Transcriptional networks coordinating luminal environment in the human epididymis: the role of the androgen receptor.

协调人类附睾管腔环境的转录网络：雄激素受体的作用。

基本信息

批准号：
10616724
负责人：
ANN HARRIS
金额：
$ 33.92万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
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 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10616724
关键词：
3-Dimensional Address Anatomy Androgen Receptor Androgen-Insensitivity Syndrome Androgens Bicarbonates Binding Biological Assay Biological Models Cell Nucleus Characteristics Chlorides Cystic Fibrosis Transmembrane Conductance Regulator Data Dependence Development Disease Duct (organ) structure Environment Epididymis Epithelial Cells Epithelium Experimental Models Fertility Study Funding Gene Expression Genes Genetic Transcription Goals Head Health Hormones Human Human Biology Individual Infertility Investigation Ion Channel Ion Transport Ions Ligands Liquid substance Male Contraceptive Agents Male Genital Organs Male Infertility Malignant neoplasm of prostate Mediating Methods MicroRNAs Modeling Molecular National Institute of Child Health and Human Development Nuclear Receptors Organoids Outcome Pathway interactions Physiology Potassium Property Prostate Protocols documentation RUNX1 gene Regulation Regulatory Element Research Research Priority Role Series Small Interfering RNA Source Sperm Maturation Sperm Motility Sperm Preservation Spermatogenesis Steroid Receptors Technology Testing Testis Therapeutic Therapeutic Intervention Tissues Vas deferens structure androgenic apical membrane basolateral membrane birth control cell type cofactor experimental study fertility preservation functional genomics gain of function mutation genome-wide human male human tissue innovation innovative technologies insight interest loss of function male male fertility member novel novel therapeutic intervention programs recruit reproductive function reproductive organ reproductive tract sertoli cell single-cell RNA sequencing small molecule inhibitor sperm cell steroid hormone technology platform transcription factor water channel zygote

项目摘要

SUMMARY: Androgens are steroid hormones with a pivotal role in determining male characteristics, which bind to the androgen receptor (AR) to direct a unique transcriptional program. Although best studied in the human prostate and in prostate cancer, the AR transcriptional network also controls normal functions of other male reproductive tract tissues, including the epididymis and the vas deferens. A critical function of the epididymis, the proximal part of the male genital ducts that transport testicular products out of the body, is in sperm maturation. Immature sperm leaving the testis acquire the capacity to fertilize an egg during their passage through the head and body of the epididymis. Many factors contribute to establishing and maintaining the specific epididymis luminal environment that is required for normal sperm maturation. These include hormones and other testicular products that enter the duct, though the epithelium lining the epididymis has a pivotal role. The proximal luminal fluid has a low pH and bicarbonate concentration to preserve sperm motility and relatively high potassium concentrations to facilitate sperm quiescence. Multiple ion channels, ion exchangers, transporters and water channels in the apical or basolateral membranes of the epididymis epithelium participate in fine-tuning the luminal fluid. Building upon a) substantial preliminary data on the AR transcriptional network in human epididymis epithelial (HEE) cells, which suggest novel co-factors for this TF are involved in the epididymis, together with b) an innovative HEE organoid model, our goal is to test the overarching hypothesis that the Androgen Receptor is pivotal to establishing and maintaining normal functions of the human epididymis epithelium. Moreover, that through the recruitment of specific co-factors, AR orchestrates a unique transcriptional program in this tissue. In the first specific aim we will determine the mechanisms through which androgens impact the differentiation and function of the human epididymis epithelium. We will use a novel epididymis organoid model for functional studies together with single cell RNA- sequencing (scRNA-seq). These experiments will provide novel and important insights into the role of androgens in the biology of the human epididymis epithelium and the transcriptional network that coordinates its functions. Studies in the second aim will determine the mechanisms of interaction of AR with other co- factors including RUNX1 and CEBPβ and how these factors integrate into the transcriptional network in the human epididymis epithelium. We will also investigate the hierarchy of dependency of AR, RUNX1 and CEBPβ using genome wide and locus-specific assays. A gene of particular interest is the cystic fibrosis transmembrane conductance regulator (CFTR), which establishes and maintains chloride and bicarbonate gradients in the epididymis lumen. CFTR recruits a unique set of cis-regulatory elements in HEE cells, some of which also bind AR. These data may facilitate the identification of novel targets for male contraceptives and also have a profound impact on therapeutic approaches to restore male fertility in epididymis disease.

