Molecular underpinnings of Prader-Willi syndrome

普瑞德威利综合征的分子基础

基本信息

批准号：
10013261
负责人：
Gordon G Carmichael
金额：
$ 59.21万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-09 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10013261
关键词：
15q Alleles Behavior ChIP-seq Chromatin Chromatin Structure Chromosomes Complex Disease Epigenetic Process Exons Failure to Thrive Gene Expression Genes Genetic Goals Hyperphagia Individual Inherited Knock-out Learning Maps Mediating Messenger RNA Methylation Methyltransferase Modeling Modification Molecular Molecular Conformation Molecular Target Morbid Obesity Muscle hypotonia Neurons Nuclear Pore Complex Nucleic Acid Regulatory Sequences Pharmacology Play Prader-Willi Syndrome Proteins RNA RNA annealing Regulation Repression Ribosomal RNA Role SETDB1 gene SNRPN Site Small Nucleolar RNA Somatic Cell Structure Techniques Therapeutic Tissues Transcript Uniparental Disomy Work Zinc Fingers differential expression experimental study gene repression histone methyltransferase histone modification human tissue imprint induced pluripotent stem cell mRNA sequencing neurogenetics recruit tool

项目摘要

ABSTRACT Prader-Willi syndrome (PWS) is a neurogenetic disorder caused by the loss of paternally inherited genes on chromosome 15q11-q13. Atypical deletions that cause PWS have narrowed the genetic region critical for the disorder to a 91kb locus including SNORD116. SNORD116 is cluster of 30 highly similar C/D box small nucleolar RNAs (snoRNAs). Canonical C/D box snoRNAs anneal to and mediate 2'O-methylation (2'-OMe) of ribosomal RNAs, however, the SNORD116 copies lack complementarity to rRNAs. Instead they are hypothesized to modify mRNAs or lncRNAs, but direct targets for SNORD116 have not been identified. Thus, their function in neurons is not known. Fortunately, every individual with PWS has an intact, but epigenetically repressed copy of the PWS critical region, including SNORD116, on their maternal allele. We have recently shown that the maternal copy of SNORD116 can be activated in PWS neurons by depleting ZNF274, a KRAB domain zinc finger protein. Not only does this suggest an intriguing therapeutic approach to PWS, but also provides a critical tool to help understand how SNORD116 is regulated in neurons. The overall goal of this proposal is to better understand the molecular underpinnings of PWS. We will determine the chromatin states and long-range chromatin interactions of active and inactive 15q11-q13 alleles in iPSCs and neuronal derivatives. We will investigate how activation of SNORD116 via ZNF274 depletion and histone methyltransferase inhibition impact the chromatin state and long-range interactions. Finally, we will identify the direct 2'-O-methylation targets of SNORD116 and determine how they influence PWS-related differentially expressed genes.

抽象的 Prader-Willi综合征（PWS）是一种神经遗传疾病 15q11-Q13染色体上的遗传基因。导致PW的非典型删除已缩小该疾病至关重要的91KB基因座（包括SNORD116）的遗传区域。 Snord116是 30个高度相似的C/D盒小核心核RNA（SNORNA）的簇。规范C/D盒 snornas向核糖体RNA的2'o-甲基化（2'-ome）退火并介导，但是 SNORD116副本缺乏与RRNA的互补性。相反，他们被假设要修改 mRNA或LNCRNA，但尚未确定SNORD116的直接靶标。因此，他们神经元的功能尚不清楚。幸运的是，每个患有PWS的人都完好无损，但是 PWS关键区域的表观遗传抑制副本，包括SNORD116 产妇等位基因。我们最近表明，可以激活Snord116的母体副本在PWS神经元中，通过耗尽ZnF274（Krab域锌指蛋白）。不仅这样做为PWS提出一种有趣的治疗方法，但也提供了一个关键的工具来帮助了解如何在神经元中调节SNORD116。该提议的总体目标是更好地了解PWS的分子基础。我们将确定染色质状态 IPSC中的活动和非活动15q11-Q13等位基因的远程染色质相互作用以及神经元素衍生物。我们将研究如何通过Znf274耗尽激活SNORD116 组蛋白甲基转移酶抑制作用影响染色质状态和远距离相互作用。最后，我们将确定SNORD116的直接2'-O-甲基化靶标，并确定如何它们影响与PW相关的差异表达基因。