title: Survival-apoptosis associated signaling in GNE myopathy-cultured myoblasts.
abstract: GNE Myopathy (GNEM) is a neuromuscular disorder caused by mutations in the GNE gene. It is a slowly progressive distal and proximal muscle weakness sparing the quadriceps. In this study, we applied our model of mutated M743T GNE enzyme skeletal muscle-cultured myoblasts and paired healthy controls to depict the pattern of signaling proteins controlling survival and/or apoptosis of the PI3K/AKT, BCL2, ARTS/XIAP pathways, examined the effects of metabolic changes/stimuli on their expression and activation, and their potential role in GNEM. Immunoblot analysis of the GNEM myoblasts indicated a notable increased level of activated PTEN and PDK1 and a trend of relative differences in the expression and activation of the examined signaling molecules with variability among the cultures. ANOVA analysis showed a highly significant interaction between the level of PTEN and the patients groups. In parallel, the interaction between the level of BCL2, BAX and PTEN with the specific PI3K/AKT inhibitor-LY294002 was highly significant for BCL2 and nearly significant for PTEN and BAX. The pattern of the ARTS/XIAP signaling proteins of GNEM and the paired controls was variable, with no significant differences between the two cell types. The response of the GNEM cells to the metabolic changes/stimuli: serum depletion and insulin challenge, as indicated by expression of selected signaling proteins, was variable and similar to the control cells. Taken together, our observations provide a clearer insight into specific signaling molecules influencing growth and survival of GNEM muscle cells.
authors.affiliation: a Goldyne Savad Institute of Gene Therapy, Hadassah-Hebrew University Medical Center , Jerusalem , Israel .
authors.forename: Avi
authors.initials: A
authors.lastname: Harazi
authors.affiliation: b Israel National Skin Bank, Laboratory of Experimental Surgery , Hadassah-Hebrew University Medical Center , Jerusalem , Israel , and.
authors.forename: Malka
authors.initials: M
authors.lastname: Chaouat
authors.affiliation: b Israel National Skin Bank, Laboratory of Experimental Surgery , Hadassah-Hebrew University Medical Center , Jerusalem , Israel , and.
authors.forename: Zippora
authors.initials: Z
authors.lastname: Shlomai
authors.affiliation: b Israel National Skin Bank, Laboratory of Experimental Surgery , Hadassah-Hebrew University Medical Center , Jerusalem , Israel , and.
authors.forename: Robina
authors.initials: R
authors.lastname: Levitzki
authors.affiliation: a Goldyne Savad Institute of Gene Therapy, Hadassah-Hebrew University Medical Center , Jerusalem , Israel .
authors.forename: Michal
authors.initials: M
authors.lastname: Becker-Cohen
authors.affiliation: c Wolfson Medical Center , Holon , Israel.
authors.forename: Menachem
authors.initials: M
authors.lastname: Sadeh
authors.affiliation: c Wolfson Medical Center , Holon , Israel.
authors.forename: Ron
authors.initials: R
authors.lastname: Dabby
authors.affiliation: b Israel National Skin Bank, Laboratory of Experimental Surgery , Hadassah-Hebrew University Medical Center , Jerusalem , Israel , and.
authors.forename: Hannah
authors.initials: H
authors.lastname: Ben-Bassat
authors.affiliation: a Goldyne Savad Institute of Gene Therapy, Hadassah-Hebrew University Medical Center , Jerusalem , Israel .
authors.forename: Stella
authors.initials: S
authors.lastname: Mitrani-Rosenbaum
mesh_terms: D000328:Adult; D017209:Apoptosis; D016022:Case-Control Studies; D002470:Cell Survival; D002478:Cells, Cultured; D049310:Distal Myopathies / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009097:Multienzyme Complexes / Q000235:genetics* / Q000378:metabolism*; D050505:Mutant Proteins / Q000235:genetics* / Q000378:metabolism*; D009154:Mutation; D032448:Myoblasts, Skeletal / Q000378:metabolism* / Q000473:pathology*; D051059:PTEN Phosphohydrolase / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D000081382:Pyruvate Dehydrogenase Acetyl-Transferring Kinase; D058112:Septins / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D051636:X-Linked Inhibitor of Apoptosis Protein / Q000378:metabolism; D055815:Young Adult
chemical_list: D009097:Multienzyme Complexes; D050505:Mutant Proteins; D051059:PTEN Phosphohydrolase; D019869:Phosphatidylinositol 3-Kinases; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D019253:Proto-Oncogene Proteins c-bcl-2; D000081382:Pyruvate Dehydrogenase Acetyl-Transferring Kinase; D058112:Septins; D051636:X-Linked Inhibitor of Apoptosis Protein; C000595707:BCL2 protein, human; C000631429:PDK1 protein, human; C482901:UDP-N-acetylglucosamine 2-epimerase - N-acetylmannosamine kinase; C496866:XIAP protein, human; C494929:PTEN protein, human; C492027:SEPTIN4 protein, human
keywords: ARTS/XIAP; BCL2; GNE; GNE myopathy (GNEM); PI3K/AKT; signaling; skeletal muscle myoblasts