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: 9351169
• 负责人: MARTIN J COHN
• 金额: $ 49.39万
• 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/9351169
• 关键词: 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; Infection; 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; 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.

项目摘要 在哺乳动物中，尿道是从体内排泄尿液的唯一途径，它是主要的 下尿路感染和性传播疾病的进入点。尿道畸形是 在人类中最常见的先天缺陷中，我们对 尿道落后的几年，甚至几十年，都落后于其他系统，例如中枢神经系统，肠道和四肢。前 尿道的后轴（A-P）轴从膀胱延伸到尿道肉类，并在尖端打开 男性的阴茎或阴道开口与女性的阴蒂之间。在细胞和组织水平上， 尿道管沿其A-P轴显示广泛的异质性。上皮结构沿着该轴变化，有 前列腺前，前列腺，膜和阴茎尿道的衬里各自具有独特的特征。 此外，形态发生过程在沿泌尿生殖器窦的离散A-P位置发生。例如， 辅助性腺体，例如前列腺和球状腺体在泌尿生殖器窦上芽 轴向水平。肌肉括约肌也在高度局部的区域发展。尽管有这种广泛的解剖学 区域化，建立这些模式的分子解剖结构尚不清楚。除此之外 发育意义，细胞的A-P认同可能会影响成人功能。在男女上有差异 沿尿道的A-P轴的细胞类型同一性和分子免疫学特征影响定殖 病原体的尿道和膀胱？在这里，我们提出了沿着细胞类型身份的规范 泌尿生殖器窦可以作为A-P区域化的基本发育问题。 类似过程已在胃肠道中进行了广泛的研究，从而导致了详细的图片 肠道区域化，括约肌发展，器官位置和身份的规范以及 控制这些器官中的细胞类型身份。以我们的GUDMAP2项目为基础，该项目的重点是 本研究旨在识别和绘制细胞的分子区域化 沿着小鼠和人类尿道的A-P轴，从膀胱到尿道肉类。我们将使用激光 捕获显微解剖以分离尿道细胞与前列腺前，前列腺，膜和阴茎的分离 小鼠和人尿道区域，并使用RNASEQ识别其转录曲线。然后我们会 对小鼠和人类尿道进行比较的原位杂交分析，整个部分 安装座，我们将使用光投影层析成像来绘制3D中基因表达的A-P模式。我们的 目标是为泌尿外科研究界所必需的基因表达数据奠定基础 研究尿道沿线的细胞类型身份如何与先天性缺陷和疾病的发展有关。