Bulk RNA-seq analysis of molecular characteristics induced by Aoah over-expression in MYC-Luc spontaneous hepatocellular carcinoma 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: We used two murine hepatocellular carcinoma models driven by MYC overexpression with (LucOS) or without (Luc) highly immunogenic OVA+SIN+SIY (OS) antigens and beta-catenin (encoded by CTNNB1) activation. Both models show resistance to anti-PD1 immunotherapy. A sterile 0.9% NaCl solution/plasmid mixture was prepared as follows: We prepared 10 µg of pT3-EF1a-MYC-IRES-luciferase (MYC-Luc), 10 µg of pT3-EF1a-MYC-IRES-luciferase-OS (MYC-LucOS), 10 µg of pT3-N90-CTNNB1 (CTNNB1), 10 µg of PKT2/CLP-Vec, 10 µg of PKT2/CLP-Aoah, and a 3:1 ratio of transposon to SB100 transposase–encoding plasmid dissolved in 2 mL of 0.9% NaCl solution and tail-vein injected 10% of the weight of each C57BL/6 mouse in volume in 5-7 seconds. To study the effects of Aoah overexpression (OE) in tumors, we performed bulk RNA-seq on the MYC-Luc spontaneous hepatocellular (HCC) tumors induced with and without OS antigens.