Unsupervised clustering reveals noncanonical myeloid cell subsets in the brain tumor microenvironment

Cancer Immunol Immunother. 2025 Jan 3;74(2):63. doi: 10.1007/s00262-024-03920-1.

Abstract

The tumor immune microenvironment (TiME) of human central nervous system (CNS) tumors remains to be comprehensively deciphered. Here, we employed flow cytometry and RNA sequencing analysis for a deep data-driven dissection of a diverse TiME and to uncover noncanonical immune cell types in human CNS tumors by using seven tumors from five patients. Myeloid subsets comprised classical microglia, monocyte-derived macrophages, neutrophils, and two noncanonical myeloid subsets: CD3+ myeloids and CD19+ myeloids. T lymphocyte subsets included double-negative (CD4- CD8-) T cells (DNTs). Noncanonical myeloids and DNTs were explored on independent datasets, suggesting that our DNT phenotype represents γδ T cells. Noncanonical myeloids were validated using orthogonal methods across 73 patients from three independent datasets. While the proportions of classical myeloids agreed with reported malignancy type-associated TiMEs, unexpectedly high lymphocyte frequencies were detected in gliosarcoma, which also showed a unique expression pattern of immune-related genes. Our findings highlight the potential of data-driven approaches in resolving CNS TiME to reveal the mosaic of immune cell types constituting TiME, warranting the need for future studies on the nonclassical immune cell subsets.

Keywords: Brain metastasis; CD19+ myeloids; CD3+ myeloids; Deconvolution; Glioblastoma; Immunophenotyping; Single-cell sequencing; Tumor microenvironment; Unsupervised clustering.

MeSH terms

  • Brain Neoplasms* / genetics
  • Brain Neoplasms* / immunology
  • Brain Neoplasms* / pathology
  • Cluster Analysis
  • Female
  • Humans
  • Male
  • Myeloid Cells* / immunology
  • Myeloid Cells* / metabolism
  • Tumor Microenvironment* / immunology