GUDMAP: Mapping molecular regionalization of cell types along the anterior-posterior axis of the urethra

GUDMAP：沿尿道前后轴绘制细胞类型的分子区域化

• 批准号: 9923343
• 负责人: MARTIN J COHN
• 金额: $ 11.2万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-15 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9923343
• 关键词: 3-Dimensional; Adult; Anatomy; Anterior; Atlases; Bartholin Glands; Bladder; Bulbourethral gland structure; Cells; Clitoris; Code; Columnar Epithelium; Communities; Congenital Abnormality; Data; Development; Disease; Distal; Epispadias; Epithelial; Excretory function; Female; Foundations; Gastrointestinal tract structure; Gene Expression; Gene Expression Profile; Genetic Transcription; Gland; Goals; Heterogeneity; Homeobox Genes; Human; Hypospadias; Immune response; Immunologics; In Situ Hybridization; Investigation; Knowledge; Lasers; Limb structure; Lower urinary tract; Male urethral structure; Mammals; Maps; Membranous Urethra; Molecular; Mus; Muscle; Neural tube; Optics; Organ; Pathway interactions; Pattern; Physiological; Polyps; Positioning Attribute; Process; Prostate; Prostatic; Prostatic Urethra; Research; Sexually Transmitted Diseases; Specific qualifier value; Sphincter; Structural defect; Structure; System; Therapeutic; Tissue Differentiation; Tissues; Transitional Epithelium; Tube; Urethra; Urinary tract infection; Urine; Urogenital Sinus; Urology; Urothelium; Utricle structure; Vagina; Vertebral column; body system; cell type; combinatorial; comparative; fly; human female; insight; laser capture microdissection; male; malformation; pathogen; penis; regional difference; sex; spine bone structure; tomography; transcription factor; transcriptome sequencing; urinary bladder neck

Project Summary In mammals, the urethra is the sole pathway for excretion of urine from the body and it is the primary point of entry for lower urinary tract infections and sexually transmitted diseases. Urethral malformations are among the most common birth defects in humans, yet our understanding of the molecular development of the urethra lags years, perhaps decades, behind other systems, such as the CNS, gut, and limbs. The anterior to posterior (A-P) axis of the urethra extends from the bladder to the urethral meatus, which opens at the tip of the penis in males or between the vaginal opening and the clitoris in females. At the cell and tissue levels, the urethral tube shows extensive heterogeneity along its A-P axis. Epithelial structure varies along this axis, with the linings of the pre-prostatic, prostatic, membranous, and penile urethra each having a distinctive character. Moreover, morphogenetic processes occur at discrete A-P positions along the urogenital sinus. For example, the accessory sex glands, such as the prostate and bulbourethral glands bud off the urogenital sinus at specific axial levels. Muscular sphincters also develop at highly localized regions. Despite this extensive anatomical regionalization, the molecular anatomy that establishes these patterns is not understood. In addition to this developmental significance, A-P identity of cells may influence adult function. Do male-female differences in cell type identity and molecular immunologic profiles along the A-P axis of the uretha influence colonization of the urethra and bladder by pathogens? Here we propose that specification of cell type identity along the urogenital sinus can be approached as a fundamental developmental problem of A-P regionalization. Analogous processes have been studied extensively in the gastrointestinal tract, resulting in a detailed picture of gut regionalization, sphincter development, specification of the positions and identities of organs, and control of cell type identities within those organs. Building on our GUDMAP2 project, which focused on the dorsoventral (D-V) axis of the LUT, this study aims to identify and map the molecular regionalization of cells along the A-P axis of the mouse and human urethra, from the bladder to the urethral meatus. We will use laser capture microdissection to isolate urethral cells from the pre-prostatic, prostatic, membranous, and penile regions of mouse and human urethra and use RNAseq to identify their transcriptional profiles. We will then carry out a comparative in situ hybridization analysis of mouse and human urethrae in sections and whole mounts, and we will use Optical Projection Tomography to map A-P patterns of gene expression in 3D. Our goal is to generate the foundation of gene expression data necessary for the urology research community to study how cell type identity along the urethra relates to development of congenital defects and to disease.

项目概要 对于哺乳动物来说，尿道是体内尿液排泄的唯一途径，也是主要的排尿途径。 下尿路感染和性传播疾病的切入点。尿道畸形有 人类最常见的出生缺陷之一，但我们对其分子发育的理解 尿道比中枢神经系统、肠道和四肢等其他系统滞后数年甚至数十年。前面的到 尿道后轴（A-P）从膀胱延伸至尿道口，尿道口开口于尿道尖端 男性的阴茎或女性的阴道口和阴蒂之间。在细胞和组织水平上， 尿道管沿其 A-P 轴显示出广泛的异质性。上皮结构沿该轴变化， 前列腺前尿道、前列腺尿道、膜状尿道和阴茎尿道的内壁各有其独特的特征。 此外，形态发生过程发生在沿泌尿生殖窦的离散 A-P 位置。例如， 附属性腺，如前列腺和尿道球腺，在特定的时间从泌尿生殖窦萌芽。 轴向水平。肌肉括约肌也在高度局部化的区域发育。尽管有如此广泛的解剖学 区域化，建立这些模式的分子解剖学尚不清楚。除此之外 发育意义，细胞的 A-P 特性可能会影响成年功能。男女有差异吗 沿尿道 A-P 轴的细胞类型识别和分子免疫学特征影响定植 尿道和膀胱是否受病原体侵害？在这里，我们建议细胞类型身份的规范 泌尿生殖窦可以被视为 A-P 区域化的基本发育问题。 胃肠道中的类似过程已被广泛研究，得出了详细的图像 肠道区域化、括约肌发育、器官位置和特性的规范，以及 控制这些器官内的细胞类型特性。以我们的 GUDMAP2 项目为基础，该项目专注于 LUT 的背腹 (D-V) 轴，本研究旨在识别和绘制细胞的分子区域化图 沿着小鼠和人类尿道的 A-P 轴，从膀胱到尿道口。我们将使用激光 捕获显微解剖，从前列腺前、前列腺、膜和阴茎中分离尿道细胞 小鼠和人类尿道区域并使用 RNAseq 来识别它们的转录谱。我们随后将 对小鼠和人类尿道的切片和整体进行比较原位杂交分析 安装，我们将使用光学投影断层扫描来绘制 3D 基因表达的 A-P 模式。我们的 目标是为泌尿学研究界生成必要的基因表达数据基础 研究尿道沿线的细胞类型特征如何与先天性缺陷和疾病的发展相关。