High Content Screening of Small Molecule Modulators Targeting Heat Shock Response Pathway

Abstract

Phenotypic high content screening (HCS) has resurged in recent years as an important platform in the drug discovery paradigm, particularly to address the serious challenges in the current target based approaches. Using the highly conserved heat shock response (HSR) pathway as a therapeutic intervention point, we established a cell based, high throughput, multiplexing, and disease relevant phenotypic screening platform for small molecule HSF1 modulators in cancer and neurodegenerative diseases. In this chapter, the authors reviewed their systematic design of methodology and workflow in detail, including the characterization of cellular phenotypic changes, image analysis and quantification, assay development and automation, screening operation and quality control, counter-screen and lead optimization, target identification and mechanism of action study. Selected compounds from the phenotypic screening, including novel HSF1 activators and HSF1 inhibitors, were discussed in regard to their chemical structures, therapeutic effects, and cytotoxicity for potential drug development. The authors also addressed the uncertainties and risks in target deconvolution, which still remains to be the most difficult hurdle in phenotypic screening. Recent HCS advances in tissues, organs and whole organisms, such as 3D tissue imaging, organ-on-a chip, IPS derived cell models, relevant animal models, etc., may bring an effective solution for delivering target-specific, first-in-class drugs in the future.