Spatial analysis identifies DC niches as predictors of pembrolizumab therapy in head and neck squamous cell cancer

Juhyun Oh; Jan Hoelzl; Jonathan C T Carlson; Ruben Bill; Hannah M Peterson; William C Faquin; Mikael J Pittet; Sara I Pai; Ralph Weissleder

doi:10.1016/j.xcrm.2025.102100

Spatial analysis identifies DC niches as predictors of pembrolizumab therapy in head and neck squamous cell cancer

Cell Rep Med. 2025 Apr 25:102100. doi: 10.1016/j.xcrm.2025.102100. Online ahead of print.

Authors

Juhyun Oh¹, Jan Hoelzl², Jonathan C T Carlson³, Ruben Bill⁴, Hannah M Peterson⁵, William C Faquin⁶, Mikael J Pittet⁷, Sara I Pai⁸, Ralph Weissleder⁹

Affiliations

¹ Center for Systems Biology, Massachusetts General Hospital, Boston, MA 02114, USA; Department of Radiology, Massachusetts General Hospital, Boston, MA 02114, USA.
² Center for Systems Biology, Massachusetts General Hospital, Boston, MA 02114, USA; Department of Medical Oncology, Heidelberg University Hospital, 69120 Heidelberg, Germany.
³ Center for Systems Biology, Massachusetts General Hospital, Boston, MA 02114, USA; Department of Medicine, Division of Oncology, Massachusetts General Hospital, Boston, MA 02114, USA; Massachusetts General Hospital Cancer Center, Boston, MA 02114, USA.
⁴ Center for Systems Biology, Massachusetts General Hospital, Boston, MA 02114, USA; Department of Medical Oncology, Inselspital, Bern University Hospital, University of Bern, 3010 Bern, Switzerland.
⁵ Center for Systems Biology, Massachusetts General Hospital, Boston, MA 02114, USA.
⁶ Massachusetts General Hospital Cancer Center, Boston, MA 02114, USA; Department of Pathology, Massachusetts General Hospital, Boston, MA 02114, USA.
⁷ Department of Pathology and Immunology, University of Geneva, 1211 Geneva, Switzerland; AGORA Cancer Research Center, and Swiss Cancer Center Leman, 1011 Lausanne, Switzerland; Department of Oncology, University of Lausanne (UNIL) and Lausanne University Hospital (CHUV), 1011 Lausanne, Switzerland; Ludwig Institute for Cancer Research, 1011 Lausanne, Switzerland.
⁸ Department of Surgery, Division of Otolaryngology-Head and Neck Surgery, Yale University School of Medicine, New Haven, CT 06510, USA.
⁹ Center for Systems Biology, Massachusetts General Hospital, Boston, MA 02114, USA; Department of Radiology, Massachusetts General Hospital, Boston, MA 02114, USA; Massachusetts General Hospital Cancer Center, Boston, MA 02114, USA; Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA. Electronic address: rweissleder@mgh.harvard.edu.

PMID: 40311615
DOI: 10.1016/j.xcrm.2025.102100

Abstract

Head and neck squamous cell carcinoma (HNSCC) shows variable response to anti-programmed cell death protein 1 (PD-1) therapy, which can be partially explained by a combined positive score (CPS) of tumor and immune cell expression of programmed death-ligand 1 (PD-L1) within the local tumor microenvironment (TME). To better define TME immune determinants associated with treatment efficacy, we conduct a study of n = 48 HNSCC tumors from patients prior to pembrolizumab therapy. Our investigation combines a rapid bioorthogonal multiplex staining method with computational analysis of whole-slide imaging to capture the single-cell spatial heterogeneity and complexity of the TME. Analyzing 6,316 fields of view (FOVs), we provide comprehensive PD-L1 phenotyping and cell proximity assays across the entirety of tissue sections. While none of the PD-L1 metrics adequately predict response, we find that the spatial organization of CCR7⁺ dendritic cells (DCs) in niches better predicts overall patient survival than CPS alone. This study highlights the importance of understanding the spatial context of immune networks for immunotherapy.

Keywords: biomarkers; head and neck cancer; immunotherapy; microscopy; spatial biology.