Collaborative Research: Development of an exosome based lipoproteoplex (E-LPP) for siRNA delivery

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NON-TECHNICAL SUMMARY The development of safe and effective means to deliver nucleic acids has become increasingly important, especially in light of the COVID pandemic. mRNA vaccines that can provide our immune system the tools to fight COVID-19 are crucial in helping us fight this devastative virus. This project supports fundamental research focused on creating better delivery agents that can better protect and carry the important nucleic acids like mRNA so it can do its job. We explore the use of exosomes as potential delivery vehicles. Exosomes are essentially little compartments that are derived from our own cells. Because of this, they have key components that allow them to enter other cells and deliver cargo. While exosomes have this unique advantage, loading them with the target therapeutic is challenging. To address this, the research team will develop small, charged proteins that can gently hold onto the negatively charged nucleic acids, which then helps improve the loading of exosomes. This project will not only train the next generation graduate student and researcher in these new approaches, but also help inspire high school students as they will be able to carry out meaningful research experience both virtually and in person. Both the PI and co-PI have a strong record of training women and underrepresented minorities and plan to continue to recruit and train them as part of this work with the goal of developing and diversifying the future scientific workforce.TECHNICAL SUMMARY Nucleic acids possess critical instructions for controlling biological processes with tangible implications in fundamental cell and organismal studies, biotechnology and medicine especially in light of delivery of vaccines for Covid-19. There is a need to develop a modular platform for the widespread delivery of nucleic acids, and exosomes have recently been identified as promising delivery agents. The mechanisms of assembly between membrane-derived lipids and recombinant proteins is poorly understood but provides an opportunity to optimize and explore the fundamental material properties of exosome-based delivery vehicles. The research team proposes to develop an engineered protein-exosome system or lipoproteoplex capable of: 1) encapsulating siRNA at a high efficiency into the exosome; 2) increasing siRNA release once inside the cell through increased endosomal escape; 3) performing efficient siRNA delivery with a favorable safety profile. For education and outreach, the team aims to focus on verified virtual platforms to provide students safe, meaningful research experiences. Courses will focus on hybrid in-person and virtual lessons with an emphasis on group-based projects to facilitate active learning. For outreach, Dr. Montclare will participate in the Applied Research Innovations in Science and Engineering (ARISE) program and engage local NYC 10th and 11th grade students in computational design of supercharged proteins. Dr. Rabbani will implement as part of the Wyss Sumer Research Fellowship, a biochemistry training that involves exosome preparation, characterization and use for transfection as well as recording protocol video and journal club discussion online. This will democratize research experience for students during the pandemic and beyond.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.