Dual IGF1R/IR inhibitors in combination with GD2-CAR T-cells display a potent anti-tumor activity in diffuse midline glioma H3K27M-mutant.

Abstract

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.

• Authors: Emmanuel de Billy, Marsha Pellegrino, Domenico Orlando, Giulia Pericoli, Roberta Ferretti, Pietro Businaro, Maria Antonietta Ajmone-Cat, Sabrina Rossi, Lucia Lisa Petrilli, Nicola Maestro, Francesca Diomedi-Camassei, Marco Pezzullo, Cristiano De Stefanis, Paola Bencivenga, Alessia Palma, Rossella Rota, Francesca Del Bufalo, Luca Massimi, Gerrit Weber, Chris Jones, Andrea Carai, Simona Caruso, Biagio De Angelis, Ignazio Caruana, Concetta Quintarelli, Angela Mastronuzzi, Franco Locatelli, Maria Vinci
• Journal: Neuro-oncology (Neuro Oncol) Vol. 24 Issue 7 Pg. 1150-1163 (07 01 2022) ISSN: 1523-5866 [Electronic] England
• PMID: 34964902 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © The Author(s) 2021. Published by Oxford University Press on behalf of the Society for Neuro-Oncology.

Chemical References

• IGF1R protein, human
• Receptor, IGF Type 1
• Receptor, Insulin

Topics

• Brain Stem Neoplasms (pathology)
• Child
• Glioma (drug therapy, genetics, pathology)
• Humans
• Immunotherapy, Adoptive
• Receptor, IGF Type 1 (antagonists & inhibitors)
• Receptor, Insulin (antagonists & inhibitors)
• T-Lymphocytes (metabolism)