High‐Throughput 3D Glioblastoma Model in Glycosaminoglycan Hydrogels for Personalized Therapeutic Screening

TL;DR

A new 3D model for glioblastoma tumors helps test personalized treatments by mimicking the complex tumor environment and capturing key biological features.

Contribution

A high-throughput 3D glioblastoma model using glycosaminoglycan hydrogels enables independent control of matrix properties and drug screening.

Findings

3D cultures recapitulate GBM molecular programs including hypoxia and immune pathways without artificial hypoxia.

Drug screening in the model reveals dose-dependent effects on tumor cell invasion and architecture.

The platform supports automated drug testing and transcriptomic profiling for personalized therapy optimization.

Abstract

Glioblastoma (GBM) is a devastating brain tumor with limited treatment success, partly because in vitro models poorly mimic in vivo complexity. This study introduces a high‐throughput 3D culture platform utilizing modular starPEG–glycosaminoglycan (GAG) hydrogels that enable independent control of extracellular matrix (ECM) cues: stiffness, cytokine affinity, matrix metalloproteinase‐responsive remodeling, and cell adhesiveness via integrin‐binding RGD peptides. This platform supports encapsulation of patient‐derived GBM cells, recreates physiologically relevant tumor microenvironments in 384‐well plates, and enables automated drug testing on primary cells. Transcriptomic analyses show that 3D cultures recapitulate primary and recurrent GBM programs‐ including hypoxia‐, immune‐, and ECM‐regulatory pathways driving growth, invasion, and resistance, without externally imposed hypoxia. The platform's versatility extends to drug screening, where single and combinatorial treatments produce reproducible cytoskeletal and transcriptomic responses. Notably, the system revealed dose‐dependent reductions in invasive filaments and spheroid architecture with 5‐fluorouracil/uridine and carmustine, demonstrating its potential for optimizing combinatorial therapies. This 3D model surpasses 2D cultures, capturing tumor‐specific molecular programs and offering a robust tool for translational research. Despite lacking vascular or immune components, its tunability, scalability, and clinical relevance make it a strong basis for advanced co‐cultures. By delivering reliable, individualized therapeutic data within a short timeframe, this model holds transformative potential for personalized GBM treatment.

Kaur–Trautmann et al. present a high‐throughput, chemically defined 3D glioblastoma model based on glycosaminoglycan hydrogels that allow orthogonal control of stiffness, adhesion, and degradability while standardizing soluble‐factor presentation. The platform supports automated imaging and transcriptomic profiling, enabling patient‐specific drug screening and revealing metabolic and microenvironmental adaptations relevant to personalized therapy development.

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