title: Efficient Transduction of Corneal Stroma by Adeno-Associated Viral Serotype Vectors for Implications in Gene Therapy of Corneal Diseases.
abstract: Corneal disease is one of the leading causes of blindness worldwide. Gene therapy is an attractive therapeutic strategy for corneal diseases, but currently underdeveloped. Recombinant adeno-associated viral (rAAV) vectors have emerged as a highly promising gene therapy platform. This study aims to identify rAAV vectors that can efficiently transduce corneal stroma for potential applications in studying pathophysiology of corneal diseases and therapeutic development. We characterized 14 rAAV serotypes expressing enhanced green fluorescent protein (EGFP), for cell specificity and transduction efficiency after either intrastromal injection or topical administration in mouse corneas in vivo. Our results show that intrastromal injections of rAAVrh.8, rAAVrh.10, rAAVrh.39, and rAAVrh.43 efficiently transduce mouse corneal stroma in vivo, and that topical administrations of rAAVrh.10 and rAAVrh.39 subsequent to epithelial scraping generate detectable transgene expression. In vivo imaging analysis revealed that transgene expression became detectable by 1 week postadministration, peaked at 2 weeks, and lasted for the duration of the study (i.e., 4 weeks). Both rAAVrh.10 and rAAVrh.39 transduced more than 50% of keratocytes, the major cell type in the corneal stroma, by intrastromal injection and 30% by topical administration. Histopathology indicated that rAAV transduction of cornea caused no morphological adverse effects. Overall, our findings suggest that some rAAV serotype vectors can efficiently transduce corneal stroma in vivo, constituting a potentially powerful and safe gene delivery platform for gene therapy of corneal diseases.
authors.affiliation: 1 Department of Ophthalmology, Shanghai First People's Hospital, School of Medicine, Shanghai Jiaotong University , Shanghai, China.|2 Shanghai Key Laboratory of Fundus Disease , Shanghai, China.|3 Horae Gene Therapy Center, University of Massachusetts Medical School , Worcester, Massachusetts.|4 Department of Microbiology and Physiological Systems, University of Massachusetts Medical School , Worcester, Massachusetts.
authors.forename: Yi
authors.initials: Y
authors.lastname: Lu
authors.affiliation: 3 Horae Gene Therapy Center, University of Massachusetts Medical School , Worcester, Massachusetts.|4 Department of Microbiology and Physiological Systems, University of Massachusetts Medical School , Worcester, Massachusetts.|5 Department of Urology, West China Hospital, Sichuan University , Chengdu, Sichuan, China.
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Ai
authors.affiliation: 3 Horae Gene Therapy Center, University of Massachusetts Medical School , Worcester, Massachusetts.|4 Department of Microbiology and Physiological Systems, University of Massachusetts Medical School , Worcester, Massachusetts.
authors.forename: Dominic
authors.initials: D
authors.lastname: Gessler
authors.affiliation: 3 Horae Gene Therapy Center, University of Massachusetts Medical School , Worcester, Massachusetts.
authors.forename: Qin
authors.initials: Q
authors.lastname: Su
authors.affiliation: 3 Horae Gene Therapy Center, University of Massachusetts Medical School , Worcester, Massachusetts.
authors.forename: Karen
authors.initials: K
authors.lastname: Tran
authors.affiliation: 6 Research and Development Department, Chengdu Kanghong Pharmaceuticals Group Company , Chengdu, Sichuan, China .
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zheng
authors.affiliation: 1 Department of Ophthalmology, Shanghai First People's Hospital, School of Medicine, Shanghai Jiaotong University , Shanghai, China.|2 Shanghai Key Laboratory of Fundus Disease , Shanghai, China.
authors.forename: Xun
authors.initials: X
authors.lastname: Xu
authors.affiliation: 3 Horae Gene Therapy Center, University of Massachusetts Medical School , Worcester, Massachusetts.|4 Department of Microbiology and Physiological Systems, University of Massachusetts Medical School , Worcester, Massachusetts.
authors.forename: Guangping
authors.initials: G
authors.lastname: Gao
mesh_terms: D000818:Animals; D003316:Corneal Diseases / Q000235:genetics / Q000628:therapy*; D003319:Corneal Stroma / Q000378:metabolism* / Q000473:pathology; D000229:Dependovirus / Q000235:genetics*; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage*; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D014161:Transduction, Genetic
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins