Over the last decade, it has become increasingly clear that a more complete understanding of human diseases requires viewing them in the context of systems biology, in particular through a comprehensive understanding of a network of molecular interactions that occur in a cell. Numerous successful applications (e.g. for disease gene prioritization) build on the fact that the local and global structures of molecular networks, mostly based on protein- protein interactions (PPIs), provide critical biological information. With the release of several large-scale systematic PPI datasets since our group produced the first proteome-scale map of the human binary interactome in 2005, integration of genetic and clinical data with interactome information has become possible and provides meaningful and critical insights towards a deeper pathophysiological understanding of diseases and the potential to revolutionize precision medicine. However, we have not reached a comprehensive map of the PPI network in any model system or in humans yet, and thus clinical applications would greatly benefit from deeper and wider explorations of the human interactome. High-throughput approaches have been developed to determine PPIs on a global scale for many organisms, but these assays remain intrinsically limited and labor intensive. A major bottleneck in screening is determining the identities of binary interacting partners. This OPTIMA project aims to eliminate that bottleneck and fill the gap in current networks by developing a novel disruptive technology for high-performance interactomics allowing en masse screening of PPIs to provide comprehensive binary PPI maps. This innovative system will result from the integration of two recently validated technologies, on the one hand the bioluminescent detection of PPIs based on complementation of a split-Nanoluc reporter and, on the other hand, one of the most sensitive optogenetically programmed promoters driving DNA barcode fusion. By leveraging en masse binary PPI detection, this new binary interaction detection strategy will dramatically enhance the overall coverage of proteome-scale interactome maps. This new high-throughput pipeline will be orthogonal to existing proteome-scale binary interaction mapping platforms, such as yeast two-hybrid followed by validation, and thus able to significantly enhance the available tools to expand existing interactomes.
Mapping the human interactome with its broad complexity including the human variome is a major biomedical challenge and the necessary keystone for better understanding of pathophysiology at a systemic level and for the next challenge of precision medicine. The innovative protein-protein interaction mapping strategy proposed here will combine the most advanced development in bioluminescence, optogenetics, DNA-barcoding and next generation sequencing. Its orthogonality with existing approaches should pave the way towards the complete mapping of human interactome network.
