Tech Note Assessing Tumor Inflammation

Tumor inflammation associated with cytotoxic T cell responses has been identified as being important in understanding response to checkpoint inhibitors. The recruitment of new T cells and clonal expansion of those which successfully recognize a tumor antigen has been shown to elicit potent anti-tumor activity. The HTG EdgeSeq Precision Immuno-Oncology Panel measures the associated with anti-tumor immune responses, including the interferon-γ mediated response illustrated below.

Expression of Inflammation Genes

16,000 NSCLC-01 Other Applications 14,000 NSCLC-03 12,000 NSCLC-09

Cytokine profiling 10,000 NSCLC-05 8,000 Immunophenotyping 6,000 Normalized Counts Normalized TILs 4,000 Immunoresistance 2,000 0 IFNG CXCL9 CXCL10 CCL5 STAT1 PRF1 GZMA GZMK pathways Elevated levels of interferon gamma is observed in cases NSCLC-03 and NSCLC-05 Immunosuppression consistent with a proinflammatory tumor microenvironment. Expression of chemokines phenotyping known to play a role in T cell and NK cell recruitment, CXCL9, CXCL10, and CCL5 are upregulated, and signaling through the STAT signal transduction pathway (represented DNA repair mechanisms by induction of STAT1) are observed. The expression of cytolytic molecules perforin and granzymes GZMA and GZMK indicate an effector-driven immune response. Drug target assessment Selected Immune Response Profiling Genes Responses Macrophages TCGA tumor subtyping Th1 Th2 Th17 MDSC M1 M2 M2 BST2 IFNG ADCY1 IL26 CCR4 IL21 ANPEP CD80 ARG1 TLR8 CCL4 IL12A BIRC5 IL4 CCR6 IL22 ARG1 CD86 CCL2 VEGFA CCR5 IL12B CDC25C IL5 IL17A IL24 CD33 IL1R1 CD14 VEGFC CD38 IL12RB2 CDC7 LAIR2 IL17B IL26 CD34 IL1R2 CD163 CD70 IL2 CENPF LIMA1 IL17C RORC FUT4 NOS2 FCGR1A_FCGR1B CSF2 IL22 CXCR6 MAF IL17F STAT3 IL10 NOS3 FCGR3A_3B CTLA4 LTA GATA3 NEIL3 IL17RA TGFB ITGAM SOCS3 IL10 CXCL10 STAT1 HELLS PMCH IL1B TGFB1 TLR2 PCNA CXCL9 STAT4 IL10 SMAD2 TLR4 PECAM1 CXCR3 TBX21 IL13 STAT6 TFRC DPP4 TNF IL25 WDHD1 TLR1

About the HTG EdgeSeq Precision Immuno-Oncology Panel The next-generation sequencing (NGS)-based HTG EdgeSeq Precision Immuno- Oncology Panel is designed to measure the immune response both inside the tumor and the surrounding microenvironment. HTG’s quantitative nuclease protection assay does not require nucleic acid extraction and is automated using the HTG EdgeSeq processor. By leveraging the high sensitivity and dynamic range of NGS instrumentation, this powerful tool interrogates 1,392 genes from a single section of formalin-fixed, paraffin- embedded (FFPE) tissue, extracted RNA, or PAXgene samples.

HTG Molecular Diagnostics, Inc. 3430 E. Global Loop, Tucson, AZ 85706, USA Call 1-877-289-2615 Fax 1-520-547-2837 For Research Use Only. Not for use in diagnostic procedures. [email protected] or [email protected] In the U.S. and other applicable jurisdictions, HTG EdgeSeq and VERI/O are Service and Support 1-877-507-3259 trademarks of HTG Molecular Diagnostics, Inc. Any other trademarks or trade names Orders [email protected] used herein are the intellectual property of their respective owners. Rev 2: 23-JAN-2019

Tech Note Assessing Tumor Inflammation