Regulatory architecture of the Androgen Receptor locus in development & evolution

正在开发的雄激素受体基因座的调控架构

• 批准号: 7939670
• 负责人: Philip L Reno
• 金额: $ 1.87万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7939670
• 关键词: 3' Flanking Region; Adult; Anatomy; Androgen Receptor; Androgen-Insensitivity Syndrome; Androgens; Architecture; Area; Bacterial Artificial Chromosomes; Biological Assay; Biological Testing; Breeding; Canis familiaris; Cartilage; Cell Lineage; Cells; Characteristics; Code; Conserved Sequence; Development; Diagnosis; Elements; Embryo; Enhancers; Event; Evolution; Face; Fatty acid glycerol esters; Functional RNA; Gene Targeting; Genes; Genetic Recombination; Genital system; Genomics; Gland; Growth; Growth and Development function; Hair; Hereditary Disease; Human; Human Biology; Human Genetics; Human Genome; Indium; Individual; Intercistronic Region; Intergenic DNA; Knock-in Mouse; Knockout Mice; Label; LacZ Genes; Location; Macaca mulatta; Malignant neoplasm of prostate; Mammals; Mammary gland; Mediating; Mus; Muscle; Nucleic Acid Regulatory Sequences; Pan Genus; Pattern; Phenotype; Play; Public Health; Receptor Gene; Regulatory Element; Reporter; Research; Role; Scanning; Sexual Development; Site; Skeleton; Skin; Specific qualifier value; Structure; Testing; Tissues; Transgenic Mice; Transgenic Organisms; Vibrissae; bone; cohort; comparative genomics; embryonic stem cell; homologous recombination; human disease; improved; insight; male; nonhuman primate; promoter; receptor; receptor expression; recombinase; research study; response; spinal and bulbar muscular atrophy; trait; vertebrate genome

DESCRIPTION (provided by applicant): Androgen Receptor (AR) is critical for the development of the sexually dimorphic phenotypes in many different mammalian tissues, including cartilage, bone, muscle, fat, skin, and secretory glands. Despite the importance of this gene in both normal development and human disease, little is currently known about the mechanisms that control where and when AR is normally expressed. The AR coding region is surrounded by over 1 MB of intergenic DNA. These flanking regions contain numerous short non-coding sequences that are highly conserved in different mammals. I have used comparative genomics to show that humans have an unusual deletion in the flanking regions of the AR gene, which removes non-coding sequences that are otherwise highly conserved in the chimpanzee, rhesus, dog, and mouse. I have used functional enhancer assays in transgenic mice to show that the homologous chimpanzee and mouse sequences drive reproducible expression in the developing facial vibrissae and genital tubercle, two anatomical locations, where humans are also morphologically distinct from most other mammals. The objectives of my research are to determine the significance of this deletion for human biology and to identify cis-regulatory mechanisms that specify the anatomical distribution of AR expression during mammalian development. I will utilize an inducible Cre-recombinase reporter assay to further characterize the expression pattern and cell lineages specified by the AR control region that is missing in humans. To rigorously test the functional consequence of the loss of this conserved sequence, I will also use a targeted gene knock-in strategy to delete this element in the mouse and replace it with that of the chimpanzee, thus replicating both the human and chimpanzee genetic conditions in this enhancer region. Finally, I will also scan a large 650 kb intergenic region surrounding the AR gene for other tissue specific and developmental enhancers, using bacterial artificial chromosome (BAC) clones and functional assays in transgenic mice. These experiments will greatly improve our understanding of the mechanisms by which AR expression is established during mammalian development and sexual differentiation, and the role it plays in specifying the unique human phenotype. This research is relevant to public health because the misexpression of AR is associated with multiple human diseases such as androgen insensitivity syndrome, spinal bulbar muscular atrophy, and prostate cancer. Identifying the mechanisms by which AR expression is regulated may aid in the long-term diagnosis and treatment of these and other human diseases.

描述（由申请人提供）：雄激素受体（AR）对于许多不同哺乳动物组织（包括软骨、骨、肌肉、脂肪、皮肤和分泌腺）中性二态表型的发育至关重要。尽管该基因在正常发育和人类疾病中都很重要，但目前人们对控制 AR 正常表达的地点和时间的机制知之甚少。 AR 编码区被超过 1 MB 的基因间 DNA 包围。这些侧翼区域包含许多短非编码序列，这些序列在不同哺乳动物中高度保守。我使用比较基因组学表明，人类 AR 基因的侧翼区域有一个不寻常的缺失，它去除了在黑猩猩、恒河猴、狗和小鼠中高度保守的非编码序列。我在转基因小鼠中使用了功能增强子测定，结果表明，黑猩猩和小鼠的同源序列在发育中的面部触须和生殖器结节中驱动可重复的表达，这两个解剖位置是人类在形态上也与大多数其他哺乳动物不同的地方。我的研究目的是确定这种缺失对人类生物学的重要性，并确定在哺乳动物发育过程中指定 AR 表达的解剖分布的顺式调控机制。我将利用诱导型 Cre 重组酶报告基因测定来进一步表征人类中缺失的 AR 控制区所指定的表达模式和细胞谱系。为了严格测试丢失这个保守序列的功能后果，我还将使用靶向基因敲入策略删除小鼠中的这个元件，并将其替换为黑猩猩的元件，从而在这个增强子区域复制人类和黑猩猩的遗传条件。最后，我还将使用细菌人工染色体 (BAC) 克隆和转基因小鼠的功能测定，扫描 AR 基因周围的一个大的 650 kb 基因间区域，寻找其他组织特异性和发育增强子。这些实验将极大地提高我们对哺乳动物发育和性分化过程中 AR 表达建立机制的理解，以及它在指定独特的人类表型中所发挥的作用。这项研究与公共健康相关，因为 AR 的错误表达与多种人类疾病有关，例如雄激素不敏感综合征、脊髓延髓肌萎缩症和前列腺癌。确定 AR 表达的调节机制可能有助于这些疾病和其他人类疾病的长期诊断和治疗。