487P - 3D-bioprinted co-cultures of glioblastoma and mesenchymal cells indicate a role for perivascular niche cells in shaping the chemotactic tumour microenvironment

Background

3D-Bioprinting has become a valuable tool for studying the biology of solid tumours, including the most aggressive and invasive primary brain tumour, glioblastoma multiforme (GBM). Intratumoural heterogeneity and the intricate tumor microenvironment (TME) communication contribute to the resistance of GBM cells to therapy and infiltration of immune effector cells. The generation of 3D-bioprinted glioblastoma models aims to more accurately reproduce the complex characteristics of GBM tissue compared to traditional cell cultures, especially when studying such a dynamic issue as chemokine secretion.

Methods

Here, we analysed publicly available bulk and single-cell RNA sequencing data to identify the cell types that secrete particular chemokines in the glioblastoma tumour-microenvironment. Next, we 3D-bioprinted cells from two glioblastoma cell lines, U251 and DK-MG, alone and as co-cultures with primary mesenchymal stromal cells, and assessed chemokine secretion using Luminex method.

Results

Our findings indicate that primary GBM tissues express multiple chemokines, whereas spherical cultures of GBM cells significantly lose this diversity. Comparative analysis of GBM spherical vs bioprinted tetra-cultures containing astrocytes, neuronal cells and macrophages, showed significant restoration of chemokine landscape diversity in 3D-bioprinted cultures. Furthermore, single cell RNA-Seq analysis showed that cells of the perivascular niche also express chemokines in the GBM TME. Analysis of our 3D-bioprinted cultures of cells from two GBM cell lines, U251 and DK-MG, alone and as co-cultures with mesenchymal stromal cells, clearly demonstrated that the interaction of tumour and mesenchymal cells results in a significant increase of the repertoire and levels of secreted chemokines.

Conclusions

Our study indicates that cells of perivascular niche may perform a substantial role in shaping the chemokine landscape in GBM tumours.

Clinical trial identification

Editorial acknowledgement

Legal entity responsible for the study

Maria Sklodowska-Curie National Research Institute of Oncology, Warsaw, Poland.

Funding

This research was funded by the National Science Centre, Poland (2020/37/B/NZ6/02191) and a subsidy via National Oncology Institute of Maria Sklodowska-Curie – National Research Institute (project BIODRUK-CAR; decision of Ministry of Science and Higher Education No. 89/WFSN/2021).

Disclosure

All authors have declared no conflicts of interest.