Caroline Cieniewski-Bernard | Université des Sciences et Technologies de Lille (Lille-1) (original) (raw)

Papers by Caroline Cieniewski-Bernard

Journal of Physiology and Biochemistry, 2016

The endocannabinoid system is dysregulated during obesity in tissues involved in the control of f... more The endocannabinoid system is dysregulated during obesity in tissues involved in the control of food intake and energy metabolism. We examined the effect of chronic exercise on the tissue levels of endocannabinoids (eCBs) and on the expression of genes coding for cannabinoid receptor 1 (CB1) and cannabinoid receptor 2 (CB2) (Cnr1 and Cnr2, respectively) in the subcutaneous (SAT) and visceral adipose tissues and in the soleus and extensor digitorim longus (EDL) muscles, in rats fed with standard or high-fat diet. Twenty-eight male Wistar rats were placed on high-fat diet or standard diet (HFD and Ctl groups, respectively) during 12 weeks whereafter half of each group was submitted to an exercise training period of 12 weeks (HFD + training and Ctl + training). Tissue levels of eCBs were measured by LC-MS while expressions of genes coding for CB1 and CB2 receptors were investigated by qPCR. High-fat diet induced an increase in anandamide (AEA) levels in soleus and EDL (p &amp;amp;amp;amp;amp;lt; 0.02). In soleus of the HFD group, these changes were accompanied by elevated Cnr1 messenger RNA (mRNA) levels (p &amp;amp;amp;amp;amp;lt; 0.05). In EDL, exercise training allowed to reduce significantly this diet-induced AEA increase (p &amp;amp;amp;amp;amp;lt; 0.005). 2-Arachidonoylglycerol (2-AG) levels were decreased and increased by high-fat diet in SAT and EDL, respectively (p &amp;amp;amp;amp;amp;lt; 0.04), but not affected by exercise training. Unlike the HFD + training group, 2-AG levels in soleus were also decreased in the HFD group compared to Ctl (p &amp;amp;amp;amp;amp;lt; 0.04). The levels of eCBs and Cnr1 expression are altered in a tissue-specific manner following a high-fat diet, and chronic exercise reverses some of these alterations.

Les maladies cardiovasculaires sont les causes majeures de la mortalité et de la morbidité dans l... more Les maladies cardiovasculaires sont les causes majeures de la mortalité et de la morbidité dans les pays développés et les mécanismes moléculaires responsables sont encore largement inconnus bien que l'expression de certain(e)s gènes/protéines soit altérés. La complexité biologique a mis en évidence le fait qu'un gène ne code pas pour une seule protéine mais pour plusieurs à cause des différents processus d'épissage alternatif, des modifications post-traductionnelles. L'analyse protéomique est une technologie qui permet d'appréhender de manière globale les modulations du niveau d'expression des protéines et d'effectuer une comparaison différentielle entre divers échantillons. L'analyse en surface-enhanced laser desorption ionization-time of flight (SELDI-TOF) est une technique, basée sur la combinaison de la chromatographie par rétention sur des puces à protéine et la spectrométrie de masse, qui permet la comparaison de profils protéiques à partir d'échantillons biologiques variés. Cette technique a été particulièrement utilisée pour l'étude du plasma/sérum. Nous aborderons l'analyse protéomique comparative du plasma de patients ayant eu un infarctus par la technique SELDI en abordant le principe de cette technique, les problèmes inhérents à l'étude du plasma, l'identification des protéines et la validation de l'approche en utilisant d'autres techniques. Ces approches se révèlent particulièrement intéressantes pour étudier les maladies cardiovasculaires dans le but de comprendre les mécanismes moléculaires impliqués, pour mettre en évidence les interactions protéiques et identifier de nouveaux facteurs/marqueurs biologiques impliqués dans les différentes pathologies cardiovasculaires. # 2007 Elsevier Masson SAS. Tous droits réservés.

