SFR TransBioMed - Université de Bordeaux

Transcription

SFR TransBioMed
2013
Bilan d’activité
Foreword
The Federative Research Structure TransBioMed was created in January 2011. It gathers about
500 people belonging to 12 laboratories.
The aim in setting up this federation was to bring together scientists involved in research on
human pathophysiology, from its very basic aspects up to clinical applications. Covered domains
include cancer, infectious diseases, immunology, cardiovascular and lung diseases, genetics and
metabolism.
The major objectives of TransBioMed are:
• To provide the community with an active scientific animation, mostly based on invited
seminars
• To establish, equip, maintain and staff state-of-the-art technical platforms
• To promote collaborations, using a specific call for projects
• To foster the career development of young scientists
This document will briefly introduce you to the organization and activities of TransBioMed and of
its constitutive teams.
Dr Jean Rosenbaum
3
contents
Flow chart..........................................................................................6
Scientific animation...........................................................................7
Call for Projects.................................................................................8
> Cancer...........................................................................................9
Genetics and Biology of Sarcomas (INSERM U916).................... 11
Breast Cancer Group (INSERM U916)............................................ 12
Molecular mechanisms of the response to treatment
(INSERM U916)............................................................................ 13
Angiogenesis and the tumor microenvironment (INSERM U1029)
(team 1)..........................................................................................
14
Angiogenesis and the tumor microenvironment (INSERM U1029)
(team 2)...................................................................................... 15
Biotherapies (INSERM U1035)...................................................... 16
Leukemic hematopoiesis (INSERM U1035).................................... 17
Dermatology group (INSERM U1035)............................................ 18
ER stress & Cancer (Inserm U1053)........................................... 19
Post-transcriptional regulation of genes, microRNAs
and liver cancer (Inserm U1053)................................................ 20
Cytoskeleton & Cancer group (Inserm U1053)............................ 21
Liver cancer (Inserm U1053)...................................................... 22
> Immunology - Microbiology............................................................ 23
Intracellular dynamic of subviral structures (CNRS UMR 5234).... 25
African Trypanosoma virulence factor and pathogenicity
(CNRS UMR 5234)........................................................................ 26
Trypanosome cytoskeleton biogenesis (CNRS UMR 5234).......... 27
Candida and Pathogenicity (CNRS UMR 5234)............................ 28
Variability, replication and mobility of viral and bacterial
genomes (CNRS UMR 5234)........................................................ 29
Pluripotency and early steps of differentiation (CNRS UMR 5164). 30
Activation of human T lymphocytes (CNRS UMR 5164)................ 31
> Biochemistry Genetics..................................................................... 33
Energy transduction systems and mitochondrial morphology
(SysTEMM) (IBGC CNRS UMR 5095)............................................ 35
Functional analysis of amyloids (IBGC CNRS UMR 5095)................ 36
Genetics of Metabolic Networks (IBGC CNRS UMR 5095)............. 37
Cellular energy metabolism (IBGC CNRS UMR 5095)......................38
Molecular genetics of mitochondrial systems
(IBGC CNRS UMR 5095)....................................................................39
Chromosome Segregation (IBGC CNRS UMR 5095)..................... 40
Mitochondria, stress and cell death (IBGC CNRS UMR 5095)........ 41
Cell Growth and Division (IBGC CNRS UMR 5095)........................ 42
4
TITRE
Mitochondrial organization and dynamics (IBGC CNRS UMR 5095).. 43
Contents
Control and Dynamics of Cell Division (IBGC CNRS UMR 5095)... 44
Cell Biology of Quiescence (IBGC CNRS UMR 5095).................... 45
Non-self recognition in fungi (IBGC CNRS UMR 5095)................... 46
Rare diseases: Genetics and Metabolism (MRGM, EA4576)....... 47
> Cardio-thoracic...........................................................................49
Bronchial remodelling (INSERM U1045)........................................51
Endothelial cell Biology (INSERM U1045).....................................52
Pathophysiology of pulmonary circulation (INSERM U1045).........53
Cardiovascular adaptation to ischemia (INSERM U1034).............54
technological platforms..................................................55
> Flow Cytometry core facility with high speed cell sorter.......58
> The Histology platform..............................................................60
> Level 3 safety laboratory...........................................................62
> Vectorology platform.................................................................64
5
Flow chart
Secrétariat Gestion
Directeur
Communication
Hélène Aouizerate
Virginie Rocher
Jean Rosenbaum
Les PlatesFormes
Les 4 Axes
Vectorologie :
Véronique
Guyonnet-Duperat
Alice Vimeney
Andreas Bikfalvi
Mojgan Mergny
Michael Kann
Julie Déchanet-Merville
Bertrand Daignan-Fornier
Didier Lacombe
Roger Marthan
Alain-Pierre Gadeau
Cytométrie :
Vincent Pitard
Santiago Gonzalez
Les Comités de ¨Pilotage de chaque axe :
Andreas Bikfalvi
Richard Iggo
Frédéric Mazurier
Muriel Priault
Hervé Bonnefoi
Michael Kann
Charlotte Behr
Derrick Robinson
Philippe Lehours
Thierry Noel
Isabelle Sagot
Anne Devin
Didier Lacombe
Roger Marthan
Cécile Duplaa
Alain-Pierre Gadeau
Thierry Letellier
Jean-François Quignard
Laboratoire P3
Michel Ventura
Histologie
Les laboratoires de chaque axe :
Histologie
Nathalie DugotSenant
Andréas BIKFALVI
INSERM U 1029
Université Bordeaux 1
Mécanismes moléculaires
de l’Angiogenèse
Michael Kann
CNRS UMR 5234
Université Bordeaux Segalen
Microbiologie cellulaire et
moléculaire et pathogénicité
Hubert DE VERNEUIL
INSERM U 1035
Université Bordeaux
Segalen
Biothérapie des maladies
génétiques et cancers
Jean-François MOREAU
CNRS UMR 5164
Composantes innées de la
réponse immunitaire et
différentiation
Jean-Philippe MERLIO
EA 2406
Segalen
Histologie et Pathologie
moléculaires des tumeurs
Francis MEGRAUD
INSERM U 853
Infection à Hélicobacter,
inflammation et cancer
Josy REIFFERS
INSERM U 916
Segalen
Validation et identification
de nouvelles cibles en
oncologie
Jean ROSENBAUM
INSERM U 1053
Segalen
Physiopathologie du
cancer du Foie
6
Bertrand DAIGNAN-FORNIER
CNRS UMR 5095
Institut de Biochimie et
Génétique Cellulaires
Roger Marthan
INSERM U 1045
Centre de Recherche
cardio-thoracique de
Bordeaux
Didier LACOMBE
Thierry Couffinhal
EA 4576
INSERM U 1034
Université Bordeaux Segalen Université Bordeaux Segalen
Maladies rares : Génétique et
Adapatation
cardiovasculaire à
métabolisme (MRGM)
l'ischémie
Scientific animation
Organizing the scientific life is one of the main tasks of the TransBioMed SFR. We offer a broad
program along several lines:
• Seminars with speakers invited by scientists from the SFR. Those are organized at least
weekly, often bi-weekly. About half of the speakers come from abroad
• A whole-day yearly seminar: this event gathers most people from the SFR and includes
guest lectures from prestigious outside speakers, talks by scientists from each of the 4
scientific axis, and a very active poster session
• Several thematic seminars have been organized by the scientific axes on MicrobiologyImmunology, Cardio-pulmonary research, and Biochemistry & Genetics
• Technical workshops: structural biology, flow cytometry.
In addition to these “generalist” events, a whole section is dedicated to the young researchers.
These events are organized by a group of young scientists nicknamed TBM docs:
• A whole-day yearly Young Scientists meeting that mixes scientific talks by young scientists
from TransBioMed and invited conferences dealing with science or career development
• Career development evening sessions where experts from recruiting committees from
INSERM, CNRS or Universities, or Biotech people, try to give the clues to recruitment
7
Call for Projects
The SFR launches a yearly call for projects for the funding of new, emerging projects, that link
teams from separate laboratories.
