Modeling correction of severe urea cycle defects in the growing murine liver using a hybrid recombinant adeno-associated virus/piggyBac transposase gene delivery system

被引：34

作者：

Cunningham, Sharon C. ^{[1
,2
,3
]}

Siew, Susan M. ^{[1
,2
]}

Hallwirth, Claus V. ^{[1
,2
]}

Bolitho, Christine ^{[1
,2
]}

Sasaki, Natsuki ^{[1
,2
]}

Garg, Gagan ^{[1
,2
,4
]}

Michael, Iacovos P. ^{[5
]}

Hetherington, Nicola A. ^{[1
,2
]}

Carpenter, Kevin ^{[6
]}

de Alencastro, Gustavo ^{[1
,2
]}

Nagy, Andras ^{[5
,7
,8
]}

Alexander, Ian E. ^{[1
,2
,9
]}

机构：

[1] Childrens Med Res Inst, Gene Therapy Res Unit, Westmead, NSW 2145, Australia

[2] Childrens Hosp, Westmead, NSW 2145, Australia

[3] Univ Sydney, Sch Med, Sydney, NSW 2006, Australia

[4] Macquarie Univ, Dept Chem & Biomol Sci, Macquarie Pk, NSW, Australia

[5] Mt Sinai Hosp, Lunenfeld Tanenbaum Res Inst, Toronto, ON M5G 1X5, Canada

[6] Childrens Hosp Westmead, Biochem Genet, Sydney, NSW, Australia

[7] Univ Toronto, Inst Med Sci, Toronto, ON, Canada

[8] Univ Toronto, Dept Obstet & Gynaecol, Toronto, ON, Canada

[9] Univ Sydney, Discipline Paediat & Child Hlth, Sydney, NSW 2006, Australia

来源：

HEPATOLOGY | 2015年 / 62卷 / 02期

基金：

加拿大健康研究院;

关键词：

VIRUS VECTOR INTEGRATION; ARGININOSUCCINATE SYNTHETASE; HEMOPHILIA-B; IN-VIVO; HEPATOCELLULAR-CARCINOMA; RETROVIRAL INTEGRATION; MAMMALIAN-CELLS; SLEEPING-BEAUTY; DEFICIENT MICE; MOUSE-LIVER;

D O I：

10.1002/hep.27842

中图分类号：

R57 [消化系及腹部疾病];

学科分类号：

100201 [内科学];

摘要：

Liver-targeted gene therapy based on recombinant adeno-associated viral vectors (rAAV) shows promising therapeutic efficacy in animal models and adult-focused clinical trials. This promise, however, is not directly translatable to the growing liver, where high rates of hepatocellular proliferation are accompanied by loss of episomal rAAV genomes and subsequently a loss in therapeutic efficacy. We have developed a hybrid rAAV/piggyBac transposon vector system combining the highly efficient liver-targeting properties of rAAV with stable piggyBac-mediated transposition of the transgene into the hepatocyte genome. Transposition efficiency was first tested using an enhanced green fluorescent protein expression cassette following delivery to newborn wild-type mice, with a 20-fold increase in stably gene-modified hepatocytes observed 4 weeks posttreatment compared to traditional rAAV gene delivery. We next modeled the therapeutic potential of the system in the context of severe urea cycle defects. A single treatment in the perinatal period was sufficient to confer robust and stable phenotype correction in the ornithine transcarbamylase-deficient Spf(ash) mouse and the neonatal lethal argininosuccinate synthetase knockout mouse. Finally, transposon integration patterns were analyzed, revealing 127,386 unique integration sites which conformed to previously published piggyBac data. Conclusion: Using a hybrid rAAV/piggyBac transposon vector system, we achieved stable therapeutic protection in two urea cycle defect mouse models; a clinically conceivable early application of this technology in the management of severe urea cycle defects could be as a bridging therapy while awaiting liver transplantation; further improvement of the system will result from the development of highly human liver-tropic capsids, the use of alternative strategies to achieve transient transposase expression, and engineered refinements in the safety profile of piggyBac transposase-mediated integration. (Hepatology 2015;62:417-428

引用

页码：417 / 428

页数：12

共 49 条

[1]

[Anonymous], 1989, Cold Spring Harbor

[2]

A pediatric patient with classical citrullinemia who underwent living-related partial liver transplantation [J].