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Retrovirus and Adenovirus-Mediated Interferon Gene Transfer into Hematopoietic Stem Cells (CD 34+) of Normal and Chronic Myelogenous Leukemia

  • N. G. Abraham
Conference paper
Part of the Haematology and Blood Transfusion / Hämatologie und Bluttransfusion book series (HAEMATOLOGY, volume 39)

Abstract

Gene therapy following gene transfer into hematopoietic cells (CD34+) is now being investigated for several genetic disorders. We have applied a similar approach using interferon gene transfer for treatment of chronic myelogenous leukemia (CML). IFN-α produces a hematologic and cytogenetic response in CML with a survival advantage for cytogenetic responders. Therefore, we examined the effect of transient overexpression of IFN-α using the adenovirus gene transfer approach. The ability of the adenovirus (Adv)-IFN-α gene construct to transfect normal and CML stem cells, CD34+, was examined. The peripheral blood mononuclear fraction from patients with CML treated with G-CSF or GM-CSF/G-CSF was enriched in CD34+ cells. Adv-cytomegalovirus (pCMV) promoter driven IFN-α at multiple doses was assessed to transfect highly purified CD34+ cells in the presence of matrix protein and in co-cultures with the stromal adherent cell layer. The use of cytokines enhances Adv-mediated IFN-α gene transfer into stem cells. Southern blot analysis demonstrated that the Adv-pCMV-IFN-α construct and IFN-α were expressed in cultured CD34+ cells. Transient expression of the IFN-a gene did not suppress proliferation of CD34+ progenitors, CFU-Meg, BFU-E or CFU-CM growth. Reverse transcriptase/polymerase chain reaction (RT/PCR) analysis of RNA from CFU-GM progenitor cells demonstrated transient IFN-α mRNA expression in CD34+ cells. Immunoassay of IFN-α shows that selective expression of IFN-α may be beneficial for CML therapy. We also report on the establishment of novel conditions which permit high efficiency of the retrovirus IFN-α gene transfer into CD34+ cells. Stem cells transfected with retrovirus were infused intravenously to irradiated mice and spleen colony forming units were evaluated for IFN-α marked clones by Southern blot analysis. These studies demonstrate that the retrovirus IFN-α gene can be used to transfect CD34+ cells and possibly for gene therapy of CML.

Keywords

Gene Transfer Chronic Myeloid Leukemia Cytogenetic Response Chronic Myelogenous Leukemia Cell Adherent Cell Layer 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag Berlin Heidelberg 1998

Authors and Affiliations

  • N. G. Abraham
    • 1
  1. 1.Department of Pharmacology/Gene TherapyNew York Medical CollegeValhallaUSA

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