Nanowired humam cardiac organoid derived exosomes for heart repair

纳米线人类心脏类器官衍生的外泌体用于心脏修复

基本信息

批准号：
10639040
负责人：
Ying Mei
金额：
$ 39.27万
依托单位：
CLEMSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-15 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10639040
关键词：
3-Dimensional Acute Affect American Attention Attenuated Biogenesis Biomimetics Blood Vessels Cardiac Cardiac Myocytes Cardiovascular system Cell Therapy Cell secretion Cells Cessation of life Coculture Techniques Data Diameter Disease Electric Conductivity Endothelial Cells Environment Fibroblasts Fibrosis Goals Harvest Heart Injuries Heart failure Human Human Engineering Hypertrophy Infarction Ischemia Lipids MicroRNAs Myocardial Infarction Myocardium Organ Organoids Pathologic Patients Persons Production Property Proteins RNA Rattus Recovery of Function Regenerative Medicine Reperfusion Injury Reperfusion Therapy Reproducibility Research Risk Signal Transduction Silicon Source Stromal Cells Structure Therapeutic Time Tissue Engineering Tissues Treatment Efficacy Vascularization angiogenesis cardiac repair cardioprotection cell replacement therapy cell type density design exosome human pluripotent stem cell immunogenicity immunoregulation implantation improved in vivo innovation large scale production mimetics nanofabrication nanomaterials nanoparticle nanowire next generation preservation public health relevance synergism three dimensional cell culture tumorigenic

项目摘要

Project Summary: Exosome therapy holds remarkable promise to treat acute infarction, a major cause of heart failure that affects over 6 million people in the US. Exosomes are nanoparticles secreted by cells to facilitate intercellular signaling through their bioactive cargos such as microRNAs (miRNAs). Compared to cell-based therapies, exosomes have distinct advantages including low immunogenicity, absence of tumorigenic risk, and amenable for large scale production and off-of-shelf storage. Among various exosomes used for cardiac regenerative medicine, exosomes from human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) have received significant attention. While 2D culture has been considered as “gold standard” for exosome production, recent studies show 3D culture promote the production of pro-reparative exosomes. To this end, we developed nanowired human cardiac organoids, which are composed of hPSC-CMs, human primary cardiac fibroblasts, endothelial cells, stromal cells, and electrically conductive silicon nanowires (e-SiNWs). Compared to 2D hPSC- CM culture, organoids provide a myocardium mimetic microtissue platform, and the e-SiNWs creates a conductive microenvironment to enhance exosome biogenesis and secretion. Compared to the unwired organoids (without e-SiNWs), the nanowired organoids showed the significantly higher ability to improve vascularization, preserve myocardium, attenuate pathological hypertrophy of the infarcted hearts and recover their contractile function. To improve the reproducibility of exosomes, we developed isogenic hPSC cardiac organoids using hPSC-CMs, -cFBs (cardiac fibroblasts), and -ECs (endothelial cells) derived from a single hPSC line. The goal of this proposal is to determine the effects of key variables of the nanowired hPSC cardiac organoids (i.e., size, cell composition and e-SiNW structures) on their exosome production and functionality. The central hypothesis of the proposal is the nanowired hPSC cardiac organoids provide an electrically conductive, biomimetic environment for hPSC-CMs, -cFBs, and -ECs to produce therapeutically potent exosomes for cardiac repair. The proposal is innovative is that, for the first time, we will synergize electrical nanomaterials with human cardiac organoids to enhance exosome production and functionality. Accordingly, we will pursue 2 Aims: 1): Determine the effects of cell seeding ratio, size and e-SiNW structure of the nanowired isogenic hPSC cardiac organoids on exosome production and function, 2): Determine the therapeutic efficacy of the exosomes derived from the optimized nanowired organoids. The proposed studies are significant in that we aim to shift the current paradigm of exosome production to develop organ-specific therapeutic exosomes by leveraging the recent advances in nanofabrication and engineered human cardiac organoids. These studies will provide a guiding principle to design and develop next generation of tissue engineering constructs that can provide a source of sustained exosome production after implantation in addition to a cell replacement therapy.

项目摘要：外泌体疗法在治疗急性梗塞（心脏病的主要原因）方面具有非凡的前景影响美国超过 600 万人的失败，外泌体是细胞分泌的纳米颗粒，以促进细胞的生长。与基于细胞的相比，通过其生物活性货物（例如 microRNA (miRNA)）进行细胞间信号传导。外泌体具有独特的优势，包括低免疫原性、无致瘤风险以及适合大规模生产和现成储存用于心脏的各种外泌体。再生医学中，来自人多能干细胞来源的心肌细胞（hPSC-CM）的外泌体具有虽然二维培养被认为是外泌体生产的“黄金标准”，最近的研究表明，3D 培养可促进修复前外泌体的产生。为此，我们开发了这种方法。纳米线人心脏类器官，由 hPSC-CM、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) 与 2D hPSC- 相比。 CM 培养、类器官提供了模拟心肌的微组织平台，而 e-SiNW 则创建了与无线相比，导电微环境可增强外泌体的生物发生和分泌。与类器官（没有 e-SiNW）相比，纳米线类器官表现出显着更高的改善能力血管化，保护心肌，减轻梗塞心脏的病理性肥大并恢复为了提高外泌体的可重复性，我们开发了同基因 hPSC 心脏。使用源自单个 hPSC 的 hPSC-CM、-cFB（心脏成纤维细胞）和 -EC（内皮细胞）制作类器官该提案的目标是确定纳米线 hPSC 心脏的关键变量的影响。类器官（即大小、细胞组成和 e-SiNW 结构）的外泌体生产和功能。该提案的中心假设是纳米线 hPSC 心脏类器官提供导电、 hPSC-CM、-cFB 和 -EC 的仿生环境，可产生具有治疗作用的心脏外泌体该提案的创新之处在于，我们首次将电气纳米材料与修复。因此，我们将追求两个目标： 1): 确定纳米线同基因 hPSC 心脏的细胞接种比例、尺寸和 e-SiNW 结构的影响类器官对外泌体产生和功能的影响，2)：确定衍生的外泌体的治疗效果所提出的研究具有重要意义，因为我们的目标是改变当前的情况。外泌体生产范例，利用最近的研究开发器官特异性治疗性外泌体纳米制造和工程人类心脏类器官的进展将提供指导。设计和开发下一代组织原理工程结构，可以提供除了细胞替代疗法之外，植入后持续产生外泌体。