title: Targeting transgene to the heart and liver with AAV9 by different promoters.
abstract: Adeno-associated virus (AAV) has become one of the most promising gene transfer tools for gene therapy. This work aims to evaluate tropism, gene transfer efficiency and safety of AAV9 vectors produced with recombinant baculovirus (rBac)-based system. AAV9-CMV-GFP and AAV9-CBA-GFP were produced using a rBac system, 1 × 10(11) particles of each vectors were administered intravenously (i.v.) into mice and animals were killed at 1, 2, 3, 4, 5 and 8 weeks after administration. The GFP expression in different organs was analyzed by fluorescence imaging and Western blot. Viral genomic quantities were measured using qPCR. In vitro transduction efficiency of AAV9 vectors in primary cardiomyocytes and hepatocytes was determined by flow cytometry. Toxicity of AAV9 vectors was evaluated by determining certain cardiac and liver injury biomarkers and renal function test in vivo and TUNEL analysis in vitro. The data showed that AAV9 viral particles packaged by the rBac system were fully functional in vivo and in vitro. The CMV promoter predominantly induced higher cardiac GFP transgene expression and DNA copy numbers while the CBA promoter resulted in robust GFP expression and high vector DNA copy numbers in mouse liver, both in a time-dependent increased manner. Such distinct preferential effects were also observed in the heart and liver as early as 3 and 5 days after co-infection. Both the AAV9-CMV and AAV9-CBA viral packages did not induce heart, liver and renal damage and cell apoptosis. These results indicated that AAV9-CMV can efficiently and safely direct cardiac gene transfer, whereas AAV9-CBA is preferential for liver gene delivery.
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi, China.
authors.forename: Bang-Dang
authors.initials: BD
authors.lastname: Chen
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi, China.|Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi, China.
authors.forename: Chun-Hui
authors.initials: CH
authors.lastname: He
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi, China.
authors.forename: Xiao-Cui
authors.initials: XC
authors.lastname: Chen
authors.affiliation: Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi, China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Pan
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi, China.
authors.forename: Fen
authors.initials: F
authors.lastname: Liu
authors.affiliation: Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Ma
authors.affiliation: Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi, China.
authors.forename: Xiao-Mei
authors.initials: XM
authors.lastname: Li
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi, China.
authors.forename: Min-Tao
authors.initials: MT
authors.lastname: Gai
authors.affiliation: Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Tao
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi, China.|Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi, China.
authors.forename: Yi-Tong
authors.initials: YT
authors.lastname: Ma
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi, China.
authors.forename: Yi-Ning
authors.initials: YN
authors.lastname: Yang
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi, China.|Baker IDI Heart and Diabetes Institute, Melbourne, Australia.
authors.forename: Xiao-Ming
authors.initials: XM
authors.lastname: Gao
mesh_terms: D000199:Actins / Q000235:genetics; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D002645:Chickens / Q000235:genetics; D003587:Cytomegalovirus / Q000235:genetics; D000229:Dependovirus / Q000235:genetics* / Q000502:physiology; D005822:Genetic Vectors / Q000235:genetics; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012449:Safety; D014161:Transduction, Genetic / Q000379:methods*; D019076:Transgenes / Q000235:genetics*; D056189:Viral Tropism
chemical_list: D000199:Actins
keywords: AAV9; CBA promoter; CMV promoter; heart; liver; rBac-based system