Finding, folding and characterizing the functions of disease-relevant RNA structures

项目概要/摘要 了解疾病背后的基本分子细节是科学的关键挑战。这项建议 旨在通过关注疾病的一个未充分研究的方面来解决这个问题：RNA结构的作用。的 该项目的总体目标是优化方法管道，以发现功能性RNA元件， 人类及其病原体的基因组，推断其结构，并描述其功能。年底 目的是更好地了解RNA结构如何与疾病有关，并利用这一点， 以新的方式提供信息，为改善人类健康做出贡献：例如，通过推进基于RNA的 治疗性治疗在这个项目的过程中，我们将通过关注感染来优化这个管道 人B细胞被EB病毒（EBV）感染。EB病毒是一种广泛存在的人类病原体，其感染超过95% 在成年人群中，并且与多种癌症和自身免疫性疾病有关。的机制 EB病毒致病性背后的原因仍然是难以捉摸的，这一提议旨在关注RNA和RNA的作用。 感染和疾病的结构。在建立终身潜伏感染的过程中，EBV既产生 其自身的RNA和失调的各种人类RNA。RNA在人类和病原体中起着核心作用 在生物学中，形成对基因表达至关重要的蛋白质编码和非蛋白质编码分子。 RNA结构起着重要的调节作用：例如，通过介导相互作用或改变RNA的可及性， 功能基序。因此，了解RNA的结构提供了大量的生物学知识， 用于从基因组中推断功能序列。我以前的工作结合了计算和 发现EBV基因组内编码的RNA结构的实验方法。其中包括一本小说 我发现了一类病毒RNA，稳定内含子序列（sis）RNA。在本提案中，我们 继续对EBV编码的RNA进行功能分析，并将我们的重点扩大到包括人类RNA。 这将通过使用我们开发的创新新方法重新访问两个基因组来实现， 目前正在开发的新方法。这将与全基因组生物化学相结合， 在体外（无细胞系统）和培养的人B中进行结构分析和功能测定 细胞这一提议的结果将是对RNA和RNA结构如何在细胞中形成的基本理解的增强。 参与EBV感染和疾病。这将有更广泛的影响超越EBV及其相关的 病理学：例如，我们的管道的一部分涉及RNA结构的保守性分析， 病原体（可能发现保守机制）。同样，许多发现的人类图案受到 EBV可能在癌症和免疫中发挥功能性作用;因此，该提案将提供一般的 对疾病的洞察。这些功能结构为新兴的RNA靶向提供了有吸引力的靶标 因此，这一建议在促进人类健康方面具有巨大潜力：例如，在新的治疗方法中， 癌症和自身免疫性疾病。

PROJECT SUMMARY/ABSTRACT Understanding the basic molecular details behind disease is a critical challenge for science. This proposal aims to approach this by focusing on an understudied aspect of disease: the roles of RNA structure. The overall goal of this project is to optimize a methodological pipeline to discover functional RNA elements in the genomes of humans and their pathogens, deduce their structures, and characterize their functions. The end objective is to gain a better understanding of how RNA structure is implicated in disease and to use this information in novel ways to contribute toward improving human health: for example, by advancing RNA-based therapeutic treatments. Over the course of this project we will optimize this pipeline by focusing on the infection of human B cells by Epstein–Barr virus (EBV). EBV is a widespread human pathogen, which infects over 95% of the adult population, and is implicated in a variety of cancers and autoimmune diseases. The mechanisms behind EBV pathogenicity remain elusive and this proposal aims to focus on the roles of RNA and RNA structure in infection and disease. Over the course of establishing lifelong latent infection, EBV both generates its own RNAs and dysregulates various human RNAs. RNA plays a central role in human and pathogen biology, forming the protein-coding and non-protein-coding molecules that are essential to gene expression. RNA structure plays important regulatory roles: e.g. by mediating interactions or altering accessibility of functional motifs. Thus, knowing the structure of RNA provides a great deal of biological knowledge and can be used to deduce functional sequences from the genome. My previous work combined computational and experimental approaches to discover RNA structures encoded within the EBV genome. This included a novel class of viral RNAs, the stable intronic sequence (sis)RNAs, which I discovered. In this current proposal, we follow up on the functional analyses of EBV-encoded RNAs and expand our focus to include human RNAs. This will be accomplished by revisiting both genomes using innovative new approaches that we developed and novel methods that are currently under development. This will be combined with genome-wide biochemical structure analyses and functional assays performed in vitro (in cell-free systems) and within cultured human B cells. The results of this proposal will be an enhanced basic understanding of how RNA and RNA structure is involved in EBV infection and disease. There will be wider implications beyond EBV and its associated pathologies: for example, part of our pipeline involves the analysis of conservation of RNA structure between pathogens (potentially finding conserved mechanisms). As well, many discovered human motifs affected by EBV will likely have functional roles in cancer and immunity; therefore, this proposal will provide general insights into disease. These functional structures provide attractive targets for emerging RNA-targeting therapies; thus, there is great potential for this proposal to advance human health: e.g. in new treatments for cancer and autoimmune disease.