title: Facile Noninvasive Retinal Gene Delivery Enabled by Penetratin.
abstract: Gene delivery to the posterior segment of the eye is severely hindered by the impermeability of defensive barriers; therefore, in clinical settings, genomic medicines are mainly administered by intravitreal injection. We previously found that penetratin could transport the covalently conjugated fluorophore to the fundus oculi by topical instillation. In this study, gene delivery systems enabled by penetratin were designed based on electrostatic binding to target the retina via a noninvasive administration route and prepared with red fluorescent protein plasmid (pRFP) and/or poly(amidoamine) dendrimer of low molecular weight (G3 PAMAM). Formulation optimization, structure confirmation, and characterization were subsequently conducted. Penetratin alone showed limited ability to condense the plasmid but had powerful uptake and transfection by corneal and conjunctival cells. G3 PAMAM was nontoxic to the ocular cells, and when introduced into the penetratin-incorporated complex, the plasmid was condensed more compactly. Therefore, further improved cellular uptake and transfection were observed. After being instilled in the conjunctival sac of rats, the intact complexes penetrated rapidly from the ocular surface into the fundus and resided in the retina for more than 8 h, which resulted in efficient expression of RFP in the posterior segment. Intraocular distribution of the complexes suggested that the plasmids were absorbed into the eyes through a noncorneal pathway during which penetratin played a crucial role. This study provides a facile and friendly approach for intraocular gene delivery and is an important step toward the development of noninvasive gene therapy for posterior segment diseases.
authors.affiliation: Key Laboratory of Smart Drug Delivery, Ministry of Education; Department of Pharmaceutics, School of Pharmacy, Fudan University , Shanghai 201203, China.
authors.forename: Chang
authors.initials: C
authors.lastname: Liu
authors.affiliation: Key Laboratory of Smart Drug Delivery, Ministry of Education; Department of Pharmaceutics, School of Pharmacy, Fudan University , Shanghai 201203, China.
authors.forename: Kuan
authors.initials: K
authors.lastname: Jiang
authors.affiliation: Key Laboratory of Smart Drug Delivery, Ministry of Education; Department of Pharmaceutics, School of Pharmacy, Fudan University , Shanghai 201203, China.|School of Pharmacy, Shenyang Pharmaceutical University , Shenyang 110016, China.
authors.forename: Lingyu
authors.initials: L
authors.lastname: Tai
authors.affiliation: Key Laboratory of Smart Drug Delivery, Ministry of Education; Department of Pharmaceutics, School of Pharmacy, Fudan University , Shanghai 201203, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Key Laboratory of Smart Drug Delivery, Ministry of Education; Department of Pharmaceutics, School of Pharmacy, Fudan University , Shanghai 201203, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Wei
authors.affiliation: Key Laboratory of Smart Drug Delivery, Ministry of Education; Department of Pharmaceutics, School of Pharmacy, Fudan University , Shanghai 201203, China.
authors.forename: Weiyue
authors.initials: W
authors.lastname: Lu
authors.affiliation: School of Pharmacy, Shenyang Pharmaceutical University , Shenyang 110016, China.
authors.forename: Weisan
authors.initials: W
authors.lastname: Pan
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D057846:Cell-Penetrating Peptides; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D008164:Luminescent Proteins / Q000235:genetics; D010957:Plasmids / Q000008:administration & dosage* / Q000737:chemistry*; D051381:Rats; D014162:Transfection; D000097573:Red Fluorescent Protein
chemical_list: D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein; C414409:penetratin
keywords: PAMAM; gene delivery; penetratin; physical complex; posterior segment of the eye