Clinical Trial

. 2021 Dec:123:105587.

doi: 10.1016/j.oraloncology.2021.105587. Epub 2021 Oct 27.

Characterization of the immune response in patients with cancer of the oral cavity after neoadjuvant immunotherapy with the IRX-2 regimen

Siyu Liu¹, Emily Bellile², Ariane Nguyen³, Katie Zarins⁴, Nisha D'Silva⁵, Laura Rozek⁴, Gregory T Wolf⁶, Maureen A Sartor⁷; INSPIRE Trial Clinical Investigators

Collaborators, Affiliations

Collaborators

INSPIRE Trial Clinical Investigators:
Jeff Moyer⁸, Mihir Patel⁹, Audrey Erman¹⁰, Wanessa A Martins¹¹, Jason Newman¹², Michael Kaplan¹³, Frabicio Oliveira¹⁴, Ana Paula Victorina¹⁵, R Bryan Bell¹⁶, Gustavo C Girotto¹⁷, Jorge Nieva¹⁸, Joseph Valentino¹⁹, Greg Krempl²⁰, Claudio R Cernea²¹, Dennis Kraus²², Kevin Higgins²³, Felipe J S M Cruz²⁴, Aru Panwar²⁵, Clodoaldo Z Campos²⁶, Jim McCaul²⁷

Affiliations

¹ Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, MI, USA.
² Department of Biostatistics, University of Michigan, Ann Arbor, MI, USA.
³ Department of Otolaryngology-Head and Neck Surgery, Michigan Medicine, Ann Arbor, MI, USA.
⁴ Department of Environmental Health Sciences, University of Michigan, Ann Arbor, MI, USA.
⁵ Department of Periodontics and Oral Medicine, University of Michigan, Ann Arbor, MI 48104, USA.
⁶ Department of Otolaryngology-Head and Neck Surgery, Michigan Medicine, Ann Arbor, MI, USA. Electronic address: gregwolf@med.umich.edu.
⁷ Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, MI, USA; Department of Biostatistics, University of Michigan, Ann Arbor, MI, USA. Electronic address: sartorma@umich.edu.
⁸ University of Michigan, Ann Arbor, MI, USA.
⁹ Emory University Winship Cancer Institute, Atlanta, GA, USA.
¹⁰ University of Arizona Medical Center, Tucson, AZ, USA.
¹¹ Instituo Goiano de Oncologia e Hematologia, Brazil.
¹² Hospital of the University of Pennsylvania, Philadelphia, PA, USA.
¹³ Stanford University Medical Center, Stanford, CA, USA.
¹⁴ Hospital Erasto Gaertner/Liga Paranaense de Combate Cancer, Brazil.
¹⁵ Instituto Nacional do Cancer, Brazil.
¹⁶ Providence Cancer Center, Providence, RI, USA.
¹⁷ Hospital de Base de Sao Jose do Rio Preto, Brazil.
¹⁸ University of Southern California, Los Angeles, CA, USA.
¹⁹ University of Kentucky Medical Center, Lexington, KY, USA.
²⁰ University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
²¹ Instituto do Cancer do Estado de Sao Paulo, Brazil.
²² Northwell Health/Lenox Hill Hospital, New York, NY, USA.
²³ Sunnybrook Research Institute, Toronto, CA, USA.
²⁴ Instituto Brasileiro de Controle ao Cancer, Brazil.
²⁵ University of Nebraska Medical Center, Omaha, NE, USA.
²⁶ Hospital do Cancer de Londrina, Brazil.
²⁷ Queen Elizabeth University Hospital, Glasgow, UK.

PMID: 34717154
PMCID: PMC8982160
DOI: 10.1016/j.oraloncology.2021.105587

Clinical Trial

Characterization of the immune response in patients with cancer of the oral cavity after neoadjuvant immunotherapy with the IRX-2 regimen

Siyu Liu et al. Oral Oncol. 2021 Dec.

