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: 7750189
• 负责人: Philip L Reno
• 金额: $ 5.01万
• 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 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7750189
• 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; embryonic stem cell; functional genomics; 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的重要性，但目前对控制AR在何时何地正常表达的机制知之甚少。AR编码区被超过1 MB的基因间DNA包围。这些侧翼区含有许多在不同哺乳动物中高度保守的短的非编码序列。我用比较基因组学的方法证明，人类AR基因的侧翼区域有一个不寻常的缺失，它删除了在黑猩猩、恒河猴、狗和小鼠中高度保守的非编码序列。我已经在转基因小鼠中使用了功能增强子检测，以表明同源黑猩猩和小鼠序列在发育中的面部触须和生殖结节中驱动可重复的表达，这两个解剖位置，人类在形态上也与大多数其他哺乳动物不同。我的研究的目标是确定这种缺失对人类生物学的意义，并确定顺式调节机制，指定在哺乳动物发育过程中AR表达的解剖分布。我将利用可诱导的Cre重组酶报告基因检测进一步表征人类缺失的AR控制区所指定的表达模式和细胞谱系。为了严格测试这种保守序列丢失的功能后果，我还将使用靶向基因敲入策略来删除小鼠中的这种元件，并用黑猩猩的元件取代它，从而在该增强子区域复制人类和黑猩猩的遗传条件。最后，我还将扫描一个大的650 kb基因间区域周围的AR基因的其他组织特异性和发育增强子，使用细菌人工染色体（BAC）克隆和功能测定转基因小鼠。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起的作用。这项研究与公共卫生有关，因为AR的错误表达与多种人类疾病有关，如雄激素不敏感综合征，脊髓延髓肌萎缩症和前列腺癌。确定调节AR表达的机制可能有助于这些和其他人类疾病的长期诊断和治疗。