Exosome Therapeutics to Dissect HFpEF Mechanisms

外泌体疗法剖析 HFpEF 机制

• 批准号: 10296255
• 负责人: EDUARDO MARBAN
• 金额: $ 83.44万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10296255
• 关键词: Animal Model; Anti-Inflammatory Agents; Arrhythmia; Attenuated; Back; Benchmarking; Biological; Biopsy; Biopsy Specimen; Cell Therapy; Cells; Chemicals; Collagen; Complex; Control Groups; Data; Disease; EFRAC; Exposure to; FRAP1 gene; Failure; Family suidae; Fibrosis; Genetic Transcription; Goals; Heart; Heart failure; Histologic; Human; Inflammation; Intervention; Mediating; Medical Device; Methods; MicroRNAs; Mining; Modeling; Molecular; Molecular Profiling; Morbidity - disease rate; Mus; Myocardial; Pathogenesis; Pathway interactions; Patients; Pharmacology; Phenotype; Post-Translational Protein Processing; Proteins; Proteome; Proteomics; RNA; RNA analysis; Rattus; Reporting; Rodent; Sorting - Cell Movement; Techniques; Testing; Therapeutic; Tissues; Translations; Treatment Efficacy; Treatment Failure; Untranslated RNA; Ventricular; base; cardioprotection; comorbidity; comparative efficacy; cost; design; disease phenotype; exosome; extracellular vesicles; improved; in vivo; mechanical properties; mortality; mouse model; novel; novel strategies; novel therapeutics; porcine model; preservation; response; single-cell RNA sequencing; targeted treatment; therapeutic candidate; transcriptome sequencing; transcriptomics; treatment response

PROJECT SUMMARY No effective pharmacological or medical device interventions are available to treat heart failure with preserved ejection fraction (HFpEF). We have pioneered the concept of cell therapy for HFpEF: Cardiosphere-derived cells (CDCs) dramatically improve diastolic function and reduce arrhythmias, while attenuating fibrosis and inflammation. Most, if not all, of these beneficial effects are mediated by exosomes secreted by CDCs (CDCEXO). Here we seek to establish detailed molecular signatures of HFpEF; to use those molecular signatures as roadmaps to identify key, potentially causal pathways by dissecting the responses to CDCEXO; and to discover novel defined molecular entities, based on CDCEXO cargo, with disease-modifying bioactivity in HFpEF. Our hypotheses, backed by strong preliminary data, are: • Underlying HFpEF are bewilderingly extensive changes in myocardial transcriptomics and proteomics. Sorting causal from associative changes presents a major challenge, but it is doable. • A subset of these HFpEF-related proteome changes are reversed by CDCs or CDCEXO and some correlated to the reversal of the key functional abnormalities of HFpEF. We posit that focusing on CDCEXO-responsive pathways will facilitate the search for causal abnormalities, enabling targeted hypothesis testing. • By mining the RNA and protein contents of CDCEXO, we have the potential to pinpoint defined factors which have disease-modifying bioactivity in HFpEF. Such defined factors may themselves be viable therapeutic candidates, or can inspire the creation of new chemical entities as therapeutic candidates. The overall goal of this proposal is to understand better the pathogenesis of HFpEF, and to develop novel cell- free approaches to treat this disease. Three aims are proposed. In Aim 1 we will perform and analyze tissue and single cell transcriptomics and proteomics (including numerous protein post-translational modifications) of HFpEF, in three different models (pig, mouse and rat) that represent different comorbidities and compare their disease signature to those in human HFpEF heart tissue. Commonalities in the OMICS responses among species will help distinguish causal versus associative pathways in HFpEF pathogenesis. Aim 2 will analyze molecular signatures of therapeutic efficacy by comparing transcriptomics and proteomics with and without exposure to CDCEXO. Because CDCs and their exosomes strikingly reverse the HFpEF phenotype, identification of CDCEXO-induced molecular changes will further refine pathway prioritization in terms of causal versus associative. Here we will study ventricular tissue and single cells from rat, mouse and pig models of HFpEF (and controls), with and without in vivo exposure to CDCEXO. In Aim 3 we will mine CDCEXO cargo to identify critical factors underlying disease-modifying bioactivity in a rat model of HFpEF. This will allow us to define and/or create specific molecular entities (either RNA species or proteins) that may be preferable to complex biologicals (cells, exosomes) in terms of mechanistic discreteness, ease of manufacturing, and therapeutic consistency.

项目摘要 没有有效的药理学或医疗器械干预可用于治疗心力衰竭， 射血分数（HFpEF）。我们开创了HFpEF细胞治疗的概念：球源性细胞 （CDC）显著改善舒张功能并减少心律失常，同时减轻纤维化和 炎症这些有益作用中的大多数（如果不是全部）是由CDC分泌的外来体（CDCEXO）介导的。 在这里，我们试图建立HFpEF的详细分子特征;使用这些分子特征作为 路线图，通过剖析对CDCEXO的反应来确定关键的、潜在的因果途径;并发现 新定义的分子实体，基于CDCEXO货物，在HFpEF中具有疾病修饰生物活性。我们 有强有力的初步数据支持的假设是： ·HFpEF的基础是心肌转录组学和蛋白质组学令人困惑的广泛变化。分选 从关联变化中产生因果关系是一个重大挑战，但这是可行的。 ·这些HFpEF相关蛋白质组变化的一个子集被CDC或CDCEXO逆转，并且一些相关的蛋白质组变化被CDCs或CDCEXO逆转。 逆转HFpEF的关键功能异常。我们认为，专注于CDCEXO响应 通路将有助于寻找因果异常，从而实现有针对性的假设检验。 ·通过挖掘CDCEXO的RNA和蛋白质含量，我们有可能精确定位定义的因素， 在HFpEF中具有改善疾病的生物活性。这些确定的因素本身可能是可行的治疗 候选物，或者可以激发产生新的化学实体作为治疗候选物。 该提案的总体目标是更好地了解HFpEF的发病机制，并开发新的细胞， 免费的治疗方法。提出了三个目标。在目标1中，我们将执行和分析组织， 单细胞转录组学和蛋白质组学（包括许多蛋白质翻译后修饰） HFpEF，在三种不同的模型（猪，小鼠和大鼠），代表不同的合并症，并比较其 疾病特征与人类HFpEF心脏组织中的那些相比较。各组织对OMICS的答复的共同点 物种将有助于区分HFpEF发病机制中的因果途径与关联途径。目标2将分析 通过比较转录组学和蛋白质组学， 暴露于CDCEXO。由于CDC和它们的外来体显著逆转HFpEF表型， 的CDCEXO诱导的分子变化将进一步细化途径的优先顺序， 联想的在这里，我们将研究HFpEF大鼠、小鼠和猪模型的心室组织和单细胞（以及 对照），有和没有体内暴露于CDCEXO。在目标3中，我们将挖掘CDCEXO货物， HFpEF大鼠模型中疾病修饰生物活性的潜在因素。这将允许我们定义和/或创建 可能优选复杂生物制品（细胞， 外泌体）在机械离散性、制造容易性和治疗一致性方面。