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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的下尿路良性泌尿系统疾病，如尿路感染、良性前列腺疾病增生、排尿功能障碍、尿失禁、间质性膀胱炎、膀胱疼痛综合征等尿道狭窄在美国造成了巨大的医疗保健负担。我们缺乏全面的分子和细胞对正常和疾病状态下这些组织的理解直接阻碍了发育针对这些病症的有效新疗法。我们建议生成高分辨率转录组以及男女一生中健康人类输尿管和膀胱组织的基因调控数据集。总体目标是构建高度集成的输尿管和输尿管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泌尿道系统的关系。为了实现这一目标，我们建立了一个强大的组织采购和加工工作流程。这使我们能够从死者身上整体恢复下尿路器官捐献者并对其进行处理，具有高细胞活力和组织完整性，用于分子测定。我们建议收集 3 个年龄组器官捐献者的 scRNA-seq 和 Visium 空间转录组学数据，反映人类一生中下泌尿道功能的不同生理状态。对于每个供体，我们将分析五个特定的解剖位置：输尿管下段、穹窿、输尿管口、膀胱颈和尿道，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，我们将执行 scATAC-seq 信息最丰富的组织位置，以进一步了解潜在的基因调控网络。这些研究将首次对人类膀胱和膀胱进行全面分析。输尿管终生。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。