Protein replacement by receptor-mediated endocytosis corrects the sensitivity of Fanconi anemia group C cells to mitomycin C

Blood. 1999 Jan 1;93(1):363-9.

Abstract

Current methods for direct gene transfer into hematopoietic cells are inefficient. Here we show that functional complementation of Fanconi anemia (FA) group C cells by protein replacement can be as efficacious as by transfection with wild-type FAC cDNA. We expressed a chimeric protein (called His-ILFAC) consisting of the mature coding portion of gibbon interleukin-3 (IL-3) and full-length FAC in Escherichia coli. The purified bacterial protein is internalized by hematopoietic cells via IL-3 receptors. The intracellular half-life of His-ILFAC is approximately 60 minutes, which is comparable to that of the transgene-encoded FAC protein. In this cell-culture model His-ILFAC completely corrects the sensitivity of FA group C cells to mitomycin C, but it has no effect on FA cells that belong to complementation groups A and B. We suggest that receptor-mediated endocytosis of cytokine-fusion proteins may be of general use to deliver macromolecules into hematopoietic progenitor cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Cycle Proteins*
  • Cell Line
  • Cell Survival / drug effects
  • Cell Survival / genetics
  • DNA-Binding Proteins*
  • Endocytosis / drug effects
  • Endocytosis / genetics*
  • Fanconi Anemia / genetics
  • Fanconi Anemia / pathology*
  • Fanconi Anemia / therapy*
  • Fanconi Anemia Complementation Group C Protein
  • Fanconi Anemia Complementation Group Proteins
  • Gene Transfer Techniques
  • Histidine / genetics
  • Humans
  • Hylobates
  • Interleukin-3 / genetics
  • Mitomycin / pharmacology*
  • Nuclear Proteins*
  • Proteins / genetics
  • Proteins / physiology*
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / physiology*
  • Receptors, Interleukin-3 / biosynthesis
  • Receptors, Interleukin-3 / metabolism
  • Recombinant Fusion Proteins / chemical synthesis
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Fusion Proteins / physiology*

Substances

  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • FANCC protein, human
  • Fanconi Anemia Complementation Group C Protein
  • Fanconi Anemia Complementation Group Proteins
  • Interleukin-3
  • Nuclear Proteins
  • Proteins
  • Receptors, Cell Surface
  • Receptors, Interleukin-3
  • Recombinant Fusion Proteins
  • Histidine
  • Mitomycin