Harnessing single-cell and multi-omics insights: STING pathway-based predictive signature for immunotherapy response in lung adenocarcinoma

BackgroundLung adenocarcinoma is the most prevalent type of small-cell carcinoma, with a poor prognosis. For advanced-stage patients, the efficacy of immunotherapy is suboptimal. The STING signaling pathway plays a pivotal role in the immunotherapy of lung adenocarcinoma; therefore, further investig...

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Main Authors: Yang Ding, Dingli Wang, Dali Yan, Jun Fan, Zongli Ding, Lei Xue
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-04-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1575084/full
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author Yang Ding
Dingli Wang
Dali Yan
Jun Fan
Zongli Ding
Lei Xue
author_facet Yang Ding
Dingli Wang
Dali Yan
Jun Fan
Zongli Ding
Lei Xue
author_sort Yang Ding
collection DOAJ
description BackgroundLung adenocarcinoma is the most prevalent type of small-cell carcinoma, with a poor prognosis. For advanced-stage patients, the efficacy of immunotherapy is suboptimal. The STING signaling pathway plays a pivotal role in the immunotherapy of lung adenocarcinoma; therefore, further investigation into the relationship between the STING pathway and lung adenocarcinoma is warranted.MethodsWe conducted a comprehensive analysis integrating single-cell RNA sequencing (scRNA-seq) data with bulk transcriptomic profiles from public databases (GEO, TCGA). STING pathway-related genes were identified through Genecard database. Advanced bioinformatics analyses using R packages (Seurat, CellChat) revealed transcriptomic heterogeneity, intercellular communication networks, and immune landscape characteristics. We developed a STING pathway-related signature (STINGsig) using 101 machine learning frameworks. The functional significance of ERRFI1, a key component of STINGsig, was validated through mouse models and multicolor flow cytometry, particularly examining its role in enhancing antitumor immunity and potential synergy with α-PD1 therapy.ResultsOur single-cell analysis identified and characterized 15 distinct cell populations, including epithelial cells, macrophages, fibroblasts, T cells, B cells, and endothelial cells, each with unique marker gene profiles. STING pathway activity scoring revealed elevated activation in neutrophils, epithelial cells, B cells, and T cells, contrasting with lower activity in inflammatory macrophages. Cell-cell communication analysis demonstrated enhanced interaction networks in high-STING-score cells, particularly evident in fibroblasts and endothelial cells. The developed STINGsig showed robust prognostic value and revealed distinct immune microenvironment characteristics between risk groups. Notably, ERRFI1 knockdown experiments confirmed its significant role in modulating antitumor immunity and enhancing α-PD1 therapy response.ConclusionThe STING-related pathway exhibited distinct expression levels across 15 cell populations, with high-score cells showing enhanced tumor-promoting pathways, active immune interactions, and enrichment in fibroblasts and IFI27+ inflammatory macrophages. In contrast, low-score cells were associated with epithelial phenotypes and reduced immune activity. We developed a robust STING pathway-related signature (STINGsig), which identified key prognostic genes and was linked to the immune microenvironment. Through in vivo experiments, we confirmed that knockdown of ERRFI1, a critical gene within the STINGsig, significantly enhances antitumor immunity and synergizes with α-PD1 therapy in a lung cancer model, underscoring its therapeutic potential in modulating immune responses.
