Advanced molecular engineering approach for understanding extracellular vesicle in vivo trafficking mechanism

ABSTRACT Deciphering extracellular vesicles (EVs) mediated cell-to-cell communications at the molecular precision has been a grand challenge. Mechanistic understanding from current laboratory investigation presents a substantial knowledge gap from clinical translation. Different cell sources derived EVs carry completely different surface receptors and regulatory molecular components. Heterogeneous populations of EVs confound the investigation of key drivers in cellular regulation, for poorly understanding mechanisms on either crossing biological barrier or extracellular matrix accumulation. To date, the molecular mechanisms on tissue targeting specificity are lacking, and the therapeutic potential of different subpopulations of EVs is unknown yet. The proposed research tackles those key challenging questions by introducing a series of advanced molecular engineering approaches for precisely predicting, defining, and in vivo testing EV surface receptor and antigenic presentation. Specifically, project #1 will develop CRISPR imaging technology for understanding molecular mechanism on EV tissue penetration and accumulation. Project #2 will develop artificial intelligent direct-flow generative adversarial model (ExoGAN) enabled construction of specific targeting EVs. Project #3 will develop microfluidic single cell single vesicle transwell system for deciphering effective MHC presenting modulators engaged in EV mediated immunity. This proposal will build a foundation to facilitate mechanistic -driven tissue targeting for advancing precision drug delivery and open broad applications to precision drug delivery.