Arginine-Modified Polymers Facilitate Poly (Lactide-Co-Glycolide)-Based Nanoparticle Gene Delivery to Primary Human Astrocytes

Jessica Proulx,1 Chaitanya Joshi,1 Sivakumar Vijayaraghavalu,2 Manju Saraswathy,2 Vinod Labhasetwar,2 Anuja Ghorpade,1 Kathleen Borgmann1,3 1Department of Microbiology, Immunology, and Genetics University of North Texas Health Science Center, Fort Worth, TX, USA; 2Department of Biomedical Engineerin...

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Autores principales:	Proulx J, Joshi C, Vijayaraghavalu S, Saraswathy M, Labhasetwar V, Ghorpade A, Borgmann K
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2020
Materias:	co-polymer transfection gene therapy astrocyte targeting nuclear entry Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/05dfd1aeb3d8465b97c1d8f9963e3b29
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spelling	oai:doaj.org-article:05dfd1aeb3d8465b97c1d8f9963e3b292021-12-02T11:38:58ZArginine-Modified Polymers Facilitate Poly (Lactide-Co-Glycolide)-Based Nanoparticle Gene Delivery to Primary Human Astrocytes1178-2013https://doaj.org/article/05dfd1aeb3d8465b97c1d8f9963e3b292020-05-01T00:00:00Zhttps://www.dovepress.com/arginine-modified-polymers-facilitate-poly-lactide-co-glycolide-based--peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Jessica Proulx,1 Chaitanya Joshi,1 Sivakumar Vijayaraghavalu,2 Manju Saraswathy,2 Vinod Labhasetwar,2 Anuja Ghorpade,1 Kathleen Borgmann1,3 1Department of Microbiology, Immunology, and Genetics University of North Texas Health Science Center, Fort Worth, TX, USA; 2Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA; 3Department of Pharmacology and Neuroscience, University of North Texas Health Science Center, Fort Worth, TX 76107, USACorrespondence: Kathleen BorgmannDepartment of Pharmacology and Neuroscience, University of North Texas Health Science Center, 3500 Camp Bowie Blvd, Fort Worth, TX 76107, USATel +1 817 735-0339Fax +1 817 735-2610Email Kathleen.Borgmann@unthsc.eduPurpose: 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 (lactide-co-glycolide) (PLGA) and polyethylenimine (PEI) are polymeric nanoparticles commonly used in gene delivery, each manifesting their own set of advantages and disadvantages. As a clinically approved polymer by the Federal Drug Administration, well characterized for its biodegradability and biocompatibility, PLGA-based nanoparticles (PLGA-NPs) are appealing for translational gene delivery systems. However, our investigations revealed PLGA-NPs were ineffective at facilitating exogenous gene expression in primary human astrocytes, despite their success in other cell lines. Furthermore, PEI polymers illustrate high delivery efficiency but induce cytotoxicity. The purpose of this study is to develop viable and biocompatible NPsystem for astrocyte-targeted gene therapy.Materials and Methods: Successful gene expression by PLGA-NPs alone or in combination with arginine-modified PEI polymers (AnPn) was assessed by a luciferase reporter gene encapsulated in PLGA-NPs. Cytoplasmic release and nuclear localization of DNA were investigated using fluorescent confocal imaging with YOYO-labeled plasmid DNA (pDNA). NP-mediated cytotoxicity was assessed via lactate dehydrogenase in primary human astrocytes and neurons.Results: Confocal imaging of YOYO-labeled pDNA confirmed PLGA-NPs delivered pDNA to the cytoplasm in a dose and time-dependent manner. However, co-staining revealed pDNA delivered by PLGA-NPs did not localize to the nucleus. The addition of AnPn significantly improved nuclear localization of pDNA and successfully achieved gene expression in primary human astrocytes. Moreover, these formulations were biocompatible with both astrocytes and neurons.Conclusion: By co-transfecting two polymeric NPs, we developed an improved system for gene delivery and expression in primary human astrocytes. These findings provide a basis for a biocompatible and clinically translatable method to regulate astrocyte function during neurodegenerative diseases and disorders.Keywords: co-polymer, transfection, gene therapy, astrocyte targeting, nuclear entryProulx JJoshi CVijayaraghavalu SSaraswathy MLabhasetwar VGhorpade ABorgmann KDove Medical Pressarticleco-polymertransfectiongene therapyastrocyte targetingnuclear entryMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 15, Pp 3639-3647 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	co-polymer transfection gene therapy astrocyte targeting nuclear entry Medicine (General) R5-920
