Cardioprotective mechanisms of novel noncoding RNA in myocardial infarction

新型非编码RNA对心肌梗死的心脏保护机制

• 批准号: 10660164
• 负责人: EDUARDO MARBAN
• 金额: $ 64.13万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10660164
• 关键词: Acute myocardial infarction; Adoptive Transfer; Antisense Oligonucleotides; Attention; Binding; Biodistribution; Biological; Biological Availability; Cardiotonic Agents; Caseins; Cell Nucleus; Cell Survival; Cell Therapy; Cells; Chemicals; Chronic; Data; Development; Dichloromethylene Diphosphonate; Disease; Duchenne muscular dystrophy; EFRAC; FDA approved; Gatekeeping; Gene Expression; Genes; Heart; Heart Diseases; Heart failure; Human Milk; In Vitro; Infarction; Inflammatory; Injury; Interleukin-10; Intravenous; Ischemia; Left; Link; Macrophage; Measures; Mediating; Mediator; Messenger RNA; Milk Proteins; Modeling; Mus; Myocardial Infarction; Necrosis; Nuclear Pore Complex Proteins; Nucleotides; Oral; Oral Administration; Outcome; Pathway interactions; Peptides; Pharmaceutical Preparations; Property; RNA; Rationalization; Reperfusion Therapy; Role; Small Interfering RNA; Testing; Thinking; Tissues; Transplantation; Untranslated RNA; Work; aged; aptamer; cardioprotection; cytokine; dosage; exosome; extracellular vesicles; in vivo; insight; mouse model; mutant; new chemical entity; novel; paracrine; preservation; prototype; regenerative cell; regenerative therapy; transcriptomics; translational potential

PROJECT SUMMARY/ABSTRACT This proposal seeks to understand the cardioprotective effects of a new noncoding RNA (ncRNA) entity, TY4, that is bioinspired by the contents of exosomes. Our focus is on one class of small ncRNA, not previously recognized as particularly relevant to cardiac disease. Y RNAs captured our attention because molecules encoded by the YRNA4 gene are exceptionally plentiful in exosomes secreted by cardiosphere-derived cells. The predominant such ncRNA, EV-YF1, has salutary properties when delivered on its own: EV-YF1 increases IL-10 secretion, is cardioprotective and anti-hypertrophic. Here we present preliminary data on TY4, a new chemical entity (NCE) bioinspired by EV-YF1. TY4 exerts global transcriptomic changes involving inflammatory and fibrotic pathways, and exerts disease-modifying bioactivity in murine models of heart failure with preserved ejection fraction, Duchenne muscular dystrophy, and MI. Remarkably, TY4, when formulated with the breast milk protein casein, works orally in all three models. In terms of mechanism, preliminary data show that TY4 binds to TPR, a nuclear pore complex protein that acts as a gatekeeper for messenger RNA export from the nucleus. We have also found that macrophage depletion abrogates TY4-mediated cardioprotection. As a small chemically- modified mutant NCE, TY4 has key features desirable in a synthetic RNA drug. But other RNA drugs, whether FDA-approved or in development, all have known mechanisms of action: they are antisense oligos, small interfering RNAs, aptamers, or miR-modifiers. TY4 fits into none of those classes and thus is the prototype for a new class of RNA drugs. The focus here is on understanding the mechanism whereby TY4 limits infarct size. We will test the following mechanistic hypothesis: TY4 is cardioprotective against MI by binding to TPR and altering gene expression in target cells (and specifically, in macrophages). We will pursue the following Aims: 1: Investigate the mechanism(s) whereby TY4 alters gene expression in macrophages. Sub-aims seek to identify TY4 binding partners, characterize the transcriptomic changes induced by TY4, and prioritize biological targets for further interrogation. 2: Optimize in vivo efficacy of oral TY4 in murine MI. Sub-aims seek to optimize the dosage paradigm for oral TY4 in acute MI, measure biodistribution of TY4 after oral delivery in mice pre- and post- acute MI, determine the window of opportunity for TY4 efficacy post-MI in acute MI, test TY4 cardioprotective efficacy in aged mice, and characterize the effects of optimized oral TY4 delivery in a chronic MI model. 3: Test whether macrophages are necessary and sufficient to explain TY4 efficacy in acute MI. Sub-aims seek to compare MI outcomes +/- TY4 with and without prior macrophage depletion, probe the relative roles of tissue-resident versus circulating macrophages, and determine whether post-MI adoptive transfer of ex vivo TY4-conditioned macrophages reduces infarct size in macrophage-depleted mice. While highly mechanistic, our proposal, focusing on a synthetic ncRNA NCE bioinspired by exosomal cargo, also offers translational value in its development of a novel cardioprotective agent that is effective even when given after reperfusion.

项目摘要/摘要 该建议旨在了解新的非编码RNA（NCRNA）实体Ty4的心脏保护作用 这是由外泌体内容物生物启发的。我们的重点是一类小NCRNA，而不是以前 被认为与心脏病特别相关。 y rnas引起了我们的注意，因为分子 由YRNA4基因编码的外泌体在心圈衍生细胞分泌的外泌体中非常丰富。 这种主要的NCRNA（EV-YF1）自行交付时具有敬意的特性：EV-YF1增加 IL-10分泌，是心脏保护性和抗嗜性的。在这里，我们介绍了Ty4的初步数据，这是一个新的 EV-YF1生物启动的化学实体（NCE）。 TY4发挥涉及炎症的全球转录组变化 和纤维化途径，并在保留的心力衰竭的鼠模型中施加疾病改良的生物活性 射血分数，杜钦（Duchenne）肌肉营养不良和MI。值得注意的是，TY4用母乳配制 蛋白质酪蛋白在所有三个模型中口头工作。在机制方面，初步数据表明TY4与 TPR是一种核孔复合蛋白，充当从细胞核导出的允许者。我们 还发现，巨噬细胞耗竭可消除TY4介导的心脏保护。作为一个小化学 修饰的突变体NCE，TY4在合成RNA药物中具有关键特征。但是其他RNA药物，是否 FDA批准或在开发中，都具有已知的作用机理：它们是反义寡素，小 干扰RNA，适体或miR型改装剂。 TY4都不适合这些类，因此是一个原型 新的RNA药物。这里的重点是理解TY4限制梗塞大小的机制。 我们将测试以下机械假设：TY4通过与TPR结合和TPR和 改变靶细胞中基因表达（特别是在巨噬细胞中）。我们将追求以下目标：1： 研究TY4在巨噬细胞中改变基因表达的机制。 sub-aims寻求 识别TY4结合伙伴，表征由TY4诱导的转录组变化，并优先考虑生物学 进一步审讯的目标。 2：优化鼠MI中口服TY4的体内功效。 sub-aims寻求 优化急性MI中口服TY4的剂量范例，在小鼠中口服递送后测量TY4的生物分布 前后MI前后，确定急性MI后TY4功效的机会窗口，测试Ty4 老年小鼠的心脏保护功效，并表征了优化口服TY4在慢性中的影响 MI模型。 3：测试巨噬细胞是否需要和足以解释急性MI中的TY4功效。 Sub-Aims试图在有或没有之前的巨噬细胞耗尽的情况下比较MI结果+/- TY4，探测相对 组织居民与循环巨噬细胞的作用 体内TY4条件巨噬细胞可减少巨噬细胞耗尽的小鼠的梗塞大小。虽然机械性高 我们的建议专注于外泌体货物启发的合成NCRNA NCE，也提供了翻译价值 在发展新颖的心脏保护剂时，即使在再灌注后也有效。