The projects are evaluated by external referees and the final selection is made by an ad hoc
committee.
Four to 6 projects are funded for an amount of 12-20 k€.
Projects funded in 2012
Applicant
Partner(s)
Title
Mélanie Bonhivers
Roger Marthan
Characterisation of SAXO proteins, a novel family of microtubule-associated proteins specific to cilia and flagella:
potential implication in bronchial ciliopathies.
Emmanuel Tetaud
Didier Lacombe
Comparative study of energetic compensation mechanisms
in human and yeast
Aurélia Cassany
Julie DéchanetMerville
Impact of core 18-27 epitope polymorphism on CD8 T cell
response in HLA-A2 patients with chronic HBV infection
Chloé James
François MoreauGaudry
Design of a new model for studying megacaryocytopoiesis
based on human iPSC (Induced Pluripotent Stem Cells)
Hélène Bœuf
François MoreauGaudry
Role of miRNA and LIF (Leukemia Inhibitory Factor) in the
plasticity of murine ES (Embryonic Stem) and iPS (induced
Pluripotent Stem) cells.
Projects funded in 2013
8
Applicant
Partner(s)
Title
Muriel Cario André
Marie-Elise Truchetet Role of pigmentary defects in the development of the fibrotic
process in systemic sclerosis
Armelle Menard
Bruno Cardinaud,
Christophe Grosset
Role of the CDT genotoxin of Helicobacter hepaticus in liver
carcinogenesis
Jean-François
Quignard
Harald Wodrich
Adenoviral infection and modification of the activity of type
T (Cav3) calcium channels through competitive recruitment
of Nedd4-2.
Christophe Richez
Cyril Goizet
Identification, quantification and characterization of extracellular mitochondrial DNA in rheumatoid arthritis
Isabelle Sagot
Violaine Moreau,
Frédéric Saltel
Actin-Bodies, a quiescence marker from yeast to human
Philippe Pourquier
Eric Chevet
Search for DNA-PKcs/Topoisomerase interaction antagonists for potentiating the action of camptothecine derivatives
CANCER
Genetics and Biology of
Sarcomas (INSERM U916)
http://www.bergonie.org/fr/basic-research/frederic-chibon.html
Frédéric Chibon
Our projects are divided into two main axes:
1) Describe and understand the genetics of sarcomas and
2) Transfer research data to patients.
Thus, we aim at 3 major objectives:
•To study, as exhaustively as possible the genome of sarcomas. To
this end, we have launched two Next generation Sequencing (NGS)
projects dealing with uterine sarcomas (complete genome) and
complex genetics sarcomas (Exome).
•Identify the mechanisms responsible for a prognosis signature identified
in the team. We study currently 4 candidate genes.
•Identify new therapeutic targets and signatures predictive for response
to chemotherapy. To this end, we have launched two projects. One aims
at characterizing, using CGH and expression arrays, 300 sarcomas
receiving a standardized adjuvant treatment in the French Sarcoma
Group and the RRePs and NetSarc networks. The second one aims at
developing a preclinical tests platform based on a panel of 25 sarcoma
cell lines established and characterized in the lab.
All these projects will be developed in the continuity of our interaction
with the pathologists and clinicians of the French Sarcoma Group for a
rapid and efficient transfer for the benefit of patients.
• Research team
Frédéric Chibon • PH-CR1 INSERM
Jean–Michel Coindre • PU-PH
Alain Aurias • DR INSERM
Binh Nguyen Bui • PH IB
Agnès Neuville • MCU-PH, IB
Antoine Italiano • CCA IB
Céline Brulard • TCH IB
Pauline Lagarde • TCH IB
Gaëlle Pérot • Ingénieur IB
• Selected Publications
Italiano A, Lagarde P, Brulard C, Terrier P, Laë M, Marques
B, Ranchere-Vince D, Michels JJ, Trassard M, Cioffi A,
Piperno-Neumann S, Chevreau C, Blay JY, Delcambre
C, Isambert N, Penel N, Bay JO, Bonvalot S, Le Cesne
A, Coindre JM, Chibon F. Genetic profiling identifies two
classes of soft-tissue leiomyosarcomas with distinct clinical
characteristics. Clin Cancer Res. 2013 Mar 1;19(5):11906. doi: 10.1158/1078-0432.CCR-12-2970.
Lagarde P, Przybyl J, Brulard C, Pérot G, Pierron G,
Delattre O, Sciot R, Wozniak A, Schöffski P, Terrier P,
Neuville A, Coindre JM, Italiano A, Orbach D, DebiecRychter M,Chibon F. Chromosome instability accounts
for reverse metastatic outcomes of pediatric and adult
synovial sarcomas. J Clin Oncol. 2013 Feb 10;31(5):60815
Pérot G, Croce S, Ribeiro A, Lagarde P, Velasco V,
Neuville A, Coindre JM, Stoeckle E, Floquet A, Macgrogan
G, Chibon F (2012). MED12 Alterations in Both Human
Benign and Malignant Uterine Soft Tissue Tumors. PLoS
One 7(6):e40015.
Lagarde P, Pérot G, Kauffmann A, Brulard C, Dapremont
V, Hostein I, Neuville A, Wozniak A, Sciot R, Schöffski P,
Aurias A, Coindre JM, Debiec-Rychter M, Chibon F (2012).
Mitotic checkpoints and chromosome instability are strong
predictors of clinical outcome in gastrointestinal stromal
tumors. Clin Cancer Res 18(3):826-38.
Gibault L, Ferreira C, Pérot G, Audebourg A, Chibon F,
Bonnin S, Lagarde P, Vacher-Lavenu MC, Terrier P, Coindre
JM, Aurias A (2012). From PTEN loss of expression to
RICTOR role in smooth muscle differentiation: complex
involvement of the mTOR pathway in leiomyosarcomas
and pleomorphic sarcomas. Mod Pathol 25(2):197-211.
Gibault L, Pérot G, Chibon F, Bonnin S, Lagarde P, Terrier
P, Coindre JM, Aurias A (2011). New insights in sarcoma
oncogenesis: a comprehensive analysis of a large series
of 160 soft tissue sarcomas with complex genomics. J
Pathol 223(1):64-71.
Pérot G, Chibon F, Montero A, Lagarde P, de Thé H, Terrier
P, Guillou L, Ranchère D, Coindre JM, Aurias A (2010).
Constant p53 pathway inactivation in a large series of
soft tissue sarcomas with complex genetics. Am J Pathol
177(4):2080-90.
Chibon F, Lagarde P, Salas S, Pérot G, Brouste V, Tirode
F, Lucchesi C, de Reynies A, Kauffmann A, Bui B, Terrier
P, Bonvalot S, Le Cesne A, Vince-Ranchère D, Blay JY,
Collin F, Guillou L, Leroux A, Coindre JM, Aurias A (2010).
Validated prediction of clinical outcome in sarcomas and
multiple types of cancer on the basis of a gene expression
signature related to genome complexity. Nat Med
16(7):781-7.
CANCER │ 11
Breast Cancer Group
(INSERM U916)
www.bergonie.org/en/basic-research/research/
Richard Iggo
• Research team
Richard Iggo • PU
Hervé Bonnefoi • PU-PH
Michel Longy • PH
Louis Mauriac • PH
Gaëtan MacGrogan • PH
Nicolas Sévenet • MCU-PH
Françoise Bonnet • IE
Elodie Richard • IE
Elodie Monceau • IE
Natalie Jones • Post-doc
Stéphanie Verbeke • Post-doc
Simon Bayle • Thesard
Audrey Rouault • Thesard
Virginie Bubien • Thesard
The breast cancer group has four principal investigators: Richard Iggo
(cell biology), Michel Longy (genetics), Gaetan MacGrogan (pathology)
and Hervé Bonnefoi (medical oncology). It conducts research on the
biology and treatment of breast cancer. The group receives funding from
multiple sources including project grants from INCa, ANR, ARC, and
the Aquitaine Region Council, and a programme grant from the Cancer
League.
Major projects in the group include:
1. Identification of breast cancer predisposition genes by next generation
sequencing of tumours and germline DNA from patients attending the
genetics clinic at the Bergonie Cancer Institute.
