# Early detection of bladder cancer through urothelial cell enrichment and DNA flow cytometry

> **NIH NIH K08** · JOHNS HOPKINS UNIVERSITY · 2024 · $279,608

## Abstract

Project Summary
 Bladder cancer is the sixth most common cancer in the United States, and according to some studies,
the most expensive cancer to manage per patient lifetime. But current screening tests have low sensitivity, high
cost, or require invasive procedures, presenting a barrier to the early detection this disease. In prior work, I led
a team showing that next-generation sequencing of bulk urinary DNA can serve as a sensitive and specific
method for the noninvasive detection of bladder cancer. I now seek to build on this work by addressing three
major obstacles to the use of urinary tumor DNA as a biomarker in a screening context: 1) Most patients
referred for bladder cancer screening present with hematuria, or blood in their urine. This results in the dilution
of tumor DNA by leukocyte DNA and decreased sensitivity of sequencing-based assays. 2) Bulk sequencing of
urinary DNA may detect mutations in cell types that do not give rise to bladder tumors (e.g. clonal
hematopoiesis), thereby decreasing assay specificity. 3) The high cost and complexity of next-generation
sequencing methods limits their accessibility. This proposal seeks to overcome all three obstacles, creating a
practical, high performance diagnostic workflow for bladder cancer screening.
 In Aim 1, I will refine a novel method for the enrichment of urothelial cells from voided urine, called Cell
Enrichment by Size and Selective Lysis (CESSL). Using urine samples from patients with known bladder
tumors, I will then investigate the achievable degree of tumor cell enrichment and generalizability of CESSL
using sequencing and microscopy. Depletion of non-urothelial cells from urine will improve the sensitivity and
specificity of downstream assays in the population that presents for screening.
 In Aim 2, I will develop a flow cytometry assay for the detection of bladder tumor cells in urine based on
the presence of aneuploidy and/or global hypomethylation, two DNA aberrations that are present in >90% of
bladder tumors and highly specific for cancer. I will then determine the analytical and clinical performance
characteristics of this assay, as well as the impact of CESSL on its performance.
 This proposal will be carried out at the Johns Hopkins Hospital under the mentorship of Bert Vogelstein,
MD. It will be guided by a scientific advisory board including experts in pathology, urology, medical oncology,
biostatistics, and flow cytometry. Through completion of this proposal, I will develop new skills in cell-based
diagnostic methods, diagnostic study design, and laboratory management. My goal is to become an
independent laboratory-based physician-scientist who develops novel molecular and cell-based diagnostic
tools for solid tumor pathology specimens and evaluates their utility in clinical trials. Successful completion of
this study will produce novel methods that I can refine and apply to other pathology sample types in my future
laboratory, as well as the data necessary to in...

## Key facts

- **NIH application ID:** 10851980
- **Project number:** 5K08CA276704-02
- **Recipient organization:** JOHNS HOPKINS UNIVERSITY
- **Principal Investigator:** Jonathan Carl Dudley
- **Activity code:** K08 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $279,608
- **Award type:** 5
- **Project period:** 2023-07-01 → 2028-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10851980

## Citation

> US National Institutes of Health, RePORTER application 10851980, Early detection of bladder cancer through urothelial cell enrichment and DNA flow cytometry (5K08CA276704-02). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10851980. Licensed CC0.

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