Spatial characterization of tertiary lymphoid structures as predictive biomarkers for immune checkpoint blockade in head and neck squamous cell carcinoma

Immune checkpoint blockade (ICB) is the standard of care for recurrent/metastatic head and neck squamous cell carcinoma (HNSCC), yet efficacy remains low. The combined positive score (CPS) for PD-L1 is the only biomarker approved to predict response to ICB and has limited performance. Tertiary Lymph...

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Main Authors: Daniel A. Ruiz-Torres, Michael E. Bryan, Shun Hirayama, Ross D. Merkin, Evelyn Luciani, Thomas J. Roberts, Manisha Patel, Jong C. Park, Lori J. Wirth, Peter M. Sadow, Moshe Sade-Feldman, Shannon L. Stott, Daniel L. Faden
Format: Article
Language:English
Published: Taylor & Francis Group 2025-12-01
Series:OncoImmunology
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Online Access:https://www.tandfonline.com/doi/10.1080/2162402X.2025.2466308
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author Daniel A. Ruiz-Torres
Michael E. Bryan
Shun Hirayama
Ross D. Merkin
Evelyn Luciani
Thomas J. Roberts
Manisha Patel
Jong C. Park
Lori J. Wirth
Peter M. Sadow
Moshe Sade-Feldman
Shannon L. Stott
Daniel L. Faden
author_facet Daniel A. Ruiz-Torres
Michael E. Bryan
Shun Hirayama
Ross D. Merkin
Evelyn Luciani
Thomas J. Roberts
Manisha Patel
Jong C. Park
Lori J. Wirth
Peter M. Sadow
Moshe Sade-Feldman
Shannon L. Stott
Daniel L. Faden
author_sort Daniel A. Ruiz-Torres
collection DOAJ
description Immune checkpoint blockade (ICB) is the standard of care for recurrent/metastatic head and neck squamous cell carcinoma (HNSCC), yet efficacy remains low. The combined positive score (CPS) for PD-L1 is the only biomarker approved to predict response to ICB and has limited performance. Tertiary Lymphoid Structures (TLS) have shown promising potential for predicting response to ICB. However, their exact composition, size, and spatial biology in HNSCC remain understudied. To elucidate the impact of TLS spatial biology in response to ICB, we utilized pre-ICB tumor tissue sections from 9 responders (complete response, partial response, or stable disease) and 11 non-responders (progressive disease) classified via RECISTv1.1. A custom multi-immunofluorescence (mIF) staining assay was applied to characterize tumor cells (pan-cytokeratin), T cells (CD4, CD8), B cells (CD19, CD20), myeloid cells (CD16, CD56, CD163), dendritic cells (LAMP3), fibroblasts (α Smooth Muscle Actin), proliferative status (Ki67) and immunoregulatory molecules (PD1). A machine learning model was employed to measure the effect of spatial metrics on achieving a response to ICB. A higher density of B cells (CD20+) was found in responders compared to non-responders to ICB (p = 0.022). The presence of TLS within 100 µm of the tumor was associated with improved overall (p = 0.04) and progression-free survival (p = 0.03). A multivariate machine learning model identified TLS density as a leading predictor of response to ICB with 80% accuracy. Immune cell densities and TLS spatial location play a critical role in the response to ICB in HNSCC and may potentially outperform CPS as a predictor of response.
