SBIR Phase I: Developing a Platform for Multiplexed Drug Profiling Using Yeast Synthetic Agglutination

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project will be to develop a preclinical drug characterization platform capable of profiling the effect of a drug candidate on whole protein-protein interaction (PPI) networks. PPIs play a pivotal role in most diseases, and are considered high-impact therapeutic targets for cancers, autoimmune diseases, infectious diseases, and more. Over 40 clinically relevant PPIs have been disrupted successfully with small molecules, and several have entered clinical trials. One recently approved cancer drug, Venetoclax, is expected to reach $2.2B in sales by 2020. Despite the enormous clinical and commercial potential of PPI disrupting drugs, preclinical characterization remains a major challenge. Pharmaceutical companies are limited by slow and laborious techniques to measure protein interactions that require each protein to be purified, and each PPI to be measured separately. As a result, only a small subset of relevant interactions are tested during preclinical drug development, which leads to a high incidence of failure during clinical trials. The proposed platform for PPI network characterization is expected to have a major commercial and societal impact by enabling more thorough preclinical screening of PPI inhibiting drugs, reducing the overall cost and time associated with drug development. This SBIR Phase I project proposes to develop and commercialize a novel platform for screening PPI disrupting drugs that provides quantitative accuracy, enables simultaneous characterization of whole PPI networks, and eliminates the need for protein purification. This platform combines the throughput of a cell-based assay with the accuracy of a bioanalytical technique by linking yeast haploid mating efficiency to the affinity of proteins displayed on the cells' surfaces. Preliminary results demonstrate that next generation sequencing of diploid cells may be used to accurately measure many PPI strengths simultaneously. The goal of this project is to demonstrate that the proposed platform can correctly recapitulate whole disease-relevant PPI networks and accurately characterize well-studied inhibitors in a format that is compatible with existing high-throughput screening workflows. To demonstrate feasibility, the well-studied BCL2 PPI network, which contains unstable proteins and considerable complexity, will be analyzed to identify each pairwise PPI and compared to known interactions from the literature. The yeast strains and assay parameters will then be optimized for screening water insoluble small molecule drugs and 96-well plate compatibility. The ultimate goal of this project is to establish a new platform technology for the preclinical characterization of PPI inhibiting drug candidates. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.