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

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，痛性膀胱综合征，以及 在美国，尿路狭窄会造成巨大的医疗负担。我们缺乏一个全面的分子和 细胞对这些组织在正常和疾病状态下的理解直接阻碍了发育 针对这些疾病的有效的新疗法。我们正在提议产生高分辨率的转录组 以及男性和女性健康的输尿管和膀胱组织的基因调控数据集。 总体目标是构建一个高度集成的输尿管和输尿管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的组成和谱系 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 采购和处理工作流程。这使得我们可以从死者身上整块地恢复下尿路 器官捐赠者，并对其进行处理，具有高细胞活性和组织完整性，用于分子分析。我们建议 收集3个年龄组的器官捐赠者的scRNA-seq和Viium空间转录组学数据，以反映 在人类一生中，下尿路功能处于不同的生理状态。对于每个 供者，我们将分析五个特定的解剖位置，输尿管下段、输尿管穹隆、输尿管开口、膀胱颈和 尿道，以全面描述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 on 信息最丰富的组织位置，以进一步深入了解潜在的基因调控网络。 这些研究将第一次提供对人类膀胱和 一生都在输尿管。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方面的努力。

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.