# Direct conversion of fibroblasts to urothelial stem cells

> **NIH NIH K08** · STANFORD UNIVERSITY · 2023 · $167,076

## Abstract

PROJECT SUMMARY
 Reconstruction of the urinary tract in urologic surgery oftentimes involves utilizing the small intestine to
replace a segment of the urinary system, and as a result, patients may suffer from the side effects of connecting
the urinary and gastrointestinal tracts. Patients that undergo urinary diversion surgery, for example, are at risk
of infection, electrolyte abnormalities, and ileus as a result of this practice. If a better substitute for the urinary
tract were available, outcomes from urinary diversion or reconstructive surgery involving the small intestine would
be drastically improved. The main goal of this proposal is to develop a source of autologous urothelial stem cells
that can potentially be used towards the development of alternative bladder or urothelial substitutes. In this
proposal we hypothesize that urothelial stem cells can be generated via direct conversion of fibroblasts and can
reconstitute the bladder urothelium in the mouse. First, we will aim to generate urothelial stem cells via direct
conversion, or transdifferentiation (Aim 1), and we will achieve this by overexpressing transcription factors
associated with bladder development and screening for suprabasal and basal urothelial markers. We will
validate our screening results with functional assays with organoids as well as multilayered assembloids.
Second, we will map the epigenetic changes that take place during urothelial stem cell differentiation to
suprabasal cells (Aim 2), and we will accomplish this by performing Omni ATAC-seq on control bladder organoids
and urothelial stem cell organoids in basal and differentiation media conditions. By identifying differences in
areas of chromatin hyperaccessibility, we will be able to identify transcription factor binding motifs enriched in
stem cell and differentiated cell states. Finally, we will develop a bladder urothelial stem cell transplant protocol
using mouse models of urothelial ablation and injury (Aim 3). We will determine if urothelial stem cell
transplantation can reconstitute all cell types within the urothelium utilizing urothelial stem cells obtained from
mouse bladders as well as those obtained from transdifferentiation, and we will test for functional outcomes. If
we are successful in these aims, we will demonstrate that autologous urothelial stem cells can be generated via
direct conversion of fibroblasts, and we will establish a source of cells for bladder substitute tissue engineering
as well as a basis for cell-based therapy for disorders of the urothelium that are typically treated with surgical
reconstruction using the gastrointestinal tract, such as severe radiation cystitis or severe interstitial cystitis. The
impact of this work on human health will be significant as this work will potentially make urinary diversion and
reconstruction surgery a much less morbid surgical option for patients with severe urothelial disorders.

## Key facts

- **NIH application ID:** 10661523
- **Project number:** 5K08DK131391-02
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** Kris Prado
- **Activity code:** K08 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $167,076
- **Award type:** 5
- **Project period:** 2022-08-01 → 2027-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10661523

## Citation

> US National Institutes of Health, RePORTER application 10661523, Direct conversion of fibroblasts to urothelial stem cells (5K08DK131391-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10661523. Licensed CC0.

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