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

• 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
• 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

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：外泌体疗法具有巨大的希望治疗急性梗死，这是心脏的主要原因 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 细胞间信号通过其生物活性肉类（如microRNA）（miRNA）。与基于细胞的相比 疗法，外泌体具有不同的优势，包括低免疫原性，缺乏肿瘤风险和 适用于大规模生产和搁置储存。在用于心脏的各种外泌体中 再生医学，来自人类多能干细胞衍生的心肌细胞（HPSC-CM）的外泌体具有 受到了极大的关注。虽然2D文化被认为是外泌体生产的“黄金标准”，但 最近的研究表明，3D培养促进了额外的外泌体的产生。为此，我们发展了 由HPSC-CM，人类原发性心脏成纤维细胞组成的纳米人心脏器官， 内皮细胞，基质细胞和导电硅纳米线（E-SINWS）。与2D HPSC相比 CM培养，类器官提供了模拟微动物平台，E-Sinws创建了一个 导电微环境以增强外泌体生物发生和分泌。与未婚 类器官（无E-sinws），纳米器官表明改善的能力明显更高 血管化，保留心肌，减弱梗塞心脏的病理肥大并恢复 他们的收缩功能。为了提高外泌体的可重复性，我们开发了等源性HPSC心脏 使用HPSC -CM，-CFBS（心脏成纤维细胞）和-ec（内皮细胞）的类器官，源自单个HPSC 线。该提案的目的是确定纳米hPSC心脏的关键变量的影响 器官（即大小，细胞组成和E-SINW结构）在其外泌体产生和功能上。这 该提案的中心假设是纳米的HPSC心脏器官提供了导电性的， HPSC -CM，-CFB和-EC的仿生环境可为心脏产生热效外外泌体 维修。该提议的创新是，我们将首次与电气纳米材料协同 人体心脏器官可增强外泌体产生和功能。根据，我们将追求2个目标： 1）：确定纳米质量HPSC心脏的细胞播种比，大小和E-sinw结构的影响 外部生产和功能上的器官，2）：确定衍生的外泌体的治疗效率 从优化的纳米器官中。拟议的研究很重要，因为我们旨在改变电流 外部生产范式通过利用最近的 纳米制作和设计的人类心脏器官的进步。这些研究将提供指导 设计和开发下一代组织工程结构的原则，可以提供 除细胞替代疗法外，植入后持续外泌体产生。