Transcriptional Networks Regulating Luminal Environment in the Epididymis

调节附睾管腔环境的转录网络

基本信息

批准号：
8508994
负责人：
ANN HARRIS
金额：
$ 27.9万
依托单位：
LURIE CHILDREN'S HOSPITAL OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-10 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8508994
关键词：
Address Androgen Receptor Bioinformatics Biological Assay Cells Chloride Channels Chromatin Chromatin Structure Chromosomes Complex Control Locus Cyclic AMP Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator DNA-Binding Proteins Data Defect Deoxyribonuclease I Deoxyribonucleases Development Disease Duct (organ) structure Ejaculation Elements Enhancers Environment Epididymis Epithelial Cells Epithelium Fertility Gene Expression Gene Expression Regulation Genes Genital system Genomics Goals Hereditary Disease Human In Vitro Inborn Genetic Diseases Infertility Inherited Insulator Elements Investigation Ion Channel Ion Transport Ions Left Link Male Genital Organs Male Infertility Maps Mediating Methods Modification Molecular Molecular Conformation Mutate Mutation Obstruction Organ Pathway interactions Play Process Proteins Recruitment Activity Regulation Regulator Genes Regulatory Element Reporter Genes Research Role Series Site Small Interfering RNA Sperm Maturation Structure Swimming System Techniques Testis Therapeutic Intervention Tissues Trans-Activators Tube Vas deferens structure Water base birth control cell type chromatin immunoprecipitation cis acting element cohesin genome-wide in vivo male malformation novel reproductive research study solute sperm cell three dimensional structure vector zygote

项目摘要

DESCRIPTION (provided by applicant): Functional inactivation (by mutation) of the cystic fibrosis transmembrane conductance regulator (CFTR), a small conductance, cAMP activated chloride ion channel, leads to developmental defects that cause male infertility in humans. The mechanism by which inactivation of the CFTR gene causes infertility is poorly understood but is believed to be due in part to malformations or pathological changes in the male genital duct system. The goals of the proposed research are to decipher the transcriptional networks that regulate luminal environment in the epididymis, by building on our current understanding of the mechanisms regulating expression of the CFTR gene. Mutations in CFTR cause the devastating inherited disorder cystic fibrosis (CF). The CFTR channel makes an essential contribution to ion transport and the luminal environment in the epididymis and its loss causes obstruction and/or loss of the male genital ducts and subsequent infertility. The epididymis forms the proximal part of the excretory duct system of the male reproductive tract and is a complex organ that secretes and absorbs ions, water and inorganic solutes. Critically, it provides the correct luminal environment for normal sperm maturation. In normal epididymis the luminal environment is established and maintained through the functional cooperation of multiple ion channels, ion exchangers, solute carriers and transporters, though the mechanisms that coordinate the expression of these genes has not been well studied. In the first specific aim we will determine how modifications in the chromatin structure of the CFTR locus control its expression in the epididymis. We will evaluate the cis-acting enhancers and insulator elements that likely establish the looped conformation of the active locus and determine how trans-acting factors such as CTCF and cohesin influence this spatial organization. Moreover, we will investigate mechanisms and DNA- binding proteins that may suppress gene expression in cells where CFTR is less active or inactive. Experiments in the second specific aim will determine the transcriptional pathways that coordinate expression of ion channels, ion exchangers, transporters and solute carriers that interact with CFTR in the epididymis epithelium to establish and maintain the luminal environment. We will use a genome-wide approach (DNase- seq) to identify the cis-acting regulatory elements for genes encoding the proteins that contribute to ion and solute transport across the epididymal epithelium. Following mapping of these elements we will use combined bioinformatic and experimental approaches to identify the trans-acting factors that mediate the coordinated regulation of gene expression. Successful conduct of these studies will provide unique translational opportunities to modify male fertility. In disease states we may be able to reinstate the normal epididymal luminal environment and thereby restore fertility. Through modulation of this environment we may also be able to inhibit normal sperm maturation and thereby develop novel methods of birth control.

描述（由申请人提供）：囊性纤维化跨膜电导调节剂（CFTR）的功能失活（通过突变），一种小电导，cAMP激活的氯离子通道，导致发育缺陷，导致男性在人类中导致男性不育症。 CFTR基因导致不育症的机制知之甚少，但被认为部分归因于男性生殖器导管系统的畸形或病理变化。拟议的研究的目标是通过我们对调节CFTR基因表达的机制的当前理解来建立对附睾中腔环境的转录网络的解密。 CFTR中的突变导致毁灭性的遗传性囊性纤维化（CF）。 CFTR通道对附睾中的离子转运和腔环境做出了重要的贡献，其损失会导致雄性生殖器管的阻塞和/或损失及随后的不育。附睾形成了男性生殖道的排泄管系统的近端部分，是一种复杂的器官，可分泌和吸收离子，水和无机溶质。至关重要的是，它为正常的精子成熟提供了正确的腔环境。在正常的附睾中，通过多个离子通道，离子交换器，溶质载体和转运蛋白的功能合作来建立和维持腔环境，尽管协调这些基因表达的机制尚未得到很好的研究。在第一个特定目的中，我们将确定CFTR基因座染色质结构中的修饰如何控制其在附睾中的表达。我们将评估可能建立活跃基因座的循环构象的顺式作用增强子和绝缘子元件，并确定诸如CTCF和粘着素等跨作用因子如何影响该空间组织。此外，我们将研究可能抑制CFTR不太活跃或不活跃的细胞中基因表达的机制和DNA结合蛋白。第二个特定目标中的实验将确定在附侧上皮中与CFTR相互作用的离子通道，离子交换器，转运蛋白和溶质载体表达的转录途径，以建立和维持腔度环境。我们将使用全基因组方法（DNase-seq）来识别编码在附加膜上皮上有助于离子和溶质转运的蛋白质的基因的顺式作用调节元件。在这些元素映射之后，我们将使用合并的生物信息学和实验方法来识别介导基因表达协调调节的跨力因素。这些研究的成功进行将为改变男性生育能力提供独特的转化机会。在疾病状态下，我们可能能够恢复正常的附睾环境，从而恢复生育能力。通过调节这种环境，我们还可以抑制正常的精子成熟，从而开发出新颖的节育方法。