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

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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.