Abstract 1312: Transcriptional signatures associated with lack of response to anti-PD-1 therapy in patients with renal cell carcinoma

• Published in: Cancer research (Chicago, Ill.) Ill.), 2015-08, Vol.75 (15_Supplement), p.1312-1312
• Main Authors: Ascierto, Maria Libera, McMiller, Tracee, Berger, Alan, Anders, Robert A., Cheadle, Chris, Hu, Haiying, Drake, Charles, Pardoll, Drew, Taube, Janis, Topalian, Suzanne L.
• Format: Article
• Language: English
• ISSN: 0008-5472; 1538-7445
• DOI: 10.1158/1538-7445.AM2015-1312

Background: The PD-1/PD-L1 immune checkpoint pathway limits host immune responses to cancer in the local tumor microenvironment. Monoclonal antibodies blocking PD-1 or PD-L1 have shown promising clinical results in a variety of advanced human cancers including renal cell carcinoma (RCC). We previously reported that response to anti-PD-1 therapy correlates with PD-L1 expression by tumor cells in pre-treatment biopsies. Although 20-30% of patients with metastatic RCC respond to anti-PD-1 therapy, many patients with PD-L1+ tumors still do not respond. The current study was undertaken to understand mechanisms underlying the failure of anti-PD-1 targeted therapies in patients with PD-L1+ RCC. Methods: The specimen cohort included formalin-fixed, paraffin-embedded (FFPE) pre-treatment tumor biopsies expressing PD-L1, derived from 13 RCC patients treated with nivolumab (anti-PD-1) at a single institution [4 responders (R), 9 non-responders (NR); RECIST]. PD-L1+ specimens were defined as those having ≥5% of tumor cells with cell surface PD-L1 expression by immunohistochemistry (IHC). RNA was isolated from PD-L1+ regions on FFPE slides. Whole genome microarray profiling with cDNA-mediated Annealing, Selection, extension and Ligation (DASL) was performed. Global gene expression analysis was profiled using BRBArrayTools. Multiplex quantitative (q)RT-PCR was used to validate differential expression of genes of interest, and IHC was used to validate protein expression from select genes, in R vs. NR. Results: Whole genome analysis revealed 234 transcripts that were differentially expressed in R vs. NR (p value ≤ 0.01, fold change ≥1.5). Ingenuity Pathway Analysis (IPA) of these transcripts showed the involvement of metabolic and immune pathways as well as genes encoding oxidation stress response molecules. Multiplex qRT-PCR for a subset of 60 differentially expressed genes validated significant over-expression of genes with metabolic functions, such as drug glucuronidation (UGT1A6/A1/A3), glucose transport (SLC23A1), and mitochondrial oxidation (AKR1C3) in NR vs. R. Conversely, R were found to overexpress immune markers such as BMP1, which has been shown to positively regulate PD-L1 expression, and CCL3 involved in leukocyte migration. Conclusions: Although tumor PD-L1 expression is associated with an increased likelihood of response to anti-PD-1/PD-L1 therapy, tumor cell-intrinsic metabolism may contribute to treatment resistance in PD-L1+ patients. Our data suggest th