Integration of spatial transcriptomics with immunofluorescence staining reveals spatial heterogeneity and plasticity of astrocytes in experimental glioblastomas

Ghosh, M.; Pilanc-Kuldek, P.; Jacek, K.; Baluszek, S.; Poleszak, K.; Szadkowska, P.; Gielniewski, B.; Ellert-Miklaszewska, A.; Kaminska, B.

doi:10.1101/2024.10.11.617740

bioRxiv · 10.1101/2024.10.11.617740

Integration of spatial transcriptomics with immunofluorescence staining reveals spatial heterogeneity and plasticity of astrocytes in experimental glioblastomas

Ghosh, M.·Pilanc-Kuldek, P.·Jacek, K.·Baluszek, S.·Poleszak, K.·Szadkowska, P.·Gielniewski, B.·Ellert-Miklaszewska, A.·Kaminska, B.

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Abstract

Astrocytes comprise [~]50% of all brain cells and present distinct morphological, molecular and functional properties in different brain regions. In glioblastoma (GBM), an aggressive primary brain tumour, tumour-associated astrocytes (TAAs) become activated and exhibit different transcriptomic profiles, morphology and functions supporting disease progression. Heterogeneity and specific roles of TAAs within various regions of tumours are poorly known. Advancements of single-cell and spatial transcriptomics allow to profile tumours at unprecedented resolution revealing cell phenotypes, hidden functionalities and spatial architecture in disease-specific context. We combined spatial transcriptomics and multiple immunofluorescent staining to visualize TAAs heterogeneity and location of various subpopulations in intracranial murine gliomas. Using distinct gene expression profiles, we identified subtypes of TAAs with distinct localization and inferred their specialized functionalities. Gene signatures associated with TAAs reflected their reprograming in the tumour microenvironment (TME), revealed their multiple roles and potential contributing factors shaping the local milieu. Using spatial correlation analysis of the spots, we inferred the interactome of Slc1a2 (encoding a glutamate transporter) with the other markers of TAAs based on segregated areas of the tumour. The designer RGD peptide blocking tumour-microglia communications, alters the spatial distribution of TAAs in experimental gliomas providing insights into potential mechanisms. Spatial transcriptomics combined with multiple staining unveils multiple functional phenotypes of TAAs and interactions within TME. It shows their distinct morphology and unveils different roles in various regions of the tumour. We demonstrate the glioma-induced heterogeneity of TAAs and their adaption to the pharmacologically-induced modification of the immunosuppressive TME.

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Ghosh, M., Pilanc-Kuldek, P., Jacek, K., Baluszek, S., Poleszak, K., Szadkowska, P., Gielniewski, B., Ellert-Miklaszewska, A., Kaminska, B.. 2024-10-12. Integration of spatial transcriptomics with immunofluorescence staining reveals spatial heterogeneity and plasticity of astrocytes in experimental glioblastomas. https://doi.org/10.1101/2024.10.11.617740

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