Leveraging the HIF-alpha pathway to improve the engraftment and therapeutic efficacy of human nanowired cardiac organoids

利用 HIF-α 途径提高人类纳米线心脏类器官的植入和治疗效果

• 批准号: 10658988
• 负责人: Ryan W Barrs
• 金额: $ 2.63万
• 依托单位: CLEMSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-16 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10658988
• 关键词: 3-Dimensional; Acceleration; Address; Advanced Development; Affect; Anastomosis - action; Animal Model; Animals; Biological Process; Blood Vessels; Brain Hypoxia-Ischemia; Cardiac; Cardiovascular Diseases; Cause of Death; Cell Survival; Cell Therapy; Cells; Cessation of life; Clinical; Data; Development; Dissociation; Echocardiography; Electric Conductivity; Electrocardiogram; Endothelial Cells; Endothelium; Engineering; Engraftment; Fibroblasts; Foundations; Genes; Genotype; Goals; Heart; Heart Diseases; Heart Injuries; Histologic; Human; Human Engineering; Hydroxylation; Hypoxia; Hypoxia Inducible Factor; Hypoxia-Inducible Factor Pathway; In Vitro; Infarction; Injections; Investigation; Ischemia; Ischemic Preconditioning; Metabolic; Metabolism; Modeling; Myocardial Infarction; Myocardial Reperfusion Injury; Myocardium; Organoids; Oxygenases; PECAM1 gene; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Principal Component Analysis; Procollagen-Proline Dioxygenase; Production; Rattus; Recovery of Function; Reperfusion Injury; Research; Role; Signal Pathway; Signal Transduction; Silicon; Source; Stromal Cells; Structure; Supporting Cell; Time; Tissue Transplantation; Tissues; Transplantation; Treatment Efficacy; United States; Vascular Endothelial Growth Factors; Vascularization; Visualization; angiogenesis; cardiac repair; cardiac tissue engineering; clinical application; clinical translation; heart damage; heart function; improved; in vivo; induced pluripotent stem cell derived cardiomyocytes; inhibitor; innovation; insight; mimetics; nanowire; pharmacologic; phase III trial; post-transplant; preconditioning; programs; public health relevance; regenerative therapy; repaired; transcription factor; transcriptome; transcriptome sequencing; transcriptomics

PROJECT SUMMARY: Heart disease accounts for nearly 1 in 4 deaths in the United States each year, highlighting the urgent need for therapies that can repair damaged hearts. Human induced pluripotent stem cell- derived cardiomyocytes (hiPSC-CMs) have emerged as a powerful cell source for cardiac repair, but their potential has been limited by poor survival and engraftment after injection. To address these challenges, our lab has pioneered the development of nanowired human cardiac organoids composed of electrically conductive silicon nanowires (e-SiNWs), hiPSC-CMs, and supporting cells. Our preliminary in vivo studies showed that nanowired cardiac organoids successfully engraft in ischemia/reperfusion (I/R) injured rat hearts and develop more organized contractile structures compared to non-nanowired cardiac organoids. Despite this progress, less than half (~30%) of injected organoids remained engrafted in infarcted hearts 7 days post-transplantation, which can be attributed to inadequate prevascularization and hypoxic/ischemic preconditioning of the organoids in vitro. To address this, we have explored pharmacological stabilization of HIF-a as a strategy to promote prevascularization and ischemic tolerance within the organoids. My preliminary in vitro data showed that treatment with Molidustat, a prolyl hydroxylase domain (PHD) inhibitor, significantly improved endothelial network and lumen formation (i.e., ~150% increase of CD31+ coverage) within the cardiac organoids. While these results are promising, further investigation is necessary to reveal phenotypic and genotypic changes in HIF-α stabilized cardiac organoids and how they correlate with transplantation efficiency. The goals of this proposal are to determine the effects of HIF-α stabilization on vascular maturation, cardiac function, cell and tissue-level metabolism, and transcriptomic changes in cardiac organoids (Aim 1), and to demonstrate therapeutic efficacy of HIF-α stabilized organoids in a rat model of myocardial I/R injury (Aim 2). The central hypothesis of this proposal is that stabilization of HIF-α signaling in cardiac organoids improves the survival and engraftment of hiPSC-CMs in infarcted myocardium and enhances their capacity to promote cardiac functional recovery in injured hearts. The proposal is innovative in that, for the first time, we will investigate how hypoxia mimetic agents precondition human engineered cardiac tissue to enhance the transplantation efficiency of hiPSC-CMs. My long-term goal is to develop clinically applicable cardiac regenerative therapies to treat cardiovascular diseases. Accordingly, we will pursue the following specific aims: 1) Determine how pharmacological HIF-α stabilization reprograms and preconditions human cardiac organoids for ischemic protection, and 2) Determine the effects of HIF-α stabilization on graft-host anastomosis, long-term engraftment, and therapeutic efficacy of nanowired human cardiac organoids in injured hearts. This research will inform emergent clinical applications of hypoxia mimetic agents to treat cardiovascular disease and will help advance our human cardiac organoid platform towards large animal studies to accelerate their clinical translation.

