Urine Biobanking: Methods, Validation, and Research Results
Jennifer L Owens, Jennifer M DiPiero, Paul Elson, Donna E Hansel. Cleveland Clinic, Cleveland, OH
Background: Bladder cancer is a significant cause of morbidity and mortality worldwide, and is difficult to detect early in disease progression, having few obvious symptoms aside from hematuria. An early urine-based diagnostic marker for low and high grade bladder cancer would be of significant value in early detection and treatment of bladder cancer.
Design: Whole urine, and centrifuged urine supernatant and cell pellets were each banked from catheterized samples taken from >200 patients being monitored for bladder cancer. Patients were stratified into groups who showed either no signs of disease, low grade bladder cancer, or high grade bladder cancer. All patient groups were confirmed by cystoscopic observations, and the latter groups were confirmed by biopsy and histopathological examination. Quantities of free RNA and DNA were compared across each urine product for 155 patients, 34 in the non-disease group, 51 in the low grade group, and 70 in the high grade group. RNA was extracted from 24 patient cell pellets; eight from non-diseased patients, eight from patients with low grade bladder cancer, and eight from patients with high grade bladder cancer. Microarray analysis and PCR validation were performed on each RNA sample, and the microarray data was analyzed for genes that were differentially expressed between patients with no disease and all patients with bladder cancer, as well as genes that were differentially expressed between patients with low grade bladder cancer and high grade bladder cancer.
Results: The cell pellet was confirmed to be the most efficient method of biobanking urine if the goal is RNA or DNA extraction. Several genes of interest were identified in the microarray analysis for possible future use as diagnostic or prognostic biomarkers.
Conclusions: Urine is a high-priority body fluid for banking, and the methods described result in excellent nucleic acid preservation. Urine-based biomarker assays for bladder cancer are a valuable addition to a care provider's arsenal for early discovery of bladder cancer.
Wednesday, March 6, 2013 9:30 AM
Poster Session V # 288, Wednesday Morning