细胞，人脂肪干细胞）到纳米hipsc上 心脏球体。我的初步数据显示，在 缺血/再灌注（I/R）受伤的老鼠心脏，宿主脉管系统迅速渗透和改善的组织 与非纳米心脏器官相比，收缩结构的发展。目标 该建议是确定类器官对HIPSC-CM的E-SINWS和OVARPASTILICE的影响 植入和整合（AIM 1）并证明了纳米人心脏的翻译潜力 修复梗塞心脏的器官（AIM 2）。该提案的中心假设是1）E-SINWS 可以改善移植器官的收缩发育，以及2）腔内的脉管系统 器官可以通过宿主心肌快速吻合。该提议具有创新性，因为 第一次，我们将协同E-SINW和血管前注射的3D心脏微动物，以开发 可扩展的平台可有效植入HIPSC-CMS并改善与成年心肌的整合。我的 长期目标是为促进基于细胞的疗法的开发做出重大贡献 修复心脏损伤。因此，我们将追求以下特定目的：1）确定电子的影响 纳米器官中的罪恶和前骨性，在收缩发育和血管整合与与 健康大鼠心脏中宿主心肌，2）确定纳米心脏的治疗功效 带有受伤的大鼠心脏的器官。拟议的研究将首次允许我们调查 E-SINWS和支持细胞类型对HIPSC-CM植入的协同作用，并在受伤的 心。这项研究将为使用纳米式人体心脏器官的基础提供大规模追求 动物研究并加速其翻译应用。