MIF/CXCR4 signaling axis contributes to survival, invasion, and drug resistance of metastatic neuroblastoma cells in the bone marrow microenvironment

Abstract

Background The bone marrow (BM) is the most common site of dissemination in patients with aggressive, metastatic neuroblastoma (NB). However, the molecular mechanisms underlying the aggressive behavior of NB cells in the BM niche are still greatly unknown. In the present study, we explored biological mechanisms that play a critical role in NB cell survival and progression in the BM and investigated potential therapeutic targets. Methods Patient-derived bone marrow (BM) primary cultures were generated using fresh BM aspirates obtained from NB patients. NB cell lines were cultured in the presence of BM conditioned media containing cell-secreted factors, and under low oxygen levels (1% O2) to mimic specific features of the BM microenvironment of high-risk NB patients. The BM niche was explored using cytokine profiling assays, cell migration-invasion and viability assays, flow cytometry and analysis of RNA-sequencing data. Selective pharmacological inhibition of factors identified as potential mediators of NB progression within the BM niche was performed in vitro and in vivo. Results We identified macrophage migration inhibitory factor (MIF) as a key inflammatory cytokine involved in BM infiltration. Cytokine profiling and RNA-sequencing data analysis revealed NB cells as the main source of MIF in the BM, suggesting a potential role of MIF in tumor invasion. Exposure of NB cells to BM-conditions increased NB cell-surface expression of the MIF receptor CXCR4, which was associated with increased cell viability, enhanced migration-invasion, and activation of PI3K/AKT and MAPK/ERK signaling pathways. Moreover, subcutaneous co-injection of NB and BM cells enhanced tumor engraftment in mice. MIF inhibition with 4-IPP impaired in vitro NB aggressiveness, and improved drug response while delayed NB growth, improving survival of the NB xenograft model. Conclusions Our findings suggest that BM infiltration by NB cells may be mediated, in part, by MIF-CXCR4 signaling. We demonstrate the antitumor efficacy of MIF targeting in vitro and in vivo that could represent a novel therapeutic target for patients with disseminated high-risk NB.

Blocking MIF secretion enhances CAR T-cell efficacy against neuroblastoma

“…Interestingly, MIF was initially recognized as a lymphocyte-secreted factor influencing macrophage motility but was later described to be a multi-functional cytokine with contextdependent functions [50][51][52]67,68 . More recently, MIF has been described in the context of several malignancies, including neuroblastoma, to correlate with tumor growth, metastases and survival 54,55,[69][70][71] . MIF upregulation in neuroblastoma can induce MYCN expression, resulting in tumor progression 72 .…”

“…MIF upregulation in neuroblastoma can induce MYCN expression, resulting in tumor progression 72 . Moreover, MIF has been described to be involved in bone marrow metastases, where the expression of MIF contributes to survival and invasion of the tumor cells as well as an immunosuppressive tumor microenvironment 69,73 . Taken together, these studies suggest an important intrinsic role for MIF in neuroblastoma growth and progression, which makes MIF an attractive target for therapeutic intervention.…”

Plant MDL proteins synergize with the cytokine MIF at CXCR2 and CXCR4 receptors in human cells

“…The MIF-CXCR4-PI3K-AKT axis is a well-studied MIF response pathway, implicated among others in cell survival, migration, and cancer development ( 57 , 74 ). A549 cells were obtained from the German Collection of Microorganisms and Cell Cultures GmbH (DSMZ) and maintained in Ham’s F-12K medium (Invitrogen/Gibco) supplemented with 10% FBS and 1% penicillin-streptomycin (Sigma-Aldrich, Deisenhofen, Germany).…”

Exploring the Role of CD74 and D-Dopachrome Tautomerase in COVID-19: Insights from Transcriptomic and Serum Analyses

“…Like MIF, D-DT is involved in various biological processes, including inflammation, tumorigenesis, and cell survival. Both MIF and D-DT have been implicated in the pathogenesis of various diseases, including autoimmune diseases, cancer, and infectious diseases [9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27].…”

“…CD74 acts as the primary receptor for MIF, whereas CXCR2 and CXCR4 act as secondary receptors. D-DT also binds to CD74 but with lower affinity than MIF [9][10][11][12][13][14][15][16][17][18][19][20][21][22].…”