Genotoxic Potential of Lineage-specific Lentivirus Vectors Carrying the β-Globin Locus Control Region

被引：67

作者：

Arumugam, Paritha I. ^{[1
]}

Higashimoto, Tomoyasu ^{[1
]}

Urbinati, Fabrizia ^{[1
]}

Modlich, Ute ^{[2
]}

Nestheide, Shawna ^{[1
]}

Xia, Ping ^{[1
]}

Fox, Catherine ^{[1
]}

Corsinotti, Andrea ^{[1
]}

Baum, Christopher ^{[1
,2
]}

Malik, Punam ^{[1
,3
]}

机构：

[1] Cincinnati Childrens Hosp, Div Expt Hematol Canc Biol, Med Ctr, Cincinnati Childrens Res Fdn, Cincinnati, OH 45229 USA

[2] Hannover Med Sch, Div Expt Hematol, D-3000 Hannover, Germany

[3] Cincinnati Childrens Hosp, Div Hematol Oncol, Cincinnati Childrens Res Fdn, Med Ctr, Cincinnati, OH 45229 USA

来源：

MOLECULAR THERAPY | 2009年 / 17卷 / 11期

关键词：

GENE-THERAPY; INTEGRATION SITE; RETROVIRAL INTEGRATION; ENHANCER BLOCKING; MOUSE MODEL; EXPRESSION; ACTIVATION; INSULATOR; DESIGN; CELLS;

D O I：

10.1038/mt.2009.183

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Insertional mutagenesis by long terminal repeat (LTR) enhancers in gamma-retrovirus-based vectors (GVs) in clinical trials has prompted deeper investigations into vector genotoxicity. Experimentally, self-inactivating (SIN) lentivirus vectors (LVs) and GV containing internal-promoters/enhancers show reduced genotoxicity, although strong ubiquitously-active enhancers dysregulate genes independent of vector type/design. Herein, we explored the genotoxicity of beta-globin (BG) locus control region (LCR), a strong long-range lineage-specific-enhancer, with/without insulator (Ins) elements in LV using primary hematopoietic progenitors to generate in vitro immortalization (IVIM) assay mutants. LCR-containing LV had similar to 200-fold lower transforming potential, compared to the conventional GV. The LCR perturbed expression of few genes in a 300 kilobase (kb) proviral vicinity but no upregulation of genes associated with cancer, including an erythroid-specific transcription factor occurred. A further twofold reduction in transforming activity was observed with insulated LCR-containing LV. Our data indicate that toxicology studies of LCR-containing LV in mice will likely not yield any insertional oncogenesis with the numbers of animals that can be practically studied.

引用

页码：1929 / 1937

页数：9

共 42 条

[1] Improved human β-globin expression from self-inactivating lentiviral vectors carrying the chicken hypersensitive site-4 (cHS4) insulator element [J].

Arumugam, Paritha I. ;

Scholes, Jessica ;

Perelman, Natalya ;

Xia, Ping ;

Yee, Jiing-Kuan ;

Malik, Punam .

MOLECULAR THERAPY, 2007, 15 (10) :1863-1871

[2] The protein CTCF is required for the enhancer blocking activity of vertebrate insulators [J].

Bell, AC ;

West, AG ;

Felsenfeld, G .

CELL, 1999, 98 (03) :387-396

[3] Alternative mRNA splicing of the novel GTPase rab28 generates isoforms with different C-termini [J].

Brauers, A ;

Schurmann, A ;

Massmann, S ;

MuhlZurbes, P ;

Becker, W ;

Kainulainen, H ;

Lie, CC ;

Joost, HG .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1996, 237 (03) :833-840

[4] Genome wide analysis of retroviral DNA integration [J].

Bushman, F ;

Lewinski, M ;

Ciuffi, A ;

Barr, S ;

Leipzig, J ;

Hannenhalli, S ;

Hoffmann, C .

NATURE REVIEWS MICROBIOLOGY, 2005, 3 (11) :848-858

[5] Recurrent retroviral vector integration at the Mds1/Evi1 locus in nonhuman primate hematopoietic cells [J].

Calmels, B ;

Ferguson, C ;

Laukkanen, MO ;

Adler, R ;

Faulhaber, M ;

Kim, HJ ;

Sellers, S ;

Hematti, P ;

Schmidt, M ;

von Kalle, C ;

Akagi, K ;

Donahue, RE ;

Dunbar, CE .

BLOOD, 2005, 106 (07) :2530-2533

[6] Hot spots of retroviral integration in human CD34+ hematopoietic cells [J].

Cattoglio, Claudia ;

Facchini, Giulia ;

Sartori, Daniela ;

Antonelli, Antonella ;

Miccio, Annarita ;

Cassani, Barbara ;

Schmidt, Manfred ;

von Kalle, Christof ;

Howe, Steve ;

Thrasher, Adrian J. ;

Aiuti, Alessandro ;

Ferrari, Giuliana ;

Recchia, Alessandra ;

Mavilio, Fulvio .

BLOOD, 2007, 110 (06) :1770-1778

[7] Thyroid hormone receptor mutations in cancer [J].

Cheng, SY .

MOLECULAR AND CELLULAR ENDOCRINOLOGY, 2003, 213 (01) :23-30

[8] Transcription Factor Binding Sites Are Genetic Determinants of Retroviral Integration in the Human Genome [J].

Felice, Barbara ;

Cattoglio, Claudia ;

Cittaro, Davide ;

Testa, Anna ;

Miccio, Annarita ;

Ferrari, Giuliana ;

Luzi, Lucilla ;

Recchia, Alessandra ;

Mavilio, Fulvio .

PLOS ONE, 2009, 4 (02)

[9] POSITION-INDEPENDENT, HIGH-LEVEL EXPRESSION OF THE HUMAN BETA-GLOBIN GENE IN TRANSGENIC MICE [J].

GROSVELD, F ;

VANASSENDELFT, GB ;

GREAVES, DR ;

KOLLIAS, G .

CELL, 1987, 51 (06) :975-985

[10] Insertional oncogenesis in 4 patients after retrovirus-mediated gene therapy of SCID-X1 [J].

Hacein-Bey-Abina, Salima ;

Garrigue, Alexandrine ;

Wang, Gary P. ;

Soulier, Jean ;

Lim, Annick ;

Morillon, Estelle ;

Clappier, Emmanuelle ;

Caccavelli, Laure ;

Delabesse, Eric ;

Beldjord, Kheira ;

Asnafi, Vahid ;

Macintyre, Elizabeth ;

Dal Cortivo, Liliane ;

Radford, Isabelle ;

Brousse, Nicole ;

Sigaux, Francois ;

Moshous, Despina ;

Hauer, Julia ;

Borkhardt, Arndt ;

Belohradsky, Bernd H. ;

Wintergerst, Uwe ;

Velez, Maria C. ;

Leiva, Lily ;

Sorensen, Ricardo ;

Wulffraat, Nicolas ;

Blanche, Stephane ;

Bushman, Frederic D. ;

Fischer, Alain ;

Cavazzana-Calvo, Marina .

JOURNAL OF CLINICAL INVESTIGATION, 2008, 118 (09) :3132-3142

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