2. Characterisation of breast cancer predisposition genes in cell lines
and transgenic mice.
3. Identification of changes acquired by breast tumours during treatment
with anti-hormonal therapy.
• Selected publications
Bertolotto C, Lesueur F, Giuliano S, Strub T, de Lichy M, Bille K, Dessen P,
d’Hayer B, Mohamdi H, Remenieras A, Maubec E, de la Fouchardière A,
Molinié V, Vabres P, Dalle S, Poulalhon N, Martin-Denavit T, Thomas L, ndryBenzaquen P, Dupin N, Boitier F, Rossi A, Perrot JL, Labeille B, Robert C,
Escudier B, Caron O, Brugieres L, Saule S, Gardie B, Gad S, Richard S,
Couturier J, Teh BT, Ghiorzo P, Pastorino L, Puig S, Badenas C, Olsson H,
Ingvar C, Rouleau E, Lidereau R, Bahadoran P, Vielh P, Corda E, Blanche
H, Zelenika D, Galan P, Aubin F, Bachollet B, Becuwe C, Berthet P, Bignon
YJ, Bonadona V, Bonafe JL, Bonnet-Dupeyron MN, Cambazard F, ChevrantBreton J, Coupier I, Dalac S, Demange L, D’Incan M, Dugast C, Faivre L,
Vincent-Fetita L, Gauthier-Villars M, Gilbert B, Grange F, Grob JJ, Humbert
P, Janin N, Joly P, Kerob D, Lasset C, Leroux D, Levang J, Limacher JM,
Livideanu C, Longy M, Lortholary A, Stoppa-Lyonnet D, Mansard S, Mansuy
L, Marrou K, Mateus C, Maugard C, Meyer N, Nogues C, Souteyrand P, VenatBouvet L, Zattara H, Chaudru V, Lenoir GM, Lathrop M, Davidson I, Avril MF,
Demenais F, Ballotti R, Bressac-de-Paillerets B. A SUMOylation-defective
MITF germline mutation predisposes to melanoma and renal carcinoma.
Nature 2011 ; 480 : 94-8.
Bertucci F, Borie N, Roché H, Bachelot T, Le Doussal JM, MacGrogan G,
Debono S, Martinec A, Treilleux I, Finetti P, Esterni B, Extra JM, Genève J,
Hermitte F, Chabannon C, Jacquemier J, Martin AL, Longy M, Maraninchi D,
Fert V, Birnbaum D, Viens P. Gene expression profile predicts outcome after
anthracycline-based adjuvant chemotherapy in early breast cancer. Breast
Cancer Res Treat 2011 ; 127 : 363-73.
Bonadona V, Bonaïti B, Olschwang S, Grandjouan S, Huiart L, Longy M,
Guimbaud R, Buecher B, Bignon YJ, Caron O, Colas C, Noguès C, LejeuneDumoulin S, Olivier-Faivre L, Polycarpe-Osaer F, Nguyen TD, Desseigne F,
Saurin JC, Berthet P, Leroux D, Duffour J, Manouvrier S, Frébourg T, Sobol H,
Lasset C, Bonaïti-Pellié C. Cancer risks associated with germline mutations
in MLH1, MSH2, and MSH6 genes in Lynch syndrome. JAMA 2011 ; 305 :
2304-10.
Bonnefoi H, Piccart M, Bogaerts J, Mauriac L, Fumoleau P, Brain E, Petit T,
Rouanet P, Jassem J, Blot E, Zaman K, Cufer T, Lortholary A, Lidbrink E, André
S, Litière S, Dal Lago L, Becette V, Cameron DA, Bergh J, Iggo R, on behalf of
12 │CANCER
the EORTC 10994/BIG 1-00 Study Investigators. TP53 status for prediction of
sensitivity to taxane versus non-taxane neoadjuvant chemotherapy in breast
cancer (EORTC 10994/BIG 1-00): a randomised phase 3 trial. Lancet Oncol
2011 ; 12 : 527-39.
Chirgwin J, Sun Z, Smith I, Price KN, Thürlimann B, Ejlertsen B, Bonnefoi
H, Regan MM, Goldhirsch A, Coates AS, for the BIG 1-98 Collaborative and
International Breast Cancer Study Groups. The advantage of letrozole over
tamoxifen in the BIG 1-98 trial is consistent in younger postmenopausal
women and in those with chemotherapy-induced menopause. Breast Cancer
Res Treat 2012 ; 131 : 295-306.
Guedj M, Marisa L, de Reynies A, Orsetti B, Schiappa R, Bibeau F, MacGrogan
G, Lerebours F, Finetti P, Longy M, Bertheau P, Bertrand F, Bonnet F, Martin
AL, Feugeas JP, Bièche I, Lehmann-Che J, Lidereau R, Birnbaum D, Bertucci
F, de Thé H, Theillet C. A refined molecular taxonomy of breast cancer.
Oncogene 2012 ; 1196-206-1206.
Houdayer C, Caux-Moncoutier V, Krieger S, Barrois M, Bonnet F, Bourdon
V, Bronner M, Buisson M, Coulet F, Gaildrat P, Lefol C, Leone M, Mazoyer
S, Muller D, Remenieras A, Revillion F, Rouleau E, Sokolowska J, Vert
JP, Lidereau R, Soubrier F, Sobol H, Sévenet N, Bressac-de-Paillerets B,
Hardouin A, Tosi M, Sinilnikova OM, Stoppa-Lyonnet D. Guidelines for splicing
analysis in molecular diagnosis derived from a set of 327 combined in silico/in
vitro studies on BRCA1 and BRCA2 variants. Hum Mutat 2012 ; 33 : 1228-38.
Rouleau E, Jesson B, Briaux A, Nogues C, Chabaud V, Demange L,
Sokolowska J, Coulet F, Barouk-Simonet E, Bignon YJ, Bonnet F, Bourdon
V, Bronner M, Caputo S, Castera L, Delnatte C, Delvincourt C, Fournier J,
Hardouin A, Muller D, Peyrat JP, Toulas C, Uhrhammer N, Vidal V, StoppaLyonnet D, Bieche I, Lidereau R. Rare germline large rearrangements in the
BRCA1/2 genes and eight candidate genes in 472 patients with breast cancer
predisposition. Breast Cancer Res Treat 2012 ; 133 : 1179-90.
Toulmonde M, Madranges N, Brouste V, Donamaria C, MacGrogan G, Durand
M, Bonnefoi H, Mauriac L, Debled M. Docetaxel rechallenge after a first
response in non-resistant metastatic breast cancer: significant activity with
manageable toxicity. Breast Cancer Res Treat 2012 ; 134 : 325-32.
Molecular mechanisms of
the response to treatment
(INSERM U916)
http://www.bergonie.org/fr/basic-research/pierre-soubeyran.html
Pierre Soubeyran
• Research team
Our group
is focusing on the identification and validation of new
therapeutic targets in 3 tumor types: colon cancers, prostate cancer,
and agressive lymphomas in aged patients. We study more particularly
calicum signaling and death receptors (see articles in PNAS and Plos
Biol 2011) as well as autophagy (Autophagy 2011), and try to identify
factros predicting the resposne to chemotherapy and targeted therapies
(publications in Pharmacogenomics, Carcinogenesis et Cancer). We
can validate targets at the clinical and functional levels, notably using a
robotized flow cytometry equipment (publications in 2011 in Am J Pathol
and Plos One). We also use multiples techniques including cell and
tissue imaging (confocal microscope and ICys laser scanning cytometer),
molecular electrophysiology, flow cytometry, especially at high throughput
(High Throughput screening), pharmacogenetics and pyrosequencing.
