Preintegration HIV-1 inhibition by a combination lentiviral vector containing a chimeric TRIM5 alpha protein, a CCR5 shRNA, and a TAR decoy.
Human immunodeficiency virus (HIV) gene therapy offers a promising alternative approach to current antiretroviral treatments to inhibit HIV-1 infection. Various stages of the HIV life cycle including pre-entry, preintegration, and postintegration can be targeted by gene therapy to block viral infection and replication. By combining multiple highly potent anti-HIV transgenes in a single gene therapy vector, HIV-1 resistance can be achieved in transduced cells while prohibiting the generation of escape mutants. Here, we describe a combination lentiviral vector that encodes three highly effective anti-HIV genes functioning at separate stages of the viral life cycle including a CCR5 short hairpin RNA (shRNA) (pre-entry), a human/rhesus macaque chimeric TRIM5 alpha (postentry/preintegration), and a transactivation response element (TAR) decoy (postintegration). The major focus on designing this anti-HIV vector was to block productive infection of HIV-1 and to inhibit any formation of provirus that would maintain the viral reservoir. Upon viral challenge, potent preintegration inhibition of HIV-1 infection was achieved in combination vector-transduced cells in both cultured and primary CD34(+) hematopoietic progenitor cell (HPC)-derived macrophages. The generation of escape mutants was also blocked as evaluated by long-term culture of challenged cells. The ability of this combination anti-HIV lentiviral vector to prevent HIV-1 infection, in vitro, warrants further evaluation of its in vivo efficacy.
Anderson
Joseph S
JS
Stem Cell Program, Department of Internal Medicine, University of California-Davis, Sacramento, California, USA. joseph.anderson@ucdmc.ucdavis.edu
Javien
John
J
Nolta
Jan A
JA
Bauer
Gerhard
G
D000818:Animals; D000090103:Antiviral Restriction Factors; D065100:CCR5 Receptor Antagonists; D002352:Carrier Proteins / Q000235:genetics*; D005434:Flow Cytometry; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage*; D006678:HIV / Q000235:genetics; D015658:HIV Infections / Q000235:genetics / Q000628:therapy* / Q000821:virology; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008253:Macaca mulatta; D008264:Macrophages / Q000378:metabolism; D011506:Proteins / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics*; D019713:Receptors, CCR5 / Q000235:genetics*; D014161:Transduction, Genetic; D019076:Transgenes / Q000502:physiology; D000070584:Tripartite Motif Proteins; D044767:Ubiquitin-Protein Ligases; D016662:Virus Integration; D054321:rev Gene Products, Human Immunodeficiency Virus / Q000037:antagonists & inhibitors / Q000502:physiology*; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000037:antagonists & inhibitors / Q000502:physiology
D000090103:Antiviral Restriction Factors; D065100:CCR5 Receptor Antagonists; D002352:Carrier Proteins; D011506:Proteins; D034741:RNA, Small Interfering; D019713:Receptors, CCR5; D000070584:Tripartite Motif Proteins; D054321:rev Gene Products, Human Immunodeficiency Virus; D054322:tat Gene Products, Human Immunodeficiency Virus; C477346:TRIM5 protein, human; C482587:TRIM5(alpha) protein, rhesus monkey; D044767:Ubiquitin-Protein Ligases
Blood. 1999 Jul 1;94(1):368-71
10381536
AIDS. 2002 Dec 6;16(18):2385-90
12461411
N Engl J Med. 2009 Feb 12;360(7):692-8
19213682
Antimicrob Agents Chemother. 2000 Sep;44(9):2475-84
10952598
Proc Natl Acad Sci U S A. 2007 Aug 7;104(32):13110-5
17670939
Nat Med. 1996 Nov;2(11):1240-3
8898752
RNA. 2002 Nov;8(11):1454-60
12458798
Mol Ther. 2000 Mar;1(3):244-54
10933940
Cell. 2008 Aug 22;134(4):577-86
18691745
Nat Biotechnol. 2002 May;20(5):500-5
11981565
Gene Ther. 2007 Sep;14(17):1287-97
17597795
J Virol. 2002 Apr;76(7):3114-24
11884536
Antivir Ther. 2000 Mar;5(1):57-63
10846594
HIV Med. 2001 Oct;2(4):231-5
11737402
Mol Ther. 2006 Oct;14(4):494-504
16844419
Gene Ther. 2009 Jan;16(1):148-53
18668146
Proc Natl Acad Sci U S A. 2005 Feb 22;102(8):2832-7
15689398
Curr HIV/AIDS Rep. 2004 Jun;1(2):82-8
16091227
Annu Rev Immunol. 1999;17:657-700
10358771
Nat Med. 2002 Jul;8(7):681-6
12042777
Mol Ther. 2004 Jun;9(6):902-13
15194057
Nature. 1998 Feb 19;391(6669):806-11
9486653
Cell. 1996 Aug 9;86(3):367-77
8756719
Oligonucleotides. 2003;13(5):281-94
15000819
Front Biosci. 2002 Feb 01;7:a15-28
11815282
Proc Natl Acad Sci U S A. 2002 Oct 29;99(22):14047-52
12376617
Proc Natl Acad Sci U S A. 2000 Sep 26;97(20):10948-53
11005867
J Virol. 1997 Jun;71(6):4707-16
9151864
Nature. 2004 Feb 26;427(6977):848-53
14985764
Nat Med. 2009 Mar;15(3):285-92
19219022
Mol Ther. 2007 Jun;15(6):1182-8
17406343
Ann N Y Acad Sci. 2000 Nov;918:318-29
11131719
Hum Gene Ther. 2008 Mar;19(3):217-28
18279037