Empty virions in AAV8 vector preparations reduce transduction efficiency and may cause total viral particle dose-limiting side effects

被引：129

作者：

Gao, Kai ^{[1
,2
]}

Li, Mengxin ^{[1
,3
,4
]}

Zhong, Li ^{[1
,3
,5
]}

Su, Qin ^{[1
,3
]}

Li, Jia ^{[1
,4
]}

Li, Shaoyong ^{[1
,4
]}

He, Ran ^{[1
,3
]}

Zhang, Yu ^{[1
,6
]}

Hendricks, Gregory ^{[7
]}

Wang, Junzhi ^{[2
]}

Gao, Guangping ^{[1
,3
,4
,8
]}

机构：

[1] Univ Massachusetts, Sch Med, Gene Therapy Ctr, Worcester, MA 01655 USA

[2] Natl Inst Food & Drug Control, Beijing, Peoples R China

[3] Univ Massachusetts, Sch Med, Viral Vector Core, Worcester, MA USA

[4] Univ Massachusetts, Sch Med, Dept Microbiol & Physiol Syst, Worcester, MA USA

[5] Univ Massachusetts, Sch Med, Dept Pediat, Div Hematol Oncol, Worcester, MA USA

[6] Guizhou Peoples Hosp, Dept Oncol, Guiyang, Guizhou, Peoples R China

[7] Univ Massachusetts, Sch Med, Electron Microscopy Core Facil, Dept Cell & Dev Biol, Worcester, MA USA

[8] Sichuan Univ, West China Hosp, Key Lab Biotherapy, Chengdu, Sichuan, Peoples R China

来源：

MOLECULAR THERAPY-METHODS & CLINICAL DEVELOPMENT | 2014年 / 1卷

基金：

美国国家卫生研究院;

关键词：

CAPSID ANTIGEN PRESENTATION; GENE-TRANSFER; ANALYTICAL ULTRACENTRIFUGATION; FACTOR-IX; IN-VIVO; VIRUS; MICE; HEMOPHILIA; TISSUE; ADENOVIRUS;

D O I：

10.1038/mtm.2013.9

中图分类号：

R-3 [医学研究方法]; R3 [基础医学];

学科分类号：

1001 ;

摘要：

Empty virions are inadvertent by-products of recombinant adeno-associated virus (rAAV) packaging process, resulting in vector lots with mixtures of full and empty virions at variable ratios. Impact of empty virions on the efficiency and side effects of rAAV transduction has not been well characterized. Here, we generated partially and completely empty AAV8 virions, fully packaged rAAV8 lots, and mixtures of empty and fully packaged virions with variable ratios of empty virions. The aforementioned dosing formulations of rAAV8 expressing either cellular (EGFP (enhanced green fluorescent protein) or nuclear-targeted (n) LacZ) or secreted (human a1-antitrypsin (hA1AT)) reporter genes were intravenously injected into two different mouse strains, followed by analyses of transgene expressions and serum alanine aminotransferase (ALT) levels at different time points. We found that addition of empty particles to the fixed doses of rAAV8 preparations repressed liver transduction up to 64% (serum hA1AT) and 44% (nLacZ) in C57BL/6 mice, respectively. The similar trend in inhibiting EGFP expression together with concurrent elevations of serum ALT levels were observed in the BALB/c mice, indicating that empty particles may also exacerbate side effects of rAAV8 EGFP transduction. Our results suggest that removal of empty particles from rAAV preparations may improve efficacy and safety of AAV in clinical applications.

引用

页数：8

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