Sustained high-level expression of human factor IX (hFIX) after liver-targeted delivery of recombinant adeno-associated virus encoding the hFIX gene in rhesus macaques

Amit C. Nathwani, Andrew M. Davidoff, Hideki Hanawa, Yunyu Hu, Fredric A. Hoffer, Alexander Nikanorov, Clive A. Slaughter, Catherine Y C Ng, Junfang Zhou, Jay N. Lozier, Timothy D. Mandrell, Elio F. Vanin, Arthur W. Nienhuis

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Abstract

The feasibility, safety, and efficacy of liver-directed gene transfer was evaluated in 5 male macaques (aged 2.5 to 6.5 years) by using a recombinant adeno-associated viral (rAAV) vector (rAAV-2 CAGG-hFIX) that had previously mediated persistent therapeutic expression of human factor IX (hFIX; 6%-10% of physiologic levels) in murine models. A dose of 4 × 10 12 vector genomes (vgs)/kg of body weight was administered through the hepatic artery or portal vein. Persistence of the rAAV vgs as circular monomers and dimers and high-molecular-weight concatamers was documented in liver tissue by Southern blot analysis for periods of up to 1 year. Vector particles were present in plasma, urine, or saliva for several days after infusion (as shown by polymerase chain reaction analysis), and the vgs were detected in spleen tissue at low copy numbers. An enzyme-linked immunosorption assay capable of detecting between 1% and 25% of normal levels of hFIX in rhesus plasma was developed by using hyperimmune serum from a rhesus monkey that had received an adenoviral vector encoding hFIX. Two macaques having 3 and 40 rAAV genome equivalents/cell, respectively, in liver tissue had 4% and 8% of normal physiologic plasma levels of hFIX, respectively. A level of hFIX that was 3% of normal levels was transiently detected in one other macaque, which had a genome copy number of 25 before abrogation by a neutralizing antibody (inhibitor) to hFIX. This nonhuman-primate model will be useful in further evaluation and development of rAAV vectors for gene therapy of hemophilia B.

Original languageEnglish (US)
Pages (from-to)1662-1669
Number of pages8
JournalBlood
Volume100
Issue number5
DOIs
StatePublished - Sep 1 2002

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Dependovirus
Factor IX
Human engineering
Macaca mulatta
Viruses
Liver
Genes
Macaca
Viral Genome
Genome
Tissue
Plasmas
Gene transfer
Hemophilia B
Gene therapy
Hepatic Veins
Viral Genes
Hepatic Artery
Polymerase chain reaction
Portal Vein

ASJC Scopus subject areas

  • Hematology

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Sustained high-level expression of human factor IX (hFIX) after liver-targeted delivery of recombinant adeno-associated virus encoding the hFIX gene in rhesus macaques. / Nathwani, Amit C.; Davidoff, Andrew M.; Hanawa, Hideki; Hu, Yunyu; Hoffer, Fredric A.; Nikanorov, Alexander; Slaughter, Clive A.; Ng, Catherine Y C; Zhou, Junfang; Lozier, Jay N.; Mandrell, Timothy D.; Vanin, Elio F.; Nienhuis, Arthur W.

In: Blood, Vol. 100, No. 5, 01.09.2002, p. 1662-1669.