Pierre Soubeyran • PU-PH
Jacques Bonnet • PU émérite
Serge Evrard • PU-PH
Jacques Robert • PU-PH
Danièle Montaudon • MCU-PH
Valérie Le Morvan • IE Université
Laurence Bresson–Bepoldin • CR1 CNRS
Mojgan Djavaheri–Mergny • CR1 INSERM
Philippe Pourquier • CR1 INSERM
Pierre Vacher • CR1 INSERM
Isabelle Mahouche • AI INSERM
Anne–Marie Vacher • AI INSERM
Gaëlle Labrunie • IE INSERM
Jean-Pierre Portail • IE INSERM
Brigitte Godard • Gestionnaire INSERM
Nadine Houédé • PH IB
Isabelle Soubeyran • PH IB
Stéphane Pedeboscq • PH CHU
Denis Smith • PH CHU
Hervé Wallerand • PH CHU
Ricardo Bellott • Ingénieur IB
Lydia Lartigue-Faustin • Post-Doc
Christophe Rey • Post-Doc
Emilie Charles • Thèse
Julie Dubois • Thèse
Barbara Lortal • Thèse
Emmanuel Roche • Thèse
Aurore Trocoli • Thèse
Djavaheri-Mergny M, Maiuri MC, Kroemer G. Cross talk between apoptosis and
autophagy by caspase-mediated cleavage of Beclin 1. Oncogene 2010 ; 29 :
1717-9.
Houédé N, Thall PF, Nguyen H, Paoletti X, Kramar A. Utility-based optimization
of combination therapy using ordinal toxicity and efficacy in phase I/II trials.
Biometrics 2010 ; 66 : 532-40.
Laroche-Clary A, Le Morvan V, Yamori T, Robert J. Cytochrome P450 1B1
(CYP1B1) gene polymorphisms as predictors of anticancer drug activity:
studies with in vitro models. Mol Cancer Ther 2010 ; 9 : 3315-21.
Pédeboscq S, Gravier D, Casadebaig F, Hou G, Gissot A, De Giorgi F, Ichas F,
Cambar J, Pometan JP. Synthesis and study of antiproliferative activity of novel
thienopyrimidines on glioblastoma cells. Eur J Med Chem 2010 ; 45 : 2473-9.
Khadra N, Bresson-Bepoldin L, Penna A, Chaigne-Delalande B, Segui B,
Levade T, Vacher AM, Reiffers J, Ducret T, Moreau JF, Cahalan M, Vacher
P, Legembre P. CD95/Fas triggers Orai1-mediated localized Ca2+ entry that
regulates recruitment of PKCß2 and prevents DISC formation. PNAS 2011 ;
108 : 19072-7.
Mathiaux J, Le Morvan V, Pulido M, Jougon J, Bégueret H, Robert J. Role of
DNA repair gene polymorphisms in the efficiency of platinum-based adjuvant
chemotherapy for non-small cell lung cancer. Mol Diagn Ther 2011 ; 15 : 159-66.
Salles G, Seymour JF, Offner F, López-Guillermo A, Belada D, Xerri L, Feugier
P, Bouabdallah R, Catalano JV, Brice P, Caballero D, Haioun C, Pedersen LM,
Delmer A, Simpson D, Leppa S, Soubeyran P, Hagenbeek A, Casasnovas O,
Intragumtornchai T, Fermé C, Gomes da Silva M, Sebban C, Lister A, Estell JA,
Milone G, Sonet A, Mendila M, Coiffier B, Tilly H. Rituximab maintenance for
2 years in patients with high tumour burden follicular lymphoma responding to
rituximab plus chemotherapy (PRIMA): a phase 3, randomised controlled trial.
Lancet 2011 ; 377 : 42-51.
Soubeyran I, Mahouche I, Grigoletto A, Leste-Lasserre T, Drutel G, Rey C,
Blanchard F, Brouste V, Pédeboscq S, Sabourin JC, Bécouarn Y, Reiffers J,
Ichas F, De Giorgi F. Tissue microarray cytometry reveals positive impact of
HIPK2 in colon cancer survival irrespective of p53 function. Am J Pathol 2011
; 178 : 1986-98.
Tauzin S, Chaigne-Delalande B, Selva E, Khadra N, Daburon S, Contin-Bordes
C, Blanco P, Le Seyec J, Ducret T, Counillon L, Moreau JF, Hofman P, Vacher
P, Legembre P. The naturally processed CD95L elicits a c-yes/calcium/PI3Kdriven cell migration pathway. PloS Biol 2011 ; 9 : e1001090.
Trocoli A, Mathieu J, Priault M, Reiffers J, Souquere S, Pierron G, Besançon F,
Djavaheri-Mergny M. ATRA-induced upregulation of Beclin 1 prolongs the life
span of mature Acute Promyelocytic Leukemia (APL) cells but is not essential
for the granulocytic differentiation process. Autophagy 2011 ; 7 : 1108-14.
Trocoli A, Djavaheri-Mergny M. The complex interplay between autophagy and
NF-B signaling pathways in cancer cells. Am J Cancer Res 2011 ; 1 : 629-49.
Vacher P, Khadra N, Vacher AM, Charles E, Bresson-Bepoldin L, Legembre
P. Does calcium contribute to the CD95 signaling pathway? Anticancer Drugs
2011 ; 22 : 481-7.
Auzanneau C, Montaudon D, Jacquet R, Puyo S, Pouysegu L, Deffieux
D, Elkaoukabi-Chaibi A, De Giorgi F, Ichas F, Quideau S, Pourquier P. The
polyphenolic ellagitannin vescalagin acts as a preferential catalytic inhibitor of
the alpha isoform of human DNA topoisomerase II. Mol Pharmacol 2012 ; 82
: 134-41.
Laroche-Clary A, Kauffmann A, Smith D, Laurand-Quancard A, Evrard S, Brunet
R, Le Morvan V, Robert J. Absence of transcriptomic signature of response to
chemotherapy in metastatic colorectal carcinoma patients. Pharmacogenomics
2012 ; 13 : 497-504.
Moisan F, Laroche-Clary A, Auzanneau C, Ricard N, Pourquier P, Robert J, Le
Morvan V. Deciphering the role of the ERCC2 gene polymorphism on anticancer
drug sensitivity. Carcinogenesis 2012 ; 33 : 962-8.
Pédeboscq S, Rey C, Petit M, Harpey C, De Giorgi F, Ichas F, Lartigue L. Nonantioxidant properties of alpha-tocopherol reduce the anticancer activity of
several protein kinase inhibitors in vitro. PLoS One 2012 ; 7 : e36811.
Soubeyran P, Fonck M, Blanc-Bisson C, Blanc JF, Ceccaldi J, Mertens C,
Imbert Y, Cany L, Vogt L, Dauba J, Palis R, Andriamampionona F, Houédé N,
Floquet A, Chomy F, Brouste V, Ravaud A, Bellera C, Rainfray M. Predictors of
early death risk in older patients treated by first-line chemotherapy for cancer. J
Clin Oncol 2012 ; 30 : 1829-34.
CANCER │ 13
Angiogenesis and the tumor
microenvironment
(INSERM U1029) (Team 1)
http://www.u-bordeaux1.fr/U1029/Site/INSERM_U1029.html
Andreas Bikfalvi
• Research team
Andreas Bikfalvi Pr
Sophie Javerzat Pr
Clotilde Billottet MCU
Patrick Auguste MCU
Sophie North MCU
Géraldine Miquel ASI
Céline Léon IGE
Nadège Pujol Tech
Cathy Quemener Post-doctoral fellow
Jessica Baud IGE
Students :
Sylvaine Guerit
Virginie Godard
Wilfried Souleyrau
Platonova N, Miquel G, Regenfuss B,
Taouji S, Cursiefen C, Chevet E, Bikfalvi
A. Evidence for the interaction of fibroblast
growth factor-2 with the lymphatic
endothelial cell marker LYVE-1. Blood.
