Single cell imaging mass spectrometry spatial immune profiling of brain metastasis from non-small-cell lung cancer patients treated with immune checkpoint inhibitors.
9 affiliations
- 1Department of Medical Oncology and Therapeutics Research, City of Hope, Duarte, CA, USA.
- 2Department of Gastrointestinal Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
- 3Integrated Genome Core, Beckman Research Institute, City of Hope, Duarte, CA, USA.
- 4Department of Pathology, City of Hope, Duarte, CA, USA.
- 5Department of Pathology, City of Hope Chicago, Chicago, IL, USA.
- 6Light Microscopy/Digital Imaging Core, Department of Developmental and Stem Cell Biology, City of Hope, Duarte, CA, USA.
- 7Department of Surgery, Division of Neurosurgery, City of Hope, Duarte, CA, USA.
- 8Department of Diagnostic Radiology, City of Hope, Duarte, CA, USA.
- 9Department of Radiation Oncology, City of Hope, Duarte, CA, USA.
Abstract
Immune checkpoint inhibitors (ICIs), alone or in combination with chemotherapy, are a primary therapy for patients with advanced non-small cell lung cancer (NSCLC) without targetable driver alterations, yet brain metastases remain a major clinical challenge and their immune microenvironment is poorly characterized. This study aimed to describe the single-cell spatial immune microenvironment of brain metastases from ICI-treated NSCLC patients using imaging mass cytometry (IMC) and to explore features associated with response and resistance. Four brain metastasis tissues were available from three patients treated with ICIs, and an eleven-marker panel was stained on formalin-fixed paraffin-embedded (FFPE) slides for IMC analysis. Spatial cell count analyses and cell neighborhood analyses were performed to describe the cell population, distribution, and spatial neighborhood in the brain metastasis tumor microenvironment (TME). Given the number of specimens available, no formal statistical comparison between response groups was performed with all analyses done with descriptive statistics and exploratory. In this study, we found enrichment of CD4+FOXP3+ (27.6%), PanCK+CD68+PD-L1+CD33+ (22.7%), and PanCK+PD-L1+CD33+ (15.6%), and weak CD68+ (12.7%) in samples from the patient who exhibited a partial response (PR) to ICIs. In samples from progressive disease (PD) patients, we observed enrichment of PanCK+ (42.5%), weak PanCK+ (28.2%) and Arginase+ (10.7%). In the PR patient with two timepoints, the earlier sample was enriched with neighborhoods 2 and 3, which consist of PanCK+ cell clusters mixed with multiple markers and overlapping T Cells. The later sample was enriched with neighborhood 0 (CD4+FOXP3+) and neighborhood 1 (weak CD68+, CD4+FOXP3+), with neighborhood 1 surrounded by neighborhood 0. Weak CD68+ cells were present in both PR and PD patients, but the majority (98.7%) were observed in the single PR patient. We investigated the spatial TME features of brain metastases from NSCLC patients and identified distinct IMC phenotypic profiles, and suggested the potential role for immunosuppressive macrophage and Tregs in ICI resistance mechanisms in brain metastasis. Given the small number of patients, the absence of orthogonal validation and the restricted marker panel, further studies of more samples with comprehensive analysis are required.
Related
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BibTeX
@article{zhao2026single,
title = {{Single cell imaging mass spectrometry spatial immune profiling of brain metastasis from non-small-cell lung cancer patients treated with immune checkpoint inhibitors.}},
author = {Dan Zhao and Haiqing Li and Isa Mambetsariev and Tamara Mirzapoiazova and Jeremy Fricke and Javier Arias-Romero and Leonidas Arvanitis and Massimo D'Apuzzo and Bradford Tan and Michelle Afkhami and Brian Armstrong and Mike Y Chen and Behnam Badie and Bihong Beth Chen and Arya Amini and Ravi Salgia},
journal = {Translational lung cancer research},
year = {2026},
month = sep,
doi = {10.21037/tlcr-2026-0839},
url = {https://europepmc.org/article/MED/42999405},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Single cell imaging mass spectrometry spatial immune profiling of brain metastasis from non-small-cell lung cancer patients treated with immune checkpoint inhibitors.
AU - Dan Zhao
AU - Haiqing Li
AU - Isa Mambetsariev
AU - Tamara Mirzapoiazova
AU - Jeremy Fricke
AU - Javier Arias-Romero
AU - Leonidas Arvanitis
AU - Massimo D'Apuzzo
AU - Bradford Tan
AU - Michelle Afkhami
AU - Brian Armstrong
AU - Mike Y Chen
AU - Behnam Badie
AU - Bihong Beth Chen
AU - Arya Amini
AU - Ravi Salgia
PY - 2026
DA - 2026/09/29
JO - Translational lung cancer research
DO - 10.21037/tlcr-2026-0839
UR - https://europepmc.org/article/MED/42999405
AB - Immune checkpoint inhibitors (ICIs), alone or in combination with chemotherapy, are a primary therapy for patients with advanced non-small cell lung cancer (NSCLC) without targetable driver alterations, yet brain metastases remain a major clinical challenge and their immune microenvironment is poorly characterized. This study aimed to describe the single-cell spatial immune microenvironment of brain metastases from ICI-treated NSCLC patients using imaging mass cytometry (IMC) and to explore features associated with response and resistance. Four brain metastasis tissues were available from three patients treated with ICIs, and an eleven-marker panel was stained on formalin-fixed paraffin-embedded (FFPE) slides for IMC analysis. Spatial cell count analyses and cell neighborhood analyses were performed to describe the cell population, distribution, and spatial neighborhood in the brain metastasis tumor microenvironment (TME). Given the number of specimens available, no formal statistical comparison between response groups was performed with all analyses done with descriptive statistics and exploratory. In this study, we found enrichment of CD4+FOXP3+ (27.6%), PanCK+CD68+PD-L1+CD33+ (22.7%), and PanCK+PD-L1+CD33+ (15.6%), and weak CD68+ (12.7%) in samples from the patient who exhibited a partial response (PR) to ICIs. In samples from progressive disease (PD) patients, we observed enrichment of PanCK+ (42.5%), weak PanCK+ (28.2%) and Arginase+ (10.7%). In the PR patient with two timepoints, the earlier sample was enriched with neighborhoods 2 and 3, which consist of PanCK+ cell clusters mixed with multiple markers and overlapping T Cells. The later sample was enriched with neighborhood 0 (CD4+FOXP3+) and neighborhood 1 (weak CD68+, CD4+FOXP3+), with neighborhood 1 surrounded by neighborhood 0. Weak CD68+ cells were present in both PR and PD patients, but the majority (98.7%) were observed in the single PR patient. We investigated the spatial TME features of brain metastases from NSCLC patients and identified distinct IMC phenotypic profiles, and suggested the potential role for immunosuppressive macrophage and Tregs in ICI resistance mechanisms in brain metastasis. Given the small number of patients, the absence of orthogonal validation and the restricted marker panel, further studies of more samples with comprehensive analysis are required.
SN - 2218-6751
SN - 2226-4477
ER -