High resolution transcriptome and gene regulatory mapping of human ureter and bladder across the lifespan

人类输尿管和膀胱整个生命周期的高分辨率转录组和基因调控图谱

• 批准号: 10355595
• 负责人: Byron H Lee
• 金额: $ 48.3万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-30 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10355595
• 关键词: Adolescent; Adult; Affect; Age; Agreement; Algorithmic Analysis; Anatomy; Area; Atlases; Bacteria; Benign; Benign Prostatic Hypertrophy; Biological Assay; Bladder; Bladder Tissue; Cell Count; Cell Survival; Cells; Characteristics; Childhood; Chromatin; Collaborations; Communities; Consent; Data; Data Analyses; Data Set; Development; Disadvantaged; Disease; Elderly; Family; Functional disorder; Gender; Gene Expression; Gene Expression Profile; Generations; Genitourinary system; Genomics; Goals; Health Expenditures; Healthcare; Histologic; Hospitals; Human; Immunofluorescence Immunologic; Individual; Institutional Review Boards; Interstitial Cystitis; Life; Location; Longevity; Lower urinary tract; Maps; Messenger RNA; Molecular; Natural regeneration; Nonprofit Organizations; Normal tissue morphology; Ohio; Operative Surgical Procedures; Organ; Organ Donor; Organ Procurements; Pathologic; Pathway interactions; Patients; Pharmacology; Physiological; Procedures; Process; Protein Isoforms; Recovery; Regulator Genes; Research; Resolution; Resources; Site; Specific qualifier value; Structure; System; Technology; Time; Tissue Procurements; Tissues; Transplantation; Transposase; Ureter; Urethra; Urethral Stricture; Urinary Incontinence; Urinary tract infection; Urologic Diseases; Viral; age group; analysis pipeline; base; care burden; cell type; cost; data hub; experience; human tissue; insight; microbiome; novel; novel marker; novel therapeutics; prevent; reconstruction; recruit; repaired; response; sex; single-cell RNA sequencing; tissue processing; transcription factor; transcriptome; transcriptomics; urinary; urinary bladder neck

Abstract Benign urologic diseases of the lower urinary tract such as urinary tract infections, benign prostatic hyperplasia, voiding dysfunction, urinary incontinence, interstitial cystitis, painful bladder syndrome, and urethral strictures incur significant health care burden in the US. Our lack of a comprehensive molecular and cellular understanding of these tissues in the normal and disease states has directly hindered the development of effective novel therapies for these conditions. We are proposing to generate high-resolution transcriptome and gene regulatory data set for healthy human ureter and bladder tissues across the lifespan in both sexes. The overall goal is to construct a highly integrated cellular and molecular anatomical map of the ureter and bladder in humans. This will enable us to comprehensively understand the cellular composition and lineage relationships of the lower urinary tract system. Towards this aim, we have established a robust tissue procurement and processing workflow. This allows us to recover the lower urinary track en bloc from deceased organ donors and process them, with high cell viability and tissue integrity, for molecular assays. We propose to collect scRNA-seq and Visium spatial transcriptomics data on organ donors in 3 age groups that reflect distinct physiological states in terms of lower urinary tract functions across the human life span. For each donor, we will analyze five specified anatomic locations, lower ureter, dome, ureteral orifice, bladder neck, and urethra, to comprehensively characterize parts of the lower urinary tract that have unique tissues structures and functions. We will use these data to understand the lineage relationships between the different cell types and validate novel markers for each novel cell type by immunostaining. Finally, we will perform scATAC-seq on the most informative tissue locations to gain further insight into the underlying gene regulatory networks. Together these studies will provide, for the first time, a comprehensive analysis of the human bladder and ureter throughout life. These molecular data will become a valuable resource to the research community and ultimately, support efforts in organ repair and regeneration.

摘要 下尿路的良性泌尿系统疾病，如尿路感染、良性前列腺 增生、排尿功能障碍、尿失禁、间质性膀胱炎、膀胱疼痛综合征，以及 在美国，尿道狭窄引起了巨大的医疗负担。我们缺乏一个全面的分子和 对这些组织在正常和疾病状态下的细胞理解直接阻碍了 有效的新疗法。我们打算用高分辨率的转录组 和基因调控数据集的健康人输尿管和膀胱组织在整个生命周期中的性别。 总体目标是构建输尿管的高度整合的细胞和分子解剖图， 人类的膀胱这将使我们能够全面了解细胞组成和谱系 下尿路系统的关系。为了实现这一目标，我们建立了一个强大的组织， 采购和处理流程。这使我们能够从死者身上完整地恢复下尿路 器官捐赠者和处理它们，具有高细胞活力和组织完整性，用于分子测定。我们提出 收集3个年龄组器官捐赠者的scRNA-seq和Visium空间转录组学数据，反映 下尿路功能在人的一生中有着不同的生理状态。为每个 供体，我们将分析五个指定的解剖位置，输尿管下段，圆顶，输尿管口，膀胱颈， 尿道，以全面表征具有独特组织结构的下尿路部分 和功能我们将使用这些数据来了解不同细胞类型之间的谱系关系 并通过免疫染色验证每种新细胞类型的新标记。最后，我们将对 信息量最大的组织位置，以进一步了解潜在的基因调控网络。 这些研究将首次提供对人类膀胱的全面分析， 一生中的输尿管。这些分子数据将成为研究界的宝贵资源， 最终，支持器官修复和再生的努力。