2013 Feb 14;121(7):1229-37
Soulet F, Kilarski WW, Roux-Dalvai F,
Herbert JM, Sacewicz I, Mouton-Barbosa
E, Bicknell R, Lalor P, Monsarrat B, Bikfalvi
A. Mapping the extracellular and membrane
proteome associated with the vasculature
and the stroma in the embryo. Mol Cell
Proteomics. 2013 May 14
The aim of the laboratory is to study the cross talk between angiogenesis
and tumor biology, and in particular metastasis, and to reinforce the
translational aspects of our research programs. While angiogenesis
is thought to promote tumor growth through the delivery of oxygen and
nutriments to the growing tumor, recent evidence suggests that additional
functions may be involved. We are interested in unraveling paracrine
effects of angiogenic vessels on primary tumor growth, tumor invasion
and metastasis. Emerging evidence also indicates that tumors can escape
angiogenic blockade. We are interested in identifying mechanisms of
escape and new therapeutic targets to prevent/treat tumor escape induced
after angiogenic blockade or radiotherapy. The aim is also to identify new
potential therapeutic targets for cancer that are related to angiogenesis
and tumor cell invasion. Furthermore, today there are no validated tests
to routinely monitor angiogenesis in patients. The conventional endpoints
used to evaluate anticancer agents, such as response rate, prolongation
of time to progression and time to death are inadequate to assess
antiangiogenic agents. Furthermore we aim to gain insights into the
formation of the prematastatic niche.
The following studies have been conducted in the last years :
- Development of new models of tumor angiogenesis in the chick embryo
- Biological role and structure-function analysis of the CXC chemokines
CXCL4L1 and CXCL4
- Role of endothelial kinesins in tumor angiogenesis
- Role of the unfolded protein response in angiogenesis and tumor cell
invasion
- Role of fibroblast growth factors (FGFs) in lymphangiogenesis and of
the lymphatic marker Lyve-1
Fergelot P, et al. The Experimental Renal
Cell Carcinoma Model in the Chick Embryo.
Angiogenesis 2013 Jan;16(1):181-94
Dubrac A, et al. Functional divergence
between two chemokines is conferred by
single amino acid change. Blood, 2010,
116(22):4703-11
Cavill R, et al. A combined metabonomic
and transcriptomic approach to investigate
metabolism during development in the chick
chorioallantoic membrane. J Proteome Res.
2010 Jun 4;9(6):3126-34]
Auf G, et al Inositol-requiring enzyme 1alpha
is a key regulator of angiogenesis and
invasion in malignant glioma. Proc Natl Acad
Sci U S A. 2010 Aug 31;107(35):15553-8.
Glioblastoma cells (green) coopting vessels (red)
14 │CANCER
Angiogenesis and the tumor
microenvironment
(INSERM U1029) (Team 2)
http://www.u-bordeaux1.fr/U1029/Site/INSERM_U1029.html
A-Majid Khatib
To attain their biological active forms, a variety of protein precursors are
processed by proteases named proprotein convertases (PCs). We are
pioneered in this field of investigation. Our previous studies were the first to
demonstrate the importance of the maturation of protein precursors such
as matrix metalloproteases, adhesion molecules, and growth factors by
these enzymes in carcinogenesis, angiogenesis and metastasis. We found
that inhibition of the PCs in various tumor cells represses their malignant
phenotypes and their ability to mediate tumor growth and angiogenesis.
We also identified MT1-MMP, PDGF-A, PDGF-B, VEGF-C and Hepcidin
as new PCs substrates. Inhibition of these molecules processing by
specific inhibitors or through directed mutagenesis blocks their functions.
For example unprocessed PDGF and VEGF-C was found to inhibit tumor
growth and the formation of tumor vascular and lymphatic vessels. Based
on these and other findings we postulate that PCs play a key role in the
growth, survival and metastatic potential of tumor cells by regulating
the activity of their cognate substrates and downstream effectors. This
multidisciplinary research program covers several interconnecting areas
from cells to small animals (zebrafish, mice) and proteins, to the basic
chemistry of inhibitors and clinical settings. The aim of our team is:
[1] Development of small molecules PC inhibitors and evaluate their
effects on angiogenesis, tumor growth and metastasis.
[2] Evaluation of the correlation between the expression/activity of PCs
and tumor progression and metastasis.
Now that the role of PCs in various diseases is well established, the
proposed investigations will not only add information on their exact mode
of actions, but will provide further evidence that expression of PCs is a
major event leading to carcinogenesis and metastasis. In the long term,
the program will provide a rationale for testing a new family of small
molecule compounds as potential anti-tumorigenic and angiogenic agents
or in conjunction with standard therapy in clinical settings.
The 4 best inhibitors (blue) docked into
the Furin active site identified by Virtual
ligand screening (VLS)
Tumor cells expressing tomato-vector were injected into eGFP-labelled, transgenic fli-1
zebrafish and tumor cells were visualisezd under UV light. Vessels are in green and tumor
cells in red. C: Higher magnification. Arrow indicates tumor cells.
• Research team
A-Majid Khatib
Martin Hagedorn
Fabienne Soulet
Claude Lalou
Jia Ma
Allain Barbara
Students :
Fatma Sfaxi
Christophe Chemineau
Scamuffa N, Metrakos P, Calvo F,
Khatib AM. Identification of the myosin
heavy polypeptide 9 as a downstream
effector of the proprotein convertases
in the human colon carcinoma HT-29
cells. Methods Mol Biol. 2011;768:207-15.
Khatib AM, Lahlil R, Scamuffa N,
Akimenko MA, Ernest S, Lomri A, Lalou
C, Seidah NG, Villoutreix BO, Calvo
F, Siegfried G. Zebrafish ProVEGF-C
expression,
proteolytic processing
and inhibitory effect of unprocessed
ProVEGF-C during fin regeneration.
PLoS One. 2010, 5:e11438.
Basak A, Chen A, Scamuffa N,
Mohottalage D, Basak S, Khatib
AM. Blockade of furin activity and
furin-induced tumor cells malignant
phenotypes
by
the
chemically
synthesized human furin prodomain.
Curr Med Chem. 2010;17:2214-21.
Scamuffa N, Basak A, Lalou C,
Wargnier A, Marcinkiewicz J, Siegfried
G, Chrétien M, Calvo F, Seidah NG,
Khatib AM. Regulation of prohepcidin
processing and activity by the subtilisinlike proprotein convertases Furin, PC5,
PACE4 and PC7. Gut. 2008. 57:157382.
Scamuffa N, Siegfried G, Bontemps
Y, Ma L, Basak A, Cherel G, Calvo
F, Seidah NG, Khatib AM. Selective
inhibition of proprotein convertases
represses the metastatic potential of
human colorectal tumor cells. J Clin
Invest. 2008,118:352-63.
CANCER │ 15
Biotherapies (INSERM U1035)
www.u1035.u-bordeaux2.fr
François Moreau-Gaudry
iPSC and porphyrias
• Research team
François Moreau-Gaudry,
PU-PH, HDR
Chercheurs et Enseignant-chercheurs
Aurélie Bedel, MCU-PH
Sandrine Dabernat, MCU-PH, HDR
Cécile Ged, MCU-PH, HDR
Evelyne Peuchant, MCU-PH, HDR
Emmanuel Richard, MCU-PH
Hubert de Verneuil, PU-PH, HDR
Jean-Marc Blouin, AHU
Personnels ITA
Magalie Lalanne, Technicienne
Bordeaux 2
Isabelle Moranvillier, Technicienne
Bordeaux 2
Corrine Bourdie, Technicienne
INSERM
Miguel Taillepierre, IE CDD
Doctorants
François Béliveau
Marie Lafitte
Yann Duchartre
Bedel A, Taillepierre M, GuyonnetDuperat V, Lippert E, Dubus P,
Dabernat S, Mautuit T, Cardinaud B,
Pain C, Rousseau B, Lalanne M, Ged C,
Duchartre Y, Richard E, de Verneuil H,
Moreau-Gaudry F. Metabolic Correction
of Congenital Erythropoietic Porphyria
with iPSCs Free of Reprogramming
Factors. Am J Hum Genet. 2012 Jul
13;91(1):109-21.