. 2021 Dec:123:105587.

doi: 10.1016/j.oraloncology.2021.105587. Epub 2021 Oct 27.

Authors

Siyu Liu¹, Emily Bellile², Ariane Nguyen³, Katie Zarins⁴, Nisha D'Silva⁵, Laura Rozek⁴, Gregory T Wolf⁶, Maureen A Sartor⁷; INSPIRE Trial Clinical Investigators

Collaborators

INSPIRE Trial Clinical Investigators:
Jeff Moyer⁸, Mihir Patel⁹, Audrey Erman¹⁰, Wanessa A Martins¹¹, Jason Newman¹², Michael Kaplan¹³, Frabicio Oliveira¹⁴, Ana Paula Victorina¹⁵, R Bryan Bell¹⁶, Gustavo C Girotto¹⁷, Jorge Nieva¹⁸, Joseph Valentino¹⁹, Greg Krempl²⁰, Claudio R Cernea²¹, Dennis Kraus²², Kevin Higgins²³, Felipe J S M Cruz²⁴, Aru Panwar²⁵, Clodoaldo Z Campos²⁶, Jim McCaul²⁷

Affiliations

¹ Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, MI, USA.
² Department of Biostatistics, University of Michigan, Ann Arbor, MI, USA.
³ Department of Otolaryngology-Head and Neck Surgery, Michigan Medicine, Ann Arbor, MI, USA.
⁴ Department of Environmental Health Sciences, University of Michigan, Ann Arbor, MI, USA.
⁵ Department of Periodontics and Oral Medicine, University of Michigan, Ann Arbor, MI 48104, USA.
⁶ Department of Otolaryngology-Head and Neck Surgery, Michigan Medicine, Ann Arbor, MI, USA. Electronic address: gregwolf@med.umich.edu.
⁷ Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, MI, USA; Department of Biostatistics, University of Michigan, Ann Arbor, MI, USA. Electronic address: sartorma@umich.edu.
⁸ University of Michigan, Ann Arbor, MI, USA.
⁹ Emory University Winship Cancer Institute, Atlanta, GA, USA.
¹⁰ University of Arizona Medical Center, Tucson, AZ, USA.
¹¹ Instituo Goiano de Oncologia e Hematologia, Brazil.
¹² Hospital of the University of Pennsylvania, Philadelphia, PA, USA.
¹³ Stanford University Medical Center, Stanford, CA, USA.
¹⁴ Hospital Erasto Gaertner/Liga Paranaense de Combate Cancer, Brazil.
¹⁵ Instituto Nacional do Cancer, Brazil.
¹⁶ Providence Cancer Center, Providence, RI, USA.
¹⁷ Hospital de Base de Sao Jose do Rio Preto, Brazil.
¹⁸ University of Southern California, Los Angeles, CA, USA.
¹⁹ University of Kentucky Medical Center, Lexington, KY, USA.
²⁰ University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
²¹ Instituto do Cancer do Estado de Sao Paulo, Brazil.
²² Northwell Health/Lenox Hill Hospital, New York, NY, USA.
²³ Sunnybrook Research Institute, Toronto, CA, USA.
²⁴ Instituto Brasileiro de Controle ao Cancer, Brazil.
²⁵ University of Nebraska Medical Center, Omaha, NE, USA.
²⁶ Hospital do Cancer de Londrina, Brazil.
²⁷ Queen Elizabeth University Hospital, Glasgow, UK.

PMID: 34717154
PMCID: PMC8982160
DOI: 10.1016/j.oraloncology.2021.105587

Abstract

Objective: IRX-2 is a homologous cell-derived multi-cytokine biologic with multifaceted immune modulatory effects that has been shown to induce increased lymphocyte infiltration into primary tumors in oral cavity carcinoma. Our objective was to characterize tumor immune gene expression and epigenomic changes after neoadjuvant IRX-2 immunotherapy in patients with squamous cell carcinoma of the oral cavity.

