Efficient retrovirus-mediated transfer and expression of a human adenosine deaminase gene in diploid skin fibroblasts from an adenosine deaminase-deficient human

Journal Article · Sun Feb 01 00:00:00 EST 1987 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.84.4.1055· OSTI ID:5878723

Palmer, T D; Hock, R A; Osborne, W R.A.; Miller, A D

Skin fibroblasts might be considered suitable recipients for therapeutic genes to cure several human genetic diseases; however, these cells are resistant to gene transfer by most methods. The authors studied the ability of retroviral vectors to transfer genes into normal human diploid skin fibroblasts. Retroviruses carrying genes for neomycin or hygromycin B resistance conferred drug resistance to greater than 50% of the human fibroblasts after a single exposure to virus-containing medium. This represents at least a 500-fold increase in efficiency over other methods. Transfer was achieved in the absence of helper virus by using amphotropic retrovirus-packaging cells. A retrovirus vector containing a human adenosine deaminase (ADA) cDNA was constructed and used to infect ADA/sup -/ fibroblasts from a patient with ADA deficiency. The infected cells produced 12-fold more ADA enzyme than fibroblasts from normal individuals and were able to rapidly metabolize exogenous deoxyadenosine and adenosine, metabolites that accumulate in plasma in ADA-deficient patients and are responsible for the severe combined immunodeficiency in these patients. These experiments indicate the potential of retrovirus-mediated gene transfer into human fibroblasts for gene therapy.

• Research Organization: Fred Hutchinson Cancer Research Center, Seattle, WA
• OSTI ID: 5878723
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 84:4
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
HEREDITARY DISEASES
THERAPY
NON-PEPTIDE C-N HYDROLASES
GENETICS
DEAMINATION
FIBROBLASTS
GENE RECOMBINATION
GENES
METABOLISM
PATIENTS
RECOMBINANT DNA
SKIN
VIRUSES
ANIMAL CELLS
BIOLOGY
BODY
CHEMICAL REACTIONS
CONNECTIVE TISSUE CELLS
DISEASES
DNA
ENZYMES
HYDROLASES
MICROORGANISMS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PARASITES
SOMATIC CELLS
550400* - Genetics