3D-Bioprinted Co-Cultures of Glioblastoma Multiforme and Mesenchymal Stromal Cells Indicate a Role for Perivascular Niche Cells in Shaping Glioma Chemokine Microenvironment
Katarzyna Zielniok,
Kinga Rusinek,
Anna Słysz,
Mieszko Lachota,
Ewa Bączyńska,
Natalia Wiewiórska-Krata,
Anna Szpakowska,
Martyna Ciepielak,
Bartosz Foroncewicz,
Krzysztof Mucha,
Radosław Zagożdżon,
Zygmunt Pojda
Affiliations
Katarzyna Zielniok
Laboratory of Cellular and Genetic Therapies, Center for Preclinical Research, Medical University of Warsaw, 02-097 Warsaw, Poland
Kinga Rusinek
Department of Regenerative Medicine, Maria Skłodowska-Curie National Research Institute of Oncology, 02-781 Warsaw, Poland
Anna Słysz
Department of Regenerative Medicine, Maria Skłodowska-Curie National Research Institute of Oncology, 02-781 Warsaw, Poland
Mieszko Lachota
Laboratory of Cellular and Genetic Therapies, Center for Preclinical Research, Medical University of Warsaw, 02-097 Warsaw, Poland
Ewa Bączyńska
Department of Regenerative Medicine, Maria Skłodowska-Curie National Research Institute of Oncology, 02-781 Warsaw, Poland
Natalia Wiewiórska-Krata
Laboratory of Cellular and Genetic Therapies, Center for Preclinical Research, Medical University of Warsaw, 02-097 Warsaw, Poland
Anna Szpakowska
Department of Regenerative Medicine, Maria Skłodowska-Curie National Research Institute of Oncology, 02-781 Warsaw, Poland
Martyna Ciepielak
Department of Regenerative Medicine, Maria Skłodowska-Curie National Research Institute of Oncology, 02-781 Warsaw, Poland
Bartosz Foroncewicz
Promix (ProteogenOmix in Medicine), Department of Clinical Immunology, Medical University of Warsaw, 02-006 Warsaw, Poland
Krzysztof Mucha
Promix (ProteogenOmix in Medicine), Department of Clinical Immunology, Medical University of Warsaw, 02-006 Warsaw, Poland
Radosław Zagożdżon
Laboratory of Cellular and Genetic Therapies, Center for Preclinical Research, Medical University of Warsaw, 02-097 Warsaw, Poland
Zygmunt Pojda
Department of Regenerative Medicine, Maria Skłodowska-Curie National Research Institute of Oncology, 02-781 Warsaw, Poland
3D bioprinting has become a valuable tool for studying the biology of solid tumors, including glioblastoma multiforme (GBM). Our analysis of publicly available bulk RNA and single-cell sequencing data has allowed us to define the chemotactic profile of GBM tumors and identify the cell types that secrete particular chemokines in the GBM tumor microenvironment (TME). Our findings indicate that primary GBM tissues express multiple chemokines, whereas spherical monocultures of GBM cells significantly lose this diversity. Subsequently, the comparative analysis of GBM spherical monocultures vs. 3D-bioprinted multicultures of cells showed a restoration of chemokine profile diversity in 3D-bioprinted cultures. Furthermore, single-cell RNA-Seq analysis showed that cells of the perivascular niche (pericytes and endocytes) express multiple chemokines in the GBM TME. Next, we 3D-bioprinted cells from two glioblastoma cell lines, U-251 and DK-MG, alone and as co-cultures with mesenchymal stromal cells (representing cells of the perivascular niche) and assessed the chemokine secretome. The results clearly demonstrated that the interaction of tumors and mesenchymal cells leads to in a significant increase in the repertoire and levels of secreted chemokines under culture in 21% O2 and 1% O2. Our study indicates that cells of the perivascular niche may perform a substantial role in shaping the chemokine microenvironment in GBM tumors.