Supplement: Defining and Controlling Protein-RNA interactions in editing and interference pathways

补充：编辑和干扰途径中蛋白质-RNA 相互作用的定义和控制

基本信息

批准号：
10807900
负责人：
PETER A. BEAL
金额：
$ 1.09万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2026-04-30
项目状态：
未结题

项目摘要

This Maximizing Investigators Research Award (MIRA) application is proposed to support research in the Beal lab at UC Davis focused on defining and controlling protein-RNA interactions in RNA editing and RNA interference pathways. The RNA editing ADAR enzymes convert adenosines (A) to inosines (I) in duplex RNA. Since I can behave similarly to guanosine (G) in RNA, this modification can have profound effects on the structure and function of the modified RNA including, but not limited to, changes in the meaning of specific codons (recoding). Mutations in the human ADAR1 gene cause the skin disorder Dyschromatosis Symmetrica Hereditaria (DSH) and the autoimmune disease Aicardi- Goutieres Syndrome (AGS). Also, ADAR1 upregulation and hyper editing has been observed in several different cancers. Despite the significance of this form of regulation of RNA structure and function, there remain key gaps in our understanding of A to I RNA editing. In addition, given ADARs’ ability to change RNA sequence, there is growing interest in harnessing this property and directing it to correct disease-associated G-to-A mutations. Key questions in this field that will be addressed in this project are: 1) What are the structures of key protein-RNA complexes in editing pathways? Structures of full length human ADAR2 bound to different RNA substrates along with structures of ADAR1 bound to RNA are necessary for a full understanding of substrate recognition and selectivity in RNA editing. 2) Can we develop potent, selective and low MW ADAR inhibitors? Such inhibitors could serve as lead compounds in the development of ADAR1-targeted cancer therapies. 3) Can we develop new strategies to evolve mutant editing enzymes and novel substrate RNAs? The results of these efforts will inform the design of highly efficient and selective reagents for directed RNA editing applications. Our laboratory also has a long standing interest in the development of chemical modifications of RNA that can control the interaction with RNA-binding proteins. Much of our recent work in this area has focused on controlling the interaction of RNA with components of siRNA-triggered or miRNA- triggered gene silencing pathways. The use of the RNAi pathway to study gene function has become a powerful tool in molecular biology and has been exploited in the development of new therapeutics. However, specific issues exist that limit its application. These issues include off-target effects that arise from the ability of an siRNA guide strand to function as a miRNA. In addition, antisense oligonucleotides targeting miRNAs (anti-miRs) have significant therapeutic potential and require chemical modification for stability and efficacy. Up to this point, the development of new chemical modifications of therapeutic RNAs has been largely an ad hoc process. The key question addressed in this aspect of the proposed project is: Can we develop an effective systematic approach to new RNA modifications that modulate protein-RNA interactions in interference pathways? The immediate impact of these studies will be to provide new modifications to siRNAs and anti-miRs that improve potency and selectivity. However, our continued refinement of an approach that uses computational screening coupled with versatile RNA modification chemistry will be generally applicable other projects that involve chemically modified RNA for therapeutics.

提出了最大化调查人员研究奖（MIRA）的申请，以支持研究 UC Davis的Beal Lab着重于定义和控制RNA编辑中的蛋白RNA相互作用和RNA干扰途径。 RNA编辑ADAR酶将腺苷转化为插插（i）在双工RNA中。由于我的行为与RNA中的鸟嘌呤（G）相似，因此这种修改可以具有对修饰的RNA的结构和功能的深远影响，包括但不限于特定密码子的含义（重新编码）的变化。人adar1基因的突变引起皮肤疾病异型症对称性遗传学（DSH）和自身免疫性疾病aicardi- Goutieres综合征（AGS）。同样，在几种不同的癌症。尽管这种调节的RNA结构和功能，我们对I对RNA编辑的理解仍然存在关键差距。此外，考虑到Adars 更改RNA序列的能力，对利用这一属性并指导它越来越兴趣纠正与疾病相关的G-TO-A突变。该领域的关键问题将在该项目是：1）在编辑途径中，关键蛋白RNA复合物的结构是什么？全长人ADAR2结构与不同的RNA底物结合与RNA结合的ADAR1对于完全理解底物识别和选择性是必要的在RNA编辑中。 2）我们可以发展潜力，选择性和低MW ADAR抑制剂吗？这样的抑制剂可以用作靶向ADAR1靶向癌症疗法的铅化合物。 3）我们可以吗？制定新的策略来发展突变编辑酶和新型底物RNA？这些努力的结果将为有向RNA的高效和选择性试剂的设计提供信息编辑应用程序。我们的实验室对化学修改的发展也很长期兴趣可以控制与RNA结合蛋白相互作用的RNA。我们最近在这一领域的大部分工作已重点是控制RNA与siRNA触发或miRNA-成分的相互作用触发基因沉默途径。 RNAi途径研究基因功能已成为一种有力的分子生物学工具，并已在新的治疗剂的开发中探索过。但是，存在限制其应用程序的特定问题。这些问题包括脱靶效应源自siRNA引导链充当miRNA的能力。另外，反义靶向miRNA的寡核苷酸（抗MIR）具有明显的治疗潜力，需要化学修改稳定性和效率。到目前为止，新化学化学的发展治疗性RNA的修改在很大程度上是一个临时过程。解决的关键问题在拟议项目的这一方面是：我们可以开发一种有效的系统方法在干扰途径中调节蛋白RNA相互作用的RNA修饰？这这些研究的直接影响将是为siRNA和抗MIR提供新的修改提高效力和选择性。但是，我们继续改进了一种使用的方法与多功能RNA修饰化学结合的计算筛选通常是适用的其他项目，涉及化学修饰的RNA进行治疗。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Impact of Disease-Associated Mutations on the Deaminase Activity of ADAR1.

疾病相关突变对 ADAR1 脱氨酶活性的影响。

DOI：
10.1021/acs.biochem.3c00405
发表时间：
2024
期刊：
Biochemistry
影响因子：
2.9
作者：
Karki,Agya;Campbell,KristenB;Mozumder,Sukanya;Fisher,AndrewJ;Beal,PeterA
通讯作者：
Beal,PeterA

Nucleoside analogs in ADAR guide strands targeting 5'-UA̲ sites.

DOI：
10.1039/d2cb00165a
发表时间：
2023-01-04
期刊：
RSC CHEMICAL BIOLOGY
影响因子：
4.1
作者：
Brinkman, Hannah F.;Jauregui Matos, Victorio;Mendoza, Herra G.;Doherty, Erin E.;Beal, Peter A.
通讯作者：
Beal, Peter A.

Oligonucleotide-directed RNA editing in primates.

DOI：
10.1016/j.ymthe.2022.04.005
发表时间：
2022-06-01
期刊：
MOLECULAR THERAPY
影响因子：
12.4
作者：
Doherty, Erin E.;Beal, Peter A.
通讯作者：
Beal, Peter A.

Library Screening Reveals Sequence Motifs That Enable ADAR2 Editing at Recalcitrant Sites.

DOI：
10.1021/acschembio.3c00107
发表时间：
2023-10-20
期刊：
ACS CHEMICAL BIOLOGY
影响因子：
4
作者：
Jacobsen, Casey S.;Salvador, Prince;Yung, John F.;Kragness, Sabrina;Mendoza, Herra G.;Mandel, Gail;Beal, Peter A.
通讯作者：
Beal, Peter A.