Adenoviral infectivity of exfoliated viable cells in urine: Implications for the detection of bladder cancer

Exfoliated cells from urine were obtained from 36 human subjects (> 40 years old). An adenovirus in which GFP expression is under control of the survivin promoter (Ad.Surv.GFP) was generated. An adenovirus in which GFP is expressed from the CMV promoter served as a control. GFP expression was analyzed by fluorescent microscopy and quantified by flow cytometry.Short-term cultures from exfoliated cells in urine could be established in 16 of 31 samples. These cultures were successfully transduced with Ad.CMV.GFP. Analysis of GFP expression following transduction with Ad.Surv.GFP, indicated that the survivin promoter was preferentially active in UM-UC-3 bladder cancer cells compared to non-malignant UROtsa cells. Interestingly, baseline levels of GFP expression in cultures from exfoliated cells in urine exhibited higher baseline levels than UROtsa following transduction with Ad.Surv.GFP.We demonstrated the feasibility of establishing and analysing short-term cultures isolated from exfoliated cells in voided urine by means of adenoviral transduction, thereby forming the foundation for future studies to determine the specificity and sensitivity of a non-invasive test based on survivin promoter activity.According to the American Cancer Society bladder cancer is the 5th highest in estimated new cases of cancers by site with 14,680 bladder cancer deaths and 70,530 new diagnoses in 2010 [1,2]. Bladder cancer can be categorized into non-muscle-invasive bladder cancer or muscle-invasive bladder cancer where 80% of the newly diagnosed cancers are non-muscle-invasive bladder cancer. Unfortunately, 70% of the patients will have recurrence of the disease and 10-30% will progress to muscle-invasive disease. Bladder cancer is detected as a result of incidental findings or by presenting hematuria. While hematuria is associated with benign conditions such as urinary tract infections and urolithiasis, 10% of the patients with gross hematuria are diagnosed with bladder cancer [3]. Contra