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Published on: October 20, 2016
Tumor-Restricted CXCR4 and MIF Signaling Axis is Associated with T Cell-Mediated Macrophage Exclusion in
Fulai Gao1, Wei Lu2, Dongqiang Zhao3
1Department of Gastroenterology, The First Hospital of Qinhuangdao, Qinhuangdao, Hebei, 066000, People's Republic of China.
Purpose:
Gastrointestinal stromal tumors (GISTs), driven by KIT/PDGFRA mutations and treated with tyrosine kinase inhibitors (TKIs), remain challenged by drug resistance, suggesting that poorly understood mechanisms sustain an immunosuppressive tumor microenvironment (TME). This study aimed to investigate the cellular and molecular basis of the immune architecture in GISTs through integrated multi-omics analysis.
Patients And Methods:
We integrated bulk RNA-sequencing data from 20 GISTs and 20 matched normal tissue samples (GSE225819) with single-cell RNA-sequencing data from 2 GISTs and 2 normal tissue samples (GSE162115) to dissect the cellular and molecular basis of TME. Weighted gene co-expression network analysis (WGCNA), immune deconvolution (CIBERSORTx), cell-cell communication inference (CellChat), and pseudotime trajectory analysis (Monocle 3) were employed. In vitro functional validation was performed using the GIST-T1 cell line, and clinical specimens were analyzed by RT-qPCR and flow cytometry.
Results:
Transcriptomic profiling of GISTs revealed an inverse correlation between activated T-cell signatures and macrophage abundance, alongside significant transcriptional remodeling (1071 upregulated and 1211 downregulated genes). scRNA-seq further identified a tumor cell subpopulation with high C-X-C Motif Chemokine Receptor 4 (CXCR4) expression that, together with activated CD4⁺ memory T cells, produced macrophage migration inhibitory factor (MIF). Ligand-receptor analysis identified the MIF pathway as the dominant intercellular signaling network, primarily targeting macrophages, thereby offering a mechanistic basis for their reduced presence in the TME. In vitro, CXCR4 knockdown suppressed MIF mRNA expression and cell proliferation. In clinical specimens, tumor tissue CXCR4 and MIF transcripts were elevated and positively correlated with T-cell-derived MIF protein, which inversely correlated with macrophage infiltration.
Conclusion:
Our study identifies a CXCR4⁺ GISTs tumor subpopulation associated with CD4⁺ T cell activation and MIF signaling amplification, which correlates with reduced macrophage infiltration. These findings nominate the CXCR4/MIF axis as a candidate therapeutic target, although further functional validation is warranted.
Insights
Gastrointestinal stromal tumors (GISTs) exhibit a CXCR4-positive subpopulation that reduces macrophage infiltration by amplifying MIF signaling. This CXCR4/MIF axis presents a potential therapeutic target for overcoming drug resistance in GISTs.
Area of Science:
- Oncology
- Immunology
- Genomics
Background:
- Gastrointestinal stromal tumors (GISTs) are driven by KIT/PDGFRA mutations and treated with tyrosine kinase inhibitors (TKIs).
- Drug resistance in GISTs is a significant challenge, potentially linked to an immunosuppressive tumor microenvironment (TME).
- Understanding the immune architecture of GISTs is crucial for developing effective therapies.
Purpose of the Study:
- To investigate the cellular and molecular basis of the immune microenvironment in GISTs.
- To identify mechanisms sustaining an immunosuppressive TME in GISTs.
- To explore potential therapeutic targets for GIST treatment.
Main Methods:
- Integrated multi-omics analysis including bulk and single-cell RNA-sequencing.
- Weighted gene co-expression network analysis (WGCNA) and immune deconvolution (CIBERSORTx).
- Cell-cell communication inference (CellChat), pseudotime trajectory analysis (Monocle 3), in vitro validation, RT-qPCR, and flow cytometry.
Main Results:
- Transcriptomic profiling revealed an inverse correlation between T-cell signatures and macrophage abundance in GISTs.
- A GIST subpopulation with high CXCR4 expression was identified, producing MIF with CD4+ T cells.
- The MIF pathway was identified as a dominant signaling network targeting macrophages, explaining reduced infiltration.
Conclusions:
- A CXCR4-positive GIST subpopulation is associated with CD4+ T cell activation and MIF signaling, correlating with reduced macrophage infiltration.
- The CXCR4/MIF axis is nominated as a candidate therapeutic target for GISTs.
- Further functional validation of the CXCR4/MIF axis is warranted for therapeutic development.
