Single-cell spatial transcriptomics unravels cell states and ecosystems associated with clinical response to immunotherapy

Background The tumor microenvironment (TME) is a complex and dynamic ecosystem that is known to influence responses to immunotherapy. We leveraged single-cell spatial transcriptomics to systematically dissect the intricate complexity of the TME, in particular the cellular heterogeneity and spatial i...

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Main Authors: Sjoerd H van der Burg, Ziena Abdulrahman, Mariette I E van Poelgeest, Roderick C Slieker, Marij J P Welters, Daniel McGuire
Format: Article
Language:English
Published: BMJ Publishing Group 2025-03-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/13/3/e011308.full
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author Sjoerd H van der Burg
Ziena Abdulrahman
Mariette I E van Poelgeest
Roderick C Slieker
Marij J P Welters
Daniel McGuire
author_facet Sjoerd H van der Burg
Ziena Abdulrahman
Mariette I E van Poelgeest
Roderick C Slieker
Marij J P Welters
Daniel McGuire
author_sort Sjoerd H van der Burg
collection DOAJ
description Background The tumor microenvironment (TME) is a complex and dynamic ecosystem that is known to influence responses to immunotherapy. We leveraged single-cell spatial transcriptomics to systematically dissect the intricate complexity of the TME, in particular the cellular heterogeneity and spatial interactions. Their collective impact on immunotherapy efficacy was studied in the context of a homogeneous group of patients with vulvar high-grade squamous intraepithelial lesions (vHSIL) treated with an immunotherapeutic tumor-specific peptide vaccine.Methods We performed single-cell spatial transcriptomics on 20 pretreatment vHSIL lesions, stratified by clinical response to immunotherapeutic vaccination into complete responders (CR), partial responders (PR) and non-responders (NR). Using a 1,000-gene panel, we mapped over 274,000 single cells in situ, identifying 18 cell clusters and 99 distinct non-epithelial cell states. Findings were validated against public single-cell transcriptomic data sets to assess their broader relevance across tumor types.Results Profound heterogeneity within the TME was detected across the response groups. CR lesions exhibited a higher ratio of immune-supportive to immune-suppressive cells—a pattern mirrored in other solid tumors following neoadjuvant checkpoint blockade. Key immune populations enriched in CRs included CD4+CD161+ effector T cells and chemotactic CD4+ and CD8+ T cells. Conversely, PRs were characterized by increased proportions of T helper 2 cells and CCL18-expressing macrophages, which are associated with the recruitment of type 2 T cells and regulatory T cells. NRs displayed preferential infiltration with immunosuppressive fibroblasts. Distinct spatial immune ecosystems further defined response groups. Although a number of immune cells were detected in all patients, type 1 effector cells dominated interactions in CRs, type 2 cells were prominently interacting in PRs, while NRs lacked organized immune cell interactions.Conclusions This study underscores the dual importance of both cellular composition and spatial organization in steering clinical response to immunotherapy.
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spelling doaj-art-c40e628fae1e4e6093a04e8c0f7cd0fa2025-08-20T01:50:06ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262025-03-0113310.1136/jitc-2024-011308Single-cell spatial transcriptomics unravels cell states and ecosystems associated with clinical response to immunotherapySjoerd H van der Burg0Ziena Abdulrahman1Mariette I E van Poelgeest2Roderick C Slieker3Marij J P Welters4Daniel McGuire5Department of Medical Oncology, Leiden University Medical Center, Leiden, The NetherlandsMedical Oncology, Oncode Institute, Leiden University Medical Center, Leiden, The NetherlandsDepartment of Gynecology, Leiden University Medical Center, Leiden, The Netherlandspostdoctoral fellowOncode Institute, Utrecht, The Netherlands4 NanoString Technologies Inc, Seattle, Washington, USABackground The tumor microenvironment (TME) is a complex and dynamic ecosystem that is known to influence responses to immunotherapy. We leveraged single-cell spatial transcriptomics to systematically dissect the intricate complexity of the TME, in particular the cellular heterogeneity and spatial interactions. Their collective impact on immunotherapy efficacy was studied in the context of a homogeneous group of patients with vulvar high-grade squamous intraepithelial lesions (vHSIL) treated with an immunotherapeutic tumor-specific peptide vaccine.Methods We performed single-cell spatial transcriptomics on 20 pretreatment vHSIL lesions, stratified by clinical response to immunotherapeutic vaccination into complete responders (CR), partial responders (PR) and non-responders (NR). Using a 1,000-gene panel, we mapped over 274,000 single cells in situ, identifying 18 cell clusters and 99 distinct non-epithelial cell states. Findings were validated against public single-cell transcriptomic data sets to assess their broader relevance across tumor types.Results Profound heterogeneity within the TME was detected across the response groups. CR lesions exhibited a higher ratio of immune-supportive to immune-suppressive cells—a pattern mirrored in other solid tumors following neoadjuvant checkpoint blockade. Key immune populations enriched in CRs included CD4+CD161+ effector T cells and chemotactic CD4+ and CD8+ T cells. Conversely, PRs were characterized by increased proportions of T helper 2 cells and CCL18-expressing macrophages, which are associated with the recruitment of type 2 T cells and regulatory T cells. NRs displayed preferential infiltration with immunosuppressive fibroblasts. Distinct spatial immune ecosystems further defined response groups. Although a number of immune cells were detected in all patients, type 1 effector cells dominated interactions in CRs, type 2 cells were prominently interacting in PRs, while NRs lacked organized immune cell interactions.Conclusions This study underscores the dual importance of both cellular composition and spatial organization in steering clinical response to immunotherapy.https://jitc.bmj.com/content/13/3/e011308.full
spellingShingle Sjoerd H van der Burg
Ziena Abdulrahman
Mariette I E van Poelgeest
Roderick C Slieker
Marij J P Welters
Daniel McGuire
Single-cell spatial transcriptomics unravels cell states and ecosystems associated with clinical response to immunotherapy
Journal for ImmunoTherapy of Cancer
title Single-cell spatial transcriptomics unravels cell states and ecosystems associated with clinical response to immunotherapy
title_full Single-cell spatial transcriptomics unravels cell states and ecosystems associated with clinical response to immunotherapy
title_fullStr Single-cell spatial transcriptomics unravels cell states and ecosystems associated with clinical response to immunotherapy
title_full_unstemmed Single-cell spatial transcriptomics unravels cell states and ecosystems associated with clinical response to immunotherapy
title_short Single-cell spatial transcriptomics unravels cell states and ecosystems associated with clinical response to immunotherapy
title_sort single cell spatial transcriptomics unravels cell states and ecosystems associated with clinical response to immunotherapy
url https://jitc.bmj.com/content/13/3/e011308.full
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