Using Chromatin Architecture to Develop of Therapeutic Pipeline for Juvenile Arthritis

利用染色质结构开发幼年关节炎的治疗管线

基本信息

批准号：
9927736
负责人：
JAMES N JARVIS
金额：
$ 21.05万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-21 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9927736
关键词：
Affect Architecture Attenuated Autoimmune Diseases Biology Budgets Cells Child Chromatin Chromatin Loop Chronic Childhood Arthritis Chronic Disease Clinical Coupled DNA Data Development Disease Drug Targeting Elements Enhancers Enzymes Epigenetic Process Gene Expression Gene Targeting Gene-Modified Genes Genetic Genetic Enhancer Element Genetic Risk Genome Genomic Segment Genomics Goals Guide RNA Haplotypes IL2RA gene Industry Interest Group Introns Lead Lymphoid Cell Methods Mutation Pathologic Patient Care Proteins Quantitative Reverse Transcriptase PCR Risk Structure Techniques Therapeutic Time Tissues Translating Untranslated RNA Work base epigenome editing epigenomics genetic information genetic variant genome sequencing genome wide association study improved new therapeutic target nuclease promoter risk variant scale up targeted treatment therapeutic target whole genome

Affect Architecture Attenuated Autoimmune Diseases Biology Budgets Cells Child Chromatin Chromatin Loop Chronic Childhood Arthritis Chronic Disease Clinical Coupled DNA Data Development Disease Drug Targeting Elements Enhancers Enzymes Epigenetic Process Gene Expression Gene Targeting Gene-Modified Genes Genetic Genetic Enhancer Element Genetic Risk Genome Genomic Segment Genomics Goals Guide RNA Haplotypes IL2RA gene Industry Interest Group Introns Lead Lymphoid Cell Methods Mutation Pathologic Patient Care Proteins Quantitative Reverse Transcriptase PCR Risk Structure Techniques Therapeutic Time Tissues Translating Untranslated RNA Work base epigenome editing epigenomics genetic information genetic variant genome sequencing genome wide association study improved new therapeutic target nuclease promoter risk variant scale up targeted treatment therapeutic target whole genome

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项目摘要

Abstract In this project, we aim to take an important step toward identifying novel targets of therapy in juvenile idiopathic arthritis (JIA). To do this, we will use genetic information that has been generated from genome- wide association studies (GWAS) and genetic fine mapping studies. These studies have shown that much of the genetic risk for JIA lies within genomic regions that are highly enriched for enhancer function, a feature that JIA shares with almost every other autoimmune disease. The underlying premise of this work is that, by identifying genetic risk variants that alter enhancer function, and then identifying the genes whose expression levels change when enhancer function is altered, we will identify promising new therapeutic targets. In this proof-of-concept project, we will focus on IL2RA, an important risk locus for JIA and multiple other autoimmune diseases. We have previously shown that the IL2RA locus contains an intronic enhancer whose function is attenuated by genetic variants that we initially identified on whole genome sequencing of children with the polyarticular form of JIA. We aim to identify the target genes of these genetic variants, i.e., the genes whose expression levels are altered when function of the intronic enhancer is attenuated. To accomplish this aim, we rely on the fact that, while enhancers may not always regulate the nearest gene, they typically regulate genes within the same chromatin loop or topologically associated domain (TAD). We will use RNA-guided epigenome editing with a deactivated nuclease coupled to an epigenome editing enzyme, Krüppel associated box (KRAB) repressor (dCAS-KRAB). We will then use conventional rtPCR approaches to identify which of the 12 genes that are incorporated within the TAD that contains the IL2RA enhancer show changes in expression level when that enhancer is attenuated. If successful, we will have established as proof-of-concept the utility of using the broader chromatin architecture that incorporates the JIA risk haplotypes and RNA-guided epigenome editing as a strategy for identify therapeutic targets in JIA. We expect that completion of this work will lead quickly to a larger project aimed at identifying all the target genes of genetically altered enhancers within the JIA risk loci. Furthermore, successful completion will also lead to the development of partnerships with key industry leaders to rapidly translate these findings to the clinical setting. !

抽象的在这个项目中，我们旨在迈出重要的一步，以确定青少年的新型治疗目标特发性关节炎（JIA）。为此，我们将使用基因组产生的遗传信息广泛的关联研究（GWAS）和遗传精细映射研究。这些研究表明，很多 JIA的遗传风险位于基因组区域内，这些区域高度丰富增强子功能，这一特征是贾与几乎所有其他自身免疫性疾病都分享。这项工作的基本前提是，识别改变增强子功能的遗传风险变异，然后识别其表达的基因当更改增强子功能时，水平会改变，我们将确定有希望的新的治疗靶标。在这个概念验证项目中，我们将重点介绍IL2RA，这是JIA和其他多个的重要风险基因座自身免疫性疾病。我们以前已经表明，IL2RA基因座包含一个内含子增强子我们最初在儿童的整个基因组测序上鉴定出的遗传变异降低了功能与jia的多关节形式。我们旨在鉴定这些遗传变异的靶基因，即基因当内含子增强子的功能减弱时，其表达水平会改变。实现这一目标目的，我们依靠以下事实：尽管增强子可能并不总是调节最近的基因，但它们通常会调节相同染色质环或拓扑相关的结构域（TAD）内的基因。我们将使用RNA引导与表观基因组编辑酶，Krüppel相关的核酸酶编辑的表观基因组编辑 Box（Krab）复制器（DCAS-KRAB）。然后，我们将使用常规的RTPCR方法来识别哪些包含IL2RA增强子的TAD中的12个基因显示出表达的变化当该增强子衰减时，水平。如果成功，我们将确定使用更广泛的染色质的效用结合JIA风险单倍型和RNA引导的表观基因组编辑的建筑作为一种策略确定JIA的治疗靶标。我们预计这项工作的完成将很快导致一个更大的项目旨在确定JIA风险基因座中普遍改变增强子的所有目标基因。此外，成功完成还将导致与主要行业领导者建立伙伴关系将这些发现转换为临床环境。呢