Immunological and Genetic Characterization of Patients With Head and Neck Cancer who Developed Recurrence

Abstract

Background/Aim: The recurrence rate of head and neck squamous cell carcinoma (HNSCC) remains high; thus the control of recurrence is a clinical problem to be challenged. To clarify the precise mechanism, specific immunological biomarkers responsible for recurrence were investigated. Patients and Methods: The expression levels of immune response-associated and Shizuoka Cancer Center 820 cancerassociated genes, and genetic mutations from whole-exome sequencing were compared between HNSCC patients who developed recurrence (n=8) and HNSCC patients who did not develop recurrence (n=19) using a volcano plot analysis. Cytokine and epithelial-mesenchymal transition marker genes were analyzed using quantitative PCR. Tumor-infiltrating lymphocytes, immune checkpoint molecules, and human papilloma virus status were investigated using immunohistochemistry (IHC). Results: Twenty-seven evaluable patients with HNSCCs received radiation therapy after surgery. Recurrence was identified in 8 patients. TP53 mutations tended to be higher in patients who developed recurrence than in those who did not develop recurrence (75% vs. 31. 6%). Gene expression profiling showed the down-regulation of T cell activation genes (ICOS, CD69 and CD83) and the upregulation of the ERBB4, EGFR, VEGF, HIF1A, TGFB1, TWIST1, IL-8, and PAX7 genes, which suggested the activation of the TP53 mutation-TGF-β1-PAX7 pathway and epithelialmesenchymal transition. Additionally, IHC indicated a tendency toward a reduction in T cell accumulation and an increase in M2-type macrophage infiltration in tumors that recurred. Conclusion: A TP53 mutation-mediated immune-suppressive state in the tumor microenvironment and TGF-β1-PAX7mediated EMT might contribute to the promotion of recurrence in patients with HNSCC after postoperative radiotherapy.Head and neck squamous cell carcinoma (HNSCC) is a common type of cancer; however, a reduction in the rate of its recurrence remains challenging; the 5-year survival rate of patients with HNSCCs is reported to be only 50-60% (1, 2). The recurrence rate is high, approximately 30%, which indicates not only high mortality and difficulty in treatment, but also the genetic complexity affecting cancer progression 4417

Genetic and Immunological Characterization of Brain Metastases from Solid Cancers

“…Gene expression profiling of immune response-associated genes and the SCC820 cancer-associated genes. The 300 immune responseassociated genes included in the panel are shown in Table I, and the SCC820 cancer-associated gene panel was reported previously (18). Briefly, expression levels of immune response-associated genes and SCC820 cancer-associated genes were compared between primary lesions and brain metastatic lesions from 9 solid cancer patients via volcano plot analysis.…”

“…Real-time PCR analysis. Real-time PCR analyses of the cytokine/chemokine gene panel and solid cancer associated gene panel were performed using the QuantStudio 12K Flex Real Time PCR System (Thermo Fisher Scientific, Waltham, MA, USA), as described previously (18). Total RNA was isolated from FFPE specimens using the High Pure RNA Paraffin Kit (Roche Diagnostics, Indianapolis, IN, USA).…”

“…Total RNA was isolated from primary and metastatic lesions of nine solid cancer patients and analyzed using the Switching Mechanisms at 5' End of RNA Template (SMARTer™) human TCRα and TCRβ profiling kits (Clontech Laboratories Inc., Mountain View, CA, USA) as described previously (18). TCR repertoire profiling was performed using MiXCR software for T-cell receptor sequencing data analysis, which is available from http://mixcr.milaboratory.com/ (19).…”

Impact of Mutations in Subunit Genes of the Mammalian SWI/SNF Complex on Immunological Tumor Microenvironment

“…Immune response-associated genes and SCC820 panel gene expression profiling. The lists of genes in the 204 immune response-associated gene panel and the SCC820 cancer-associated panel have been shown previously (22). Briefly, expression levels of 204 immune response-associated genes and the SCC820 cancerassociated genes of SMARCA4-mutant cancers were compared between the SMARCA4 expression-high (above median) group and SMARCA4 expression-low (below median) group with volcano plot analysis.…”