spellingShingle	co-polymer transfection gene therapy astrocyte targeting nuclear entry Medicine (General) R5-920 Proulx J Joshi C Vijayaraghavalu S Saraswathy M Labhasetwar V Ghorpade A Borgmann K Arginine-Modified Polymers Facilitate Poly (Lactide-Co-Glycolide)-Based Nanoparticle Gene Delivery to Primary Human Astrocytes
description	Jessica Proulx,1 Chaitanya Joshi,1 Sivakumar Vijayaraghavalu,2 Manju Saraswathy,2 Vinod Labhasetwar,2 Anuja Ghorpade,1 Kathleen Borgmann1,3 1Department of Microbiology, Immunology, and Genetics University of North Texas Health Science Center, Fort Worth, TX, USA; 2Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA; 3Department of Pharmacology and Neuroscience, University of North Texas Health Science Center, Fort Worth, TX 76107, USACorrespondence: Kathleen BorgmannDepartment of Pharmacology and Neuroscience, University of North Texas Health Science Center, 3500 Camp Bowie Blvd, Fort Worth, TX 76107, USATel +1 817 735-0339Fax +1 817 735-2610Email Kathleen.Borgmann@unthsc.eduPurpose: 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 (lactide-co-glycolide) (PLGA) and polyethylenimine (PEI) are polymeric nanoparticles commonly used in gene delivery, each manifesting their own set of advantages and disadvantages. As a clinically approved polymer by the Federal Drug Administration, well characterized for its biodegradability and biocompatibility, PLGA-based nanoparticles (PLGA-NPs) are appealing for translational gene delivery systems. However, our investigations revealed PLGA-NPs were ineffective at facilitating exogenous gene expression in primary human astrocytes, despite their success in other cell lines. Furthermore, PEI polymers illustrate high delivery efficiency but induce cytotoxicity. The purpose of this study is to develop viable and biocompatible NPsystem for astrocyte-targeted gene therapy.Materials and Methods: Successful gene expression by PLGA-NPs alone or in combination with arginine-modified PEI polymers (AnPn) was assessed by a luciferase reporter gene encapsulated in PLGA-NPs. Cytoplasmic release and nuclear localization of DNA were investigated using fluorescent confocal imaging with YOYO-labeled plasmid DNA (pDNA). NP-mediated cytotoxicity was assessed via lactate dehydrogenase in primary human astrocytes and neurons.Results: Confocal imaging of YOYO-labeled pDNA confirmed PLGA-NPs delivered pDNA to the cytoplasm in a dose and time-dependent manner. However, co-staining revealed pDNA delivered by PLGA-NPs did not localize to the nucleus. The addition of AnPn significantly improved nuclear localization of pDNA and successfully achieved gene expression in primary human astrocytes. Moreover, these formulations were biocompatible with both astrocytes and neurons.Conclusion: By co-transfecting two polymeric NPs, we developed an improved system for gene delivery and expression in primary human astrocytes. These findings provide a basis for a biocompatible and clinically translatable method to regulate astrocyte function during neurodegenerative diseases and disorders.Keywords: co-polymer, transfection, gene therapy, astrocyte targeting, nuclear entry
format	article
author	Proulx J Joshi C Vijayaraghavalu S Saraswathy M Labhasetwar V Ghorpade A Borgmann K
author_facet	Proulx J Joshi C Vijayaraghavalu S Saraswathy M Labhasetwar V Ghorpade A Borgmann K
author_sort	Proulx J
title	Arginine-Modified Polymers Facilitate Poly (Lactide-Co-Glycolide)-Based Nanoparticle Gene Delivery to Primary Human Astrocytes
title_short	Arginine-Modified Polymers Facilitate Poly (Lactide-Co-Glycolide)-Based Nanoparticle Gene Delivery to Primary Human Astrocytes
title_full	Arginine-Modified Polymers Facilitate Poly (Lactide-Co-Glycolide)-Based Nanoparticle Gene Delivery to Primary Human Astrocytes
title_fullStr	Arginine-Modified Polymers Facilitate Poly (Lactide-Co-Glycolide)-Based Nanoparticle Gene Delivery to Primary Human Astrocytes
title_full_unstemmed	Arginine-Modified Polymers Facilitate Poly (Lactide-Co-Glycolide)-Based Nanoparticle Gene Delivery to Primary Human Astrocytes
title_sort	arginine-modified polymers facilitate poly (lactide-co-glycolide)-based nanoparticle gene delivery to primary human astrocytes
publisher	Dove Medical Press
publishDate	2020
url	https://doaj.org/article/05dfd1aeb3d8465b97c1d8f9963e3b29
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Arginine-Modified Polymers Facilitate Poly (Lactide-Co-Glycolide)-Based Nanoparticle Gene Delivery to Primary Human Astrocytes

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