Poly lactic-co-glycolic acid (PLGA) mediated gene delivery to astrocytes requires arginine-modified polyethylenimine (PEI) polymer to facilitate gene expression
0000-0002-5452-0461 (Proulx, Jessica)
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Purpose: The global burden of neurodegenerative diseases and disorders devastate not only their victims but also their loved ones. Astrocyte dysfunction is a hallmark of central nervous system injury or infection. As a primary contributor to neurodegeneration, astrocytes are an ideal therapeutic target to combat neurodegenerative conditions. Gene therapy has arisen as an innovative technique that provides excellent prospect for disease intervention. Poly lactic-co-glycolic-acid (PLGA) and polyethylenimine (PEI) are polymeric nanoparticles commonly used in gene delivery, each manifesting their own set of advantages and disadvantages. While PLGA nanoparticles are FDA approved and well established for their biocompatibility, they fail to facilitate exogenous gene expression in primary human astrocytes. Furthermore, PEI polymers illustrate high delivery efficiency but induce cytotoxicity. The purpose of this study is to develop viable and biocompatible nanoparticles for astrocyte-targeted gene therapy. Methods: Successful gene expression by PLGA nanoparticles alone or in combination with arginine-modified PEI polymers (A5P10) was accessed by a luciferase reporter gene encapsulated in PLGA nanoparticles. Cytoplasmic release and nuclear localization were investigated using fluorescent confocal imaging with YOYO-labeled DNA. Nanoparticle-mediated cytotoxicity was assessed via lactate dehydrogenase (LDH) in primary human astrocytes and neurons. Results: Confocal imaging of YOYO-labeled DNA confirmed PLGA nanoparticles delivered DNA to the cytoplasm in a dose and time dependent manner. However, co-staining revealed DNA delivered by PLGA did not localize to the nucleus. The addition of A5P10 improved nuclear localization and successfully achieved gene expression in primary human astrocytes. Moreover, these formulations were biocompatible with both astrocytes and neurons. Conclusion: By integrating two polymeric nanoparticles, we developed an improved system for gene delivery and expression in primary human astrocytes. These findings provide a biocompatible and clinically translatable method to regulate astrocyte function during neurodegenerative diseases and disorders.