Médecine Nucléaire, 2007

Frontiers in Physiology, 2014

Frontiers in Endocrinology, 2014

The cellular diversity of proteins results in part from their post-translational modifications. A... more The cellular diversity of proteins results in part from their post-translational modifications. Among all of them, the O-GlcNAcylation is an atypical glycosylation, more similar to phosphorylation than classical glycosylations. Highly dynamic, reversible, and exclusively localized on cytosolic, nuclear, and mitochondrial proteins, O-GlcNAcylation is known to regulate almost all if not all cellular processes. Fundamental for the cell life, O-GlcNAcylation abnormalities are involved in the etiology of several inherited diseases. Assessing to O-GlcNAcylation pattern will permit to get relevant data about the role of O-GlcNAcylation in cell physiology.To get understanding about the role of O-GlcNAcylation, as also considering its interplay with phosphorylation, the O-GlcNAc profiling remains a real challenge for the community of proteomists/glycoproteomists. The development of multiplexed proteomics based on fluorescent detection of proteins permits to go further in the understanding of the proteome complexity. We propose herein a multiplexed proteomic strategy to detect O-GlcNAcylated proteins, phosphoproteins, and the whole proteome within the same bidimensional gel. In particular, we investigated the phosphoproteome through the ProQ Diamond staining, while the whole proteome was visualized through Sypro Ruby staining, or after the labeling of proteins with a T-Dye fluorophore. The O-GlcNAcome was revealed by the way of the Click chemistry and the azide-alkyne cycloaddition of a fluorophore on GlcNAc moieties. This method permits, after sequential image acquisition, the direct in-gel detection of O-GlcNAcome, phosphoproteome, and whole proteome.

FEBS Journal, 2005

Myosin isoforms are major determinants of the contractile properties of skeletal muscle fibres [1... more Myosin isoforms are major determinants of the contractile properties of skeletal muscle fibres [1], and the neural discharge pattern has an important role in the regulation of myosin isoform expression. This has been demonstrated by cross-innervation [2], denervation and chronic low-frequency stimulation (CLFS) experiments. In particular, after some weeks, denervation and CLFS cause changes in the myosin heavy chain (MHC) distribution in slow and fast muscles, which validates the view that the pattern

PROTEOMICS, 2008

Recent improvements in therapeutic strategies did not prevent left ventricular remodeling (LVR), ... more Recent improvements in therapeutic strategies did not prevent left ventricular remodeling (LVR), which remains a common event (30%) after acute myocardial infarction (AMI). We report the use of a systematic approach, based on comparative proteomics, to select circulating biomarkers that may be associated with LVR. We selected 93 patients enrolled in a prospective study. These patients with anterior wall Q-wave AMI underwent echocardiographic follow-up at hospitalization, 3 months and 1 year after AMI. They were divided into three groups (no, low, or high remodeling). Plasma samples of these patients (day 5 of hospitalization) were processed and stored at -80 degrees C within 2 h and analyzed using SELDI-TOF protein chip technology. This systematic approach allowed to select candidate proteins modulated by LVR: post-translational variants of alpha1-chain of haptoglobin (Hpalpha1) corresponding to m/z 9493, 9565, and 9623, which were more elevated in remodeling patients. The peak 9493 m/z was shown having a receiving-operating characteristic (ROC) value of 0.71 between non- and remodeling patients. SELDI-TOF approach may lead to the identification of circulating proteins associated with LVR. Whether these candidate proteins will help to identify patients who are at high risk of heart failure after AMI will have to be tested in future studies.

PLoS ONE, 2012

O-N-acetylglucosaminylation is a reversible post-translational modification which presents a dyna... more O-N-acetylglucosaminylation is a reversible post-translational modification which presents a dynamic and highly regulated interplay with phosphorylation. New insights suggest that O-GlcNAcylation might be involved in striated muscle physiology, in particular in contractile properties such as the calcium activation parameters. By the inhibition of O-GlcNAcase, we investigated the effect of the increase of soleus O-GlcNAcylation level on the contractile properties by establishing T/pCa relationships. We increased the O-GlcNAcylation level on soleus biopsies performing an organ culture of soleus treated or not with PUGNAc or Thiamet-G, two O-GlcNAcase inhibitors. The enhancement of O-GlcNAcylation pattern was associated with an increase of calcium affinity on slow soleus skinned fibers. Analysis of the glycoproteins pattern showed that this effect is solely due to O-GlcNAcylation of proteins extracted from skinned biopsies. We also characterized the O-GlcNAcylated contractile proteins using a proteomic approach, and identified among others troponin T and I as being O-GlcNAc modified. We quantified the variation of O-GlcNAc level on all these identified proteins, and showed that several regulatory contractile proteins, predominantly fast isoforms, presented a drastic increase in their O-GlcNAc level. Since the only slow isoform of contractile protein presenting an increase of O-GlcNAc level was MLC2, the effect of enhanced O-GlcNAcylation pattern on calcium activation parameters could involve the O-GlcNAcylation of sMLC2, without excluding that an unidentified O-GlcNAc proteins, such as TnC, could be potentially involved in this mechanism. All these data strongly linked O-GlcNAcylation to the modulation of contractile activity of skeletal muscle.

Pflügers Archiv - European Journal of Physiology, 2014

Although calcium is the major regulator of excitation-contraction coupling, myofilament function ... more Although calcium is the major regulator of excitation-contraction coupling, myofilament function can also be modulated through post-translational modifications. In particular, phosphorylation and O-GlcNAcylation are key modulators of calcium activation parameters. Among the regulatory proteins of skeletal muscle contraction, the myosin light chain 2 (MLC2) can undergo both types of post-translational modification. During aging or physical inactivity, the phosphorylation status of the slow isoform of MLC2 (sMLC2) does not correlate with calcium sensitivity, suggesting that the O-GlcNAcylation might modulate sMLC2 activity. To increase understanding of the contractile dysfunction associated with muscle atrophy, we studied the phosphorylation/O-GlcNAcylation interplay on the sMLC2. We demonstrate a two-fold decrease of O-GlcNAcylation level on sMLC2 in a rat model of skeletal muscle atrophy (hindlimb unloading), while phosphorylation increased. Both post-translational modifications were mutually exclusive. Their interplay reversed during reloading. The expression of enzymes involved in the phosphorylation and O-GlcNAcylation interplay on sMLC2 was modified on whole protein pattern as well as on myofilament, and was load-dependent. All enzymes were colocalized on the contractile apparatus. Finally, we describe a multienzymatic complex which might finely modulate the phosphorylation/dephosphorylation and O-GlcNAcylation/de-O-GlcNAcylation of sMLC2 that could be involved in the contractile dysfunction of atrophied muscle. Importantly, this complex was localized at the Z-disk, a nodal point of signalling in skeletal muscle.

Neuropathology and Applied Neurobiology, 2008

Although modifications of the survival motor neurone gene are responsible for most spinal muscula... more Although modifications of the survival motor neurone gene are responsible for most spinal muscular atrophy (SMA) cases, the molecular pathophysiology and the muscular target proteins involved are still unknown. The aim of this study was to compare the expression of contractile and regulatory protein isoforms in quadriceps muscles from SMA children with age-matched control quadriceps. Methods: The isoform patterns of myosin heavy chains (MHC), troponin subunits (T, C and I) and tropomyosin were determined by immunoblotting, reverse transcription-polymerase chain reaction and mass spectrometry analyses. Depending on the disease severity, their expression levels were followed in specific variants of SMA populations (types I, II and III), with comparison with agematched control muscles. Results: The isoform transitions in SMA muscles were different from the fast-to-faster transitions occurring in normal muscles from children aged 1 month to 5 years old. Moreover, the expression of the neonatal MHC isoform was not repressed in SMA muscles.

