OBJECTIVE: To test the hypothesis that in vivo transgene expression mediated by single intra-articular injection of adeno-associated virus serotype 2 (AAV2) persists within intra-articular tissues 1 year post-injection and can be externally controlled using an AAV2-based tetracycline-inducible gene regulation system containing the tetracycline response element (TRE) promoter. METHODS: Sprague Dawley rats received intra-articular injections of AAV2-cytomegalovirus (CMV)-enhanced green fluorescent protein (GFP) and AAV2-CMV-luciferase (Luc) into their right and left knees, respectively. Luciferase expression was evaluated over 1 year using bioluminescence imaging. After sacrifice, tissues were analyzed for GFP+ cells by fluorescent microscopy. To study external control of intra-articular AAV-transgene expression, another set of rats was co-injected with AAV2-TRE-Luc and AAV2-CMV-reverse-tetracycline-controlled transactivator (rtTA) into the right knees, and AAV2-CMV-Luc and AAV2-CMV-rtTA into the left knees. Rats received oral doxycycline (Dox), an analog of tetracycline, for 7 days. Luciferase expression was assessed by bioluminescence imaging. RESULTS: Luciferase expression was localized to the injected joint and persisted throughout the 1-year study period. Abundant GFP+ cells were observed within intra-articular soft tissues. Transgene expression in AAV2-TRE-Luc injected joints was upregulated by oral administration of Dox, and downregulated following its removal, at 14 days and 13 months post-AAV injection. CONCLUSIONS: This longitudinal in vivo study shows that sustained and stable AAV-mediated intra-articular transgene expression can be achieved through a single intra-articular injection and can be controlled using a tetracycline-controlled inducible AAV system in a normal rat knee model. Highly regulatable long-term intra-articular transgene expression is of potential clinical utility for development of treatment strategies for chronic intra-articular disease processes such as inflammatory and degenerative arthritis.
OBJECTIVE: To test the hypothesis that in vivo transgene expression mediated by single intra-articular injection of adeno-associated virus serotype 2 (AAV2) persists within intra-articular tissues 1 year post-injection and can be externally controlled using an AAV2-based tetracycline-inducible gene regulation system containing the tetracycline response element (TRE) promoter. METHODS:Sprague Dawley rats received intra-articular injections of AAV2-cytomegalovirus (CMV)-enhanced green fluorescent protein (GFP) and AAV2-CMV-luciferase (Luc) into their right and left knees, respectively. Luciferase expression was evaluated over 1 year using bioluminescence imaging. After sacrifice, tissues were analyzed for GFP+ cells by fluorescent microscopy. To study external control of intra-articular AAV-transgene expression, another set of rats was co-injected with AAV2-TRE-Luc and AAV2-CMV-reverse-tetracycline-controlled transactivator (rtTA) into the right knees, and AAV2-CMV-Luc and AAV2-CMV-rtTA into the left knees. Rats received oral doxycycline (Dox), an analog of tetracycline, for 7 days. Luciferase expression was assessed by bioluminescence imaging. RESULTS: Luciferase expression was localized to the injected joint and persisted throughout the 1-year study period. Abundant GFP+ cells were observed within intra-articular soft tissues. Transgene expression in AAV2-TRE-Luc injected joints was upregulated by oral administration of Dox, and downregulated following its removal, at 14 days and 13 months post-AAV injection. CONCLUSIONS: This longitudinal in vivo study shows that sustained and stable AAV-mediated intra-articular transgene expression can be achieved through a single intra-articular injection and can be controlled using a tetracycline-controlled inducible AAV system in a normal rat knee model. Highly regulatable long-term intra-articular transgene expression is of potential clinical utility for development of treatment strategies for chronic intra-articular disease processes such as inflammatory and degenerative arthritis.
Authors: Elvire Gouze; Jean-Noel Gouze; Glyn D Palmer; Carmencita Pilapil; Christopher H Evans; Steven C Ghivizzani Journal: Mol Ther Date: 2007-04-17 Impact factor: 11.454
Authors: M R Hasanjani Roushan; M J Soleimani Amiri; N Janmohammadi; M Sadeghi Hadad; M Javanian; M Baiani; A Bijani Journal: J Antimicrob Chemother Date: 2010-03-09 Impact factor: 5.790
Authors: Iñigo Izal; Carlos Alberto Acosta; Purificación Ripalda; Mikel Zaratiegui; Juan Ruiz; Francisco Forriol Journal: Arch Orthop Trauma Surg Date: 2007-07-28 Impact factor: 3.067
Authors: G L Hung; J Galea-Lauri; G M Mueller; H I Georgescu; L A Larkin; M K Suchanek; M H Tindal; P D Robbins; C H Evans Journal: Gene Ther Date: 1994-01 Impact factor: 5.250
Authors: Philip J Mease; Nathan Wei; Edward J Fudman; Alan J Kivitz; Joy Schechtman; Robert G Trapp; Kathryn F Hobbs; Maria Greenwald; Antony Hou; Stephen A Bookbinder; Galen E Graham; Craig W Wiesenhutter; Larry Willis; Eric M Ruderman; Joseph Z Forstot; Michael J Maricic; Kathryn H Dao; Charles H Pritchard; Darrell N Fiske; Francis X Burch; H Malin Prupas; Pervin Anklesaria; Alison E Heald Journal: J Rheumatol Date: 2009-12-23 Impact factor: 4.666
Authors: Jeffrey B Mason; Brittney L Gurda; Julie B Engiles; Kurt D Hankenson; James M Wilson; Dean W Richardson Journal: Hum Gene Ther Methods Date: 2013-06 Impact factor: 2.396
Authors: Hannah H Lee; Amgad M Haleem; Veronica Yao; Juan Li; Xiao Xiao; Constance R Chu Journal: Tissue Eng Part A Date: 2011-05-11 Impact factor: 3.845
Authors: Hannah H Lee; Michael J O'Malley; Nicole A Friel; Karin A Payne; Chunping Qiao; Xiao Xiao; Constance R Chu Journal: Hum Gene Ther Date: 2013-04 Impact factor: 5.695