The role of NLRP7 and related genes in hydatidiform moles and reproductive failur

NLRP7及相关基因在葡萄胎和生殖障碍中的作用

• 批准号: 7882072
• 负责人: IGNATIA B VAN DEN VEYVER
• 金额: $ 0.75万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7882072
• 关键词: 19q13.42; Address; Affect; Affinity Chromatography; Apoptosis; Biological Assay; Candidate Disease Gene; Cell Culture System; Chromatin; Chromosomes; Chromosomes, Human, Pair 13; Complete Hydatidiform Moles; Complex; Conceptions; CpG Islands; Cultured Cells; DNA; DNA Methylation; Data; Defect; Discipline of obstetrics; Disease; Family; Female; Fetus; Funding Mechanisms; Gene Silencing; Genes; Genetic Materials; Genome; Genomic Imprinting; Goals; Growth; Human; Hydatidiform Mole; Hyperplasia; Immune; Immune response; Inherited; Karyotype; Length; Leucine-Rich Repeat; Maintenance; Methods; Methylation; Mus; Mutate; Mutation; Natural Immunity; Oocytes; Oogenesis; Organ; Pathology; Pathway interactions; Pattern; Placenta; Play; Pregnancy; Pregnancy Complications; Pregnancy loss; Premature Birth; Proteins; Recurrence; Regulation; Reproductive Health; Research; Risk; Rodent; Role; Secondary to; Spontaneous abortion; Tissues; Woman; X Chromosome; Yeasts; base; chromatin immunoprecipitation; demethylation; genome-wide; imprint; in vitro Assay; insight; member; natural Blastocyst Implantation; new therapeutic target; novel; overexpression; protein function; reproductive; research study; trophoblast; yeast two hybrid system

Hydatidiform moles (HM) are abnormally developing pregnancies with hyperproliferative trophoblast and absence of a fetus. The more common sporadic complete hydatidiform moles are 46,XX or 46,XY androgenetic conceptions, in which all the genetic material is paternally derived (AnCHM). Rare recurrent hydatidiform moles are clinically and pathologically identical to AnCHM, but have normal biparental inheritance (BiHM) and their molar trophoblast tissues show abnormal expression of imprinted genes and abnormal methylation of CpG islands at imprinting control regions (ICRs). Recently, autosomal recessive mutations in NLRP7, encoding a protein (NLRP7) with a putative role in innate immunity and apoptosis, were identified in women with recurrent BiHM pregnancies. Prior to this surprising finding, we and others hypothesized that the gene mutated in women with BiHM should be a major regulator of imprinting. However, it is currently not known whether and how NLRP7 might carry out this additional function. Furthermore, it has not yet been studied whether NLRP7 interacts directly with DNA or proteins in chromatin complexes that regulate establishment or maintenance of imprinting marks. Therefore, the overarching hypothesis for this project is that NLRP7 regulates reprogramming and/or maintenance of imprinting marks that are set during human oogenesis by direct interaction with DNA and/or chromatin modifying factors at ICRs. Because NLRP7 has no rodent orthologue, inactivating mutations in mice cannot be generated. Hence, we propose to use in vitro assays and cell culture systems to explore this hypothesis in three specific aims. In specific aim 1 we will investigate by two complementary methods, electromobility shift assays and chromatin immunoprecipitation, whether NLRP7 associates with methylated and unmethylated CpG sequences at ICRs. For specific aim 2 we will perform yeast-two-hybrid interaction studies and co-affinity purification experiments of NLRP7 with candidate proteins that participate in reprogramming and maintenance of imprinting. For Specific aim 3 we will search for novel NLRP7 interactors by a saturated yeast-two-hybrid screen with the full-length NLRP7 protein and the NLRP7-leucine-rich repeat region. For all three specific aims, we will first focus our analysis on known or novel interactors that play a role in imprinting. However, if the data do not support a direct role in imprinting, the experiments, will be able to address an alternate hypothesis, which is that the imprinting defects seen in BiHM are secondary to disruption of a more general role of NLRP7 in immune response within reproductive organs and/or the developing oocyte by focusing on candidate proteins for these pathways. Overall, our goal is to explore new pathways, centered on NLRP7 that are important for imprinting and for reproductive health. Because some women with recurrent BiHM have rare non-molar pregnancies affected by miscarriage, intra-uterine growth retardation or preterm delivery, we predict that this project will uncover candidate genes for these common reproductive disorders, and potentially novel therapeutic targets. We will investigate a new function of a gene, NLRP7, found to be mutated in women who have rare recurrent hydatidiform moles (a severe pregnancy complication with hyperplastic placenta and absent fetus), and sometimes other pregnancy complications. Because genetic imprinting is abnormal in these hydatidiform moles, we propose to determine whether this gene play a role in the regulation of imprinting. This project has the potential to result in new understanding of genetic imprinting disorders and causes of obstetrics complications and pregnancy loss in general.

