Implementation of a High-Throughput Pilot Screen in Peptide Hydrogel-Based Three-Dimensional Cell Cultures

Area of Science:

• Biotechnology
• Drug Discovery
• Oncology

Background:

• High-throughput drug screening (HTS) is crucial for drug discovery.
• Integrating complex cell culture systems with HTS is challenging due to mechanistic and material limitations.
• There is a need for more clinically applicable results from HTS.

Purpose of the Study:

• To develop and implement a peptide-based hydrogel scaffold for three-dimensional (3D) cell culture in HTS.
• To demonstrate the feasibility of using this 3D hydrogel system within an automated HTS setup.
• To conduct a pilot drug screen targeting medulloblastoma using the integrated 3D HTS system.

Main Methods:

• Utilized a peptide-based shear-thinning hydrogel (MAX8) with RGDS sequence for synthetic extracellular scaffold creation.
• Encapsulated cells within the hydrogel for 3D culture.
• Performed automated HTS in a 384-well format, screening 2202 compounds against cells in hydrogel and 2D plastic.

Main Results:

• Eighty-two compounds initially passed the screening at a single concentration.
• A 16-point dose-response assay validated 17 of these compounds.
• Successfully demonstrated the integration of the peptide-based hydrogel into an automated HTS workflow.

Conclusions:

• Three-dimensional (3D) cell-based HTS using engineered hydrogels can provide more clinically applicable data.
• This approach allows fine-tuning of the cellular microenvironment for improved screening outcomes.
• The study successfully integrated a peptide-based hydrogel into a high-throughput screening format.