Poster Presentation Hunter Cell Biology Meeting 2025

An optimized explant organoid method for preclinical brain tumour research (#117)

Helen M. Palethorpe 1 , Chloe Shard 1 , Kaitlin G. Scheer 1 , Conor P. Ryan 1 , Anahita Fouladzadeh 1 , Daniel Wenholz 2 , Quach Truong 2 , Niclas Skarne 3 4 , Erica C.F. Yeo 1 , Annika R. Mascarenhas 5 , Vanesa M. Tomatis 5 , Rebecca J. Ormsby 6 , Parul Mittal 7 , Clifford Young 7 , Manuela Klingler-Hoffmann 7 , Peter Hoffmann 7 , Anna Brown 8 , Joe McConnell 8 , Dominik Kaczorowski 8 , Santosh Poonnoose 5 6 , Lisa M. Ebert 1 9 10 , Tessa Gargett 1 9 10 , Kimberley L. Alexander 11 12 13 , Naresh Kumar 14 , Hui K. Gan 15 16 17 , Bryan W. Day 3 4 18 , Ashwini Patil 19 , John Wilkinson 2 , Guillermo A. Gomez 1
  1. Centre for Cancer Biology, University of South Australia and SA Pathology, Adelaide, SA 5000, Australia
  2. Noxopharm, Castle Hill, NSW 2154, Australia
  3. QIMR Berghofer Medical Research Institute, Brisbane, QLD 4006, Australia
  4. The University of Queensland, Brisbane, QLD 4072, Australia
  5. Department of Neurosurgery, Flinders Medical Centre, Adelaide, SA 5042, Australia
  6. Flinders University, College of Medicine and Public Health, Flinders Health and Medical Research Institute, Adelaide, SA 5042, Australia
  7. Clinical & Health Sciences, University of South Australia, Adelaide, SA 5000, Australia
  8. Department of Genetics and Molecular Pathology, SA Pathology, Adelaide, SA 5000, Australia
  9. Adelaide Medical School, University of Adelaide, Adelaide, SA 5000, Australia
  10. Cancer Clinical Trials Unit, Royal Adelaide Hospital, Adelaide, SA 5000, Australia
  11. Brain Cancer Research, Neurosurgery Department, Chris O'Brien Lifehouse, Camperdown, NSW 2050, Australia
  12. School of Medical Sciences, The University of Sydney, Camperdown, NSW 2050, Australia
  13. Department of Neuropathology, Royal Prince Alfred Hospital, Camperdown, NSW 2050, Australia
  14. School of Chemistry, The University of New South Wales, UNSW Sydney, NSW 2052, Australia
  15. Cancer Therapies and Biology Group, Centre of Research Excellence in Brain Tumors, Olivia Newton-John Cancer Wellness and Research Centre, Austin Hospital, Heidelberg, Victoria 3084, Australia
  16. La Trobe University School of Cancer Medicine, Heidelberg, Victoria 3084, Australia
  17. Department of Medicine, University of Melbourne, Heidelberg, Victoria 3084, Australia
  18. School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD 4072, Australia
  19. Combinatics, Chiba, 272-0824, Japan

Aim: Tumour heterogeneity presents a significant obstacle to identifying effective treatments for brain cancer, explaining the ineffectiveness of standard treatment protocols. Patient-derived brain tumour explant organoids (GBOs) replicate this heterogeneity and can predict a patient's response to therapy, constituting a critical tool in precision Neuro-oncology. However, current methods to generate GBOs are time-consuming, costly, and technically challenging. These limitations preclude the development of larger diverse organoid biobanks that capture the true spectrum of the disease and impede rapid patient-personalised preclinical testing.

Methods: By introducing semi-automated tissue processing, size exclusion chromatography and immediate cryopreservation, we optimized current methods1 for the generation and biobanking of tumour pieces (from low-grade glioma, high-grade glioma and primary/recurrent glioblastoma) that are suitable for GBO culture, reducing processing time from up to 6 hours to less than 60 minutes. We used this biobank to compare the effects of novel idronoxil-conjugated benzopyran compounds (NX786, NX904) on GBO growth, viability, cell death, invasion and senescence utilizing a combination of live and fixed imaging and multi-omics.

Results: We used our optimized protocol to generate 24 GBOs from 33 samples of diverse brain tumour types (low-grade glioma, high-grade glioma and primary/recurrent glioblastoma) and tested the effect of NX786 and NX904 on 9 of these. NX786 and NX904 significantly reduced GBO growth (≥50%) and invasion. Low-grade gliomas were more resistant to these treatments. NX904 induced significant cell death (>60%), while NX786 increased cellular senescence and induction of the mesenchymal-like state, with its effects being reversible.

Conclusions: We developed an efficient method for processing and cryopreserving brain tumour tissues suitable for GBO growth, enabling the capture of more patient samples and the establishment of an extensive biobank collection. We successfully used this biobank for targeted drug screening, highlighting its potential for patient-personalised preclinical testing. We found that through the inhibition of crucial brain cancer hallmarks, including proliferation and invasion, together with the induction of a less aggressive senescence phenotype, NX786 and NX904 have the potential for development as new therapeutic agents for brain cancer.

References:

  1. Jacob, F., Salinas, R.D., Zhang, D.Y., Nguyen, P.T.T., Schnoll, J.G., Wong, S.Z.H., Thokala, R., Sheikh, S., Saxena, D., Prokop, S., et al. (2020). A Patient-Derived Glioblastoma Organoid Model and Biobank Recapitulates Inter- and Intra-tumoral Heterogeneity. Cell 180, 188-204 e122. 10.1016/j.cell.2019.11.036.