An engineered glioblastoma model yields macrophage-secreted drivers of invasion
Erin A. Akins, Dana Wilkins, Zaki Abou-Mrad, Kelsey Hopland, Robert C. Osorio, Kenny K.H. Yu, Manish K. Aghi, Sanjay Kumar
Erin A. Akins, Dana Wilkins, Zaki Abou-Mrad, Kelsey Hopland, Robert C. Osorio, Kenny K.H. Yu, Manish K. Aghi, Sanjay Kumar
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Research Article Immunology Oncology

An engineered glioblastoma model yields macrophage-secreted drivers of invasion

Abstract

While the accumulation of tumor-associated macrophages (TAMs) in glioblastoma (GBM) has been well documented, targeting TAMs has thus far yielded limited clinical success in slowing GBM progression due, in part, to an incomplete understanding of TAM function. Using an engineered 3D hydrogel–based model of the brain tumor microenvironment (TME), we show that M2-polarized macrophages stimulate transcriptional and phenotypic changes in GBM stem cells (GSCs) closely associated with the highly aggressive and invasive mesenchymal subtype. By combining proteomics with GBM patient single-cell transcriptomics, we identify multiple TAM-secreted proteins with putative proinvasive functions and validate TGF-β induced (TGFBI, also known as BIGH3) as a targetable TAM-secreted tumorigenic factor. Our work highlights the utility of coupling multiomics analyses with engineered TME models to investigate TAM–cancer cell crosstalk and offers insights into TAM function to guide TAM-targeting therapies.

Authors

Erin A. Akins, Dana Wilkins, Zaki Abou-Mrad, Kelsey Hopland, Robert C. Osorio, Kenny K.H. Yu, Manish K. Aghi, Sanjay Kumar

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Figure 1

An engineered biomaterials platform to enable investigation of GBM-macrophage crosstalk.

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An engineered biomaterials platform to enable investigation of GBM-macro...
(A) Schematic of HA-based hydrogel. (B) Schematic of mono- or coculture platforms to study GBM invasion. (C) Schematic of THP-1–derived monocyte-to-macrophage differentiation and M1/M2 polarization protocol. (D and E) GSC-295 invasion assay with direct M2 macrophage coculture and M2 macrophage–conditioned media (CM) (n = 19 spheres) showing (D) quantification and (E) representative phase images. (F and G) GSC-295 invasion assay with CM from monocytes (mono.), M1 macrophages, and M2 macrophages (n = 30 spheres) showing (F) quantification and (G) representative phase images. (H and I) GSC-268 invasion assay with M2 CM (n = 31 spheres) showing (H) quantification and (I) representative phase images. (J and K) GSC-11 invasion assay with M2 CM (n = 39 spheres) showing (J) quantification and (K) representative phase images. (L and M) GSC-268 invasion assays with GBM patient–derived macrophage CM (n = 18-59 spheres) showing (L) quantification and (M) representative phase images for Patient A. Spheroid invasion results were pooled across at least 2–3 independent replicates. Statistical significance was analyzed using 1-way ANOVA followed by Tukey’s multiple-comparison test (D, F, and L) or an unpaired, 2-sided Student’s t test (H and J). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Scale bars: 200 μm.