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The neurofibromin-1 tumor suppressor gene ( NF1 ) is altered in approximately 20% of sporadic glioblastoma (GBM) cases.  NF1  deficient GBM frequently shows a mesenchymal gene expression signature, suggesting a relationship between  NF1  status and the tumor microenvironment. To identify changes in the production of secreted cytokines/chemokines in  NF1  deficient glioma, we applied cytokine arrays to conditioned media from a panel of three GBM cell lines after siRNA-mediated  NF1  knockdown. We identified increased secretion of platelet-derived growth factor AA (PDGF-AA), chitinase-3-like protein 1 (CHI3L1), interleukin-8 (IL-8), and endoglin (ENG) in different subsets of these cell lines. Secretion was associated with induction of the corresponding messenger RNA, suggesting a mechanism involving transcriptional upregulation. By contrast, in non-transformed immortalized normal human astrocytes, PDGF-AA secretion was increased upon  NF1  knockdown, while secreted CHI3L1, ENG, and IL-8 were reduced or unchanged. Analysis of The Cancer Genome Atlas confirmed a relationship between glioma  NF1  status and  ENG  and  CHI3L1  in tumor samples. Overall, this study identifies candidate changes in secreted proteins from  NF1  deficient glioma cells that could influence the tumor microenvironment, and suggests a direct link between  NF1  loss and increased tumor cell production of CHI3L1 and endoglin, two factors implicated in mesenchymal identity in glioblastoma.

Neurofibromin knockdown in glioma cell lines is associated with changes in cytokine and chemokine secretion in vitro