Multi-omics and spatial analysis of microgravity-grown glioblastoma organoids reveal superior modeling of advanced disease after long-term spaceflight.

Creating accurate organoid models remains a challenge, particularly for diseases like glioblastoma (GBM), an aggressive, treatment-resistant brain cancer with an immunosuppressive microenvironment. To overcome limitations of Earth-based models (sedimentation and disaggregation) and leverage the unique advantages of space (accelerated disease progression and immune dysregulation), this study utilizes a panel of GBM-myeloid organoids cultured for 40 d aboard the International Space Station. Spaceflight-grown GBM organoids have more uniform morphology compared to ground controls. We observed a context-dependent response to spaceflight, with the inclusion of macrophages and monocytes altering the bulk and spatial transcriptional response. While GBM-alone organoids have transcriptional markers of less aggressive disease in microgravity, organoids containing GBM cells + monocytes have increased expression of genes associated with disease progression and poor patient outcomes. Gene expression patterns are more spatially organized, with mesenchymal-related genes enriched in the core and inflammation-related genes enriched at the periphery, mimicking GBM tumor architecture. Microgravity-grown GBM+monocyte organoids exhibit increased secretion of proteins such as CXCL12 and LOX-1, associated with more aggressive disease. Infrared chemical imaging confirms spatial chemical gradients of DNA, protein, and lipid species in both space- and terrestrially-grown organoids. In summary, this work presents not only an advanced model of glioblastoma for basic and translational research but also demonstrates methods to acquire diverse data from organoids compatible with the unique constraints of in-space systems, helping establish a working model for orbital oncology.