Developing Long Noncoding RNA Therapy for Precision Cardiac Repair

项目摘要 心脏病是美国的主要死因。大约875，000美国人死于心脏病 冠心病（CAD）是全球每年最严重的心脏病，其中50-60%的死亡是由冠心病（CAD）引起的。CAD，最 心脏病是一种常见的心脏病，可导致心脏病发作和猝死。虽然存活的患者可以 进展为慢性心肌病和以心肌纤维化为特征的心力衰竭（HF）。 心肌纤维化与不良结局相关。目前，尚无针对心脏病的特效药。 纤维化干细胞或外泌体治疗已成为纤维化疾病的一种有前途的选择。但 质量控制和GMP级大规模生产的挑战以及难以捉摸的机制， 阻碍了翻译的应用。应对这些挑战的一个解决方案是识别特定的分子 这是外来体抗纤维化机制的基础，并设计纳米颗粒载体来递送它们。在我们 初步研究形成的建议的基础上，我们发现，转录因子Tcf 21是至关重要的， 抑制肌成纤维细胞活化，外泌体货物长非编码RNA（lncRNA）-TARID起作用， Tcf 21基因表达的上游增强子。最近，脂质纳米颗粒（LNP）的应用已经得到了广泛的应用。 在核酸递送的大范围临床试验中扩大。在这项提案中，我们的目标是开发lncRNA LNP用于治疗心脏纤维化。总体假设是负载lncRNA-TARID的LNP （LNP-TARID）可上调Tcf 21，抑制心肌纤维化，改善心功能。我们原来的 格兰特专注于外泌体中的microRNA货物，如miR-21和miR-146。从测序 在实验中，我们意外地在外泌体中遇到了其他非编码RNA，这些RNA发挥着重要的作用。 在心脏修复中的作用LncRNA TARID就是其中之一。我们的初步研究证实了 心肌纤维化中的LncRNA TARID。我们的更新提交将集中在这种RNA代理，使用LNP作为 送货员目的1：LNP-TARID制剂的优化和体外表征;毒性 在幼稚动物身上的研究目的2：确定LNP-TARID治疗在小鼠模型中的安全性和有效性 急性和慢性心肌梗死目的3：将LNP-TARID疗法转化为临床相关的猪模型， 心脏I/R损伤。我们的建议是机制驱动和产品导向。我们的研究结果 将为开发新的非编码RNA疗法来治疗心肌梗死铺平道路， 帮助我们获得Tcf 21调节心脏纤维化途径的机制见解。

PROJECT SUMMARY Heart disease is the leading cause of death in the United States. About 875,000 Americans die from heart disease each year, of which 50-60% deaths are caused by coronary artery disease (CAD). CAD, the most common type of heart disease, can lead to heart attack and sudden death. While the survival patients can progress into chronic cardiomyopathy and heart failure (HF) that is featured with myocardial fibrosis. Myocardial fibrosis is associated with adverse outcomes. Currently, there is no specific drugs for cardiac fibrosis. Stem cell or exosome therapy has been a promising option for fibrotic disease. However, the challenges in quality control and GMP-grade large-scale production as well as the elusive mechanisms have hindered the translational application. One solution to address the challenges is to identify specific molecules that underlie the antifibrotic mechanisms of exosomes, and design nanoparticle carriers to deliver them. In our preliminary studies that form the basis of proposal, we found that transcription factor Tcf21 is crucial in suppressing myofibroblast activation, and exosome cargo long noncoding RNA (lncRNA)-TARID played as the upstream enhancer of Tcf21 gene expression. Recently, the application of lipid nanoparticles (LNPs) has expanded in a large scope of clinical trials for nucleic acid delivery. In this proposal, we aim to develop lncRNA LNP for treatment of cardiac fibrosis. The overarching hypothesis is that LNPs loaded with lncRNA-TARID (LNP-TARID) can upregulate Tcf21 to suppress myocardial fibrosis and improve cardiac functions. Our original grant focuses on microRNA cargos in the exosomes, such as miR-21 and miR-146. From the sequencing experiments, we serendipitously encountered other non-coding RNAs in the exosomes that play an essential role in cardiac repair. LncRNA TARID is one of them. Our preliminary studies confirmed the therapeutic role of LncRNA TARID in cardiac fibrosis. Our renewal submission will be focusing on this RNA agent, using LNP as the delivery carrier. Aim 1: Optimization of LNP-TARID formulations and in vitro characterization; toxicity studies in naïve animals. Aim 2: Determine the safety and efficacy of LNP-TARID treatment in mouse models of acute and chronic MI. Aim 3: Translate the LNP-TARID therapy into a clinically-relevant pig model of cardiac I/R injury. Our proposal is both mechanism-driven and product-oriented. The results from our research will pave the ground for the development of new non-coding RNA therapies to treat myocardial infarction and help us gain mechanistic insights on the Tcf21-regulated cardiac fibrosis pathways.