title: Evaluation of energy metabolism and calcium homeostasis in cells affected by Shwachman-Diamond syndrome.
abstract: Isomorphic mutation of the SBDS gene causes Shwachman-Diamond syndrome (SDS). SDS is a rare genetic bone marrow failure and cancer predisposition syndrome. SDS cells have ribosome biogenesis and their protein synthesis altered, which are two high-energy consuming cellular processes. The reported changes in reactive oxygen species production, endoplasmic reticulum stress response and reduced mitochondrial functionality suggest an energy production defect in SDS cells. In our work, we have demonstrated that SDS cells display a Complex IV activity impairment, which causes an oxidative phosphorylation metabolism defect, with a consequent decrease in ATP production. These data were confirmed by an increased glycolytic rate, which compensated for the energetic stress. Moreover, the signalling pathways involved in glycolysis activation also appeared more activated; i.e. we reported AMP-activated protein kinase hyper-phosphorylation. Notably, we also observed an increase in a mammalian target of rapamycin phosphorylation and high intracellular calcium concentration levels ([Ca(2+)]i), which probably represent new biochemical equilibrium modulation in SDS cells. Finally, the SDS cell response to leucine (Leu) was investigated, suggesting its possible use as a therapeutic adjuvant to be tested in clinical trials.
authors.affiliation: DIFAR-Biochemistry Lab., Department of Pharmacy, University of Genova, 16132 Genova, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Ravera
authors.affiliation: Haematology Unit, Istituto Giannina Gaslini, 16148 Genova, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Dufour
authors.affiliation: Oncoematologia Pediatrica, Azienda Ospedaleira universitaria Integrata, Verona, Italy.
authors.forename: Simone
authors.initials: S
authors.lastname: Cesaro
authors.affiliation: Department of Medical Sciences, University of Trieste, Trieste, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Bottega
authors.affiliation: Institute for Maternal and Child Health - IRCCS Burlo Garofolo, Trieste, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Faleschini
authors.affiliation: S. C. Mutagenesis, IRCCS AOU San Martino - IST (Istituto Nazionale per la Ricerca sul Cancro), CBA Torre A2, 16123 Genova, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Cuccarolo
authors.affiliation: Centro Diagnostica Genetica e Biochimica Malattie Metaboliche, Istituto Giannina Gaslini, 16148 Genova, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Corsolini
authors.affiliation: Institute of Biophysics, National Research Council, 16149 Genova, Italy.
authors.forename: Cesare
authors.initials: C
authors.lastname: Usai
authors.affiliation: SC Laboratory of Musculoskeletal Cell Biology, IOR, Bologna, Italy.
authors.forename: Marta
authors.initials: M
authors.lastname: Columbaro
authors.affiliation: Cystic Fibrosis Centre, Azienda Ospedaliera Universitaria, Piazzale Stefani, 1-37126 Verona, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Cipolli
authors.affiliation: Institute for Maternal and Child Health - IRCCS Burlo Garofolo, Trieste, Italy.|Department of Medical Sciences, University of Trieste, Trieste, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Savoia
authors.affiliation: S. C. Mutagenesis, IRCCS AOU San Martino - IST (Istituto Nazionale per la Ricerca sul Cancro), CBA Torre A2, 16123 Genova, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Degan
authors.affiliation: Haematology Unit, Istituto Giannina Gaslini, 16148 Genova, Italy.
authors.forename: Enrico
authors.initials: E
authors.lastname: Cappelli
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000172:deficiency; D001854:Bone Marrow Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D001855:Bone Marrow Diseases / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D002118:Calcium / Q000378:metabolism*; D030401:Cytochrome-c Oxidase Deficiency / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D003576:Electron Transport Complex IV / Q000235:genetics / Q000378:metabolism; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects / Q000235:genetics; D010188:Exocrine Pancreatic Insufficiency / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D005786:Gene Expression Regulation; D006019:Glycolysis / Q000235:genetics; D006801:Humans; D007930:Leucine / Q000494:pharmacology; D008068:Lipomatosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D009154:Mutation; D010766:Phosphorylation; D061251:Primary Cell Culture; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D012270:Ribosomes / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D000081003:Shwachman-Diamond Syndrome; D015398:Signal Transduction; D058570:TOR Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism
chemical_list: D011506:Proteins; D017382:Reactive Oxygen Species; C470521:SBDS protein, human; D000255:Adenosine Triphosphate; D003576:Electron Transport Complex IV; C546842:MTOR protein, human; D058570:TOR Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases; D007930:Leucine; D002118:Calcium