ID #1017 Microphysiological Culture of Paediatric Low-Grade Glioma Slices: Towards Establishing a more Predictive Preclinical Model
Esmee Broekhuizen, Tesi Liu, Benjamin Thierry, Marnie Winter, Jasmine Kilyen-Coles, Kirk Jensen, Jordan Hansford, Lisa EbertAbstract
Background
Paediatric low-grade glioma (pLGG) is the most common brain tumour in children. Despite a favourable prognosis compared to paediatric high-grade glioma (pHGG), pLGG patients suffer substantial tumour- and treatment-associated morbidities. This warrants the need for novel treatment strategies. While pLGG has benefited from major advances in molecular profiling, uncovering key alterations driving pLGG biology and identifying putative therapeutic targets, translation of these discoveries into broadly effective treatments remains slow. A recognised bottleneck in the development of new treatment options for these tumours is the failure of current preclinical model systems to recapitulate the molecular complexity, tumour microenvironment (TME), and indolent biology of pLGG, which greatly limits their translational relevance.
Objectives
Develop and assess a preclinical model of pLGG designed to preserve the TME and phenotypical heterogeneity of parent tumours, towards addressing a critical bottleneck in paediatric neuro-oncology and accelerating the translation of novel pLGG treatments into the clinic.
Methods
A pumpless microphysiological platform was used for organotypic tissue slice (400 µm thickness) culture. Adult GBM tissue was used for optimisation, as pLGG tissue is scarce. Rocker-induced gravity perfusion enabled continuous nutrient and oxygen delivery to the tissue slice. Functional assays included 3D CellTiter-Glo, cleaved caspase-3, Glucose-Glo, and Lactate-Glo. In addition, a 3D pathology workflow was applied to tissue slices. For this, samples were optically cleared and stained using eosin and TO-PRO-3, enabling high-resolution volumetric light sheet microscopy imaging of the tissue structure over culture time.
Results
Tissue slices better maintained viability under perfused conditions compared to static conditions, with high Ki-67 and low cleaved-caspase 3, as well as metabolic activity and cytoarchitecture, based on H&E staining. 3D Pathology further confirmed well-preserved tissue structure under perfusion, while static conditions resulted in nuclear condensation and structural degradation in long term culture.
Conclusions
This microphysiological system enables perfusion of brain tumour tissue slices and preserves the TME. Its ongoing application to pLGG tissue is anticipated to provide greater physiological relevance compared to current standard models for studying pLGG biology and therapeutic responses.