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学科主题: 临床医学
题名:
Enhanced Factor VIII Heavy Chain for Gene Therapy of Hemophilia A
作者: Chen, Lingxia1,2; Lu, Hui1; Wang, Jinhui1; Sarkar, Rita1; Yang, Xiao1; Wang, Hongli3; High, Katherine A.1,4; Xiao, Weidong1
刊名: MOLECULAR THERAPY
发表日期: 2009-03-01
DOI: 10.1038/mt.2008.292
卷: 17, 期:3, 页:417-424
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biotechnology & Applied Microbiology ; Genetics & Heredity ; Medicine, Research & Experimental
研究领域[WOS]: Biotechnology & Applied Microbiology ; Genetics & Heredity ; Research & Experimental Medicine
关键词[WOS]: RECOMBINANT ADENOASSOCIATED VIRUS ; COAGULATION-FACTOR VIII ; VON-WILLEBRAND-FACTOR ; FACTOR-IX ; LIGHT-CHAIN ; BLOOD-COAGULATION ; AAV VECTORS ; INTRAMUSCULAR INJECTION ; SUSTAINED EXPRESSION ; PACKAGING CAPACITY
英文摘要:

Hemophilia A gene therapy using recombinant adenovirus-associated virus (AAV) vectors has been hampered by the size of the factor VIII (FVIII) cDNA. Previously, splitting the FVIII coding sequence into a heavy-chain (HC) fragment and a light-chain (LC) fragment for dual recombinant AAV vector delivery has been successfully explored. However, the main disadvantage of this approach is a "chain imbalance" problem in which LC secretion is similar to 1-2 logs higher than that of HC, and therefore, the majority of protein synthesized is nonfunctional. To improve HC secretion, we constructed alternate FVIII HCs based on our observation that LC facilitates HC secretion. To our surprise, most of the new HC molecules exhibited enhanced expression over the traditional HC molecule (HC(745)). The optimized HC mutein, HC(HL), including additional acidic-region-3 (ar3) sequences, exhibited three-to fivefold higher activity in both enzyme-linked immunosorbent assay (ELISA) and activated partial thromboplastin time (aPTT) assay in in vitro testing. Further characterization suggested ar3 sequences increased HC secretion, rather than promoting HC synthesis. Intravenous delivery of AAV8-HC(HL)+AAV8-LC or AAV8-HC(745)+AAV8-LC achieved phenotypic correction in hemophilia A mice. Mice receiving AAV8-HC(HL)+AAV8-LC achieved three-to fourfold higher HC expression than AAV8-HC(745)+AAV8-LC, consistent with the FVIII functional assays. HC(HL) should be substituted for HC(745) in a dual AAV vector strategy due to its enhanced expression.

语种: 英语
所属项目编号: HL080789 ; HL084381
项目资助者: National Institutes of Health
WOS记录号: WOS:000263788200009
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/57148
Appears in Collections:北京大学第二临床医学院_期刊论文

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作者单位: 1.Univ Penn, Med Ctr, Dept Pediat, Philadelphia, PA 19104 USA
2.Peking Univ, Peoples Hosp, Beijing, Peoples R China
3.Shanghai Jiao Tong Univ, Ruijin Hosp, Sch Med, Shanghai 200030, Peoples R China
4.Childrens Hosp Philadelphia, Howard Hughes Med Inst, Philadelphia, PA 19104 USA

Recommended Citation:
Chen, Lingxia,Lu, Hui,Wang, Jinhui,et al. Enhanced Factor VIII Heavy Chain for Gene Therapy of Hemophilia A[J]. MOLECULAR THERAPY,2009,17(3):417-424.
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