Abstract 3729: An individualized approach to bladder cancer treatment using patient-derived cell lines to predict response to chemotherapeutic agents

Abstract

Abstract Introduction: Chemotherapy (both intravesical and systemic) can reduce the risk of recurrence and progression in various stages of bladder cancer. However, recurrence after treatment failure is associated with an increased risk of progression. There are currently no established methods for predicting patient-specific responses to treatment prior to drug selection. In our studies, we have developed a novel protocol for efficient establishment of cell lines from primary human bladder tumors, which enables in vitro drug sensitivity assays using chemotherapeutic agents. Methods: Using an Institutional Review Board-approved tissue acquisition protocol, informed consent was obtained prior to specimen acquisition for all samples. Specimens were obtained during standard transurethral resection of papillary bladder tumors. Following generation of a single-cell suspension, epithelial cells were isolated using immunomagnetic cell separation and used for establishment of adherent cell cultures using a novel protocol. We performed immunohistochemistry on parental tissue as well as cultured cells to confirm that the urothelial cancer phenotype was maintained during serial passaging. For sensitivity assays, cultured cells were passaged and treated with chemotherapeutic agents, followed by assessment of cell viability using MTT assays. Results: To date, seven specimens from patients with papillary urothelial carcinoma have been obtained, resulting in the establishment of six independent adherent cell lines. All established lines have been serially passaged (as high as P10) without significant decline in growth rate, and maintained expression of CK7, uroplakin III, p53, and Ki67 in patterns similar to parental tissue. Cells from line #7 were treated with mitomycin C, docetaxel, gemcitabine, and rapamycin at three different equivalent concentrations, resulting in a unique sensitivity profile that was reproduced in a replicate experiment performed at a subsequent passage. Conclusions: We have established a novel protocol for culture and rapid expansion of primary cells from human bladder tumors for assays of drug response. Ultimately, we envision that this approach will provide a basis for the design of patient-specific therapeutic regimens for bladder cancer. Citation Format: LaMont Barlow, Chee Wai Chua, Ming Lei, G. Joel DeCastro, Ketan Badani, Mitchell Benson, James McKiernan, Michael Shen. An individualized approach to bladder cancer treatment using patient-derived cell lines to predict response to chemotherapeutic agents. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3729. doi:10.1158/1538-7445.AM2014-3729