Expression of human factor IX in rat capillary endothelial cells: Toward somatic gene therapy for hemophilia B
- Univ. of Michigan, Ann Arbor (United States)
In aiming to develop a gene therapy approach for hemophilia B, the authors expressed and characterized human factor IX in rat capillary endothelial cells (CECs). Moloney murine leukemia virus-derived retrovirus vectors that contain human factor IX cDNA linked to heterologous promoters and the neomycin-resistant gene were constructed and employed to prepare recombinant retroviruses. Rat CECs and NIH 3T3 cells infected with these viruses were selected with the neomycin analogue, G418 sulfate, and tested for expression of factor IX. A construct with the factor IX cDNA under direct control by long terminal repeat gave the highest level of expression as quantitated by immunoassays as well as clotting activity assays. A single RNA transcript of 4.4 kilobases predicted by the construct and a recombinant factor IX were found. The recombinant human factor IX produced showed full clotting activity, demonstrating that CECs have an efficient mechanism for posttranslational modifications, including {gamma}-carboxylation, essential for its biological activity. These results, in addition to other properties of the endothelium, including large number of cells, accessibility, and direct contact with the circulating blood, suggest that CECs can serve as an efficient drug delivery vehicle producing factor IX in a somatic gene therapy for hemophilia B.
- OSTI ID:
- 5820969
- Journal Information:
- Proceedings of the National Academy of Sciences of the United States of America; (United States), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (United States) Vol. 88:18; ISSN 0027-8424; ISSN PNASA
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
ANIMAL TISSUES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOSYNTHESIS
BLOOD COAGULATION FACTORS
BLOOD VESSELS
BODY
CAPILLARIES
CARDIOVASCULAR SYSTEM
CHROMOSOMES
COAGULANTS
DAYS LIVING RADIOISOTOPES
DISEASES
DRUGS
ENDOTHELIUM
HEMATOLOGIC AGENTS
HEMIC DISEASES
HEMOPHILIA
HEREDITARY DISEASES
HETEROCHROMOSOMES
HUMAN CHROMOSOMES
HUMAN X CHROMOSOME
ISOTOPES
LEUKEMIA VIRUSES
LIGHT NUCLEI
MICROORGANISMS
NUCLEI
ODD-ODD NUCLEI
ONCOGENIC VIRUSES
ORGANIC COMPOUNDS
ORGANS
PARASITES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PROTEINS
RADIOISOTOPES
SYNTHESIS
THERAPY
TISSUES
VIRUSES
X CHROMOSOME