摘要：雄激素是立体激素，在确定男性特征方面具有关键作用，与雄激素受体（AR）结合以指导独特的转录程序。虽然最好的研究人类前列腺和前列腺癌中，AR转录网络还控制着其他的正常功能男性生殖道组织，包括附睾和VAS延期。一个关键功能附子症是雄性生殖管的近端部分，将验证产物从体内运送到体内精子成熟。未成熟的精子离开睾丸，可以在其卵子期间获得卵的施肥通过附加膜的头部和身体。许多因素有助于建立和维护正常精子成熟所需的特定附睾环境。这些包括骑马和其他进入管道的有证据的产品，尽管附加膜的上皮有一个关键作用。近端腔液具有低pH值和碳酸氢盐浓度以保持精子运动钾的浓度相对较高，以促进精子静止。多个离子通道，离子附睾的顶端或基底外侧膜中的交换器，转运蛋白和水道上皮参与微调腔流体。基于A）AR的大量初步数据人附睾上皮（HEE）细胞中的转录网络，该细胞建议该TF的新型辅助因子参与附睾，以及b）创新的HEE器官模型，我们的目标是测试总体假设，即雄激素受体建立和维持正常功能是关键的人附睾上皮。此外，通过招募特定的共同因素AR 在该组织中策划了独特的转录程序。在第一个具体目的中，我们将确定雄激素会影响人类附子症的分化和功能的机制上皮。我们将使用一种新型的附生膜有机模型与单细胞RNA-一起进行功能研究测序（SCRNA-SEQ）。这些实验将提供新颖的重要见解人附子上皮的生物学中的雄激素和坐标的转录网络它的功能。第二个目标的研究将确定AR与其他共同共同的相互作用机制包括runx1和cebpβ以及这些因素如何整合到转录网络中的因素人附子上皮。我们还将研究AR，Runx1和 CEBPβ使用基因组宽和基因座特异性测定法。特别感兴趣的基因是囊性纤维化跨膜电导调节剂（CFTR），建立和维持氯化物和碳酸氢盐附睾腔中的梯度。 CFTR在HEE细胞中招募了一套独特的顺式调节元素，其中一些也结合AR。这些数据可能有助于确定男性避孕药的新目标，并还对恢复附睾疾病中男性生育能力的热方法有深远的影响。

项目成果

期刊论文数量（37）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

An organoid model to assay the role of CFTR in the human epididymis epithelium.

DOI：
10.1007/s00441-020-03208-7
发表时间：
2020-08
期刊：
Cell and tissue research
影响因子：
3.6
作者：
Leir SH;Yin S;Kerschner JL;Xia S;Ahmadi S;Bear C;Harris A
通讯作者：
Harris A

Krüppel-Like Factor 5 Regulates CFTR Expression Through Repression by Maintaining Chromatin Architecture Coupled with Direct Enhancer Activation.

DOI：
10.1016/j.jmb.2022.167561
发表时间：
2022-05-30
期刊：
JOURNAL OF MOLECULAR BIOLOGY
影响因子：
5.6
作者：
Paranjapye, Alekh;NandyMazumdar, Monali;Harris, Ann
通讯作者：
Harris, Ann

A genome-wide analysis of open chromatin in human tracheal epithelial cells reveals novel candidate regulatory elements for lung function.

DOI：
10.1136/thoraxjnl-2011-200880
发表时间：
2012-05
期刊：
Thorax
影响因子：
10
作者：
Bischof JM;Ott CJ;Leir SH;Gosalia N;Song L;London D;Furey TS;Cotton CU;Crawford GE;Harris A
通讯作者：
Harris A

Hepatocyte nuclear factor 1 coordinates multiple processes in a model of intestinal epithelial cell function.

肝细胞核因子 1 协调肠上皮细胞功能模型中的多个过程。