Abstract

EVs and CDVs: Nanovesicle Therapeutics and Drug Delivery Systems

Mario Gimona, Head of Manufacturing-GMP, Paracelsus Medical University Salzburg

Nanovesicle-based therapy is increasingly being pursued as a safe, cell-free strategy to combat various immunological, musculoskeletal and neurodegenerative diseases. Small secreted extracellular vesicles (sEVs) obtained from multipotent mesenchymal stromal cells (MSCs) are of particular interest for therapeutic use since they convey anti-inflammatory, anti-scarring and neuroprotective activities to the recipient cells. A novel drug delivery platform technology that is based on cell-derived vesicles (CDVs) produced by a proprietary extrusion process generates nanosized vesicles that are surrounded by a lipid bilayer membrane with a correct membrane topology and that display biological activities similar to MSC-derived EVs. Translation of nanovesicle-based therapeutics into clinical application requires quantitative, and reproducible analysis of bioactivity and stability. The GPI-anchored ecto-5’-nucleotidase CD73 is abundantly present on MSC-derived EVs and CDVs and efficiently converts AMP to Adenosine. In an in-vitro assay the enzymatic activity per particle correlated with CD73 abundance and we reproducibly obtained 2 - 8 x 10^-8 mM Adenosine production / particle in both sEV and CDV preparations. Nucleotides can work as trophic, differentiating, and immunomodulatory molecules in many physiological and pathological situations through autocrine and paracrine mechanisms, and purinergic ligands are potent candidates to mediate cellular crosstalk and to promote cell growth and survival, regulate inflammation and contribute to local tissue homeostasis and repair. Our data suggest that CD73-mediated Adenosine signaling supports the creation of an anti-inflammatory and pro-regenerative environment that can pave the road to tissue repair and functional regeneration.