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spelling doaj-art-32af956872c8428ea194801ace4e6b3d2025-08-20T01:56:56ZengTaylor & Francis GroupOncoImmunology2162-402X2025-12-0114110.1080/2162402X.2025.2466308Spatial characterization of tertiary lymphoid structures as predictive biomarkers for immune checkpoint blockade in head and neck squamous cell carcinomaDaniel A. Ruiz-Torres0Michael E. Bryan1Shun Hirayama2Ross D. Merkin3Evelyn Luciani4Thomas J. Roberts5Manisha Patel6Jong C. Park7Lori J. Wirth8Peter M. Sadow9Moshe Sade-Feldman10Shannon L. Stott11Daniel L. Faden12Department of Otolaryngology-Head and Neck Surgery, Massachusetts Eye and Ear, Boston, MA, USADepartment of Otolaryngology-Head and Neck Surgery, Massachusetts Eye and Ear, Boston, MA, USADepartment of Otolaryngology-Head and Neck Surgery, Massachusetts Eye and Ear, Boston, MA, USADepartment of Medicine, Harvard Medical School, Boston MA, USAKrantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center, Boston, MA, USADepartment of Medicine, Harvard Medical School, Boston MA, USADepartment of Medicine, Harvard Medical School, Boston MA, USADepartment of Medicine, Harvard Medical School, Boston MA, USADepartment of Medicine, Harvard Medical School, Boston MA, USADepartment of Medicine, Harvard Medical School, Boston MA, USAKrantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center, Boston, MA, USAKrantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center, Boston, MA, USADepartment of Otolaryngology-Head and Neck Surgery, Massachusetts Eye and Ear, Boston, MA, USAImmune checkpoint blockade (ICB) is the standard of care for recurrent/metastatic head and neck squamous cell carcinoma (HNSCC), yet efficacy remains low. The combined positive score (CPS) for PD-L1 is the only biomarker approved to predict response to ICB and has limited performance. Tertiary Lymphoid Structures (TLS) have shown promising potential for predicting response to ICB. However, their exact composition, size, and spatial biology in HNSCC remain understudied. To elucidate the impact of TLS spatial biology in response to ICB, we utilized pre-ICB tumor tissue sections from 9 responders (complete response, partial response, or stable disease) and 11 non-responders (progressive disease) classified via RECISTv1.1. A custom multi-immunofluorescence (mIF) staining assay was applied to characterize tumor cells (pan-cytokeratin), T cells (CD4, CD8), B cells (CD19, CD20), myeloid cells (CD16, CD56, CD163), dendritic cells (LAMP3), fibroblasts (α Smooth Muscle Actin), proliferative status (Ki67) and immunoregulatory molecules (PD1). A machine learning model was employed to measure the effect of spatial metrics on achieving a response to ICB. A higher density of B cells (CD20+) was found in responders compared to non-responders to ICB (p = 0.022). The presence of TLS within 100 µm of the tumor was associated with improved overall (p = 0.04) and progression-free survival (p = 0.03). A multivariate machine learning model identified TLS density as a leading predictor of response to ICB with 80% accuracy. Immune cell densities and TLS spatial location play a critical role in the response to ICB in HNSCC and may potentially outperform CPS as a predictor of response.https://www.tandfonline.com/doi/10.1080/2162402X.2025.2466308Head and neck cancerimmune checkpoint blockadespatial biologytertiary lymphoid structures
spellingShingle Daniel A. Ruiz-Torres
Michael E. Bryan
Shun Hirayama
Ross D. Merkin
Evelyn Luciani
Thomas J. Roberts
Manisha Patel
Jong C. Park
Lori J. Wirth
Peter M. Sadow
Moshe Sade-Feldman
Shannon L. Stott
Daniel L. Faden
Spatial characterization of tertiary lymphoid structures as predictive biomarkers for immune checkpoint blockade in head and neck squamous cell carcinoma
OncoImmunology
Head and neck cancer
immune checkpoint blockade
spatial biology
tertiary lymphoid structures
title Spatial characterization of tertiary lymphoid structures as predictive biomarkers for immune checkpoint blockade in head and neck squamous cell carcinoma
title_full Spatial characterization of tertiary lymphoid structures as predictive biomarkers for immune checkpoint blockade in head and neck squamous cell carcinoma
title_fullStr Spatial characterization of tertiary lymphoid structures as predictive biomarkers for immune checkpoint blockade in head and neck squamous cell carcinoma
title_full_unstemmed Spatial characterization of tertiary lymphoid structures as predictive biomarkers for immune checkpoint blockade in head and neck squamous cell carcinoma
title_short Spatial characterization of tertiary lymphoid structures as predictive biomarkers for immune checkpoint blockade in head and neck squamous cell carcinoma
title_sort spatial characterization of tertiary lymphoid structures as predictive biomarkers for immune checkpoint blockade in head and neck squamous cell carcinoma
topic Head and neck cancer
immune checkpoint blockade
spatial biology
tertiary lymphoid structures
url https://www.tandfonline.com/doi/10.1080/2162402X.2025.2466308
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