PROJECT SUMMARY Bladder cancer (BC) is the costliest cancer (per case) among all cancer types and yet among the top 10 cancers it is the most underfunded cancer by NCI. There has been no significant improvement in overall survival and prognosis over the last thirty years except for the recent development of immunotherapy. At initial diagnosis, approximately 75% of cases are diagnosed at the non-muscle-invasive stages, and are usually treated with transurethral resection (TUR) followed by intravesical instillation of therapeutic agents into the bladder cavity in high risk patients. This approach is associated with cancer recurrence of over 60% at two years and progression into advanced stages in up to 25% of patients. Therefore, almost all patients will need long-term expensive cystoscopy. If a sensitive but less expensive method to detect cancer were available, it would improve the treatment outcomes and decrease the cost. For locally advanced BC, neoadjuvant chemotherapy is associated with a complete response of less than 40%. For metastatic BC, the response rate for the first-line platinum-based chemotherapy is approximately 50%. It is less than 20% for second-line chemotherapy. Currently no method is available to predict which patients will respond to therapy. Liquid biopsies are non-invasive methods that may be applicable for cancer precision medicine. Circulating factors, including circulating tumor cells (CTCs), cfDNAs, RNAs (miRNAs, long non-coding RNAs [lncRNAs], mRNAs), cell-free proteins, peptides, or exosomes et al., are derived from cells in human body. However, it is still unclear where and at what tumor stage these circulating molecules are coming from. There are still too many technical issues that limit the study of CTC biology and their applications. Our latest data show that we are able to generate conditionally reprogrammed cell cultures (CRC) from urine samples of 60 BC patients. Therefore, we will focus on CR technology for generating urine-derived cultures to predict efficacy of treatment and recurrence of BC. We will also determine if urine-derived cell cultures are a simpler and potentially more sensitive technique to monitor bladder cancer recurrence. Urine samples from 70-90 BC cases (prior to initial transurethral resection and chemotherapy) will be used to generate bladder cancer cultures. Following therapy, the patients will be monitored every 3 months (for a total of 2 years) by cystoscopy, and urine cell culture. By comparing the detection of recurrence by each of these techniques, we can determine whether cell culture and/or mutant hTERT detection can provide sufficient sensitivity and specificity to replace the expensive and invasive cystoscopy procedure. We will also use cell cultures derived from pre-surgery patients to evaluate for sensitivity to standard of care drugs and compared with the clinical response of the patient to these same drugs. Last, we will use CR cultures from non-responding patients or recurrent patients in unbiased high-throughput screening to identify and validate new potential therapies for BC.

PROJECT NARRATIVE: The development of accurate biomarkers for the non-invasive detection of bladder cancer (BC) will improve clinical treatment by reducing reliance on cystoscopy, which is burdensome for patients and expensive for healthcare providers, and by providing new information on tumor recurrence potential and response to drug therapy.