Lafitte M, Rousseau B, Moranvillier I,
Taillepierre M, Peuchant E, GuyonnetDupérat V, Bedel A, Dubus P, de
Verneuil H, Moreau-Gaudry F, Dabernat
S. In vivo gene transfer targeting in
pancreatic adenocarcinoma with cell
surface antigens. Mol Cancer. 2012 Oct
22;11:81
16 │CANCER
Erythropoietic porphyrias are genetic diseases linked to an enzymatic defect in the
heme biosynthetic pathway, leading to the toxic accumulation of porphyrins in the
bone marrows, red blood cells, spleen and liver. They are associated with a cutaneous
photosensitivity of variable severity according to the type of porphyria. The oncly
curative treatment for severe porphyrias is allogeneic bone marrow transplantation,
which requires a HLA-compatible donor. We are developing gene therapy approaches
for erythropoietic porphyrias using lentiviral-based gene transfer into autologous
hematopoietic stem cells. We are currently evaluating the potential of human and
murine induced pluripotent stem cells obtained through reprogramming somatic
stem cells and their differentiation into the hematopoietic and hepatic lineage for the
treatment of these diseases.
Hepatocyte
differentiation
from
human
iPSCs:
physiopathological studies and therapeutic applications for
inherited liver diseases.
Transplantation of allogenic or genetically modified autologous hepatocytes may be
an alternative to whole-liver transplantation for the treatment of inherited liver diseases
(ILD). Several aspects can actually limit the clinical use of hepatocyte transplantation
such as poor in vitro amplification, limited engraftment of transplanted hepatocytes
in vivo, and the necessity for immune suppression (allogenic graft). Furthermore,
relevant ILD-patients-specific models of hepatocytes would strongly enhance our
knowledge of the pathophysiological mechanism responsible for ILD. In this project, we
propose to generate induced pluripotent stem cells (iPSCs) from patients with primary
hyperoxaluria type-1 (PH1). PH1 is an inherited disorder due alanine-glyoxylateamino transferase (AGT) deficiency and characterized by overproduction of oxalate
by the liver leading to sever nephrocalcinosis, nephrolithiasis and end-stage renal
failure. PH1-iPSCs-derived hepatocytes would represent faithfully models to support
pathophysiological studies and therapeutic drug screening. We will also evaluate a
targeted gene therapy approach in PH1-iPSCs-derived hepatocytes using TALENmediated gene correction at the endogenous human AGXT locus. This strategy would
offer a safe and broad genetic correction together with physiological regulation of the
therapeutic transgene. A similar approach is considered for acute hepatic porphyrias
to develop patient-specific hepatocytes as models for porphyrinogenic drug screening.
Our goal is to improve the development of iPSCs-derived hepatocytes as valuable
models of ILD and develop new therapeutic approaches in regenerative medicine.
Molecular targets and gene therapy of pancreas
adenocarcinoma
Pancreas cancer is among the 10 most common cancers and is the 4th cause of
cancer death in occidental countries. Ductal pancreas adenocarcinoma is by far the
most deadly since its diagnosis is difficult. Indeed, it lacks early and specific clinical
features, and there is also a lack of biological markers that can be used for prevention
or early detection. Thus, only few patients can benefit from a curative surgery. Other
treatments are limited since this cancer is very much radio- and chemo-resistant.
A major focus in research in this field deals in the understanding of the molecular
mechanisms involved in tumor development or aggressiveness, with the aim of finding
new targets. Alternately, one could target genes known to be specifically deregulated
in this disease, in order to slow down tumor progression, or induce tumor cell death.
Within this frame, we develop several projects: (1) is the FGFR3 receptor a possible
target to control pancreas adenocarcinoma progression, (2) can we use targeted gene
therapy against this disease, (3) can we use an association of several natural products
with anti-cancer properties in order to reduce the progression of this cancer?
Leukemic hematopoiesis
(INSERM U1035)
François-Xavier Mahon
Chronic myeloid leukemia
We study chronic myeloid leukemia (CML: www.lmc-cml.org) as a model
disease. This hematologic malignancy is characterized by a marker
allowing to recognize leukemic cells. This marker, the Philadelphia
chromosome, has a molecular equivalent, the BCR-ABL gene. The
chimeric BCR-ABL protein has a deregulated tyrosine kinase (TK) activity.
This TK is the “engine” of leukemic cells as it activates proliferation, antiapoptotic pathways and genetic instability. This hematologic malignancy
has greatly contributed to the advent of targeted therapies and our team
has contributed to the pre-clinical and clinical development of an original
drug, STI571, better known today as imatinib (or Glivec°). It became
the leader of a new class of drugs known as TK inhibitors (TKI). We
have shown in vitro and in vivo the emergence of resistances that led
the pharmaceutical industry to develop 2nd and 3rd generation TKIs
on which we are now focusing our research. One of the key questions
nowadays is more the persistence of leukemic cells than the resistance.
Indeed a proportion of tumor cells, likely the more immature, are persisting
despite a long-term treatment. We are thus investigating the molecular
mechanism that underlie oncogene independence or the cooperation with
other kinases.
Other myeloid malignancies
- Clinical biology of myeloproliferative syndromes (MPS): within european
and international networks (European Leukemia Net, MPN&MPNreuronet), we participate to the collaborative efforts aiming at developing
and standardizing tests for the detection and quantification of mutations
found in SMP (JAK2V617F, exon 12, MPLW515, PDGFR) or related
diseases (hereditary erythrocytosis and thrombocytosis). We also
characterize the functional consequences of genetic alterations that we
find
- Pathogenesis of des myeloid malignancies: this research is also in close
link with the clinic. We investigate how genetic/cytogenetic alterations
participate to leukemogenesis. We work nowadays especially on two
models: 1) we have recently described an acute myeloid leukemia
characterized by a fusion gene MYB-GATA1 that defines a new
cytogenetic entity, and are refining the functional characterization of the
fusion gene and its relationship with basophil hematopoiesis. 2) mixed
myeloproliferative and myelodysplastic syndromes, especially atypical
CML, which pathophysiology is poorly understood. We are especially
interested in expression and epigenetic regulation of micro-ARN in this
model.
• Research team
François-Xavier MAHON, PU-PH
Chercheurs et Enseignant-chercheurs
Françis BELLOC, Ingénieur, CHU de
Bordeaux, HDR
Bruno CARDINAUD, MC, Institut
Polytechnique de Bordeaux
Francis Lacombe, PH, CHU de
Bordeaux
Eric LIPPERT, MCU-PH
Jean-Max PASQUET, CR1 INSERM
Arnaud PIGNEUX, PH, CHU de
Bordeaux
Béatrice TURCQ, CR1 CNRS, HDR
Stéphanie DULUCQ, PH, CHU de
Bordeaux
Personnels ITA
Valérie PROUZET-MAULEON, IE
Bordeaux 2
Valérie LAGARDE, Technicienne
Bordeaux 2
Post-Doctorant
Romain GIOIA, CDD INCA
Doctorantes
Kelly AIRIAU
Marina JOSSELIN
Gioia R, Leroy C, Drullion C, Lagarde V,
Etienne G, Dulucq S, Lippert E, Roche
S, Mahon FX, Pasquet JM. Quantitative
phosphoproteomics revealed interplay
between Syk and Lyn in the resistance
to nilotinib in chronic myeloid leukemia
cells.Blood. 2011 Aug 25;118(8):221121.
Rea D, Etienne G, Nicolini F, ConyMakhoul P, Johnson-Ansah H, Legros
L, Huguet F, Tulliez M, Gardembas M,
Bouabdallah K, Rousselot P, Cayuela
JM, Guilhot F, Mahon FX. First-line
imatinib mesylate in patients with newly
diagnosed accelerated phase-chronic
myeloid leukemia. Leukemia. 2012 Mar
30. doi: 10.1038/leu.2012.92
CANCER │ 17
Dermatology group
(INSERM U1035)
www.u1035.u-bordeaux2.fr • www.maladiesrarespeau.fr
Alain Taieb
• Research team
Alain TAIEB, PU-PH, HDR
Annie BERARD, MCU-PH, HDR
Katia BONIFACE, MC
Muriel CARIO-ANDRE, Ingénieur,
CHU de Bordeaux, HDR
Vincent CASOLI, PU-PH, HDR
Khaled EZZEDINE, PH, CHU de
Bordeaux
Thomas JOUARY, PH, CHU de
Bordeaux
Frédéric MAZURIER, DR2 INSERM
Djavad MOSSALAYI, PU, HDR
Daniel MOYNET, MC
Hamid REZVANI, CR2 INSERM
Isabelle LAMRISSI-GARCIA,
Technicienne Bordeaux 2
Denis THIOLAT, AGT Bordeaux 2
Catherine PAIN, Technicienne CDI
contrat privé
Walid MAHFOUF, IE CDD
Post-Doctorants
François Moisan
Houssam Raad
Martin SERRANO-SANCHEZ
Doctorants
Mohsen HASSEINI
Sorilla PREY
Abir ZEBIAN
XPC silencing in normal human
keratinocytes
triggers
metabolic
alterations that drive the formation of
squamous cell carcinomas.Rezvani
HR, Kim AL, Rossignol R, Ali N, Daly
M, Mahfouf W, Bellance N, Taïeb A, de
Verneuil H, Mazurier F, Bickers DR. J
Clin Invest. 2011 Jan;121(1):195-211.
