Multiplex spatial analysis reveals increased CD137 expression and m-MDSC neighboring tumor cells in refractory classical Hodgkin Lymphoma

Oncoimmunology. 2024 Aug 10;13(1):2388304. doi: 10.1080/2162402X.2024.2388304. eCollection 2024.

Abstract

The Hodgkin and Reed - Sternberg (HRS) cells in classical Hodgkin Lymphoma (cHL) actively modify the immune tumor microenvironment (TME) attracting immunosuppressive cells and expressing inhibitory molecules. A high frequency of myeloid cells in the TME is correlated with an unfavorable prognosis, but more specific and rare cell populations lack precise markers. Myeloid-derived suppressor cells (MDSCs) have been identified in the peripheral blood of cHL patients, where they appear to be correlated with disease aggressiveness. TNFRSF9 (CD137) is a T cell co-stimulator expressed by monocytic and dendritic cells. Its expression has also been described in HRS cells, where it is thought to play a role in reducing antitumor responses. Here, we perform qualitative and quantitative analyses of lymphocytic and MDSC subtypes and determine the CD137 cell distribution in cHL primary tumors using multiplex immunofluorescence and automated multispectral imaging. The results were correlated with patients' clinical features. Cells were stained with specific panels of immune checkpoint markers (PD-1, PD-L1, CD137), tumor-infiltrating T lymphocytes (CD3, PD-1), and monocytic cells/MDSCs (CD68, CD14, CD33, Arg-1, CD11b). This approach allowed us to identify distinct phenotypes and to analyze spatial interactions between immune subpopulations and tumor cells. The results confirm CD137 expression by T, monocytic and HRS cells. In addition, the expression of CD137, T exhausted cells, and monocytic MDSCs (m-MDSCs) in the vicinity of malignant HRS cells were associated with a worse prognosis. Our findings reveal new elements of the TME that mediate immune escape, and confirm CD137 as a candidate target for immunotherapy in cHL.

Keywords: Classical Hodgkin lymphoma; multiplex immunofluorescence; myeloid-derived suppressor cells (MDSCs); tumor microenvironment; tumor necrosis factor receptor 9 (TNFRSF9/CD137).

Plain language summary

CD137-expressing immune cells and HRS cells are more abundant and in closer proximity in refractory patients than in responders.Monocytic myeloid-derived suppressor cells (m-MDSCs) are associated with unfavorable outcomes and relapse in cHL, unlike granulocytic MDSCs (g-MDSCs), which are located far from HRS cells in non-responders.The cHL tumor microenvironment promotes immune escape in refractory patients by holistically driving polarization and/or recruitment of several cell types with increased expression of CD137 and PD-L1 checkpoints.

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Biomarkers, Tumor / metabolism
  • Female
  • Hodgkin Disease* / immunology
  • Hodgkin Disease* / metabolism
  • Hodgkin Disease* / pathology
  • Humans
  • Lymphocytes, Tumor-Infiltrating / immunology
  • Lymphocytes, Tumor-Infiltrating / metabolism
  • Male
  • Middle Aged
  • Myeloid-Derived Suppressor Cells* / immunology
  • Myeloid-Derived Suppressor Cells* / metabolism
  • Myeloid-Derived Suppressor Cells* / pathology
  • Prognosis
  • Reed-Sternberg Cells* / metabolism
  • Reed-Sternberg Cells* / pathology
  • Spatial Analysis
  • Tumor Microenvironment* / immunology
  • Tumor Necrosis Factor Receptor Superfamily, Member 9* / metabolism
  • Young Adult

Substances

  • Tumor Necrosis Factor Receptor Superfamily, Member 9
  • Biomarkers, Tumor
  • TNFRSF9 protein, human

Grants and funding

This work was supported by Instituto de Salud Carlos III (ISCIII), projects PI19/00083 and PI22/00556 (co-funded by the European Union), and a Roche Foundation Research Grant. V.M. is a recipient of a Predoctoral Grant from ISCIII FIS (PFIS ref. FI20/00184) and the MD Anderson Foundation scholarship for projects in translational oncology 2022.