Molecular underpinnings of Prader-Willi syndrome

普瑞德威利综合征的分子基础

• 批准号: 10199881
• 负责人: Gordon G Carmichael
• 金额: $ 58.03万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 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/10199881
• 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

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 普瑞德威利综合征 (PWS) 是一种因失去父系亲缘关系而引起的神经遗传性疾病 遗传基因位于染色体 15q11-q13 上。导致 PWS 的非典型缺失已缩小 对该疾病至关重要的遗传区域为 91kb 基因座，包括 SNORD116。 SNORD116 是 由 30 个高度相似的 C/D 盒小核仁 RNA (snoRNA) 组成的簇。佳能C/D盒 snoRNA 退火并介导核糖体 RNA 的 2'O-甲基化 (2'-OMe)，然而， SNORD116 拷贝缺乏与 rRNA 的互补性。相反，他们被假设要修改 mRNA 或 lncRNA，但 SNORD116 的直接靶标尚未确定。因此，他们的 神经元的功能尚不清楚。幸运的是，每个患有 PWS 的人都有一个完整的、但 PWS关键区域的表观遗传抑制副本，包括SNORD116，在其 母体等位基因。我们最近证明 SNORD116 的母体副本可以被激活 通过消耗 ZNF274（一种 KRAB 结构域锌指蛋白），在 PWS 神经元中发挥作用。这不仅 提出了一种有趣的 PWS 治疗方法，同时也提供了一个重要的工具来帮助 了解 SNORD116 在神经元中的调节方式。该提案的总体目标是 更好地了解 PWS 的分子基础。我们将确定染色质状态 iPSC 中活性和非活性 15q11-q13 等位基因的长程染色质相互作用 神经元衍生物。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会影响染色质状态和长程相互作用。 最后，我们将确定 SNORD116 的直接 2'-O-甲基化靶点并确定如何 它们影响 PWS 相关的差异表达基因。