title: Persistence, localization, and external control of transgene expression after single injection of adeno-associated virus into injured joints.
abstract: A single intra-articular injection of adeno-associated virus (AAV) results in stable and controllable transgene expression in normal rat knees. Because undamaged joints are unlikely to require treatment, the study of AAV delivery in joint injury models is crucial to potential therapeutic applications. This study tests the hypotheses that persistent and controllable AAV-transgene expression are (1) highly localized to the cartilage when AAV is injected postinjury and (2) localized to the intra-articular soft tissues when AAV is injected preinjury. Two AAV injection time points, postinjury and preinjury, were investigated in osteochondral defect and anterior cruciate ligament transection models of joint injury. Rats injected with AAV tetracycline response element (TRE)-luciferase received oral doxycycline for 7 days. Luciferase expression was evaluated longitudinally for 6 months. Transgene expression was persistent and controllable with oral doxycycline for 6 months in all groups. However, the location of transgene expression was different: postinjury AAV-injected knees had luciferase expression highly localized to the cartilage, while preinjury AAV-injected knees had more widespread signal from intra-articular soft tissues. The differential transgene localization between preinjury and postinjury injection can be used to optimize treatment strategies. Highly localized postinjury injection appears advantageous for treatments targeting repair cells. The more generalized and controllable reservoir of transgene expression following AAV injection before anterior cruciate ligament transection (ACLT) suggests an intriguing concept for prophylactic delivery of joint protective factors to individuals at high risk for early osteoarthritis (OA). Successful external control of intra-articular transgene expression provides an added margin of safety for these potential clinical applications.
authors.affiliation: Cartilage Restoration Center, Department of Orthopaedic Surgery, University of Pittsburgh, PA 15313, USA.
authors.forename: Hannah H
authors.initials: HH
authors.lastname: Lee
authors.forename: Michael J
authors.initials: MJ
authors.lastname: O'Malley
authors.forename: Nicole A
authors.initials: NA
authors.lastname: Friel
authors.forename: Karin A
authors.initials: KA
authors.lastname: Payne
authors.forename: Chunping
authors.initials: C
authors.lastname: Qiao
authors.forename: Xiao
authors.initials: X
authors.lastname: Xiao
authors.forename: Constance R
authors.initials: CR
authors.lastname: Chu
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics*; D015316:Genetic Therapy; D005822:Genetic Vectors; D007270:Injections, Intra-Articular; D007592:Joint Diseases / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D007596:Joints / Q000293:injuries* / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D019076:Transgenes
chemical_list: D008156:Luciferases