[In vivo delivery of Caspase 8 or Fas siRNA improves the survival of septic mice]
>>Blood First Edition Paper, prepublished online June 7, 2005; DOI 10.1182/blood-2004-10-4086.
Submitted October 22, 2004
Accepted May 26, 2005
In vivo delivery of Caspase 8 or Fas siRNA improves the survival of septic mice
Doreen E Wesche-Soldato, Chun-Shiang Chung, Joanne Lomas-Neira, Lesley A Doughty, Stephen H Gregory, and Alfred Ayala*
Surgical Research, Department of Surgery, Brown University School of Medicine and Rhode Island Hospital, Providence, RI, USA
Department of Pediatrics, Brown University School of Medicine, Providence, RI, USA
Department of Medicine, Brown University School of Medicine, Providence, RI, USA
* Corresponding author; email: AAyala@Lifespan.org.
While studies have shown increased evidence of death receptor-driven apoptosis in intestinal lymphoid cells, splenocytes, and the liver following the onset of polymicrobial sepsis, little is known about the mediators controlling this process or their pathological contribution. We, therefore, attempted to test the hypothesis that the hydrodynamic administration of siRNA against the death receptor, Fas or Caspase 8 should attenuate the onset of morbidity and mortality seen in sepsis, as produced by cecal ligation and puncture (CLP). We initially show that in vivo administration of GFP siRNA in GFP transgenic mice results in a decrease in GFP fluorescence in most tissues. Subsequently, we also found that treating septic non-transgenic mice with siRNA targeting Fas or Caspase 8 but not GFP (used as a control here) decreased the mRNA, in a sustained fashion up to 10 days, and protein expression of Fas and Caspase 8, respectively. In addition, TUNEL and active Caspase 3 analyses revealed a decrease in apoptosis in the liver, spleen, but not the thymus following siRNA treatment. Indices of liver damage were also decreased. Finally, the injection of Fas or Caspase 8 given not only thirty minutes but up to twelve hours after CLP significantly improved the survival of septic mice.<<
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