Endothelial cells promote 3D invasion of GBM by IL-8-dependent induction of cancer stem cell properties.

Rapid growth and perivascular invasion are hallmarks of glioblastoma (GBM) that have been attributed to the presence of cancer stem-like cells (CSCs) and their association with the perivascular niche. However, the mechanisms by which the perivascular niche regulates GBM invasion and CSCs remain poorly understood due in part to a lack of relevant model systems. To simulate perivascular niche conditions and analyze consequential changes of GBM growth and invasion, patient-derived GBM spheroids were co-cultured with brain endothelial cells (ECs) in microfabricated collagen gels. Integrating these systems with 3D imaging and biochemical assays revealed that ECs increase GBM invasiveness and growth through interleukin-8 (IL-8)-mediated enrichment of CSCs. Blockade of IL-8 inhibited these effects in GBM-EC co-cultures, while IL-8 supplementation increased CSC-mediated growth and invasion in GBM-monocultures. Experiments in mice confirmed that ECs and IL-8 stimulate intracranial tumor growth and invasion in vivo. Collectively, perivascular niche conditions promote GBM growth and invasion by increasing CSC frequency, and IL-8 may be explored clinically to inhibit these interactions.

Collagen Fiber Orientation Regulates 3D Vascular Network Formation and Alignment.

Alignment of collagen type I fibers is a hallmark of both physiological and pathological tissue remodeling. However, the effects of collagen fiber orientation on endothelial cell behavior and vascular network formation are poorly understood because of a lack of model systems that allow studying these potential functional connections. By casting collagen type I into prestrained (0, 10, 25, 50% strain), poly(dimethylsiloxane) (PDMS)-based microwells and releasing the mold strain following polymerization, we have created collagen gels with varying fiber alignment as confirmed by structural analysis. Endothelial cells embedded within the different gels responded to increased collagen fiber orientation by assembling into 3D vascular networks that consisted of thicker, more aligned branches and featured elevated collagen IV deposition and lumen formation relative to control conditions. These substrate-dependent changes in microvascular network formation were associated with altered cell division and migration patterns and related to enhanced mechanosignaling. Our studies indicate that collagen fiber alignment can directly regulate vascular network formation and that culture models with aligned collagen may be used to investigate the underlying mechanisms ultimately advancing our understanding of tissue development, homeostasis, and disease.