Investigating the Interactions of Glioma Stem Cells in the Perivascular Niche at Single‐Cell Resolution using a Microfluidic Tumor Microenvironment Model

Abstract

The perivascular niche (PVN) is a glioblastoma tumor microenvironment (TME) that serves as a safe haven for glioma stem cells (GSCs), and acts as a reservoir that inevitably leads to tumor recurrence. Understanding cellular interactions in the PVN that drive GSC treatment resistance and stemness is crucial to develop lasting therapies for glioblastoma. The limitations of in vivo models and in vitro assays have led to critical knowledge gaps regarding the influence of various cell types in the PVN on GSCs behavior. This study developed an organotypic triculture microfluidic model as a means to recapitulate the PVN and study its impact on GSCs. This triculture platform, comprised of endothelial cells (ECs), astrocytes, and GSCs, is used to investigate GSC invasion, proliferation and stemness. Both ECs and astrocytes significantly increased invasiveness of GSCs. This study futher identified 15 ligand‐receptor pairs using single‐cell RNAseq with putative chemotactic mechanisms of GSCs, where the receptor is up‐regulated in GSCs and the diffusible ligand is expressed in either astrocytes or ECs. Notably, the ligand–receptor pair SAA1‐FPR1 is demonstrated to be involved in chemotactic invasion of GSCs toward PVN. The novel triculture platform presented herein can be used for therapeutic development and discovery of molecular mechanisms driving GSC biology.

Harmonized single-cell landscape, intercellular crosstalk and tumor architecture of glioblastoma

“…The OPC/NPC layer near the blood vessels (Fig. 6a, boxes I, II) is in accordance with the published description of a perivascular niche that is enriched for cancer stem-like cells (Adjei-Sowah et al, 2022;Jung et al, 2021).…”

3D Biomimetic Models to Reconstitute Tumor Microenvironment In Vitro: Spheroids, Organoids, and Tumor‐on‐a‐Chip

“…To create a realistic and complex microenvironment model, Adjei‐Sowah et al proposed an organotypic triculture microfluidic model containing ECs, ACs, and GSCs. [ 155 ] Using single‐cell RNA sequencing, 15 ligand–receptor pairs were identified to be related to the chemotactic invasion of GSCs toward the perivascular niche.…”

Bridging the gap between tumor-on-chip and clinics: a systematic review of 15 years of studies

“…108 Properly designed pillars induce surface tension to hold the hydrogels in a given area of the chip, while nutrients are supplied through media in adjacent channels. Adjei-Sowah et al 109 (Fig. 4 top left panel) developed a tri-culture of endothelial cells, astrocytes and glioma stem cells utilizing a pillar compartmentalized chip and focused on the identification of ligand-receptor pairs.…”

“…Almost half of the ToC belong to the "compartmentalized chips" category. 76,[101][102][103][104][105][106][107][108][109] Compartmentalization of cells embedded in hydrogels can be achieved by various physical means. Several studies use co-flow patterning flowing side by side to two different hydrogel solutions.…”

“…Fig.4Pie chart illustrating the proportion of different toc designs used to mimic cell architecture: compartmentalized chip 39%, micro-wells array 18%, 2D chip 16%, membrane chip 11%, lumen chip 8%, other 8%. (Top left panel) The glioblastoma toc model is a compartmentalized chip with three concentric cell culture regions separated by pillars: namely, the vasculature, stroma, and tumor regions surrounded by media channels 109. (Top right) In this micro-well array ToC, cancer and stromal cells are encapsulated in Matrigel and seeded into the bottom layer, while flow is applied in the top layer 116.…”