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

Abstract

Cell-based high-throughput drug screening (HTS) is a common starting point for the drug discovery and development process. Currently, there is a push to combine complex cell culture systems with HTS to provide more clinically applicable results. However, there are mechanistic requirements inherent to HTS as well as material limitations that make this integration challenging. Here, we used the peptide-based shear-thinning hydrogel MAX8 tagged with the RGDS sequence to create a synthetic extracellular scaffold to culture cells in three dimensions and showed a preliminary implementation of the scaffold within an automated HTS setup using a pilot drug screen targeting medulloblastoma, a pediatric brain cancer. A total of 2202 compounds were screened in the 384-well format against cells encapsulated in the hydrogel as well as cells growing on traditional two-dimensional (2D) plastic. Eighty-two compounds passed the first round of screening at a single point of concentration. Sixteen-point dose–response was done on those 82 compounds, of which 17 compounds were validated. Three-dimensional (3D) cell-based HTS could be a powerful screening tool that allows researchers to finely tune the cell microenvironment, getting more clinically applicable data as a result. Here, we have shown the successful integration of a peptide-based hydrogel into the high-throughput format.

3D spheroid models of paediatric SHH medulloblastoma mimic tumour biology, drug response and metastatic dissemination

“…There has been limited research on the use of hydrogels for the culture of medulloblastoma cell lines. Three publications have described the use of the MAX8 β-hairpin self-assembling peptide hydrogels as a suitable scaffold-based model for the culture of the DAOY and ONS76 cell lines 53 – 55 . Singh et al also described the growth of DAOY single cells on a cellulose-derived hydrogel 56 .…”

Proteinaceous Hydrogels for Bioengineering Advanced 3D Tumor Models

“…[ 277 ] This platform was also employed for automated drug screening, enabling researchers to screen 2202 candidate therapeutic molecules simultaneously. [ 278 ]…”

Microrheology for biomaterial design

“…For example, a high-throughput screening tool was developed for malignant pediatric brain drug screening in 3D in vitro cancer models using a peptide-based hydrogel as a 3D scaffold. 61 Incorporating microrheology into this screening could reveal other mechanical changes to these scaffolds during drug treatment, which could provide information relevant to tumor pathology. 62 …”