Background: We have identified a bladder cancer-associated signature which has been confirmed in several large cohorts, specifically noting its ability to identify smaller tumors (~1cm), which had an elevation at the time of diagnosis in one or more of the 10 biomarkers but not to the same extent as we see in the studies with primary de novo tumors (~3 cm). Thus, we have reported, as the tumor grows, as the grade increases and as the stage increases, there is a corresponding increase in the absolute levels of one or more of the biomarkers that comprise the bladder cancer-associated signature. Currently, the bladder cancer-associated signature is being tested in 4 large prospective studies; i) bladder cancer detection in subjects with gross hematuria, ii) bladder cancer detection in subjects with microscopic hematuria, iii) tumor surveillance in patients with a history of bladder cancer and iv) predicting response to intravesical BCG. In the tumor surveillance study, subjects are followed for 2 years to monitor for tumor recurrence. After enrolling 300 subjects into this study as of May 2020, to date, we have noted tumors recurrences in the 21-24 months follow-up period in 20 subjects. This group was picked to generate the current preliminary data because they would have had several serial urine samples available for analysis. Interestingly of these 20 subjects (57 voided urine samples), the multiplex immunoassay noted an elevation of our bladder cancer-associated signature as early as 18 months prior to the clinical diagnosis of bladder cancer and an actual positive immunoassay in all 20 subjects at 12 months prior to the clinical diagnosis of bladder cancer. Trial Design: This is a single arm pilot study evaluating the feasibility of conducting a longitudinal study monitoring a high-risk population for the development of bladder cancer. Note–This pilot study is design to determine feasibility of recruiting, retaining, and testing a very high-risk cohort in hopes of noting a positive signal from our test. The subsequent definitive study would be expanded to include woman >50 years of age. Eligibility Criteria: Participants must be age 50 years or older, males, >30 pack year history of tobacco exposure, free of any malignancy, willing and able to give written informed consent, willing to provide voided urine sample, and able and willing to complete annual research clinic visits for 4 years. Trial Objectives: 1) To assess the ability to recruit and retain an appropriate high-risk group of subjects into a 4-year longitudinal study monitoring them for the development of bladder cancer. 2) To report the sensitivity and specificity of the multiplex immunoassay ability to detect bladder cancer in patients with high risk of developing bladder cancer: males, >50 years and >30 pack year history of tobacco exposure. 3) To develop a prediction risk calculator from this cohort using cutting edge machine learning techniques (e.g., random forest) incorporating biomarker data and clinical data. Accrual: The target enrollment is approximately 150 patients.

Citation Format: Hideki Furuya, Charles J. Rosser. A novel multiplex immunoassay for the early detection of bladder cancer. [abstract]. In: Proceedings of the AACR Special Conference: Precision Prevention, Early Detection, and Interception of Cancer; 2022 Nov 17-19; Austin, TX. Philadelphia (PA): AACR; Can Prev Res 2023;16(1 Suppl): Abstract nr P032.