Bax siRNA promotes survival of cultured and allografted granule cell precursors through blockade of caspase-3 cleavage

Cell Death Differ. 2008 Jun;15(6):1042-53. doi: 10.1038/cdd.2008.29. Epub 2008 Mar 7.

Abstract

Transplantation of neuronal precursor cells (NPCs) into the central nervous system could represent a powerful therapeutical tool against neurodegenerative diseases. Unfortunately, numerous NPCs die shortly after transplantation, predominantly due to caspase-dependent apoptosis. Using a culture of cerebellar neuronal precursors, we have previously demonstrated protective effect of the neuropeptide PACAP, which suppresses ceramide-induced apoptosis by blockade of the mitochondrial apoptotic pathway. The main objective of this study was to determine whether Bax repression can promote survival of NPCs allotransplanted into a host animal. In vivo and ex vivo experiments revealed that C2-ceramide increases Bax expression, while PACAP reverses this effect. In vitro tests using cerebellar NPCs demonstrated that the Bax-specific small interfering RNA (siRNA) could reduce their death and caspase-3 cleavage within the first 24 h. BrdU-labelled NPCs were subjected to transfection procedure with or without siRNA introduction before using for in vivo transplantation. Twenty-four hours after, the allografted NPCs containing siRNA showed significantly reduced level of caspase-3 cleavage, and the volume of their implants was almost twofold higher than in the case of empty-transfected precursors. These data evidence an important role of Bax in life/death decision of grafted NPCs and suggest that RNA interference strategy may be applicable for maintaining NPCs survival within the critical first hours after their transplantation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Caspase 3 / metabolism
  • Caspase Inhibitors*
  • Cell Survival
  • Cells, Cultured
  • Cerebellum / cytology*
  • Neurons / cytology*
  • Pituitary Adenylate Cyclase-Activating Polypeptide / pharmacology
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • Rats
  • Rats, Wistar
  • Sphingosine / analogs & derivatives
  • Sphingosine / pharmacology
  • Stem Cell Transplantation*
  • Stem Cells / drug effects
  • Stem Cells / enzymology
  • Stem Cells / metabolism*
  • Transplantation, Homologous
  • bcl-2-Associated X Protein / antagonists & inhibitors*
  • bcl-2-Associated X Protein / genetics
  • bcl-2-Associated X Protein / metabolism

Substances

  • Caspase Inhibitors
  • N-acetylsphingosine
  • Pituitary Adenylate Cyclase-Activating Polypeptide
  • RNA, Small Interfering
  • bcl-2-Associated X Protein
  • Caspase 3
  • Sphingosine