Sustained multilineage gene persistence and expression in dogs transplanted with CD34(+) marrow cells transduced by RD114-pseudotype oncoretrovirus vectors.

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  • Additional Information
    • Source:
      Publisher: Elsevier Country of Publication: United States NLM ID: 7603509 Publication Model: Print Cited Medium: Print ISSN: 0006-4971 (Print) Linking ISSN: 00064971 NLM ISO Abbreviation: Blood Subsets: MEDLINE
    • Publication Information:
      Publication: 2021- : [New York] : Elsevier
      Original Publication: New York, Grune & Stratton [etc.]
    • Subject Terms:
      Graft Survival*/genetics; Antigens, CD34/*analysis ; Bone Marrow Cells/*immunology ; Bone Marrow Transplantation/*methods ; Cell Lineage/*genetics ; Genetic Vectors/*standards ; Transduction, Genetic/*methods; Amino Acid Transport System ASC/genetics ; Amino Acid Transport System ASC/metabolism ; Animals ; Blood Cells/metabolism ; Bone Marrow Cells/metabolism ; Cats ; Dogs ; Endogenous Retroviruses/genetics ; Follow-Up Studies ; Gene Expression ; Green Fluorescent Proteins ; Hematopoiesis ; Leukemia Virus, Gibbon Ape/genetics ; Luminescent Proteins/genetics ; Receptors, Virus/genetics ; Receptors, Virus/metabolism ; Retroviridae Proteins/genetics ; Retroviridae Proteins/metabolism ; Survival Rate ; Transduction, Genetic/standards
    • Abstract:
      Previous studies have shown that the choice of envelope protein (pseudotype) can have a significant effect on the efficiency of retroviral gene transfer into hematopoietic stem cells. This study used a competitive repopulation assay in the dog model to evaluate oncoretroviral vectors carrying the envelope protein of the endogenous feline virus, RD114. CD34-enriched marrow cells were divided into equal aliquots and transduced with vectors produced by the RD114-pseudotype packaging cells FLYRD (LgGLSN and LNX) or by the gibbon ape leukemia virus (GALV)-pseudotype packaging cells PG13 (LNY). A total of 5 dogs were studied. One dog died because of infection before sustained engraftment could be achieved, and monitoring was discontinued after 9 months in another animal that had very low overall gene-marking levels. The 3 remaining animals are alive with follow-ups at 11, 22, and 23 months. Analyses of gene marking frequencies in peripheral blood and marrow by polymerase chain reaction revealed no significant differences between the RD114 and GALV-pseudotype vectors. The LgGLSN vector also contained the enhanced green fluorescent protein (GFP), enabling us to monitor proviral expression by flow cytometry. Up to 10% of peripheral blood cells expressed GFP shortly after transplantation and approximately 6% after the longest follow-up of 23 months. Flow cytometric analysis of hematopoietic subpopulations showed that most of the GFP-expressing cells were granulocytes, although GFP-positive lymphocytes and monocytes were also detected. In summary, these results show that RD114-pseudotype oncoretroviral vectors are able to transduce hematopoietic long-term repopulating cells and, thus, may be useful for human stem cell gene therapy.
    • Grant Information:
      DK47754 United States DK NIDDK NIH HHS; DK56465 United States DK NIDDK NIH HHS; HL36444 United States HL NHLBI NIH HHS
    • Accession Number:
      0 (Amino Acid Transport System ASC)
      0 (Antigens, CD34)
      0 (Luminescent Proteins)
      0 (Receptors, Virus)
      0 (Retroviridae Proteins)
      0 (leukemia virus receptor, gibbon ape)
      147336-22-9 (Green Fluorescent Proteins)
    • Publication Date:
      Date Created: 20010925 Date Completed: 20011204 Latest Revision: 20210216
    • Publication Date:
      20231215
    • Accession Number:
      10.1182/blood.v98.7.2065
    • Accession Number:
      11567991