Bladder cancer (BC) remains a common malignancy, with muscle-invasive bladder cancer (MIBC) comprising 20 % of cases and a poor 5-year survival rate of ∼50 %. While neoadjuvant chemotherapy (NAC) followed by radical cystectomy is the standard treatment for locally advanced disease, NAC is limited by toxicity and non-response in many patients.
Predictive biomarkers are urgently needed to guide treatment decisions. We developed and analytically validated a multiplex qPCR array to measure mRNA expression of 10 NAC response-associated biomarkers in MIBC. RNA from 8 formalin-fixed paraffin-embedded (FFPE) and 4 fresh-frozen (FF) MIBC specimens was analyzed to assess the effects of tissue type, necrosis, and pre-analytical variables on assay performance. The assay showed high concordance between FF and FFPE samples, with stable performance using FFPE curls stored at ≤4 °C for up to two weeks. It remained robust across RNA input levels (5-100 ng) and required a minimum quality threshold (DV200 >15 %). Necrosis had minimal impact on gene expression, and reproducibility was confirmed across technicians and time points. In conclusion, this qPCR array-based assay offers a reliable platform for evaluating key biomarkers in FFPE tissues and holds promise as a potential clinical tool to predict NAC response in MIBC patients.
Translational oncology. 2025 Sep 09 [Epub ahead of print]
Toru Sakatani, Sunao Tanaka, Kaoru Murakami, Francisco Aguilar, Owen T M Chan, Catherine Bresee, Daniel J Luthringer, Charles J Rosser, Wayne Hogrefe, Hideki Furuya
Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Department of Urology, Kyoto University, Kyoto, Japan. Electronic address: ., Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA. Electronic address: ., Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Department of Urology, Kyoto University, Kyoto, Japan. Electronic address: ., Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA. Electronic address: ., Clinical and Translational Research Program University of Hawaii Cancer Center, Honolulu, HI, USA. Electronic address: ., Biostatistics Shared Resources, Cedars-Sinai Medical Center, Los Angeles, CA, USA. Electronic address: ., Department of Pathology, Cedars-Sinai Medical Center, Los Angeles, CA, USA. Electronic address: ., Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Department of Urology, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Nonagen Bioscience Corp., Los Angeles, CA, USA. Electronic address: ., Nonagen Bioscience Corp., Los Angeles, CA, USA., Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Department of Biomedical Sciences Cedars-Sinai Medical Center, Los Angeles, CA, USA. Electronic address: .
PubMed http://www.ncbi.nlm.nih.gov/pubmed/40930009