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：
10491331
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/10491331
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 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 gene regulatory network human tissue insight microbiome novel novel marker novel therapeutics organ regeneration organ repair 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.

抽象的下尿路的良性泌尿科疾病，例如尿路感染，良性前列腺增生，空隙功能障碍，尿失禁，间质性膀胱炎，疼痛膀胱综合征和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中这些组织的理解直接阻碍了发展这些条件有效的新型疗法。我们建议生成高分辨率转录组以及两性寿命的健康人输尿管和膀胱组织的基因调节数据集。总体目标是构建输尿管的高度集成的细胞和分子解剖图和人类膀胱。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，我们建立了坚固的组织采购和处理工作流程。这使我们能够从死者中恢复下泌尿道器官捐献者并以高细胞活力和组织完整性处理分子测定。我们建议收集有关3个年龄段的器官捐献者的SCRNA-SEQ和vistium空间转录组数据，以反映在整个人类生命周期中，在较低的尿路功能方面，不同的生理状态。每个捐助者，我们将分析五个指定的解剖位置，下输尿管，圆顶，输尿管孔，膀胱颈和尿道，全面表征了具有独特组织结构的下尿路的一部分和功能。我们将使用这些数据来了解不同单元类型之间的谱系关系并通过免疫染色来验证每种新型细胞类型的新标记。最后，我们将在最有用的组织位置，以进一步了解潜在的基因调节网络。这些研究将首次提供对人类膀胱的全面分析和一生中输尿管。这些分子数据将成为研究界的宝贵资源，并最终，支持器官维修和再生方面的努力。