CD271/p75NTR is a novel diagnostic marker, prognostic indicator and therapeutic target in SHH medulloblastoma

The extensive heterogeneity both between and within the medulloblastoma (MB) subgroups underscores a critical need for variant-specific biomarkers and therapeutic strategies. We previously identified a role for the CD271/p75 neurotrophin receptor (p75NTR) in regulating stem/progenitor cells in the SHH MB subgroup. Here, we demonstrate the utility of CD271 as a novel diagnostic and prognostic marker for SHH MB using immunohistochemical analysis as well as transcriptome data across 763 primary tumors. Characterization of CD271+ and CD271- cells by RNA sequencing revealed that these two subpopulations are molecularly distinct, co-existing cellular subsets both in vitro and in vivo. MAPK/ERK signaling is upregulated in the CD271+ population and inhibiting this pathway reduced CD271 levels, stem/progenitor cell proliferation and cell survival as well as cell migration in vitro. Importantly, the MEK inhibitor selumetinib extends survival and reduces CD271 levels in vivo. Our study demonstrates the clinical utility of CD271 as both a diagnostic and prognostic tool for SHH MB tumors and reveals a novel role for MEK inhibitors in targeting CD271+ SHH MB cells. Overall design: UI226 tumorspheres were dissociated and counted using an automated cell counter. Cells were stained with CD271 and 7-Aminoactinomycin D (7AAD) viability dye (Beckman Coulter, Inc., Mississauga ON, Canada) and then FACS sorted on the basis of CD271+ and CD271- expression using a MoFlo XDP cell sorter (Beckman Coulter, Inc.) at the University of Manitoba. Analysis was performed using FlowJo software (Tree Star Inc., Ashland, OR, USA). Sorted CD271+ and CD271- cells were collected and RNA was isolated using the Norgen RNA extraction kit (Norgen Biotek) according to manufacturer's instructions for mammalian cultured cells