Q1 · MEDICINE
Article
Author: De Angelis, Biagio ; Caruso, Simona ; Carai, Andrea ; Pellegrino, Marsha ; Ajmone-Cat, Maria Antonietta ; Ferretti, Roberta ; Businaro, Pietro ; Jones, Chris ; Rossi, Sabrina ; Rota, Rossella ; de Billy, Emmanuel ; De Stefanis, Cristiano ; Palma, Alessia ; Pezzullo, Marco ; Del Bufalo, Francesca ; Caruana, Ignazio ; Vinci, Maria ; Petrilli, Lucia Lisa ; Orlando, Domenico ; Locatelli, Franco ; Bencivenga, Paola ; Massimi, Luca ; Pericoli, Giulia ; Quintarelli, Concetta ; Maestro, Nicola ; Weber, Gerrit ; Diomedi-Camassei, Francesca ; Mastronuzzi, Angela
Background:Diffuse midline gliomas (DMG) H3K27M-mutant, including diffuse intrinsic pontine glioma (DIPG), are pediatric brain tumors associated with grim prognosis. Although GD2-CAR T-cells demonstrated significant anti-tumor activity against DMG H3K27M-mutant in vivo, a multimodal approach may be needed to more effectively treat patients. We investigated GD2 expression in DMG/DIPG and other pediatric high-grade gliomas (pHGG) and sought to identify chemical compounds that would enhance GD2-CAR T-cell anti-tumor efficacy.
Methods:Immunohistochemistry in tumor tissue samples and immunofluorescence in primary patient-derived cell lines were performed to study GD2 expression. We developed a high-throughput cell-based assay to screen 42 kinase inhibitors in combination with GD2-CAR T-cells. Cell viability, western blots, flow-cytometry, real time PCR experiments, DIPG 3D culture models, and orthotopic xenograft model were applied to investigate the effect of selected compounds on DIPG cell death and CAR T-cell function.
Results:GD2 was heterogeneously, but widely, expressed in the tissue tested, while its expression was homogeneous and restricted to DMG/DIPG H3K27M-mutant cell lines. We identified dual IGF1R/IR antagonists, BMS-754807 and linsitinib, able to inhibit tumor cell viability at concentrations that do not affect CAR T-cells. Linsitinib, but not BMS-754807, decreases activation/exhaustion of GD2-CAR T-cells and increases their central memory profile. The enhanced anti-tumor activity of linsitinib/GD2-CAR T-cell combination was confirmed in DIPG models in vitro, ex vivo, and in vivo.
Conclusion:Our study supports the development of IGF1R/IR inhibitors to be used in combination with GD2-CAR T-cells for treating patients affected by DMG/DIPG and, potentially, by pHGG.