FMSG: Enabling Technologies for Biomanufacturing EV-Based Therapeutics

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Extracellular vesicles (EVs) are cell-made particles that provide a natural mechanism of information and material transfer between cells. There is growing interest in large-scale production of EVs that can be used as therapeutics due to their ability to communicate signals from producer cells and their potential use as carriers for delivery of drug molecules. EV therapeutics are being developed for treatment of a wide range of diseases including metabolic disorders, cancer, and neurodegeneration. Despite the excitement generated by several early-stage EV biotech companies, the technology to produce mass quantities of purified EVs with tunable properties is still in its infancy. The long-term goal of this project is to enable production of "designer EVs" that can be packaged with desired cargo molecules, decorated with tunable surface ligands, and secreted in high yield from specific producer cell types. The overall objective of the current project is to identify cellular processes that can be engineered to control the production, content, and in vivo trafficking of therapeutic EVs. Making this new class of drugs available to the public has potential to improve the health and quality of life of millions of patients in the US and around the world. The project will also provide the unique educational opportunity for trainees to engage in collaborative research with industry scientists. Other broader impacts will be accomplished through engaging undergraduate and high-school students in EV research, and through integration of the project with the Vanderbilt Program for EV Research and bioengineering courses led by the PIs. The investigators will focus their research program on loading and delivery of small RNAs, which are promising drug molecules that are not delivered efficiently to recipient cells using established nanoparticle-based carriers. The central hypothesis is that developing tissue-specific producer cells with tuned expression of EV-associated proteins and RNAs will enable researchers to maximize the product yield of specific EVs and the efficiency of RNA delivery from EVs to recipient cells. First, they will boost targeting of miRNAs to extracellular vesicles through modulating specific molecules at ER-contact sites. Second, they will optimize cargo delivery to target cells by engineering EVs for efficient endosomal escape and macrophage evasion properties. Third, they will develop a scalable platform for manufacturing tissue-specific EVs by differentiating induced pluripotent stem cells (iPSCs) to producer cells in 3D suspension cultures. The proposed research is innovative because it applies a multidisciplinary approach to address several critical barriers to commercial production of therapeutic EVs: cargo loading, cargo delivery, and scalable manufacturing. EVs secreted by MSCs are recognized as a viable alternative to overcome the potential risks from transplantation of primary MSCs. By developing the tools and strategies to customize and maximize EV production from iPSC-derived MSCs, the researchers will establish a flexible EV manufacturing platform that can expand to other relevant organ and tissue systems.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.