Rezvani HR, Ali N, Serrano-Sanchez
M, Dubus P, Varon C, Ged C, Pain C,
Cario-André M, Seneschal J, Taïeb
A, de Verneuil H, Mazurier F. Loss of
epidermal hypoxia-inducible factor1α accelerates epidermal aging and
affects re-epithelialization in human and
mouse. J Cell Sci. 2011 Dec 15;124(Pt
24):4172-83
18 │CANCER
Relationships between genomic mutations, alteration of energy
metabolism and increase in reactive oxygen species during cancer
initiation
Key words: Xeroderma pigmentosum, DNA repair, Warburg effect, skin
cancers
Study of the role of HIF factors in hematopoietic and epidermic cells
Key words: Hypoxia, HIF, hematopoiesis, epidermis, aging, cancer
Beta-adrenergic signalization and angiogenesis
Key words: propranolol, hemangioma, vasculogenesis, antiangiogenic
therapy
Vitiligo pathophysiology
Key words: Vitiligo, melanocyte, dermis, paracrine factors, inflammation
A description of our projects can be found on our website:
ER stress & Cancer
(INSERM U1053)
www.gref-bordeaux.fr/fr/ER_stress
Eric Chevet
The «ER stress and cancer» team focuses on the understanding of
adpatation mechanisms set in place by the normal and pathological cells
to cope with their environment. The team is particularly interested in the
role of the first sub-cellular compartment of the secretory pathway, the
endoplasmic reticulum (ER). This compartment, which can represent
up to 50% of the total cellular membranes, plays instrumental roles in the
maintenance of calcium, lipides, and protein homeostasis.
• Research team
Eric Chevet, DR2
S Taouji, IE Inserm
N Dejeans, post doc
S Lhomond, Thèse
When ER homeostasis is perturbed, the imbalance is sensed in the lumen
of this compartment and transmitted to the cytosol and the nucleus. Three
sensors have been identified thus far and named protein kinase RNA-like
endoplasmic reticulum kinase (PERK), Activating Transcription Factor 6
(ATF6) and Inositol Requiring Enzyme 1 (IRE1). The signalling pathways
activated downstream of these three ER stress sensors are collectively
called the Unfolded Protein Response (UPR). The UPR aims at restoring
ER homeostasis but if this fails, it triggers programmed cell death.
The UPR is activated in numerous pathologies including diabetes,
degenerative disorders ro cancer. Our team focuses on elucidating the
molecular mechanisms of ER stress in cancers (the models studied
in the laboratory are glioblastoma, hepatocellular carcinoma and
cholangiocarcinoma). We demonstrated that the IRE1 arm of the UPR
significantly contributes to tumor growth in vitro and in vivo through the
activation of its cytosolic endoribonuclease activity. The currrent projects
aim at dissecting IRE1 signalling networks in cancer and identifying natural
and synthetic molecules susceptible to alter IRE1 functions. On a longer
term, the experimental strategies currently developed in the laboratory will
be applied to the other ER stress sensors PERK and ATF6.
Huber AL, Lebeau J, Guillaumot P, Pétrilli V, Malek M, Chilloux
J, Fauvet F, Payen L, Kfoury A, Renno T, Chevet E, Manié SN.
p58(IPK)-Mediated Attenuation of the Proapoptotic PERK-CHOP
Pathway Allows Malignant Progression upon Low Glucose. Mol
Cell. 2013
Dejeans N, Pluquet O, Lhomond S, Grise F, Bouchecareilh M,
Juin A, Meynard-Cadars M, Bidaud-Meynard A, Gentil C, Moreau
V, Saltel F, Chevet E. Autocrine control of glioma cells adhesion/
migration through Inositol Requiring enzyme 1α (IRE1α)mediated cleavage of Secreted Protein Acidic Rich in Cysteine
(SPARC) mRNA. J Cell Sci. 2012 125:4278-87
Guichard C, Amaddeo G, Imbeaud S, Ladeiro Y, Pelletier L,
Ben Maad I, Calderaro J, Bioulac-Sage P, Letexier M, Degos F,
Clément B, Balabaud C, Chevet E, Laurent A, Couchy G, Letouzé
E, Calvo F, Zucman-Rossi J. Integrated analysis of somatic
mutations and focal copy-number changes identifies key genes
and pathways in hepatocellular carcinoma. Nature Genetics,
2012;44(6):694-8
Yi P, Higa A, Taouji S, Bexiga MG Marza E, Arma D, Castain C Le
Bail B, Simpson JC, Rosenbaum J, Balabaud C, Bioulac-Sage P,
Blanc JF, Chevet E. Sorafenib–mediated targeting of the AAA+
ATPase p97/VCP leads to disruption of the secretory pathway,
endoplasmic reticulum stress and hepatocellular cancer cell
death. Mol Cancer Ther 2012 11:2610-20
Bouchecareilh M, Higa A, Fribourg S, Moenner M, Chevet E.
Peptides derived from the bifunctional kinase/RNase enzyme
IRE1{alpha} modulate IRE1{alpha} activity and protect cells from
endoplasmic reticulum stress. FASEB J. 2011 25: 3115-29.
Yi P, Thang Nguyên D, Higa-Nishiyama A, Auguste P,
Bouchecareilh M, Dominguez M, Bielmann R, Palcy S, Liu JF and
Chevet E. MAPK scaffolding by the Bcl2-inhibitor of transcription
1 (BIT1) in the Golgi apparatus modulates stress resistance. J
Cell Science 2010; 23 :1060-72
CANCER │ 19
Post-transcriptional
regulation of genes,
microRNAs and liver cancer
(INSERM U1053)
www.gref-bordeaux.fr/en/microRNA
Christophe Grosset
• Research team
Christophe GROSSET, CR1 INSERM
Francis SAGLIOCCO, Assistant
Professor, Bordeaux Segalen
University
Sarah LESJEAN, AI INSERM
Flora Cartier, Post-doc
1- Maurel, M. et al. MicroRNA-1291mediated silencing of IRE1α enhances
Glypican-3 expression. RNA, 2013,
19(6): 778-788.
2- Maurel, M. et al. A functional
screening identifies five miRNAs
controlling Glypican-3: Role of miR1271 down-regulation in hepatocellular
carcinoma. Hepatology, 2013, 57 (1):
195-204.
3- Jalvy-Delvaille, S. et al. Molecular
basis of differential target regulation by
miR-96 and miR-182. The Glypican-3
as a model. Nucl Acids Res, 2012, 40
(3), 1356-1365.
4- Simon, D. et al. A mutation in the 3’UTR of the HDAC6 gene abolishing the
post-transcriptional regulation mediated
by hsa-miR-433 is linked to a new form
of dominant X-linked chondrodysplasia,
Hum Mol Genet, 2010, 19, 2015-2027.
5- Laloo, B., et al. Analysis of posttranscriptional
regulations
by
a
functional, integrated, and quantitative
method. Mol and Cel Prot, 2009,
8(8):1777-1788.
My team works on the post-transcriptional regulation of genes by
microRNAs (miRNAs), small non-coding RNAs which control translation
and messenger RNA (mRNA) stability. Our favourite gene is the Glypican-3
(GPC3), a gene overexpressed in liver cancer and which participates
in liver tumorigenesis through the Wnt/β-catenin pathway. We study the
miRNAs regulating GPC3 expression and evaluate the role played by
these regulations in the growth of cancerous hepatic cells.
