Using a Cardiac Microtissue System to Evaluate and Replicate Clinical Therapy Responses using Patient Cell-Derived Exosomes

使用心脏微组织系统评估和复制患者细胞衍生的外泌体的临床治疗反应

基本信息

批准号：
9924691
负责人：
Camila Hochman-Mendez
金额：
$ 20.38万
依托单位：
TEXAS HEART INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-01 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9924691
关键词：
Action Potentials Allogenic Architecture Autologous Basement membrane Beds Biological Bone Marrow Cardiac Cardiac Myocytes Cardiovascular Diseases Cardiovascular system Cell Proliferation Cell Survival Cell Therapy Cell Transplantation Cell physiology Cells Characteristics Clinical Clinical Trials Clinical Trials Network Congestive Heart Failure Data Development Endothelial Cells Environment Exposure to Extracellular Matrix Extracellular Matrix Proteins Fibroblasts Frequencies Future Genes Glucose Heart Human Hypoxia In Vitro Individual Infarction Injury Institutes Laboratories Lead Left Ventricular Ejection Fraction Link Mediator of activation protein Mesenchymal Stem Cells Modeling Molecular Myocardial Myocardial Infarction Myocardium National Heart, Lung, and Blood Institute Natural regeneration Organ Outcome Oxygen Consumption Patients Phenotype Physiological Placebos Play Process Property Proteins RNA Resistance Role Sampling Stress System Testing Texas Therapeutic Studies Tissue Engineering Vesicle base biobank cardiac regeneration cardiac repair cardiac tissue engineering cell behavior clinical effect design exosome experience functional improvement high throughput screening improved in vitro Model in vivo individual patient induced pluripotent stem cell injured innovation insight interest interstitial loss of function novel novel strategies paracrine patient response prevent regenerative repaired response stem cell differentiation stem cells success tissue repair

项目摘要

Summary Cell therapy for post-infarct cardiac repair has shown limited and mixed results, most likely due to patients' individual cell characteristics or to the infarct environment experienced by the transplanted cells. Identifying the potential molecular and cellular components that contribute to patient-specific responses and ways to minimize the environmental stress on transplanted cells may lead to the development of new strategies for cardiac repair. Additionally, numerous studies have shown that the effects of at least some cell-based therapies can be attributed primarily to secreted cellular factors that are packaged inside exosomes. These exosomes are considered critical mediators of intercellular information and play a direct role in injury-induced tissue repair processes in multiple physiological systems. Our group is particularly interested in the characterization of exosomes from cardiovascular disease patients who improved in a cell therapy study and in the utilization of these exosome cargos in the development of tissue-engineered cardiac patches for optimal functional myocardial repair. We are also interested in using our in vitro cardiac microtissue system as a testbed to evaluate patient cell-exosome interactions and associations with clinical trial responses. We will derive bone marrow mesenchymal stem cells (MSCs), MSC exosomes, and human induced pluripotent stem cells (hiPSCs) from healthy individuals and patients in the Cardiovascular Cell Therapy Research Network (CCTRN)-FOCUS clinical trial categorized as improvers at 6 months, [i.e., improved left ventricular ejection fraction, end-systolic volume and maximal oxygen consumption], or as non-improvers (declined in the 3 outcomes). Cardiac microtissues, containing cardiac cells derived from healthy control hiPSCs, cultured in native and infarct-like conditions will be treated with improvers' exosomes. Changes in microtissue function, cardiomyocyte maturation, cell survival and proliferation will be compared among improvers, placebo and non-improvers exosomes. Meanwhile, high throughput assays will be used to identify differences in the exosome cargos of the top 3 clinical improvers and non-improvers and correlated with in vitro responses. We expect that improver's exosomes will contain potent beneficial factors to enhance cell and microtissue maturation and function under infarct-like stress. Next, patient- specific microtissues containing hiPSC-derived cardiac cells generated from bone marrow of the top 3 improvers and non-improvers will be treated with autologous or allogenic (improver, non-improver or, healthy) exosomes under infarct-like conditions. By exposing patient-specific “infarcted” cardiac microtissues to autologous or allogeneic exosomes, we can evaluate patient-specific cell-exosome interactions in a controlled in vitro setting that mimics the clinical condition. Ultimately, these studies will provide new insights into cell-exosome interactions in an infarct as well as their relative potency and association with clinical trial responses, thus providing an innovative new approach to developing cardiac patches with superior regenerative properties.

概括可感染后心脏修复的细胞疗法显示出有限和混杂的结果，很可能是由于患者的单个细胞特征或移植细胞所经历的基础设施环境。识别潜在的分子和细胞成分有助于患者特异性反应以及最小化方法移植细胞的环境压力可能导致发展心脏修复的新策略。此外，许多研究表明，至少某些基于细胞的疗法的影响可以是主要归因于外泌体内包装的分泌细胞因子。这些外泌体是被认为是细胞间信息的关键介体，并在损伤引起的组织修复中发挥直接作用多个物理系统中的过程。我们的小组对特征特别感兴趣在细胞疗法研究中改善的心血管疾病患者的外泌体和这些外泌体的兑现在组织工程心脏斑块的开发中，以实现最佳功能心肌修复。我们也有兴趣使用我们的体外心脏微动物系统作为测试床来评估患者细胞 - 异位相互作用以及与临床试验反应的关联。我们将得出骨髓间充质干细胞（MSC），MSC外泌体和人类诱导的多能干细胞（HIPSC）从心血管细胞疗法研究网络（CCTRN）的健康个体和患者 - 对焦临床试验类别为6个月，[即改善左心室射血分数，末端节音量和最大氧的消耗]，或作为非注射剂（在3个结果中降低）。心脏微动物，含有源自健康对照hipsc的心脏细胞，在天然和梗死条件下培养的心脏细胞将是接受改进的外泌体治疗。微作用功能，心肌细胞成熟，细胞存活和将在改善，安慰剂和非凹痕外泌体之间进行比较。同时，高吞吐量测定将用于识别前3名临床改进的外泌体羧化的差异和非凹口，与体外反应相关。我们预计改进剂的外泌体将包含有效的在梗塞样压力下增强细胞和微动物成熟和功能的有益因素。接下来，患者 - 特定的微动物，这些微动物含有从前3个改进的骨髓产生的HIPSC衍生心脏细胞并将用自体或同种异体（改良剂，非僵局或健康）外泌体治疗非注射剂在类似梗塞的条件下。通过将特定于患者的“梗塞”心脏微动物暴露于自体或同种异体外泌体，我们可以在受控的体外环境中评估患者特异性的细胞 - 异位相互作用模仿临床状况。最终，这些研究将提供对细胞诊断的新见解梗塞中的相互作用以及它们的相对效力和与临床试验反应的关联，因此提供了一种创新的新方法，用于开发具有出色再生特性的心脏斑块。