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Tissue Engineering

Tissue Engineering - Poster Session C

Poster L18 - Investigating Mediators of Interstitial Fluid Flow Enhanced Invasion in Glioblastoma

Friday, October 25, 2024
10:00 AM – 11:00 AM EST
Location: Exhibit Hall E, F & G

Introduction:  Glioblastoma is a malignant cancer found in the central nervous system that invades healthy tissue via interstitial fluid flow (IFF) at the tumor border. IFF-enhanced invasion is driven by molecular mediators such as sphingosine-1-phosphate receptor 3 (S1PR3), wherein preliminary work in the lab indicates that targeting S1PR3 reduces invasion in a mouse glioma model. The glioblastoma tumor microenvironment (TME) is supported by glial cells such as astrocytes and microglia, which sensitize glioblastoma stem cells (GSCs) to S1PR3 inhibitor TY52156. The TME mediates molecular processes of invasion, but the cytokines that aid in this mediation are yet to be known. One promising cytokine, TGF-β, is linked to invasion in multiple cancer types, including glioblastoma. Preliminary work demonstrates that astrocytes and microglia sheared with TGF-β inhibitor IM412 lowered chemotaxis in G34 GSCs.

Materials and

Methods:  Correlations between S1PR3 expression in glia and flow paths were found in mouse glioma model GL261 using immunohistochemistry and time-series MRI. Mice were injected with GL261, and gadolinium was used to track fluid flow within and adjacent to the tumor. Pixel flow metrics generated by the software Lymph4D were correlated to immunohistochemistry data with custom code. Brain slices were stained for glial cell markers and S1PR3 positive cells were identified using a machine learning model. To examine this correlation in a 3D culture system that replicates IFF in the TME, collagen hyaluronan hydrogels were seeded with glial cells and exposed to a fluid pressure head (flow). To look at GSC invasion, the same 3D culture transwell system consisting of collagen hyaluronan hydrogels was used. The hydrogels were seeded with either G34 cells alone or G34s and astrocytes combined, and then exposed to either static or flow with the TGF-β inhibitor IM412 or vehicle. Invasion was quantified by counting the number of invaded cells within a given area at the center of each well membrane and calculating the percentage of invaded cells on the total membrane.

Results, Conclusions, and Discussions: The percentage of S1PR3 positive astrocytes in the peritumoral region of the mouse glioma model is strongly positively correlated to flow originating from inside the tumor (Figure 1). Preliminary assays suggest that S1PR3 RNA may be moderately increased in the presence of glia in flow, but future assays will validate this work in other models of flow. Results from the TGF-β invasion assay will be presented at this meeting.

Acknowledgements (Optional):