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Intramucosal Inoculation of Squamous Cell Carcinoma Cells in Mice for Tumor Immune Profiling and Treatment Response Assessment
Published on: April 22, 2019
The immunologic landscape of HRAS-mutant head and neck squamous-cell carcinoma
P Economopoulou1, T Rampias2, A Spathis3
1Oncology Unit, 2nd Department of Internal Medicine, National and Kapodistrian University of Athens, Attikon University Hospital, Haidari, Greece.
Background:
HRAS mutations define a distinct biologic subset of head and neck squamous-cell carcinoma (HNSCC). There are limited data regarding HRAS-mutant (mut) tumors' sensitivity to immunotherapy. We sought to evaluate the mutational landscape and transcriptional profile, as well as analyze the tumor microenvironment (TME) of HRAS-mut tumors to provide the conceptual framework for combinatorial treatment approaches.
Materials And Methods:
We analyzed mutational and transcriptome data from The Cancer Genome Atlas (TCGA). In addition, genomic DNA from baseline tumor biopsies was targeted for sequencing. Our study included 10 patients with HRAS-mut and 40 with HRAS-wild-type (WT) HNSCC. Programmed death-ligand 1 (PD-L1) expression in formalin-fixed paraffin-embedded tumor samples was assessed using the PD-L1 IHC 22C3 pharmDx assay. We characterized subpopulations of exhausted CD8(+) T cells by measuring the expression of T-cell factor-1 (TCF1) and programmed cell death protein 1 (PD-1) in both the center and the periphery of the tumors using multiplex immunohistochemistry, followed by analysis using a manually trained algorithm in QuPath software.
Results:
The analysis of TCGA HNSCC mutation and mRNA expression data demonstrated that 6% of HNSCCs harbor mutant HRAS. Transcriptome analysis showed that HRAS-mut HNSCCs are infiltrated by immune cells (CD8A, CD8B, CD2) and have higher expression levels of CXCL11, CXCL10, CXCL9 and CCL4 chemokines. Moreover, the percentage of HRAS-mut samples increased in higher PD-L1 score groups (11% versus 20% versus 100% in tumor positive scores <1%, 1%-49% and ≥50%, respectively, P = 0.006). The analysis of TME showed that HRAS-mut tumors have a statistically significant higher number of total immune cells (5123.17/mm2 versus 3527.93/mm2, P = 0.002) and a higher percentage of pre-exhausted CD8(+) PD-1(+) TCF1(+) T cells in the periphery (384.67/mm2 versus 51.18/mm2, P = 0.040) than HRAS-WT tumors.
Conclusions:
HRAS-mut HNSCCs are characterized by a significantly increased number of pre-exhausted PD-1(+) TCF1(+) T cells and PD-L1 expression, suggesting a potential sensitivity to immunotherapy.
Insights
HRAS mutations in head and neck cancer are linked to increased immune cell infiltration and PD-L1 expression. These findings suggest HRAS-mutant tumors may respond well to immunotherapy treatments.
Area of Science:
- Oncology
- Immunology
- Genetics
Background:
- HRAS mutations identify a specific subgroup of head and neck squamous-cell carcinoma (HNSCC).
- Limited data exist on the immunotherapy sensitivity of HRAS-mutant (mut) tumors.
- Understanding the tumor microenvironment (TME) of HRAS-mut tumors is crucial for developing combinatorial treatments.
Purpose of the Study:
- To evaluate the mutational landscape and transcriptional profile of HRAS-mut tumors.
- To analyze the TME of HRAS-mut tumors.
- To establish a framework for combinatorial treatment strategies in HRAS-mut HNSCC.
Main Methods:
- Analysis of The Cancer Genome Atlas (TCGA) HNSCC mutation and transcriptome data.
- Targeted sequencing of genomic DNA from tumor biopsies (10 HRAS-mut, 40 HRAS-wild-type).
- Assessment of programmed death-ligand 1 (PD-L1) expression and characterization of exhausted CD8(+) T cells (TCF1, PD-1) using multiplex immunohistochemistry.
Main Results:
- HRAS mutations were found in 6% of HNSCCs.
- HRAS-mut HNSCCs showed increased immune cell infiltration and higher expression of chemokines (CXCL11, CXCL10, CXCL9, CCL4).
- HRAS-mut tumors exhibited significantly higher total immune cell counts and a greater percentage of pre-exhausted CD8(+) PD-1(+) TCF1(+) T cells in the tumor periphery compared to HRAS-wild-type tumors.
Conclusions:
- HRAS-mutant HNSCCs are characterized by a higher density of pre-exhausted PD-1(+) TCF1(+) T cells.
- Elevated PD-L1 expression in HRAS-mut tumors suggests potential sensitivity to immunotherapy.
- These findings support further investigation into immunotherapy approaches for HRAS-mutant HNSCC.
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