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.

项目概要/摘要 该提案旨在了解新的非编码 RNA (ncRNA) 实体 TY4 的心脏保护作用， 这是受到外泌体内容物的生物启发。我们的重点是一类小型 ncRNA，而不是以前的 被认为与心脏病特别相关。 Y RNA 引起了我们的注意，因为分子 由 YRNA4 基因编码的外泌体在心脏来源的细胞分泌的外泌体中异常丰富。 主要的 ncRNA EV-YF1 在单独递送时具有有益的特性：EV-YF1 增加 IL-10 分泌，具有心脏保护和抗肥厚作用。在这里，我们提供了 TY4 的初步数据，这是一种新的 受 EV-YF1 启发的化学实体 (NCE)。 TY4 发挥涉及炎症的全局转录组变化 和纤维化途径，并在保留了心力衰竭的小鼠模型中发挥疾病修饰生物活性 射血分数、杜氏肌营养不良症和 MI。值得注意的是，当用母乳配制时，TY4 蛋白质酪蛋白，在所有三种模型中都口服起作用。在机制方面，初步数据表明TY4结合 TPR 是一种核孔复合蛋白，充当信使 RNA 从细胞核输出的看门人。我们 还发现巨噬细胞耗竭会消除 TY4 介导的心脏保护作用。作为一个小型化学- TY4 是经过修饰的 NCE 突变体，具有合成 RNA 药物所需的关键特征。但其他RNA药物，无论是 FDA 批准或正在开发的药物均具有已知的作用机制：它们是反义寡核苷酸、小分子 干扰 RNA、适体或 miR 修饰物。 TY4 不属于这些类别，因此是 新型RNA药物。这里的重点是了解 TY4 限制梗塞面积的机制。 我们将测试以下机制假设：TY4 通过与 TPR 结合而具有针对 MI 的心脏保护作用 改变靶细胞（特别是巨噬细胞）中的基因表达。我们将追求以下目标： 1： 研究 TY4 改变巨噬细胞基因表达的机制。子目标旨在 识别 TY4 结合伴侣，表征 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。图2：优化口服TY4在小鼠MI中的体内功效。子目标旨在 优化急性 MI 中口服 TY4 的剂量范例，测量小鼠口服给药后 TY4 的生物分布 急性 MI 之前和之后，确定 TY4 在急性 MI 后 MI 疗效的机会窗口，测试 TY4 老年小鼠的心脏保护功效，并表征了优化口服 TY4 递送对慢性疾病的影响 米模型。图 3：测试巨噬细胞是否是解释 TY4 在急性 MI 中的功效所必需和充分的。 子目标旨在比较有和没有先前巨噬细胞耗竭的 MI 结果 +/- TY4，探讨相对 组织驻留巨噬细胞与循环巨噬细胞的作用，并确定前心肌梗死后是否过继转移 体内 TY4 条件巨噬细胞可减少巨噬细胞耗竭小鼠的梗死面积。虽然高度机械化， 我们的提案专注于受外泌体货物生物启发的合成 ncRNA NCE，也提供了转化价值 开发一种即使在再灌注后也有效的新型心脏保护剂。