Sequences controlling H19 Gene Imprinting

控制 H19 基因印记的序列

• 批准号: 9534673
• 负责人: MARISA S. BARTOLOMEI
• 金额: $ 40.25万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9534673
• 关键词: Affect; Alleles; Angelman Syndrome; Ants; Beckwith-Wiedemann Syndrome; Behavior; Cells; Development; Disease; Enhancers; Epigenetic Process; Etiology; Gene Expression; Genes; Genetic; Genetic Diseases; Genomic Imprinting; Growth; H19 gene; Homeostasis; Human; Human Genetics; IGF2 gene; Individual; Link; Malignant Neoplasms; Mammals; Mediating; Methylation; MicroRNAs; Modeling; Mus; Mutation; Nephroblastoma; Nuclear; Parents; Play; Prader-Willi Syndrome; Process; Regulation; Role; Silver-Russell syndrome; Site; Syndrome; Untranslated RNA; experimental study; imprint; microdeletion; mouse model; offspring; sex; transmission process; tumor progression

A subset of genes in mammals is regulated by genomic imprinting, a process that results in unequal expression of the maternal and paternal alleles of this class of genes. Imprinted genes are hypothesized to expiain why nuclear contributions from both parents are required for normal mammalian development. Furthermore, imprinting plays a role in the transmission of a number of human genetic diseases, including Beckwith-Wiedemann Syndrome (BWS), Silver-Russell Syndrome (SRS), Prader-Willi and Angelman Syndrome, in that the sex ofthe parent that transmits the affected gene(s) determines whether offspring will develop the disease. Aben-ant imprinted gene expression is also involved in the establishment or progression of cancers, inciuding Wilms tumors. The objective of this proposal is to investigate the- mechanism by which parental identity of imprinted genes is established and maintained. The studies will employ the conserved H19/lgf2 locus. The imprinting of H19, which produces a non-coding RNA from the maternally-derived allele, and the linked and oppositely imprinted growth-promoting Igf2 gene is mediated through the 2 kb imprinting control region (ICR) and shared enhancers. The ICR acts as a methylation- senstitive, CTCF-dependent insulator. When unmethylated on the maternal allele, the insulator allows H19 exclusive access to the enhancers. In contrast, a methylated paternal insulator enables Igf2 to engage the enhancers. This proposal will investigate the mechanism of H19/lgf2 imprinting through the following experiments. Individuals with BWS, sporadic Wilms tumors and SRS have been identified that have microdeletions and epimutations in the human ICR and aberrant imprinted regulation of H19 and Igf2. We wiil generate mouse models and human IPS cells with these mutations and study the mechanism of loss of imprinting. We will also investigate the function of a conserved non-coding RNA located between H19 and Igf2 and the H19 micro RNA, miR-675, using mutations constructed at the mouse locus. We will additionally explore the role of TET1 in the establishment and erasure of ICR methylation. Last, the imprinting mechanism will be studied at the Grb10 locus, which may employ aspects of H19 imprinted regulation. RELEVANCE (See insfruc:tions): Imprinted genes are critical for normal mammalian development, behavior and energy homeostasis. This genes have genetic or epigenetic mutations in a number of human syndromes and cancer. The experiments in this proposal will model such newly identified mutations in individuals with Beckwith-Wiedemann Syndrome and Silver-Russell Syndrome, providing a better understanding ofthe etiology ofthe disease. PROJECT/PERFORIVIANCE SITE(S) (if additional space Is needed, use

哺乳动物的一部分基因受到基因组印记的调节，这一过程导致不平等 此类基因的母本和父本等位基因的表达。印记基因被假设为 解释为什么哺乳动物的正常发育需要父母双方的核贡献。 此外，印记在许多人类遗传疾病的传播中发挥着作用，包括 Beckwith-Wiedemann 综合征 (BWS)、Silver-Russell 综合征 (SRS)、Prader-Willi 和 Angelman 综合症，传播受影响基因的父母的性别决定了后代是否会 发展疾病。 Aben-ant 印记基因表达也参与建立或 癌症的进展，包括肾母细胞瘤。该提案的目的是调查—— 建立和维持印记基因亲本同一性的机制。这些研究将 采用保守的 H19/lgf2 基因座。 H19 的印记，产生非编码 RNA 母源等位基因，并且介导相连且相反印记的生长促进 Igf2 基因 通过 2 kb 印记控制区 (ICR) 和共享增强子。 ICR 充当甲基化- 敏感的、依赖于 CTCF 的绝缘体。当母体等位基因上未甲基化时，绝缘体允许 H19 独家访问增强器。相反，甲基化的父系绝缘子使 Igf2 能够与 增强剂。该提案将通过以下方面研究H19/lgf2印记机制 实验。患有 BWS、散发性肾母细胞瘤和 SRS 的个体已被确定为 人类 ICR 中的微缺失和表观突变以及 H19 和 Igf2 的异常印记调节。我们 将生成具有这些突变的小鼠模型和人类 IPS 细胞，并研究这些突变的丢失机制 印记。我们还将研究位于 H19 和 H19 之间的保守非编码 RNA 的功能。 Igf2 和 H19 微 RNA miR-675，使用在小鼠基因座构建的突变。我们还将另外 探讨 TET1 在 ICR 甲基化的建立和消除中的作用。最后，印记 将在 Grb10 基因座上研究机制，该基因座可能采用 H19 印记调控的各个方面。 相关性（参见说明）： 印记基因对于哺乳动物的正常发育、行为和能量稳态至关重要。这 在许多人类综合症和癌症中，基因具有遗传或表观遗传突变。实验 该提案将对贝克威斯-维德曼个体中新发现的突变进行建模 综合征和西尔弗-拉塞尔综合征，有助于更好地了解该疾病的病因。 项目/绩效场地（如果需要额外空间，请使用