葡萄胎（HM）是一种异常发育的妊娠，具有过度增殖的滋养细胞， 没有胎儿。较常见的散发性完全性葡萄胎是46，XX或46，XY 雄激素发生的概念，其中所有的遗传物质是父系衍生（AnCHM）。罕见复发 葡萄胎在临床和病理上与AnCHM相同，但具有正常的双亲遗传 （BiHM）及其磨牙滋养层组织显示印迹基因表达异常， 在印迹控制区（ICR）的CpG岛的甲基化。最近，常染色体隐性突变， NLRP 7编码一种蛋白质（NLRP 7），在先天免疫和细胞凋亡中具有假定的作用， 反复BiHM妊娠的女性。在这一令人惊讶的发现之前，我们和其他人假设， BiHM女性中突变的基因应该是印记的主要调节器。然而，目前还没有 NLRP 7是否以及如何执行这一额外功能。此外，还没有 研究了NLRP 7是否直接与DNA或染色质复合物中的蛋白质相互作用， 建立或维持印记。因此，本项目的总体假设是 NLRP 7调节在人类发育过程中设定的印记标记的重编程和/或维持， 卵子发生通过在ICR处与DNA和/或染色质修饰因子直接相互作用。因为NLRP 7没有 在啮齿动物直系同源物中，不能在小鼠中产生失活突变。因此，我们建议在体外使用 分析和细胞培养系统，以探索这一假设在三个具体的目标。在具体目标1中，我们将 通过两种互补方法，电迁移率变动分析和染色质免疫沉淀， NLRP 7是否与ICR处的甲基化和未甲基化CpG序列相关。具体目标2，我们 将进行酵母双杂交相互作用研究和NLRP 7与 参与重编程和维持印记的候选蛋白质。具体目标3： 通过用全长NLRP 7蛋白进行饱和酵母双杂交筛选来寻找新的NLRP 7相互作用物 和NLRP 7-富含亮氨酸的重复区。对于所有这三个具体目标，我们将首先集中分析已知的 或者是在印记中起作用的新的相互作用者。然而，如果数据不支持印记的直接作用， 实验，将能够解决一个替代假设，这是印迹缺陷中看到的， BiHM是继发于NLRP 7在生殖系统内免疫应答中的更一般作用的破坏。 器官和/或发育中的卵母细胞，通过关注这些途径的候选蛋白质。 总的来说，我们的目标是探索新的途径，以NLRP 7为中心，这对印记和 生殖健康。因为一些患有复发性BiHM的女性患有罕见的非磨牙妊娠， 流产，宫内发育迟缓或早产，我们预测，这个项目将揭示， 这些常见生殖疾病的候选基因，以及潜在的新治疗靶点。我们将研究一种新的基因NLRP 7的功能，该基因在患有罕见复发性乳腺癌的女性中发现突变， 葡萄胎（一种严重的妊娠并发症，伴有胎盘增生和胎儿缺如），以及 有时候是其他的妊娠并发症。因为遗传印记在这些棘球蚴中是异常的 摩尔，我们建议确定是否这个基因在调节印记中发挥作用。这个项目 可能导致对遗传印记疾病和产科原因的新理解 并发症和流产。