An altered redox balance mediates the hypersensitivity of Cockayne syndrome primary fibroblasts to oxidative stress
Barbara Pascucci
Istituto di Cristallografia, Consiglio Nazionale delle Ricerche, Via Salaria, Km 29,300, 00016 Monterotondo Stazione, Rome, Italy
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
These authors equally contributed to this work.
Search for more papers by this authorTiziana Lemma
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
These authors equally contributed to this work.
Search for more papers by this authorEgidio Iorio
Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorSara Giovannini
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorBruno Vaz
Istituto di Genetica Molecolare, Consiglio Nazionale delle Ricerche, Via Abbiategrasso 207, 27100 Pavia, Italy
Search for more papers by this authorIvano Iavarone
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorAngelo Calcagnile
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorLaura Narciso
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorPaolo Degan
IRCCS Azienda Ospedaliera Universitaria San Martino – IST – Istituto Nazionale per la Ricerca sul Cancro, Largo Rosanna Benzi 10, 16132, Genova, Italy
Search for more papers by this authorFranca Podo
Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorVera Roginskya
Department of Pharmacology and Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA
Hillman Cancer Center, The University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA
Search for more papers by this authorBratislav M. Janjic
Hillman Cancer Center, The University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA
Search for more papers by this authorBennett Van Houten
Department of Pharmacology and Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA
Hillman Cancer Center, The University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA
Search for more papers by this authorMiria Stefanini
Istituto di Genetica Molecolare, Consiglio Nazionale delle Ricerche, Via Abbiategrasso 207, 27100 Pavia, Italy
Search for more papers by this authorEugenia Dogliotti
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorMariarosaria D’Errico
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorBarbara Pascucci
Istituto di Cristallografia, Consiglio Nazionale delle Ricerche, Via Salaria, Km 29,300, 00016 Monterotondo Stazione, Rome, Italy
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
These authors equally contributed to this work.
Search for more papers by this authorTiziana Lemma
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
These authors equally contributed to this work.
Search for more papers by this authorEgidio Iorio
Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorSara Giovannini
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorBruno Vaz
Istituto di Genetica Molecolare, Consiglio Nazionale delle Ricerche, Via Abbiategrasso 207, 27100 Pavia, Italy
Search for more papers by this authorIvano Iavarone
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorAngelo Calcagnile
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorLaura Narciso
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorPaolo Degan
IRCCS Azienda Ospedaliera Universitaria San Martino – IST – Istituto Nazionale per la Ricerca sul Cancro, Largo Rosanna Benzi 10, 16132, Genova, Italy
Search for more papers by this authorFranca Podo
Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorVera Roginskya
Department of Pharmacology and Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA
Hillman Cancer Center, The University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA
Search for more papers by this authorBratislav M. Janjic
Hillman Cancer Center, The University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA
Search for more papers by this authorBennett Van Houten
Department of Pharmacology and Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA
Hillman Cancer Center, The University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA
Search for more papers by this authorMiria Stefanini
Istituto di Genetica Molecolare, Consiglio Nazionale delle Ricerche, Via Abbiategrasso 207, 27100 Pavia, Italy
Search for more papers by this authorEugenia Dogliotti
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorMariarosaria D’Errico
Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Search for more papers by this authorSummary
Cockayne syndrome (CS) is a rare hereditary multisystem disease characterized by neurological and development impairment, and premature aging. Cockayne syndrome cells are hypersensitive to oxidative stress, but the molecular mechanisms involved remain unresolved. Here we provide the first evidence that primary fibroblasts derived from patients with CS-A and CS-B present an altered redox balance with increased steady-state levels of intracellular reactive oxygen species (ROS) and basal and induced DNA oxidative damage, loss of the mitochondrial membrane potential, and a significant decrease in the rate of basal oxidative phosphorylation. The Na/K-ATPase, a relevant target of oxidative stress, is also affected with reduced transcription in CS fibroblasts and normal protein levels restored upon complementation with wild-type genes. High-resolution magnetic resonance spectroscopy revealed a significantly perturbed metabolic profile in CS-A and CS-B primary fibroblasts compared with normal cells in agreement with increased oxidative stress and alterations in cell bioenergetics. The affected processes include oxidative metabolism, glycolysis, choline phospholipid metabolism, and osmoregulation. The alterations in intracellular ROS content, oxidative DNA damage, and metabolic profile were partially rescued by the addition of an antioxidant in the culture medium suggesting that the continuous oxidative stress that characterizes CS cells plays a causative role in the underlying pathophysiology. The changes of oxidative and energy metabolism offer a clue for the clinical features of patients with CS and provide novel tools valuable for both diagnosis and therapy.
Supporting Information
Fig. S1 CS-A and CS-B primary fibroblasts present increased levels of intracellular ROS as detected by EPR.
Fig. S2 Mitochondrial function declines in CS-A and CS-B cells.
Fig. S3 CS-A and CS-B primary fibroblasts have decreased mitochondrial function.
Fig. S4 CS-A and CS-B cells display higher steady-state levels of ATP.
Fig. S5 CS-A and CS-B primary fibroblasts exposed to a ROS-inducing agent present increased levels of intracellular ROS and decreased mitochondrial membrane potential.
Fig. S6 CS-A and CS-B cells are hypersensitive to the killing effects of H2O2.
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