title: Impact of lysosomal storage disorders on biology of mesenchymal stem cells: Evidences from in vitro silencing of glucocerebrosidase (GBA) and alpha-galactosidase A (GLA) enzymes.
abstract: Lysosomal storage disorders (LDS) comprise a group of rare multisystemic diseases resulting from inherited gene mutations that impair lysosomal homeostasis. The most common LSDs, Gaucher disease (GD), and Fabry disease (FD) are caused by deficiencies in the lysosomal glucocerebrosidase (GBA) and alpha-galactosidase A (GLA) enzymes, respectively. Given the systemic nature of enzyme deficiency, we hypothesized that the stem cell compartment of GD and FD patients might be also affected. Among stem cells, mesenchymal stem cells (MSCs) are a commonly investigated population given their role in hematopoiesis and the homeostatic maintenance of many organs and tissues. Since the impairment of MSC functions could pose profound consequences on body physiology, we evaluated whether GBA and GLA silencing could affect the biology of MSCs isolated from bone marrow and amniotic fluid. Those cell populations were chosen given the former's key role in organ physiology and the latter's intriguing potential as an alternative stem cell model for human genetic disease. Our results revealed that GBA and GLA deficiencies prompted cell cycle arrest along with the impairment of autophagic flux and an increase of apoptotic and senescent cell percentages. Moreover, an increase in ataxia-telangiectasia-mutated staining 1 hr after oxidative stress induction and a return to basal level at 48 hr, along with persistent gamma-H2AX staining, indicated that MSCs properly activated DNA repair signaling, though some damages remained unrepaired. Our data therefore suggest that MSCs with reduced GBA or GLA activity are prone to apoptosis and senescence due to impaired autophagy and DNA repair capacity.
authors.affiliation: Department of Experimental Medicine, Biotechnology and Molecular Biology Section, University of Campania "Luigi Vanvitelli", Naples, Italy.|Institute of Bioscience and Bioresources, National Research Council, Naples, Italy.
authors.forename: Tiziana
authors.initials: T
authors.lastname: Squillaro
authors.affiliation: Laboratory of Molecular Genetics, Department of Psychological, Health and Territorial Sciences, School of Medicine and Health Sciences, G. d'Annunzio University, Chieti-Pescara, Italy.
authors.forename: Ivana
authors.initials: I
authors.lastname: Antonucci
authors.affiliation: Department of Experimental Medicine, Biotechnology and Molecular Biology Section, University of Campania "Luigi Vanvitelli", Naples, Italy.
authors.forename: Nicola
authors.initials: N
authors.lastname: Alessio
authors.affiliation: Department of Experimental Medicine, Biotechnology and Molecular Biology Section, University of Campania "Luigi Vanvitelli", Naples, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Esposito
authors.affiliation: Department of Experimental Medicine, Biotechnology and Molecular Biology Section, University of Campania "Luigi Vanvitelli", Naples, Italy.
authors.forename: Marilena
authors.initials: M
authors.lastname: Cipollaro
authors.affiliation: Department of Medical, Surgical, Neurological, Metabolic Sciences, and Aging; Division of Neurology and InterUniversity Center for Research in Neurosciences, University of Campania "Luigi Vanvitelli", Naples, Italy.
authors.forename: Mariarosa Anna Beatrice
authors.initials: MAB
authors.lastname: Melone
authors.affiliation: Institute of Bioscience and Bioresources, National Research Council, Naples, Italy.
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Peluso
authors.affiliation: Laboratory of Molecular Genetics, Department of Psychological, Health and Territorial Sciences, School of Medicine and Health Sciences, G. d'Annunzio University, Chieti-Pescara, Italy.
authors.forename: Liborio
authors.initials: L
authors.lastname: Stuppia
authors.affiliation: Department of Experimental Medicine, Biotechnology and Molecular Biology Section, University of Campania "Luigi Vanvitelli", Naples, Italy.
authors.forename: Umberto
authors.initials: U
authors.lastname: Galderisi
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D017209:Apoptosis; D001343:Autophagy; D001854:Bone Marrow Cells / Q000201:enzymology* / Q000473:pathology; D002469:Cell Separation; D002478:Cells, Cultured; D016922:Cellular Senescence; D002648:Child; D004260:DNA Repair; D000795:Fabry Disease / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D005260:Female; D005776:Gaucher Disease / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D005962:Glucosylceramidase / Q000172:deficiency* / Q000235:genetics; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000201:enzymology* / Q000473:pathology; D034622:RNA Interference; D016160:Retinoblastoma Protein / Q000235:genetics / Q000378:metabolism; D059807:S Phase Cell Cycle Checkpoints; D015398:Signal Transduction; D055153:Stem Cell Niche; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism; D000519:alpha-Galactosidase / Q000235:genetics / Q000378:metabolism*
chemical_list: D016160:Retinoblastoma Protein; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; C576330:GLA protein, human; D000519:alpha-Galactosidase; D005962:Glucosylceramidase
keywords: autophagy; fabry disease; gaucher disease; lysosomal storage disorders; mesenchymal stem cells; senescence