Long-term safety and efficacy following systemic administration of a self-complementary AAV vector encoding human FIX pseudotyped with serotype 5 and 8 capsid proteins.
Adeno-associated virus vectors (AAV) show promise for liver-targeted gene therapy. In this study, we examined the long-term consequences of a single intravenous administration of a self-complementary AAV vector (scAAV2/ 8-LP1-hFIXco) encoding a codon optimized human factor IX (hFIX) gene in 24 nonhuman primates (NHPs). A dose-response relationship between vector titer and transgene expression was observed. Peak hFIX expression following the highest dose of vector (2 × 10(12) pcr-vector genomes (vg)/kg) was 21 ± 3 µg/ml (~420% of normal). Fluorescent in-situ hybridization demonstrated scAAV provirus in almost 100% of hepatocytes at that dose. No perturbations of clinical or laboratory parameters were noted and vector genomes were cleared from bodily fluids by 10 days. Macaques transduced with 2 × 10(11) pcr-vg/kg were followed for the longest period (~5 years), during which time expression of hFIX remained >10% of normal level, despite a gradual decline in transgene copy number and the proportion of transduced hepatocytes. All macaques developed serotype-specific antibodies but no capsid-specific cytotoxic T lymphocytes were detected. The liver was preferentially transduced with 300-fold more proviral copies than extrahepatic tissues. Long-term biochemical, ultrasound imaging, and histologic follow-up of this large cohort of NHP revealed no toxicity. These data support further evaluation of this vector in hemophilia B patients.
Department of Hematology, University College London Cancer Institute, London, UK. a.nathwani@ucl.ac.uk
Amit C
AC
Nathwani
Cecilia
C
Rosales
Jenny
J
McIntosh
Ghasem
G
Rastegarlari
Devhrut
D
Nathwani
Deepak
D
Raj
Sushmita
S
Nawathe
Simon N
SN
Waddington
Roderick
R
Bronson
Scott
S
Jackson
Robert E
RE
Donahue
Katherine A
KA
High
Federico
F
Mingozzi
Catherine Y C
CY
Ng
Junfang
J
Zhou
Yunyu
Y
Spence
M Beth
MB
McCarville
Marc
M
Valentine
James
J
Allay
John
J
Coleman
Susan
S
Sleep
John T
JT
Gray
Arthur W
AW
Nienhuis
Andrew M
AM
Davidoff
D000818:Animals; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D000229:Dependovirus / Q000235:genetics*; D005164:Factor IX / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D057809:HEK293 Cells; D002836:Hemophilia B / Q000235:genetics / Q000628:therapy*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008099:Liver / Q000378:metabolism; D008251:Macaca; D051379:Mice
D036022:Capsid Proteins; D005164:Factor IX
Blood. 2004 Nov 1;104(9):2714-21
15231566
Gene Ther. 2000 Feb;7(3):183-95
10694794
Toxicol Pathol. 1993;21(2):206-18
8210943
Blood. 2003 Feb 15;101(4):1359-66
12393743
Proc Natl Acad Sci U S A. 1997 Oct 14;94(21):11563-6
9326649
J Virol. 2004 Mar;78(6):3110-22
14990730
Science. 2007 Jul 27;317(5837):477
17656716
Development. 1998 Jul;125(13):2451-6
9609828
Blood. 2007 Oct 1;110(7):2334-41
17609423
Gene Ther. 2009 Jan;16(1):60-9
18701909
Blood. 2001 Mar 1;97(5):1258-65
11222368
Mol Ther. 2005 Jun;11(6):875-88
15922958
Curr Opin Microbiol. 2000 Apr;3(2):144-8
10744998
J Virol. 2001 Aug;75(15):6969-76
11435577
Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3490-6
8170935
J Virol. 1999 Oct;73(10):8549-58
10482608
Hum Gene Ther. 2008 Nov;19(11):1317-23
19866493
Lab Anim. 1976 Oct;10(10):419-42
979138
Cancer Genet Cytogenet. 2006 May;167(1):92-4
16682295
J Virol. 2005 Mar;79(6):3606-14
15731255
Hum Gene Ther. 2004 Jul;15(7):709-15
15298029
Blood. 2006 Apr 1;107(7):2653-61
16322469
Blood. 1998 Jul 1;92(1):168-74
9639513
Br J Haematol. 2007 Aug;138(3):305-15
17614818
Vox Sang. 2004 Aug;87(2):73-81
15355497
Proc Natl Acad Sci U S A. 2002 Sep 3;99(18):11854-9
12192090
J Virol. 2005 Dec;79(23):14793-803
16282479
Blood. 2002 Sep 1;100(5):1662-9
12176886
Blood. 2010 Jun 10;115(23):4678-88
20335222
Mol Ther. 2005 Aug;12(2):299-306
16043099
Blood. 2003 Apr 15;101(8):2963-72
12515715
Exp Cell Res. 2002 Mar 10;274(1):1-8
11855851
Haemophilia. 2004 Jul;10(4):309-18
15230943
Nat Med. 2006 Aug;12(8):967-71
16845388
Cell. 1994 Jul 1;77(7):993-1002
8020105
Nat Genet. 1998 Jan;18(1):56-9
9425901
Mol Ther. 2010 Jan;18(1):126-34
19888196
Mol Ther. 2006 Jul;14(1):34-44
16682254
Nat Med. 2006 Mar;12(3):342-7
16474400
Blood. 2007 Feb 15;109(4):1414-21
17090654
J Virol. 2005 Jan;79(1):214-24
15596817
Gene Ther. 2008 Jun;15(11):840-8
18418418
Proc Natl Acad Sci U S A. 1989 Dec;86(24):10095-9
2481310