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: 10380131
• 负责人: Matthew D Disney
• 金额: $ 138.75万
• 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/10380131
• 关键词: Affect; Alleles; Award; Base Pairing; Biology; Chemicals; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Defect; Dementia; Disease; Engineering; Ensure; Foundations; Fragile X Syndrome; Funding; Gene Silencing; Genes; Genetic Polymorphism; Health; Human; In Situ; Introns; Investments; Mediating; Methods; Microsatellite Repeats; Modality; Neuromuscular Diseases; Oligonucleotides; Open Reading Frames; Pathology; Pathway interactions; Patients; Pharmaceutical Preparations; RNA; RNA Decay; Research; Site; Structure; Therapeutic; Tissues; Toxin; Transcript; Untranslated Regions; Work; base; design; drug action; flexibility; human disease; in vivo; innovation; molecular recognition; novel; nuclease; preclinical development; programs; recruit; small molecule; tool

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：一个极具挑战性的问题是开发针对缺陷的通用方法 或选择性导致疾病的功能失常的 RNA 是当前针对 RNA 的治疗策略的基础。 寡核苷酸对特定序列的识别然而，许多人类疾病是由高度引起的。 传统碱基配对不易靶向的结构化 RNA，特别是 RNA 重复扩展 导致或促成超过 30 种无法治愈的神经肌肉疾病和遗传性痴呆。 通过针对多态性开发了针对这些微卫星疾病的特定 ASO 模式 这种方法的后果是只有具有重复序列的患者。 多态性受益于治疗，并且必须为每种疾病开发 ASO，即使是由这种疾病引起的 如果毒素在这些疾病中通过相同的重复序列扩大，就可以被靶向。 选择性地使用结构特异性的小分子，那么单一模式可以是治疗或化学药物 针对多种疾病和所有患者进行探测。 在过去的 14 年里，我们已经证明小分子可以选择性地靶向 RNA 结构 事实上，我们已经设计了针对许多化合物的化合物。 RNA 重复扩增可选择性地识别目标结构并挽救与疾病相关的疾病 此外，这些化学探针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，该途径在脆弱的 X 中起作用 这些研究以及我们在疾病部位合成药物和治疗的创新策略。 设计具有新颖活性的小分子，包括反义或类似 CRISPR 的作用模式，奠定了 我们拟议的研究计划的基础。 在此，我们提出了一种研究 RNA 重复扩增的分子识别的综合策略 通过小分子原位和体内，能够建立新的化学生物学框架来靶向 我们的研究涉及多种类型的 RNA 使用小分子和临床前候选药物的开发。 重复扩展，序列和基因背景（内含子、非翻译区或开放阅读）都不同 我们设计了创新策略来：（i）利用 RNA 重复结构来诱导疾病 - (ii) 将小分子与天然 RNA 衰减和 QC 途径连接起来； (iii) 用小分子将内源核酸酶招募到重复序列中。我们不仅会提供证明- 概念小分子可在原位和体内挽救与疾病相关的缺陷，但又取得了新发现 因此，我们提出的关于如何使用小分子对 RNA 进行药物治疗的工作将得到一个很好的支持。 R35 奖项，因为该奖项赋予的灵活性对于确保可持续的长期资助确实是必要的 以及开发新代码所需的投资，以了解分子如何在健康和疾病中与 RNA 相互作用。