Expression of human factor IX in rabbit hepatocytes by retrovirus-mediated gene transfer: Potential for gene therapy of hemophilia B

Journal Article · Wed Aug 01 00:00:00 EDT 1990 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

OSTI ID:5736168

Thompson, A R ^[1]; Darlington, G ^[2]; Armentano, D; Woo, S L.C.

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1. Univ. of Washington, Seattle (USA) Puget Sound Blood Center, Seattle, WA (USA)
2. Baylor College of Medicine, Houston, TX (USA)

Hemophilia B (Christmas disease) is a chromosome X-linked blood clotting disorder which results when factor IX is deficient or functionally defective. The enzyme is synthesized in the liver, and the existence of animal models for this genetic disease will permit the development of somatic gene therapy protocols aimed at transfer of the functional gene into the liver. The authors report the construction of an N2-based recombinant retroviral vector, NCMVFIX, for efficient transfer and expression of human factor IX cDNA in primary rabbit hepatocytes. In this construct the human cytomegalovirus immediate early promoter directs the expression of factor IX. Hepatocytes were isolated from 3-week-old New Zealand White rabbits, infected with the recombinant virus, and analyzed for secretion of active factor IX. The infected rabbit hepatocytes produced human factor IX that is indistinguishable from enzyme derived from normal human plasma. The recombinant protein is sufficiently {gamma}-carboxylated and is functionally active in clotting assays. These results establish the feasibility of using infected hepatocytes for the expression of this protein and are a step toward the goal of correcting hemophilia B by hepatic gene transfer.

• OSTI ID: 5736168
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Vol. 87:16; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
62 RADIOLOGY AND NUCLEAR MEDICINE
BLOOD COAGULATION FACTORS
RECOMBINANT DNA
SECRETION
HEMOPHILIA
THERAPY
HEREDITARY DISEASES
RABBITS
GENETIC ENGINEERING
SERINE PROTEINASES
GENE RECOMBINATION
GENE REGULATION
HUMAN X CHROMOSOME
LIVER CELLS
PATHOGENESIS
PHOSPHORUS 32
TRANSCRIPTION
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHROMOSOMES
COAGULANTS
DAYS LIVING RADIOISOTOPES
DISEASES
DNA
DRUGS
ENZYMES
HEMATOLOGIC AGENTS
HEMIC DISEASES
HETEROCHROMOSOMES
HYDROLASES
ISOTOPES
LIGHT NUCLEI
MAMMALS
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PEPTIDE HYDROLASES
PHOSPHORUS ISOTOPES
PROTEINS
RADIOISOTOPES
SOMATIC CELLS
VERTEBRATES
X CHROMOSOME
550201* - Biochemistry- Tracer Techniques
550601 - Medicine- Unsealed Radionuclides in Diagnostics