A human iPSC-based neural spheroid platform for modelling glioblastoma infiltration using high-content imaging

Area of Science:

• Neuroscience
• Oncology
• Stem Cell Biology

Background:

• Glioblastoma (GBM) is an aggressive brain tumor with poor prognosis.
• Therapy resistance and recurrence are driven by diffuse tumor cell infiltration.
• Existing models do not fully replicate the complex tumor microenvironment and axonal interactions.

Purpose of the Study:

• To develop a physiologically relevant co-culture model for studying glioblastoma infiltration.
• To identify therapeutic vulnerabilities by screening drugs targeting GBM cell migration.
• To understand gene expression profiles associated with GBM invasion and drug sensitivity.

Main Methods:

• Co-culture of patient-derived glioblastoma cell lines with human induced pluripotent stem cell-derived neural spheroids.
• High-content live imaging and endpoint assays to assess cell migration and infiltration.
• Bulk RNA sequencing to analyze gene expression.
• Proof-of-principle drug screening.

Main Results:

• Glioblastoma cells demonstrated directional migration along axons and extensive spheroid infiltration.
• Non-cancerous neural stem cells did not exhibit invasive behavior.
• PF-573228 (FAK inhibitor) and motixafortide (CXCR4 inhibitor) suppressed GBM cell infiltration.
• Gene expression profiles correlated with invasive behavior and drug sensitivity.

Conclusions:

• The developed co-culture model accurately mimics glioblastoma infiltration along axons.
• Tumor cell migration is a measurable phenotype for therapeutic screening.
• This platform can identify novel therapeutic strategies targeting glioblastoma invasion.