Automated High‐Throughput Drug Discovery in Peptide Hydrogel‐Based 3D Cell Cultures

Automated cell‐based high‐throughput screening (HTS) is a powerful tool in drug discovery and currently, there is a push to combine complex cell culture systems with HTS to provide more clinically applicable results. Despite the increasing recognition that three‐dimensional (3D) models provide more in vivo ‐like cell culture environments, there are mechanistic requirements inherent to HTS as well as material limitations that make this integration for high‐throughput drug discovery challenging. We used the peptide‐based hydrogel MAX8 tagged with the RGDS sequence to create a synthetic extracellular scaffold to culture cells in 3D, and show a preliminary implementation of the scaffold for 3D HTS with a pilot drug screen targeting the pediatric brain cancer medulloblastoma. MAX8 undergoes assembly under physiological conditions and ambient temperature into a hydrogel with a well‐defined, nanofibrillar matrix, desired porosity and stiffness to provide a biocompatible cell culture matrix. Due to the physical gel properties such as shear‐thinning and the fast gelation kinetics, cells can be homogenously encapsulated into evenly dispensed gel‐cell constructs using automatic distribution of cell‐peptide mixtures into 384‐well assay plates with standard liquid handling equipment. 2,202 compounds were screened in the 384‐well format against cells encapsulated in the hydrogel as well as cells growing on traditional 2D plastic. 82 compounds passed the first round of screening at a single point of concentration. 16‐point dose response was done on those 82 compounds, of which 17 compounds validated. Three‐dimensional 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. Support or Funding Information This work was supported by funds from the NIH IDeA program, with grants from the National Institute of General Medical Sciences NIGMS (P20‐GM103464, P30‐GM114736, and U54‐GM104941), the DO Believe Foundation and the Nemours Foundation. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .