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Combination of optison with ultrasound and electroporation increases albumin and thrompoietin transgene expression whilst elongation factor promoter prolongs its duration

By Nagy Habib, Roman Havlik, Long Jiao, Arpna Kiri, Steen Jensen, Joanna Nicholls, Catherine Sarraf, Geoffrey Goldspink, Anthony Davies, Ludek Slavik, Chris Wood, Michele Tiraby, Daniel Drocourt, Jean-Paul Rynes, Claude Smadja and Jean Gerard Tiraby

Abstract

Hypoalbuminaemia and thrombocytopaenia are two clinical problems frequently encountered in patients with chronic liver failure or cancer following treatment with chemotherapy. The current study was designed to assess the\ud magnitude and duration of thrombopoietin and albumin transgene expression hoping to increase the production of\ud albumin and platelets. Immunocompetent and immunocompromised (nude) mice were injected intramuscularly\ud with plasmids expressing either human serum albumin or human thrombopoietin. The therapeutic expression cassette of the plasmids was driven by either CMV or elongation factor 1- promoters respectively. In order to achieve muscle specific expression both gene constructs included the myosin light chain enhancer. The experiment was conducted in a group of mice which were injected with the transgene plasmid either in normal saline or plasmid\ud followed by electroporation, ultrasound, optison and a combination of all three to increase transgene expression.\ud The result showed that plasmids with the CMV promoter induced the highest transgenic expression lasting for one\ud week whilst plasmids with the elongation factor 1-alpha promoter produced a weaker expression lasting for a longer and more stable duration of expression up to 3 months in both immunocompetent and nude mice. The combination of electroporation and ultrasound with Optison TM provided the highest transgene expression. We concluded that it would be possible to increase albumin and platelets production by an intramuscular injection of plasmids expressing human albumin and thromopoietin. A combination of electroporation and ultrasound with Optison TM can increase their expression.\u

Topics: UOW2
OAI identifier: oai:westminsterresearch.wmin.ac.uk:3228
Provided by: WestminsterResearch

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Citations

  1. (1999). A gene therapy for cancer using intramuscular injection of plasmid DNA encoding interferon alpha.
  2. (1990). A putative truncated cytokine receptor gene transduced by the myeloproliferative leukemia virus immortalizes hematopoietic progenitors.
  3. (1994). Accumulation of human apolipoprotein-E in rat plasma after in vivo intramuscular injection of naked DNA.
  4. (1998). Alila HW
  5. (1998). Bacterial DNA and immunostimulatory CpG oligonucleotides trigger maturation and activation of murine dendritic cells.
  6. (1998). Constitutive expression of phVEGF165 after intramuscular gene transfer promotes collateral vessel development in patients with critical limb ischemia.
  7. (1995). CpG motifs
  8. (1996). CpG motifs present in bacteria DNA rapidly induce lymphocytes to secrete interleukin 6, interleukin 12, and interferon gamma.
  9. (2002). CpG-depleted plasmid DNA vectors with enhanced safety and long-term gene expression in vivo.
  10. (2002). Development of safe and efficient novel nonviral gene transfer using ultrasound: enhancement of transfection efficiency of naked plasmid DNA in skeletal muscle.
  11. (1990). Direct gene transfer into mouse muscle in vivo.
  12. (2000). Dose response to a single intramuscular injection of recombinant adeno-associated viruserythropoietin in monkeys.
  13. (2000). Gene electrotransfer results in a high-level Habib et al: Intramuscular transgene expression transduction of rat skeletal muscle and corrects anemia of renal failure.
  14. (1999). Gene therapy for renal anemia in mice with polycystic kidney using an adenovirus vector encoding the human erythropoietin gene.
  15. (1997). Gene transfer and expression of human alpha-galactosidase from mouse muscle in vitro and in vivo.
  16. (1993). Heterologous protection against influenza by injection of DNA encoding a viral protein. Sc ence 259,
  17. (2001). High and sustained transgene expression in vivo from plasmid vectors containing a hybrid ubiquitin promoter.
  18. (2000). Improvement of erythropoiesis in beta-thalassemic mice by continuous erythropoietin delivery from muscle.
  19. (1996). In vivo gene electroinjection and expression in rat liver.
  20. (2002). In vivo gene ra sfer into muscle via electro-sonoporation.
  21. (2001). Increased persistence of lung gene expression using plasmids containing the ubiquitin C or elongation factor 1a promoter.
  22. (1998). Induction of antigen-specific cytotoxic T lymphocytes in humans by a malaria DNA vaccine.
  23. (1996). Induction of NK activity in murine and human cells by CpG motifs in ligodeoxynucleotides and bacterial DNA.
  24. (1997). Intramuscular injection of expression plasmid DNA is an effective means of long-tern systemic delivery of interleukin-0.
  25. (2002). LCR-mediated, long-term tissue-specific gene expression within replicating episomal plasmid and cosmid vectors.
  26. (1996). Long-term expression of erythropoietin in the systemic circulation of mice after intramuscular injection of plasmid DNA vector.
  27. (1997). Macrophages sense pathogens via DNA motifs: induction of tumour necrosis factor-alphamediated shock.
  28. (1995). Modulation of disease activity in murine systemic lupus erythermatosus by cytokine gene delivery.
  29. (1992). Molecular cloning and characterization of MPL, the human homolog of the v-mpl oncogene: identification of a member of the hematopoietic growth factor receptor superfamily.
  30. (1989). Myosin light chain enhancer activates muscle-specific developmentally regulated gene expression in transgenic mice.
  31. (1997). Promoter attenuation in gene therapy: interferon gamma and tumor necrosis factor-alpha inhibit transgene expression.
  32. (2001). Rabbit EPO gene and cDNA: expression of rabbit EPO after intramuscular injection of pDNA.
  33. (1993). Systemic immunological effects of cytokine genes injected into skeletal muscle.
  34. (2001). Time course of gene expression after plasmid DNA gene transfer to the liver.
  35. (2000). Trans-splicing vectors expand the utility of adeno-associated virus for gene therapy.

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