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spelling doaj-art-dcecffdb2b4c4b8293f5c87d3d0c01252025-08-20T03:53:39ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-04-011610.3389/fimmu.2025.15750841575084Harnessing single-cell and multi-omics insights: STING pathway-based predictive signature for immunotherapy response in lung adenocarcinomaYang Ding0Dingli Wang1Dali Yan2Jun Fan3Zongli Ding4Lei Xue5Department of Pathology, Nanjing Drum Tower Hospital Group Suqian Hospital, Suqian, ChinaDepartment of Lung Cancer, Tianjin Lung Cancer Center, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin’s Clinical Research Center for Cancer, Tianjin Medical University Cancer Institute and Hospital, Tianjin, ChinaDepartment of Oncology, The Affiliated Huai’an Hospital of Xuzhou Medical University and the Second People’s Hospital of Huai’an, Huai’an, ChinaDepartment of Thoracic Surgery, The First Affiliated Hospital with Nanjing Medical University, Nanjing, ChinaDepartment of Geriatrics, The Affiliated Huai’an Hospital of Xuzhou Medical University and the Second People’s Hospital of Huai’an, Huai’an, ChinaDepartment of Thoracic Surgery, The First Affiliated Hospital with Nanjing Medical University, Nanjing, ChinaBackgroundLung adenocarcinoma is the most prevalent type of small-cell carcinoma, with a poor prognosis. For advanced-stage patients, the efficacy of immunotherapy is suboptimal. The STING signaling pathway plays a pivotal role in the immunotherapy of lung adenocarcinoma; therefore, further investigation into the relationship between the STING pathway and lung adenocarcinoma is warranted.MethodsWe conducted a comprehensive analysis integrating single-cell RNA sequencing (scRNA-seq) data with bulk transcriptomic profiles from public databases (GEO, TCGA). STING pathway-related genes were identified through Genecard database. Advanced bioinformatics analyses using R packages (Seurat, CellChat) revealed transcriptomic heterogeneity, intercellular communication networks, and immune landscape characteristics. We developed a STING pathway-related signature (STINGsig) using 101 machine learning frameworks. The functional significance of ERRFI1, a key component of STINGsig, was validated through mouse models and multicolor flow cytometry, particularly examining its role in enhancing antitumor immunity and potential synergy with α-PD1 therapy.ResultsOur single-cell analysis identified and characterized 15 distinct cell populations, including epithelial cells, macrophages, fibroblasts, T cells, B cells, and endothelial cells, each with unique marker gene profiles. STING pathway activity scoring revealed elevated activation in neutrophils, epithelial cells, B cells, and T cells, contrasting with lower activity in inflammatory macrophages. Cell-cell communication analysis demonstrated enhanced interaction networks in high-STING-score cells, particularly evident in fibroblasts and endothelial cells. The developed STINGsig showed robust prognostic value and revealed distinct immune microenvironment characteristics between risk groups. Notably, ERRFI1 knockdown experiments confirmed its significant role in modulating antitumor immunity and enhancing α-PD1 therapy response.ConclusionThe STING-related pathway exhibited distinct expression levels across 15 cell populations, with high-score cells showing enhanced tumor-promoting pathways, active immune interactions, and enrichment in fibroblasts and IFI27+ inflammatory macrophages. In contrast, low-score cells were associated with epithelial phenotypes and reduced immune activity. We developed a robust STING pathway-related signature (STINGsig), which identified key prognostic genes and was linked to the immune microenvironment. Through in vivo experiments, we confirmed that knockdown of ERRFI1, a critical gene within the STINGsig, significantly enhances antitumor immunity and synergizes with α-PD1 therapy in a lung cancer model, underscoring its therapeutic potential in modulating immune responses.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1575084/fullnon-small cell lung cancerlung adenocarcinomasingle-cell RNA sequencingprognosis signaturemachine learning
spellingShingle Yang Ding
Dingli Wang
Dali Yan
Jun Fan
Zongli Ding
Lei Xue
Harnessing single-cell and multi-omics insights: STING pathway-based predictive signature for immunotherapy response in lung adenocarcinoma
Frontiers in Immunology
non-small cell lung cancer
lung adenocarcinoma
single-cell RNA sequencing
prognosis signature
machine learning
title Harnessing single-cell and multi-omics insights: STING pathway-based predictive signature for immunotherapy response in lung adenocarcinoma
title_full Harnessing single-cell and multi-omics insights: STING pathway-based predictive signature for immunotherapy response in lung adenocarcinoma
title_fullStr Harnessing single-cell and multi-omics insights: STING pathway-based predictive signature for immunotherapy response in lung adenocarcinoma
title_full_unstemmed Harnessing single-cell and multi-omics insights: STING pathway-based predictive signature for immunotherapy response in lung adenocarcinoma
title_short Harnessing single-cell and multi-omics insights: STING pathway-based predictive signature for immunotherapy response in lung adenocarcinoma
title_sort harnessing single cell and multi omics insights sting pathway based predictive signature for immunotherapy response in lung adenocarcinoma
topic non-small cell lung cancer
lung adenocarcinoma
single-cell RNA sequencing
prognosis signature
machine learning
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1575084/full
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