Design of precision small molecules targeting RNA repeating transcripts to manipulate and study disease biology

设计针对 RNA 重复转录本的精密小分子，以操纵和研究疾病生物学

• 批准号: 10595458
• 负责人: Matthew D Disney
• 金额: $ 72.54万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10595458
• 关键词: Design; precision; small; molecules; targeting

PROJECT SUMMARY: An extraordinarily challenging problem is to develop general methods to target defective or malfunctioning RNAs that cause disease selectively. Current therapeutic strategies to target RNAs are based on specific sequence recognition by oligonucleotides. However, many human disorders are caused by highly structured RNAs not readily targetable by conventional base pairing, in particular RNA repeat expansions that cause or contribute to >30 incurable neuromuscular diseases and genetically defined dementia. Thus, allele- specific ASOs modalities for these microsatellite disorders have been developed by targeting polymorphisms outside of the repeating sequence. The consequences of this approach are that only patients with the polymorphisms benefit from treatment and that an ASO has to be developed for each disease, even if caused by the same repeating sequence. If the toxin in these diseases, the expanded repeat, could be targeted selectively with a structure-specific small molecule, then a single modality could be a therapeutic or chemical probe for multiple diseases and for all patients. Over the past 14 years, we have shown that RNA structures can be targeted selectively with small molecules in situ and in vivo, more selectively than oligonucleotides. Indeed, we have designed compounds against many RNA repeat expansions that selectively recognize the target’s structure and rescue disease-associated pathobiology in situ and in vivo. Further, these chemical probes have elucidated new mechanisms of disease, including a previously unknown RNA-mediated transcriptional silencing pathway that operates in fragile X syndrome. These studies, along with our innovative strategies to synthesize drugs at the site of disease and to engineer small molecules with novel activities, including antisense- or CRISPR-like modes of action, lay the foundation for our proposed research program. Herein, we propose a comprehensive strategy to study the molecular recognition of RNA repeat expansions by small molecules in situ and in vivo, enabling the establishment of new chemical biology frameworks to target RNA using small molecules and the development of preclinical candidates. Our studies span many types of repeat expansions, differing in both sequence and gene contexts (intron, untranslated region, or open reading frame). We have devised innovative strategies to: (i) exploit the structures of RNA repeats to coax the disease- causing RNA to synthesize its own drug; (ii) interface small molecules with natural RNA decay and QC pathways; and (iii) recruit endogenous nuclease to the repeats with small molecules. We will not only deliver proof-of- concept small molecules that rescue disease-associated defects in situ and in vivo, but make new discoveries about how to drug RNA using small molecules. Our proposed work would therefore be well supported by an R35 award, as the flexibility conferred by this award is truly necessary to ensure sustainable, long-term funding and the investment required to develop a new code for how molecules interface with RNA in health and disease.

项目摘要：一个非常具有挑战性的问题是开发针对缺陷的通用方法 或者是有选择性地导致疾病的故障RNA。目前靶向RNA的治疗策略是基于 寡核苷酸对特定序列的识别。然而，许多人类疾病是由高度 不容易通过常规碱基配对靶向的结构化RNA，特别是 导致或促成> 30种无法治愈的神经肌肉疾病和遗传性痴呆。因此，等位基因- 针对这些微卫星疾病的特异性ASO模式已经通过靶向多态性而被开发出来 在重复序列之外。这种方法的后果是，只有患有 多态性从治疗中受益，并且必须为每种疾病开发ASO，即使是由基因多态性引起的。 相同的重复序列。如果这些疾病中的毒素，即扩展的重复序列， 选择性地与结构特异性小分子结合，那么单一形式可以是治疗剂或化学制剂。 适用于多种疾病和所有患者的探头。 在过去的14年里，我们已经证明，RNA结构可以选择性地与小分子靶向 在原位和体内，比寡核苷酸更有选择性。事实上，我们已经设计了针对许多 RNA重复序列扩增，选择性地识别靶结构并挽救疾病相关的 原位和体内病理生物学。此外，这些化学探针已经阐明了疾病的新机制， 包括以前未知的RNA介导的转录沉默途径， 综合征这些研究，沿着我们的创新策略，在疾病部位合成药物， 工程小分子具有新的活性，包括反义或CRISPR样作用模式，奠定了 为我们提出的研究计划奠定基础。 在此，我们提出了一个全面的策略来研究RNA重复序列扩增的分子识别 通过原位和体内的小分子，能够建立新的化学生物学框架来靶向 使用小分子的RNA和临床前候选物的开发。我们的研究涵盖了许多类型的 重复扩增，在序列和基因背景（内含子、非翻译区或开放阅读）上都不同 框架）。我们设计了创新的策略：（i）利用RNA重复序列的结构来诱导疾病- 使RNA合成其自身的药物;（ii）将小分子与天然RNA衰变和QC途径连接; 和（iii）用小分子将内源性核酸酶募集到重复序列中。我们不仅会提供证据- 概念小分子，拯救原位和体内疾病相关的缺陷，但新的发现 关于如何用小分子给RNA加药因此，我们建议的工作将得到 R35奖，因为该奖项所赋予的灵活性对于确保可持续的长期资金确实是必要的 以及开发一种新的密码所需的投资，以了解分子如何与RNA在健康和疾病中相互作用。