title: A novel gene therapy vector based on hyaluronic acid and solid lipid nanoparticles for ocular diseases.
abstract: The introduction of therapeutic genes in target tissues is considered as a novel tool for the treatment of several diseases. We have developed nanoparticles consisting on SLNs, protamine (P) and hyaluronic acid (HA) as carrier for gene therapy. Stable complexes positively charged and with a particle size ranging from 240 nm to 340 nm were obtained. Transfection studies in ARPE-19 and HEK-293 cells showed the versatility of vectors to efficiently transfect cells with different division rate, widening the potential applications of SLN-based vectors. In ARPE 19cells, the incorporation of P and HA induced almost a 7-fold increase in the transfection capacity of SLNs. The CD44 inhibition studies suggested the participation of this receptor in the internalization of the vectors in this cell line. The intracellular disposition of DNA showed that the HA is able to modulate the high degree of condensation of DNA due to the protamine inside the cells; an important fact, if the vector is uptaken via non-degradative endocytosis. Besides, the therapeutic plasmid which encodes the protein retinoschisin was employed achieving a positive transfection in ARPE-19 cells, showing a promising application of this new non-viral system for the treatment of X-linked juvenile retinoschisis by gene therapy.
authors.affiliation: Pharmacokinetic, Nanotechnology and Gene Therapy Group (PharmaNanoGene), Faculty of Pharmacy, Centro de investigación Lascaray ikergunea, University of the Basque Country UPV/EHU, Vitoria-Gasteiz, Spain.
authors.forename: Paola Stephanie
authors.initials: PS
authors.lastname: Apaolaza
authors.affiliation: Pharmacokinetic, Nanotechnology and Gene Therapy Group (PharmaNanoGene), Faculty of Pharmacy, Centro de investigación Lascaray ikergunea, University of the Basque Country UPV/EHU, Vitoria-Gasteiz, Spain.
authors.forename: Diego
authors.initials: D
authors.lastname: Delgado
authors.affiliation: Pharmacokinetic, Nanotechnology and Gene Therapy Group (PharmaNanoGene), Faculty of Pharmacy, Centro de investigación Lascaray ikergunea, University of the Basque Country UPV/EHU, Vitoria-Gasteiz, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: del Pozo-Rodríguez
authors.affiliation: Pharmacokinetic, Nanotechnology and Gene Therapy Group (PharmaNanoGene), Faculty of Pharmacy, Centro de investigación Lascaray ikergunea, University of the Basque Country UPV/EHU, Vitoria-Gasteiz, Spain.
authors.forename: Alicia Rodríguez
authors.initials: AR
authors.lastname: Gascón
authors.affiliation: Pharmacokinetic, Nanotechnology and Gene Therapy Group (PharmaNanoGene), Faculty of Pharmacy, Centro de investigación Lascaray ikergunea, University of the Basque Country UPV/EHU, Vitoria-Gasteiz, Spain. Electronic address: marian.solinis@ehu.es.
authors.forename: M Ángeles
authors.initials: MÁ
authors.lastname: Solinís
mesh_terms: D004705:Endocytosis; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D006820:Hyaluronic Acid / Q000737:chemistry / Q000378:metabolism*; D050356:Lipid Metabolism; D008055:Lipids / Q000737:chemistry; D053758:Nanoparticles; D010316:Particle Size; D011479:Protamines / Q000378:metabolism; D055213:Retinal Pigment Epithelium / Q000378:metabolism; D041441:Retinoschisis / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D013997:Time Factors; D014162:Transfection / Q000379:methods*
chemical_list: C497483:CD44 protein, human; D005136:Eye Proteins; D018960:Hyaluronan Receptors; D008055:Lipids; D011479:Protamines; C108726:RS1 protein, human; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D006820:Hyaluronic Acid
keywords: Gene therapy; Hyaluronic acid; Protamine; Solid lipid nanoparticles; Transfection; X linked juvenile retinoschisis