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: 7446912
• 负责人: IGNATIA B VAN DEN VEYVER
• 金额: $ 22.7万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-24 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7446912
• 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; Disruption; 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; Other Finding; Pathology; Pathway interactions; Pattern; Placenta; Play; Polymerase Chain Reaction; Pregnancy; Pregnancy Complications; Pregnancy loss; Premature Birth; Protein Overexpression; Proteins; Public Health; Recurrence; Regulation; Reproductive Health; Research; Risk; Rodent; Role; Secondary to; Spontaneous abortion; Tissues; Woman; X Chromosome; Yeasts; base; chromatin immunoprecipitation; demethylation; imprint; in vitro Assay; insight; member; natural Blastocyst Implantation; novel; novel therapeutics; protein function; reproductive; research study; therapeutic target; trophoblast; yeast two hybrid system

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: 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.

描述（由申请人提供）：氢化痣（HM）是异常发育的怀孕，具有超增强的滋养细胞和缺乏胎儿。较常见的零星完整的氢摩尔痣是46，XX或46，XY雄激素概念，其中所有遗传物质都是亲子得出的（锚定）。罕见的复发性氢摩尔鼠在临床和病理上与锚固均相同，但具有正常的两出生遗传（BIHM），其摩尔滋养细胞组织在印刷控制区（ICRS）表现出印迹基因的异常表达和CPG岛上CPG岛的异常表达。最近，在复发性BIHM妊娠的女性中发现了NLRP7中的常染色体隐性突变，该蛋白质（NLRP7）在先天免疫和凋亡中具有假定作用。在这一令人惊讶的发现之前，我们和其他人假设BIHM女性突变的基因应该是印迹的主要调节者。但是，目前尚不清楚NLRP7是否以及如何执行此额外功能。此外，尚未研究NLRP7是否与调节印迹标记的建立或维持的染色质复合物中的DNA或蛋白质直接相互作用。因此，该项目的总体假设是NLRP7通过与DNA和/或ICRS的Chromatin修饰因子直接相互作用来调节在人类卵子发生过程中设定的印迹标记的重编程和/或维护。由于NLRP7没有啮齿动物直系同源物，因此无法产生小鼠的灭活突变。因此，我们建议使用体外测定和细胞培养系统以三个特定目标探索这一假设。在特定目标1中，我们将通过两种互补方法，电动性转移测定和染色质免疫沉淀进行研究，无论NLRP7是否与ICRS处的甲基化和未甲基化的CpG序列相关。对于特定目标2，我们将与参与印迹的重编程和维护的候选蛋白进行NLRP7的酵母 - 杂交相互作用研究和NLRP7的共亲密纯化实验。对于特定目标3，我们将通过具有全长NLRP7蛋白和NLRP7-柠檬氨酸富含重复区域的饱和酵母杂种筛网搜索新型的NLRP7相互作用器。对于所有三个特定目标，我们将首先将分析重点放在已知或新颖的互动者上，这些相互作用者在烙印中发挥了作用。但是，如果数据不支持在烙印中的直接作用，那么实验将能够解决另一种假设，即BIHM中看到的烙印缺陷是继发于NLRP7在生殖器官中的免疫反应中更普遍的作用的继发性作用，并通过专注于候选蛋白质来促进这些peTAREDERETE的蛋白质。总体而言，我们的目标是探索以NLRP7为中心的新途径，这些途径对于印迹和生殖健康很重要。由于一些复发性BIHM的妇女患有流产，灭菌内生长迟缓或早产递送的罕见非摩尔妊娠，因此我们预测该项目将发现这些常见的生殖疾病的候选基因，并可能具有新的新型治疗靶标。 公共卫生相关性：我们将研究一个基因NLRP7的新功能，该功能在罕见的复发性氢化痣（患有增生的胎盘和缺席的胎儿），有时以及其他妊娠并发症的女性中被发现突变。由于遗传印迹在这些氢化痣中是异常的，因此我们建议确定该基因是否在烙印的调节中起作用。该项目有可能导致对遗传烙印疾病的新了解以及妇产并发症的原因和妊娠丧失。