Barcoded Nanoparticles for Rapid Directed Evolution Modeling

In this innovative Phase I effort, nSI will construct and demonstrate an empirically driven directed evolution model that rapidly selects for BBB-crossing NPs in vitro, as a proof of concept. Leveraging cutting-edge NP formulation advances, we will (1) synthesize a library of DNA-barcoded NPs, (2) administer the library in multiplex to in vitro BBB models, (3) use the BBB infilitration data collected to drive the evolution of the library, and (4) repeat to optimize. Critically, both the library, barcoding method and directed evolution model will also work in vivo. Anticipating future FDA approval, we will utilize only polymers and lipids that are endogenous or in clinical use. This constraint will not negatively impact our design space; multiple investigators have demonstrated exquisite organ- and cell-specific tropism by varying single components of lipid/PEG-lipid/cholesterol formulations. The result will be a multiplexed, barcoded NP library and accompanying directed evolution model that enables rapid screening and iterative selection of in vivo BBB crossing in Phase II. Our approach avoids both the high cost of singleton in vivo trials and the intractable complexity of a priori computer modeling that plague this problem space.