CD8+ and FoxP3+ T-Cell Cellular Density and Spatial Distribution After Programmed Death-Ligand 1 Check Point Inhibition

Joseph Curry; Angela Alnemri; Ramez Philips; Michele Fiorella; Sarah Sussman; Robert Stapp; Charalambos Solomides; Larry Harshyne; Andrew South; Adam Luginbuhl; Madalina Tuluc; Ubaldo Martinez-Outschoorn; Athanassios Argiris; Alban Linnenbach; Jennifer Johnson

doi:10.1002/lary.30389

CD8+ and FoxP3+ T-Cell Cellular Density and Spatial Distribution After Programmed Death-Ligand 1 Check Point Inhibition

Laryngoscope. 2023 Aug;133(8):1875-1884. doi: 10.1002/lary.30389. Epub 2022 Sep 20.

Authors

Joseph Curry¹, Angela Alnemri¹, Ramez Philips¹, Michele Fiorella¹, Sarah Sussman¹, Robert Stapp², Charalambos Solomides², Larry Harshyne^{3

4}, Andrew South⁵, Adam Luginbuhl¹, Madalina Tuluc², Ubaldo Martinez-Outschoorn³, Athanassios Argiris³, Alban Linnenbach⁵, Jennifer Johnson³

Affiliations

¹ Department of Otolaryngology-Head and Neck Surgery, Thomas Jefferson University, Philadelphia, Pennsylvania, U.S.A.
² Department of Pathology, Thomas Jefferson University, Philadelphia, Pennsylvania, U.S.A.
³ Department of Medical Oncology, Thomas Jefferson University, Philadelphia, Pennsylvania, U.S.A.
⁴ Department of Cancer Biology, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania, U.S.A.
⁵ Department of Dermatology and Cutaneous Biology, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania, U.S.A.

PMID: 36125263
DOI: 10.1002/lary.30389

Abstract

Objectives: To analyze CD8+ and FoxP3+ T-cell cellular density (CD) and intercellular distances (ID) in head and neck squamous cell carcinoma (HNSCC) samples from a neoadjuvant trial of durvalumab +/- metformin.

Methods: Paired pre- and post-treatment primary HNSCC tumor samples were stained for CD8+ and FoxP3+. Digital image analysis was used to determine estimated mean CD8+ and FoxP3+ CDs and CD8+-FoxP3+ IDs in the leading tumor edge (LTE) and tumor adjacent stroma (TAS) stratified by treatment arm, human papillomavirus (HPV) status, and pathologic treatment response. A subset of samples was characterized for T-cell related signatures using digital spatial genomic profiling.

Results: Post-treatment analysis revealed a significant decrease in FoxP3+ CD and an increase in CD8+ CDs in the TAS between patients receiving durvalumab and metformin versus durvlaumab alone. Both treatment arms demonstrated significant post-treatment increases in ID. Although HPV+ and HPV- had similar immune cell CDs in the tumor microenvironment, HPV+ pre-treatment samples had 1.60 times greater ID compared with HPV- samples, trending toward significance (p = 0.05). At baseline, pathologic responders demonstrated a 1.16-fold greater CD8+ CDs in the LTE (p = 0.045) and 2.28-fold greater ID (p = 0.001) than non-responders. Digital spatial profiling revealed upregulation of FoxP3+ and cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) in the TAS (p = 0.006, p = 0.026) in samples from pathologic responders.

Conclusions: Analysis of CD8+ and FoxP3+ detected population differences according to HPV status, pathologic response, and treatment. Greater CD8+-FoxP3+ ID was associated with pathologic response. CD8+ and FoxP3+ T-cell distributions may be predictive of response to immune checkpoint inhibition.

Clinicaltrials: gov (Identifier NCT03618654).

Level of evidence: 3 Laryngoscope, 133:1875-1884, 2023.

Keywords: CD8; FoxP3; digital spatial transcriptomics; head and neck cancer; immunotherapy; tumor microenvironment.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CD8-Positive T-Lymphocytes
Head and Neck Neoplasms*
Humans
Lymphocytes, Tumor-Infiltrating
Metformin*
Papillomavirus Infections*
Squamous Cell Carcinoma of Head and Neck
T-Lymphocytes
Tumor Microenvironment

Substances

CD274 protein, human
Metformin

Associated data

ClinicalTrials.gov/NCT03618654