Multiplexed autoantibody (AA) profiling of patients (pts) with metastatic urothelial carcinoma (mUC) receiving immune checkpoint inhibitors or platinum-based chemotherapy.
Authors: Sonpavde Guru P. , Freeman Dory , Adib Elio , El Zarif Talal , Thomas Jonathan , Nuzzo Pier Vitale , Ravi Arvind , Tuff Martha , Mantia Charlene , McGregor Bradley Alexander , Berchuck Jacob E , Budde Petra , Rupieper Elena , Gajewski Jana , McDaid Ronan , Schbuert Ann-Sophie , Bräutigam Manuel , Zucht Hans-Dieter , Ravi Praful 
Published: February 2022

558

Background: The AA profile may be altered in malignancies and provide insights into tumor biology and the immune state. We hypothesized that the longitudinal AA profiling of mUC pts receiving an immune checkpoint inhibitor (ICI) may provide insights into the immune response, which may be associated with immune events and help discover new therapeutic targets. Methods: We utilized serum from mUC pts receiving an ICI or platinum-based chemotherapy (PBC) at the Dana-Farber Cancer Institute. Age and sex matched healthy controls were also studied. The SeroTag immuno-oncology discovery array (Oncimmune) was utilized, with quantification of the AA reactivity towards 1150 antigens. Bound autoantibodies were detected using an anti-IgG-specific detection antibody conjugated to the fluorescent reporter dye phycoerythrin. The AA reactivity was reported as the median fluorescence intensity (MFI) for each color and sample using a Luminex FlexMAP3D analyzer. A significance analysis of microarrays was performed to identify AAs with elevated levels in bladder cancer compared to matched healthy controls (HCs). AAs with > 1.5 increase between pre- and post-treatment were reported. Scatter and box-whisker plots were reported for all pts and antigens, respectively. Results: Pre- (n = 66) and post treatment (n = 65) serum samples were available from mUC pts receiving pembrolizumab (n = 25), atezolizumab (n = 21), nivolumab (n = 5), avelumab (n = 1), durvalumab + tremelimumab (n = 1), nivolumab plus vaccine (n = 1), and 12 pts who received PBC (cisplatin n = 8, carboplatin n = 4). The median duration between the pre- and post-therapy samples was 6 months, median age was 67.7 years (range 40-91) with 51 men (77.3%). Overall, significant heterogeneity of AAs between pts was observed with 37 AAs showing higher reactivity in pre-treatment mUC pts vs. 47 HCs, notably anti-CTAG1 (NY-ESO-1), CTAG2 (NY-ESO-2), MAGE B-18, KRAS, GRB2, RARRES2, HSP72 and FGFR3 (all p < 0.05). Pre- and post-therapy AA profiles were similar with unique changes seen in each patient. Notably, 3 pts receiving an ICI developed AAs to NY-ESO-1. Pts receiving PBC less frequently developed new AAs, although pts treated with cisplatin appeared to develop AAs more frequently compared to carboplatin-treated pts. Conclusions: This is the first report of a comprehensive AA profile using a novel platform in mUC pts. The study identified multiple elevated AAs in mUC pts vs. HCs, most notably NY-ESO-1, which also developed in some pts following ICIs. Pts treated with PBC did not develop new AAs frequently, although there appeared to be a difference between cisplatin and carboplatin-based chemotherapy. Further development of this platform is warranted to provide data that is orthogonal to genomic/transcriptomic profiling and shed insights on potential therapeutically actionable antigens.