Molecular & Cellular Proteomics, 2004

) is a regulatory post-translational modification of nucleo-cytoplasmic proteins that has a compl... more ) is a regulatory post-translational modification of nucleo-cytoplasmic proteins that has a complex interplay with phosphorylation. O-GlcNAc has been described as a nutritional sensor, the level of UDP-GlcNAc that serves as a donor for the uridine diphospho-N-acetylglucosamine:polypeptide ␤-N-acetyl-glucosaminyltransferase being regulated by the cellular fate of glucose. Because muscular contraction is both dependent on glucose metabolism and is highly regulated by phosphorylation/dephosphorylation processes, we decided to investigate the identification of O-GlcNAc-modified proteins in skeletal muscle using a proteomic approach. Fourteen proteins were identified as being O-GlcNAc modified. These proteins can be classified in three main classes: i) proteins implicated in the signal transduction and in the translocation between the cytoplasm and the nucleus or structural proteins, ii) proteins of the glycolytic pathway and energetic metabolism, and iii) contractile proteins (myosin heavy chain). A decrease in the O-GlcNAc level was measured in the slow postural soleus muscle after 14-day hindlimb unloading, a model of functional atrophy characterized by a decrease in the force of contraction. These results strongly suggest that O-GlcNAc modification may serve as an important regulation system in skeletal muscle physiology.

Journal of Proteome Research, 2008

The development of chronic heart failure (CHF) following myocardial infarction is characterized b... more The development of chronic heart failure (CHF) following myocardial infarction is characterized by progressive alterations of left ventricle (LV) structure and function called left ventricular remodeling (LVR), but the mechanism of LVR remains still unclear. Moreover, information concerning the global alteration protein pattern during the LVR will be helpful for a better understanding of the process. We performed differential proteomic analysis of whole LV proteins using an experimental model of CHF in which myocardial infarction was induced in adult male rats by left coronary ligation. Among 1000 protein spots detected in 2D-gels, 49 were differentially expressed in LV of 2-month-old CHF-rats, corresponding to 27 different identified proteins (8 spots remained unidentified), classified in different functional groups as being heat shock proteins, reticulum endoplasmic stress proteins, oxidative stress proteins, glycolytic enzymes, fatty acid metabolism enzymes, tricarboxylic acid cycle proteins and respiratory chain proteins. We validated modulation of selected proteins using Western blot analysis. Our data showed that proteins involved in cardiac metabolism and oxidative stress are modulated during LVR. Interestingly, proteins of stress response showed different adaptation pathways in the early and late phase of LVR. Expression of four proteins, glyceraldehyde-3-phosphate dehydrogenase, alphaB-crystallin, peroxiredoxin 2, and isocitrate dehydrogenase, was linked to echographic parameters according to heart failure severity.

Journal of Muscle Research and Cell Motility, 2009

There is growing evidence that O-linked N-acetyl-D-glucosaminylation, more simply termed O-GlcNAc... more There is growing evidence that O-linked N-acetyl-D-glucosaminylation, more simply termed O-GlcNAcylation or O-GlcNAc, is a post-translational modification involved in many cellular processes from transcription to modulation of protein properties. O-GlcNAc is a dynamic and reversible glycosylation and therefore quite similar to the phosphorylation/dephosphorylation process, with which O-GlcNAc can interplay. Since O-GlcNAc serves as a glucose sensor by the way of hexosamine biosynthesis pathway, this glycosylation is often associated with glucose toxicity and development of insulin resistance. In this way, O-GlcNAc could be involved in muscle pathological consequences of diabetes. Nevertheless, in regards of several studies performed in healthy striated muscles, O-GlcNAc seems to exert protective effects against different types of injuries. Recent new insights suggest a key implication of O-GlcNAc in skeletal and cardiac muscles contractile activity, in particular by O-GlcNAc modification of motor as well as regulating contractile proteins. While evidence linked O-GlcNAc to the regulation of calcium activation properties, its exact role remains to be defined as well as the existence of potential interference with phosphorylation. The better understanding of the exact function of OGlcNAc in this physiological process could contribute to the determination of newly markers of skeletal dysfunctions.

Journal of Biological Chemistry, 2007

O-Linked N-acetylglucosaminylation termed O-GlcNAc is a dynamic cytosolic and nuclear glycosylati... more O-Linked N-acetylglucosaminylation termed O-GlcNAc is a dynamic cytosolic and nuclear glycosylation that is dependent both on glucose flow through the hexosamine biosynthesis pathway and on phosphorylation because of the existence of a balance between phosphorylation and O-GlcNAc. This glycosylation is a ubiquitous post-translational modification, which probably plays an important role in many aspects of protein functions. We have previously reported that, in skeletal muscle, proteins of the glycolytic pathway, energetic metabolism, and contractile proteins were O-GlcNAc-modified and that O-Glc-NAc variations could control the muscle protein homeostasis and be implicated in the regulation of muscular atrophy.

Journal of Applied Physiology, 2006

O-GlcNAc level variations are associated with the development of skeletal muscle atrophy. linked ... more O-GlcNAc level variations are associated with the development of skeletal muscle atrophy. linked N-acetylglucosaminylation (O-GlcNAc) is a regulatory posttranslational modification of nucleocytoplasmic proteins, which consists of the attachment of N-acetylglucosamine to serine or threonine residues of a protein. This glycosylation is a ubiquitous posttranslational modification, which probably plays important roles in many aspects of protein function. Our laboratory has previously reported that, in skeletal muscle, proteins of the glycolytic pathway and energetic metabolism and contractile proteins were O-GlcNAc modified (Cieniewski-Bernard C, Bastide B, Lefebvre T, Lemoine J, Mounier Y, and Michalski JC. Mol Cell Proteomics 3: [577][578][579][580][581][582][583][584][585] 2004). O-GlcNAc has been recently demonstrated to play a role in modulating cellular function in response to nutrition and also in stress conditions. Therefore, we have investigated here the implication of the glycosylation/deglycosylation process in the development of atrophy in rat skeletal muscle after hindlimb unloading. The high O-GlcNAc level found in control soleus [compared with control extensor digitorum longus (EDL)] becomes lower in atrophied soleus. On the opposite side, the low rate of O-GlcNAc in control EDL reaches higher levels in EDL, not atrophied after hindlimb unloading. These variations in O-GlcNAc level are correlated with a variation of the O-GlcNAc process enzyme activities and could be associated with a differential expression of heat shock proteins. Our results suggest that O-GlcNAc variations could control the muscle protein homeostasis and be implicated in the regulation of muscular atrophy.

Glycobiology, 2004

This article reports the first rigorous evidence for the existence of N-glycans in Giardia intest... more This article reports the first rigorous evidence for the existence of N-glycans in Giardia intestinalis, a parasite that is a widespread human pathogen, being a major cause of enteric disease in the world. Excreted/secreted molecules of G. intestinalis are known to stimulate the immune system. Structural strategies based on MALDI and electrospray mass spectrometry were employed to examine the excreted/secreted molecules for their N-glycan content. These revealed that the major oligosaccharides released by peptide N-glycosidase F are complex-type structures and correspond to bi-, and triantennary structures without core (␣1,6) fucosylation. The major nonreducing epitopes in these complex-type glycans are: Gal␤1-4GlcNAc (LacNAc) and NeuAc ␣2-6Gal␤1-4GlcNAc (sialylated LacNAc).

Clinical and Experimental Pharmacology and Physiology, 2008

1. Cardiovascular diseases are a major cause of morbidity and mortality in western countries. The... more 1. Cardiovascular diseases are a major cause of morbidity and mortality in western countries. The molecular mechanisms responsible for heart dysfunction are still largely unknown, except in cases of genetic defects or alteration of genes and proteins.

Archives of Cardiovascular Diseases, 2009

Le remodelage ventriculaire est un processus complexe qui intervient après un infarctus du myocar... more Le remodelage ventriculaire est un processus complexe qui intervient après un infarctus du myocarde chez 30 % des patients en dépit des meilleurs traitements connus actuellement. AÀ n d'en identiÀ er les déterminants moléculaires et d'en comprendre les mécanismes, nous avons étudié les modiÀ cations post-traductionnelles des protéines du ventricule gauche et en particulier, la phosphorylation et la O-N-acétylglucosaminylation (O-GlcNAc).