Research output: Contribution to journalArticle

Nathwani, AC, Davidoff, AM, Hanawa, H, Hu, Y, Hoffer, FA, Nikanorov, A, Slaughter, CA, Ng, CYC, Zhou, J, Lozier, JN, Mandrell, TD, Vanin, EF & Nienhuis, AW 2002, 'Sustained high-level expression of human factor IX (hFIX) after liver-targeted delivery of recombinant adeno-associated virus encoding the hFIX gene in rhesus macaques', Blood, vol. 100, no. 5, pp. 1662-1669. https://doi.org/10.1182/blood-2002-02-0589
Nathwani, Amit C. ; Davidoff, Andrew M. ; Hanawa, Hideki ; Hu, Yunyu ; Hoffer, Fredric A. ; Nikanorov, Alexander ; Slaughter, Clive A. ; Ng, Catherine Y C ; Zhou, Junfang ; Lozier, Jay N. ; Mandrell, Timothy D. ; Vanin, Elio F. ; Nienhuis, Arthur W. / Sustained high-level expression of human factor IX (hFIX) after liver-targeted delivery of recombinant adeno-associated virus encoding the hFIX gene in rhesus macaques. In: Blood. 2002 ; Vol. 100, No. 5. pp. 1662-1669.
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abstract = "The feasibility, safety, and efficacy of liver-directed gene transfer was evaluated in 5 male macaques (aged 2.5 to 6.5 years) by using a recombinant adeno-associated viral (rAAV) vector (rAAV-2 CAGG-hFIX) that had previously mediated persistent therapeutic expression of human factor IX (hFIX; 6{\%}-10{\%} of physiologic levels) in murine models. A dose of 4 × 10 12 vector genomes (vgs)/kg of body weight was administered through the hepatic artery or portal vein. Persistence of the rAAV vgs as circular monomers and dimers and high-molecular-weight concatamers was documented in liver tissue by Southern blot analysis for periods of up to 1 year. Vector particles were present in plasma, urine, or saliva for several days after infusion (as shown by polymerase chain reaction analysis), and the vgs were detected in spleen tissue at low copy numbers. An enzyme-linked immunosorption assay capable of detecting between 1{\%} and 25{\%} of normal levels of hFIX in rhesus plasma was developed by using hyperimmune serum from a rhesus monkey that had received an adenoviral vector encoding hFIX. Two macaques having 3 and 40 rAAV genome equivalents/cell, respectively, in liver tissue had 4{\%} and 8{\%} of normal physiologic plasma levels of hFIX, respectively. A level of hFIX that was 3{\%} of normal levels was transiently detected in one other macaque, which had a genome copy number of 25 before abrogation by a neutralizing antibody (inhibitor) to hFIX. This nonhuman-primate model will be useful in further evaluation and development of rAAV vectors for gene therapy of hemophilia B.",
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AU - Davidoff, Andrew M.

AU - Hanawa, Hideki

AU - Hu, Yunyu

AU - Hoffer, Fredric A.

AU - Nikanorov, Alexander

AU - Slaughter, Clive A.

AU - Ng, Catherine Y C

AU - Zhou, Junfang

AU - Lozier, Jay N.

AU - Mandrell, Timothy D.

AU - Vanin, Elio F.

AU - Nienhuis, Arthur W.

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N2 - The feasibility, safety, and efficacy of liver-directed gene transfer was evaluated in 5 male macaques (aged 2.5 to 6.5 years) by using a recombinant adeno-associated viral (rAAV) vector (rAAV-2 CAGG-hFIX) that had previously mediated persistent therapeutic expression of human factor IX (hFIX; 6%-10% of physiologic levels) in murine models. A dose of 4 × 10 12 vector genomes (vgs)/kg of body weight was administered through the hepatic artery or portal vein. Persistence of the rAAV vgs as circular monomers and dimers and high-molecular-weight concatamers was documented in liver tissue by Southern blot analysis for periods of up to 1 year. Vector particles were present in plasma, urine, or saliva for several days after infusion (as shown by polymerase chain reaction analysis), and the vgs were detected in spleen tissue at low copy numbers. An enzyme-linked immunosorption assay capable of detecting between 1% and 25% of normal levels of hFIX in rhesus plasma was developed by using hyperimmune serum from a rhesus monkey that had received an adenoviral vector encoding hFIX. Two macaques having 3 and 40 rAAV genome equivalents/cell, respectively, in liver tissue had 4% and 8% of normal physiologic plasma levels of hFIX, respectively. A level of hFIX that was 3% of normal levels was transiently detected in one other macaque, which had a genome copy number of 25 before abrogation by a neutralizing antibody (inhibitor) to hFIX. This nonhuman-primate model will be useful in further evaluation and development of rAAV vectors for gene therapy of hemophilia B.

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