Nanowired human isogenic cardiac organoids to treat acute myocardial ischemia/reperfusion injuries

项目摘要：在美国，每年有超过735,000例心肌梗塞(MI)。而当 经皮冠状动脉介入治疗(PCI)显著减少了急性不良反应，长期 缺血/再灌注(I/R)后患者的预后仍然很差。由于可再生能力有限， 人心脏、人多能干细胞来源的心肌细胞(hPSC-CMS)已经收到了显著的 由于它们被证明有能力在移植到受损心脏后恢复收缩功能而引起的关注 各种哺乳动物模型，导致多项正在进行的临床试验。然而，目前的移植 方法主要依赖于游离的hPSC-CMS，导致细胞存活率低，功能适度改善， 导致心律失常的风险，可伸缩性差。为了应对这些挑战，我们的实验室开发了纳米线、预制 由hPSC-CMS、人原代心脏成纤维细胞、 内皮细胞、基质细胞和导电硅纳米线(e-SiNW)。血管内皮细胞 用于诱导有机体内的血管形成，添加e-SiNW以产生电子 促进hPSC-CM收缩发育及其电整合的导电微环境 宿主心肌。我们的初步体内数据显示，纳米线有机类化合物显示出强大的hPSC- CM植入术，功能恢复良好。它们在临床翻译中的主要障碍包括：1) 动物蛋白在细胞和有机培养中的使用以及2)缺乏功能益处的证明 大型动物模型。用同基因hPSC来源的细胞替代人原代细胞制造类器官 将减少批次之间的差异，并通过主要的组织相容性增强免疫兼容性 复杂(MHC)配对hPSC供者和人类受者。此外，虽然目前的hPSC-CM 植入策略一直专注于心肌内注射，开发了一种有效的治疗方法 有机化合物的冠状动脉内输送将加速它们的临床转化。这项提议的目标是 开发临床级hPSC心脏器官并在大型动物身上展示其功能益处 模型以生成用于IND提交的启用数据。这一提议的中心假设是纳米线 等基因hPSC心脏有机体提供了一种可扩展的系统，以高效和有效地植入hPSC-CMS 用于心脏修复。这项提议的创新之处在于，我们将1)在无异种、 开发用于植入的临床级心脏有机化合物的化学定义条件和2)利用 器物的大小和内皮腔状结构发展有效的冠状动脉内给药 策略。因此，我们将追求以下两个目标：1)构建和表征纳米线人类 在无异种、化学定义的条件下使用来自hPSCs的同基因心脏细胞的心脏器官， 2)用猪I/R模型检测纳米线等基因hPSC心脏器官的治疗效果 (缺血/再灌注)模型。

Project Summary: In the U.S., there are more than 735,000 myocardial infarctions (MI) each year. While percutaneous coronary intervention (PCI) has significantly reduced acute adverse repones, the long-term prognosis for post-ischemia/reperfusion (I/R) patients remains poor. Due to the limited regenerative capacity of human hearts, human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) have received significant attention due to their proven capacity to restore contractile function upon transplantation to injured hearts in various mammalian models, leading to multiple ongoing clinical trials. However, the current transplantation approach mainly relies on dissociated hPSC-CMs, leading to low cell survival, moderate functional improvement, arrhythmogenic risk, and poor scalability. To address these challenges, our lab developed nanowired, pre- vascularized human cardiac organoids composed of hPSC-CMs, human primary cardiac fibroblasts, endothelial cells, stromal cells, and electrically conductive silicon nanowires (e-SiNWs). Endothelial cells are used to induce vasculature formation within the organoids, and e-SiNWs are added to create an electrically conductive microenvironment to facilitate hPSC-CM contractile development and their electrical integration with the host myocardium. Our preliminary in vivo data showed that nanowired organoids illustrated robust hPSC- CM engraftment and superior functional recovery. The major barriers in their clinical translation include: 1) the use of animal proteins in the cell and organoid culture and 2) the lack of functional benefit demonstration in a large animal model. Replacing human primary cells with isogenic hPSC-derived cells for organoid fabrication would reduce batch-to-batch variations and enhance immune compatibility through Major Histocompatibility Complex (MHC) matching hPSC donors with human recipients. In addition, while the current hPSC-CM implantation strategy has been focused on intramyocardial injection, developing an effective approach for intracoronary delivery of the organoids will accelerate their clinical translation. The goal of this proposal is to develop clinical-grade hPSC cardiac organoids and demonstrate their functional benefits with a large animal model to generate enabling data for IND submission. The central hypothesis of this proposal is the nanowired isogenic hPSC cardiac organoids provide a scalable system to both efficiently and effectively implant hPSC-CMs for cardiac repair. The proposal is innovative in that we will 1) derive isogenic hPSC-derived cells in xeno-free, chemically defined conditions to develop clinical-grade cardiac organoids for implantation and 2) leverage the size and the endothelial lumen-like structures in the organoids to develop an effective intracoronary delivery strategy. Accordingly, we will pursue the following 2 aims: 1) Fabricate and characterize nanowired human cardiac organoids using isogenic cardiac cells derived from hPSCs in xeno-free, chemically defined conditions, and 2) Determine the therapeutic efficacy of the nanowired isogenic hPSC cardiac organoids with a porcine I/R (ischemia/reperfusion) model.