项目摘要：在美国，每年有近四分之一的人死于心脏病， 强调迫切需要能够修复受损心脏的疗法。 衍生心肌细胞（hiPSC-CM）已成为心脏修复的强大细胞来源，但它们 为了应对这些挑战，我们的潜力受到注射后存活率和植入不良的限制。 实验室率先开发了由导电材料组成的纳米线人类心脏器官 我们的初步体内研究表明，硅纳米线 (e-SiNW)、hiPSC-CM 和支持细胞。 纳米线心脏类器官成功植入缺血/再灌注（I/R）损伤的大鼠心脏并发育 与非纳米心脏类器官相比，其收缩结构更有组织性，尽管取得了这些进展，但仍较少。 移植后 7 天，超过一半（约 30%）的注射类器官仍植入梗塞心脏中，这 可归因于体外类器官的预血管化和缺氧/缺血预处理不足。 为了解决这个问题，我们探索了 HIF-a 的药理稳定性作为促进 我的初步体外数据表明，类器官内的血管前形成和缺血耐受性。 Molidustat（一种脯氨酰羟化酶结构域 (PHD) 抑制剂）治疗可显着改善内皮网络 心脏类器官内的腔形成（即 CD31+ 覆盖率增加约 150%）。 有希望，需要进一步研究以揭示 HIF-α 稳定的表型和基因型变化 心脏类器官及其与移植效率的关系 该提案的目标是 确定 HIF-α 稳定性对血管成熟、心脏功能、细胞和组织水平的影响 心脏类器官的代谢和转录组变化（目标 1），并证明治疗效果 HIF-α 稳定类器官在心肌 I/R 损伤大鼠模型中的作用（目标 2）。 该提议认为，心脏类器官中 HIF-α 信号的稳定可提高心脏类器官的存活和植入 hiPSC-CM 参与梗死心肌并增强其促进心脏功能恢复的能力 该提案的创新之处在于，我们将首次研究如何模拟缺氧。 试剂预处理人类工程心脏组织以提高 hiPSC-CM 的移植效率。 我的长期目标是开发临床适用的心脏再生疗法来治疗心血管疾病 因此，我们将追求以下具体目标： 1) 确定 HIF-α 的药理学作用。 稳定重编程和预处理人类心脏类器官以实现缺血保护，以及 2) 确定 HIF-α 稳定性对移植物-宿主吻合、长期植入和治疗效果的影响 这项研究将为受伤心脏中的纳米线人类心脏类器官提供信息。 低氧模拟剂治疗心血管疾病并将有助于推进我们的人类心脏类器官 大型动物研究平台，以加速其临床转化。

项目成果

Transplantation of Human Pluripotent Stem Cell-Derived Cardiomyocytes for Cardiac Regenerative Therapy.

移植人类多能干细胞来源的心肌细胞用于心脏再生治疗。

• 发表时间: 2021
• 作者: Silver, Sophia E;Barrs, Ryan W;Mei, Ying
• 通讯作者: Mei, Ying

Nanowired human cardiac organoid transplantation enables highly efficient and effective recovery of infarcted hearts.

纳米线人类心脏类器官移植能够高效且有效地恢复梗塞心脏。

• 发表时间: 2023-08-04
• 影响因子: 13.6
• 作者: Tan, Yu;Coyle, Robert C;Barrs, Ryan W;Silver, Sophia E;Li, Mei;Richards, Dylan J;Lin, Yiliang;Jiang, Yuanwen;Wang, Hongjun;Menick, Donald R;Deleon;Tian, Bozhi;Mei, Ying
• 通讯作者: Mei, Ying