Bulk RNA-seq analysis of molecular characteristics induced by Aoah over-expression in B16 subcutaneous implanted tumors

Secreted proteins are central mediators of intercellular communications and can serve as therapeutic targets in diverse diseases. The ~1903 human genes encoding secreted proteins are difficult to study through common genetic approaches. To address this hurdle and, more generally, to discover cancer therapeutics, we developed the Cancer Immunology Data Engine (CIDE, https://cide.ccr.cancer.gov), which incorporates 90 omics datasets spanning 8575 tumor profiles with immunotherapy outcomes from 17 solid tumor types. CIDE systematically identifies all genes associated with immunotherapy outcomes. Then, we focused on secreted proteins prioritized by CIDE without known cancer roles and validated regulatory effects on immune checkpoint blockade for AOAH, CR1L, COLQ, and ADAMTS7 in mouse models. The top hit, AOAH (Acyloxyacyl Hydrolase), potentiates immunotherapies in multiple tumor models by sensitizing T-cell receptors to weak antigens and protecting dendritic cells through depleting immunosuppressive arachidonoyl phosphatidylcholines and oxidized derivatives. Overall design: To compare genetic differences between B16-mhgp100-Vec and B16-mhgp100-Aoah (and B16F10-Vec vs. B16F10-Aoah) subcutaneous tumors grown on C57BL/6 mice in response to anti-PD1 treatment (and anti-PD1+ anti-CTLA4 treatments for B16F10 models): 10 days (6 days for B16F10 cells) after the inoculation of tumor cells, anti-PD1 (BioXCell, #BE0146, 200 µg/dose) or anti-PD1+ anti-CTLA4 (BioXCell, #BE0131, 200 µg/dose) was given to the tumor-bearing mice twice per week for a total of 4 doses. One day after the last dose, tumor samples were harvested from the mice and used for RNA extraction.