Poster Presentation 27th Annual Lorne Proteomics Symposium 2022

Quantitative proteomic profiling of diffuse midline glioma cell lines treated with ONC201 identifies increased expression and surface localisation of members of the antigen presentation pathway (#167)

Mika Persson 1 2 , Evangeline R. Jackson 1 2 , Liesl Bramberger 3 , Ryan J. Duchatel 1 2 , Martin R. Larsen 4 , Nicholas A. Vitanza 5 6 , Pouya Faridi 3 , Matthew Dun 1 2
  1. Cancer Signalling Research Group, School of Biomedical Sciences & Pharmacy, College of Health, University of Newcastle, Callaghan, NSW, Australia
  2. Precision Medicine Program, Hunter Medical Research Institute, New Lambton Heights, NSW, Australia
  3. Department of Medicine, School of Clinical Sciences, Monash University, Melbourne, VIC, Australia
  4. Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark
  5. Ben Towne Center for Childhood Cancer Research, Seattle Children’s Research Institute, Seattle, WA, USA
  6. Division of Pediatric Hematology/Oncology, Department of Pediatrics, Seattle Children’s Hospital, Seattle, WA, USA

Diffuse midline glioma (DMG) is a paediatric high-grade glioma diagnosed along the critical structures of the midline of the central nervous system and harbours a median overall survival (OS) of 9-11 months, with <10% of patients surviving 2-years. Phase I/II clinical trial testing the small molecule imipridone ‘ONC201’ reports increased 20–month OS for patients with brainstem lesions. In vitro testing of ONC201 using 14 patient derived DMG cell line models revealed a heterogeneous response. Employing proteomic profiling of ONC201 resistant DMGs, we aimed to determine the mechanisms underpinning resistance. Using TMT-16plex quantitation we assessed the proteome of DMG cells -/+ ONC201 alone, and in combination with the PI3K inhibitor paxalisib for 24-hrs, by HR-nRPLC-MS/MS followed by peptide analysis using Proteome Discoverer 2.5, and de novo peptide sequencing using PEAKS. Proteomic profiling of ONC201 resistant SU-DIPG-VI cells, identified significantly increased expression of proteins mapping to the cytoprotective antioxidant response element axis following ONC201 treatment, including NQO1, as well as proteins explicitly linked to antigen presentation, including beta-2-microglobulin (B2M), HLA-A and HLA-B. Overexpression of NQO1 drives resistance to cytotoxic chemotherapies in multiple cancers, with loss of B2M correlated with resistance to immune checkpoint inhibitors (ICI) in metastatic melanoma, suggesting that B2M expression is crucial for immune initiated tumour cell death.1 Intriguingly, NQO1 inhibition by beta-lapachone has been shown to overcome ICI resistance in mouse models of cancer, suggesting that ONC201 treatment combined with NQO1 inhibition could make DMG more sensitive to ICIs.2 Using tissues resected from a ONC201 +/- paxalisib treated DMG patient-derived mouse model, we have confirmed the increased expression of NQO1 and B2M, highlighting the novel connection between ONC201 and immune modulation in DMG. Future work is focused on developing a multimodal and metronomic treatment paradigm, including radiotherapy, ONC201 and beta-lapachone to increase response of DMGs to ICIs.

  1. Sade-Feldman M, et al. Nat Commun. 2017. PMC5656607. 10.1038/s41467-017-01062-w
  2. Li X, et al. Nat Commun. 2019. PMC6642086. 10.1038/s41467-019-11238-1