title: SBDS-deficient cells undergo accelerated apoptosis through the Fas-pathway.
abstract: BACKGROUND
  Shwachman-Diamond syndrome is an inherited multisystem disorder characterized by bone marrow and pancreatic dysfunction as well as metaphyseal dysostosis. Ninety percent of the patients have mutations in the Shwachman-Bodian-Diamond syndrome gene (SBDS). The relationship between SBDS and cell survival is unknown. In this study we investigated whether deficiency of the SBDS protein can cause increased apoptosis and, if so, what pathways are involved in this process.
  
  
  METHODS
  To determine whether accelerated apoptosis of Shwachman-Diamond syndrome cells is caused by a deficiency in SBDS we generated two SBDS-knockdown cell clones. We then evaluated, Fas expression and levels of the intracellular proteins, BAX, BCL-2 and BCL-X(L) and determined the effects of apoptosis inhibitors. Using oligonucteotide-microarrays we also analyzed apoptosis-related gene expression in Shwachman-Diamond syndrome marrow cells.
  
  
  RESULTS
  We found that knocking down SBDS by short interfering hairpin RNA in HeLa cells resulted in a prominent increase in cell death. The mechanism for the accelerated apoptosis was related to marked hypersensitivity to Fas stimulation, and increased Fas expression. In contrast, there was no increase in the expression ratio of the pro-apoptotic factor, BAX, to the pro-survival factors, BCL2 and BCL-X(L) in the SBDS-knockdown cells and in the patients' marrow cells. Furthermore, inhibition of Fas and caspase 8, but not caspase 9, significantly improved the defective cell growth phenotype.
  
  
  CONCLUSIONS
  Our work provides new data about the pro-survival properties of SBDS, whose inhibition results in accelerated apoptosis through the Fas pathway.
authors.affiliation: Division of Haematology/Oncology, The Hospital for Sick Children, 555 University Avenue, Toronto, Ontario, Canada.
authors.forename: Piya
authors.initials: P
authors.lastname: Rujkijyanont
authors.forename: Ken-Ichiro
authors.initials: K
authors.lastname: Watanabe
authors.forename: Chhaya
authors.initials: C
authors.lastname: Ambekar
authors.forename: Hanming
authors.initials: H
authors.lastname: Wang
authors.forename: Aaron
authors.initials: A
authors.lastname: Schimmer
authors.forename: Joseph
authors.initials: J
authors.lastname: Beyene
authors.forename: Yigal
authors.initials: Y
authors.lastname: Dror
mesh_terms: D017209:Apoptosis / Q000235:genetics / Q000502:physiology*; D001853:Bone Marrow / Q000378:metabolism; D001855:Bone Marrow Diseases / Q000235:genetics; D053181:Caspase 8 / Q000096:biosynthesis / Q000235:genetics; D053453:Caspase 9 / Q000096:biosynthesis / Q000235:genetics; D002460:Cell Line / Q000378:metabolism; D004392:Dwarfism / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics; D018390:Gene Targeting; D019254:Genes, bcl-2; D006367:HeLa Cells; D006801:Humans; D010182:Pancreatic Diseases / Q000235:genetics; D011506:Proteins / Q000235:genetics / Q000502:physiology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis; D034741:RNA, Small Interfering / Q000494:pharmacology; D015398:Signal Transduction / Q000235:genetics; D013577:Syndrome; D051028:bcl-2-Associated X Protein / Q000096:biosynthesis / Q000235:genetics; D051020:bcl-X Protein / Q000096:biosynthesis / Q000235:genetics; D019014:fas Receptor / Q000037:antagonists & inhibitors / Q000502:physiology*
chemical_list: C494826:BAX protein, human; C494811:BCL2L1 protein, human; C491901:FAS protein, human; D011506:Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D034741:RNA, Small Interfering; C470521:SBDS protein, human; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein; D019014:fas Receptor; C505530:CASP8 protein, human; D053181:Caspase 8; D053453:Caspase 9