Developing Long Noncoding RNA Therapy for Precision Cardiac Repair

项目总结 心脏病是美国人的主要死因。约87.5万美国人死于心脏 每年有50-60%的死亡是由冠心病(CAD)引起的。CAD，最多的 常见类型的心脏病，可导致心脏病发作和猝死。而幸存的患者可以 进展为以心肌纤维化为特征的慢性心肌病和心力衰竭(HF)。 心肌纤维化与不良结局相关。目前，还没有治疗心脏病的特效药。 纤维化症。干细胞或外切体疗法一直是治疗纤维化疾病的一种有前途的选择。然而， 在质量控制和GMP级大规模生产方面的挑战以及难以捉摸的机制 阻碍了翻译的应用。应对挑战的一种解决方案是识别特定的分子 这是外切体抗纤维化机制的基础，并设计了纳米颗粒载体来运送它们。在我们的 形成建议基础的初步研究，我们发现转录因子TCF21在 抑制肌成纤维细胞的激活，并在外体携带长非编码RNA(LncRNA)-Tarid中发挥作用 TCF21基因表达的上游增强子。近年来，脂质纳米粒(LNPs)的应用已经 扩大了核酸输送的大范围临床试验。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。最重要的假设是携带lncRNA-Tarid的LNPs (LNP-Tarid)可上调TCF21，抑制心肌纤维化，改善心功能。我们的原创 格兰特专注于外体中的microRNA货物，如miR-21和miR-146。从测序结果看 实验中，我们偶然在外体中遇到了其他非编码RNA，它们发挥着必不可少的作用 在心脏修复中的作用。IncRNA Tarid就是其中之一。我们的初步研究证实了它的治疗作用。 心肌纤维化中的IncRNA Tarid。我们的续订提交将集中在此RNA代理上，使用LNP作为 送货承运人。目的1：LNP-Tarid处方的优化和体外表征；毒性 对幼稚动物的研究。目的2：确定LNP-Tarid治疗小鼠模型的安全性和有效性 急性和慢性心肌梗塞。目的3：将LNP-Tarid疗法转化为临床相关的猪糖尿病模型 心脏I/R损伤。我们的建议既是机制驱动的，也是产品导向的。我们的研究结果 将为开发新的非编码RNA疗法治疗心肌梗死和 帮助我们从机制上深入了解TCF21调控的心脏纤维化途径。

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.