Methods: A randomized phase II trial was conducted of the IRX regimen 3 weeks prior to surgery for previously untreated patients with Stage II-IV oral cavity carcinoma. The treatment regimen consisted of low dose (300 mg/m²) cyclophosphamide (day 1) followed by 10 days of regional perilymphatic IRX-2 cytokine injections and daily oral indomethacin, zinc and omeprazole (Regimen 1) compared to the identical regimen without the IRX-2 cytokines (Regimen 2). The NanoString immune panel (730 genes) and Infinium MethylationEPIC BeadChip were performed to assess the gene expression and DNA methylation signatures, respectively, in pre- and post-immunotherapy tumor samples.

Results: A total of 51 and 79 immune-related genes were found upregulated and downregulated, respectively, in the samples from Regimen 1 patients after treatment, while 51 and 56 were found upregulated and downregulated in the samples for Regimen 2. When comparing the changes between the two regimens, we identified 9 genes significantly different, including DMBT1, a potential tumor suppressor, functioning in tumor invasion of head and neck cancer. The exploration of DNA methylation showed slight overall hypermethylation after treatment in both regimens, especially for Regimen 1 immune responders, and methylation-based cell type deconvolution demonstrated high concordance with tumor infiltrating T lymphocyte cell counts.

Conclusion: While a consistent patient response after treatment was observed, most changes were similar between regimens, indicating a subtle, targeted, or patient-specific effect of IRX-2 cytokines. Change in DMBT1 expression was a unique finding that will require further study to better understand its significance.

Keywords: Cytokine; Immunotherapy; NanoString; Neoadjuvant; Oral cavity carcinoma; Tumor infiltrating lymphocytes.

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Figures

**Figure 1:**
Immune response genes changed similarly in both Regimen 1 and Regimen 2. (A) Venn diagrams showing the total number of differentially expressed genes, the number of upregulated genes, and the number of downregulated genes after treatment that are overlapped between Regimen 1 and Regimen 2, respectively. (B) Heatmap showing the log₂ fold change of gene expression before versus after treatment (log2FC, resection/biopsy) of all 153 differentially expressed immune genes in either regimen. Samples are ordered in columns by regimen and immune responder status, and hierarchical clustering was performed on genes only, which are displayed as rows.

**Figure 2:**
A total of 9 genes showed significantly different changes after treatment between the two regimens. (A) Heatmap showing the log₂ fold change (resection/biopsy) of the 9 genes whose difference before vs after treatment is significantly different between Regimen 1 and Regimen 2. (B) Box plot of the DMBT1 gene expression at biopsy and resection in the two regimens separately. Spaghetti plot showed the change of DMBT1 expression in patients of different immune response groups.

**Figure 3:**
The expression levels of CD274 (PDL1) and PDCD1 (PD1) in patients (A) with and (B) without immune response groups of the two regimens.

**Figure 4:**
The average DNA methylation levels in both regimens at (A) LINE-1 and LINE-2 elements and (B) immune response gene promoters.

**Figure 5:**
Cell type deconvolution using DNA methylation data revealed higher CD8 in immune responders of Regimen 1. (A) Bar plot showing the proportion of different cell types from 48 patients in different immune response groups. (B) Dot plot indicating the correlation between MethylCIBERSORT CD8 proportion and TMA density (r = 0.51), and the correlation between MethylCIBERSORT CD8 proportion change (resection - biopsy) andTMA CD8 density change (r = 0.31), respectively.

See this image and copyright information in PMC

References

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Characterization of the immune response in patients with cancer of the oral cavity after neoadjuvant immunotherapy with the IRX-2 regimen

Collaborators

Affiliations

Characterization of the immune response in patients with cancer of the oral cavity after neoadjuvant immunotherapy with the IRX-2 regimen

Authors

Collaborators

Affiliations

Abstract

Figures

References

Publication types

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full Text Sources

Medical