We developed a system, named Dual-Fluorescence-FunREG (Figure 1),
which allows the screening of miRNA (and siRNA) libraries in human cells
and the identification of miRNAs functionally active on a specific gene.
Figure 1: Principle of the DF-FunREG
screening methodology. Target cells
expressing the Reference «Tomato»
and Test «GFP + 3’UTR of interest»
transgenes are transfected by each
miRNA from the library. The miRNAs
inducing a significant variation of the
GFP/Tomato ratio compared to the
control RNA (reference value « A »)
are selected as «hits». PRO: promoter
EF1-alpha-long (Ref. 2).
Using Dual-Fluorescence-FunREG, we identified 5 miRNAs regulating
positively or negatively GPC3 expression in tumoral hepatic cells through
its 3’ untranslated region (Figure 2; Ref. 1 to 3). MiR-96 and miR-1271
down-regulated GPC3 expression through a direct microRNA:3’UTR
recognition process. The 3 others, miR-129-1-3p, miR-1291 and miR1303, enhanced GPC3 expression, likely by an indirect mechanism. This
assumption was further demonstrated with miR-1291 which favours GPC3
expression by inhibiting the expression of the endoplasmic reticulum (ER)resident stress sensor IRE1α through a specific site located in its 5′ UTR.
Finally IRE1α cleaves GPC3 mRNA at a 3′ UTR consensus site, thereby
prompting its decay and GPC3 downregulation.
Figure 2: Summary of GPC3 3’UTRassociated post-transcriptional regulations mediated by 5 miRNAs and the
endoribonuclease IRE1α. Red line:
negative effect. Green arrow: positive
(and likely indirect) effect (Ref. 1 to 3).
In conclusion, our data show that GPC3 expression is regulated by
several miRNAs and the endoribonuclease IRE1α (Figure 2) illustrating
the fascinating notions of collaborative post-transcriptional gene regulation
network. Finally, the altered expression of some of these factors
(especially miR-1271) in liver cancer favours GPC3 expression and thus,
the expansion of cancerous liver cells. Therefore, these factors constitute
bona fide molecular targets in liver cancer.
20 │CANCER
Cytoskeleton & Cancer group
(INSERM U1053)
www.gref-bordeaux.fr/en/cytoSk-cancer-group
Violaine Moreau et Frédéric Saltel
Reorganization of the actin cytoskeleton is important for cell migration,
cell shape change, and interactions with other cells and the environment.
Abnormal cell migration and invasion is a characteristic of malignant cancer
cells and is one component of metastasis, the major clinical problem in
cancer. Consequently, alterations in cytoskeletal signalling pathways
are increasingly being recognized as important for cancer invasion and
metastasis. Our group uses a wide range of biochemical, cell biological
methods and animal models to study how actin cytoskeleton is regulated
and involved in hepatocellular carcinoma cell invasion.
We mainly developed two axes:
Role of Rho GTPases in the development of hepatocellular carcinoma
(Violaine Moreau)
Our recent work focused on the atypical Rho GTPase, RhoE/Rnd3. Using
patient samples and cultured cells, we have documented the frequent
down-regulation of RhoE/Rnd3 in hepatocellular carcinoma (HCC) and
shown that it behaves as a suppressor of invasion (Hepatology, 2012).
Left-hand side: Phylogenetic tree of the Rho GTPase family (from Grise et al., BBA-Reviews
in Cancer, 2009). Rnd3/RhoE is shown in red. Right-hand side: Rnd3 is down-regulated in
tumor liver. Immunohistochemistry of Rnd3 in HCC versus cirrhotic tissue. Tissues were
stained with anti-Rnd3 antibody.
Involvement of invadosomes in hepatocellular carcinoma (Frédéric
Saltel)
Invadosomes are F-actin structures involved in extracellular matrix
degradation. Our last study established the existence of a new linear
organization of invadosomes specifically induced by type I collagen fibrils
(MBoC, 2012). This physiological organization of type I collagen is present
in liver endothelial and hepatic cancer cells. Our future objective will be to
determine the impact of linear invadosomes in HCC cell invasion.
• Research team
Violaine MOREAU, CR1 INSERM
Frédéric SALTEL, CR1 INSERM
Valérie LAGREE, MCU
Lisa PAYSAN, Thèse
Julie DI MARTINO, Thèse
Caroline GEST, Post-doc
Bidaud-Meynard A*, Arma D*, Taouji S*,
Laguerre M, Dessolin J, Rosenbaum J,
Chevet E# & Moreau V#. A novel smallmolecule screening strategy identifies
mitoxantrone as a RhoGTPase inhibitor.
Biochem. J., 2013 Feb 15;450(1):5562.
Juin A, Planus E, Guillemot F, Horakova
P, Albiges-Rizo C, Génot E, Rosenbaum
J, Moreau V, Saltel F. Extracellular
matrix rigidity controls podosome
induction in microvascular endothelial
cells. Biol Cell. 2013 Jan;105(1):46-57.
Grise F*, Sena S*, Bidaud-Meynard
A, Baud J, Hiriart J-B, Makki K, DugotSenant N, Staedel C, Bioulac-Sage
P, Zucman-Rossi J, Rosenbaum J
& Moreau V. Rnd3/RhoE is downregulated in hepatocellular carcinoma
and involved in cellular invasion.
Hepatology 2012, 55:1766-1775.
Juin A, Billottet C*, Moreau V*, Destaing
O, Albiges-Rizo C, Rosenbaum J,
Génot E# & Saltel F#. Physiological
type I collagen organization induces
the formation of a novel class of linear
invadosomes. Mol. Biol. Cell 2012,
23(2):297-309.
Confocal images of the formation of linear invadosomes induced by type I collagen fibrils
(in red), and visualized using the staining of a specific marker (Tks5 in green). These linear
invadosomes are able to promote degradation of the gelatin (in grey) underneath, the
hallmark of invadosomes.
CANCER │ 21
Liver cancer (INSERM U1035)
www.gref-bordeaux.fr/en/liver-cancer
Jean Rosenbaum
• Research team
Jean Rosenbaum, DR1 INSERM
Patrick Lestienne, DR2 INSERM
Paulette Bioulac-Sage, PU-PH
Jean-Frédéric Blanc, PU-PH
Aksam Merched, professor
Véronique Neaud, adjoint technique
UBS
Nathalie Allain-Courtois, technicienne
UBS
Anne-Aurélie Raymond, post-doc
Samira Benhamouche, post-doc
Hepatocellular carcinoma is the most frequent primary liver cancer. It is
one of the most frequent and deadly cancers worldwide.
We have identified the overexpression in HCC of Reptin and Pontin, two
members of the AAA+ family, and shown that were important for HCC
progression. We aim at better understanding their mechanisms of action
and target them for therapy. We have notably generated a new conditional
knock-out mouse for Reptin that will allow in-depth analysis of its role in a
relevant setting.
Senescent tumor cells with senescenceassociated
heterochromatin
foci
following Reptin silencing
A
B
C
D
Identification
of
Small
molecule
inhibitors
of
Pontin ATPase activity
1.Grigoletto A, Neaud V, Allain-Courtois N, Lestienne P,
Rosenbaum J. The ATPase activity of Reptin is required for
its effects on tumor cell growth and viability in hepatocellular
carcinoma. Mol Cancer Res, 2013, 11(2) : 133–9
2.Guichard C, Amaddeo G, Imbeaud S, Ladeiro Y, Pelletier
L, Ben Maad I, Calderaro J, Bioulac-Sage P, Letexier M,
Degos F, Clément B, Balabaud C, Chevet E, Laurent
A, Couchy G, Letouzé E, Calvo F, Zucman-Rossi J.
Integrated analysis of somatic mutations and focal copynumber changes identifies key genes and pathways in
hepatocellular carcinoma. Nature Genetics, 2012, May
6;44(6):694-8
3.Ménard L, Taras D, Grigoletto A, Haurie V, Nicou A,
Dugot-Senant N, Costet P, Rousseau B, Rosenbaum J. In
vivo silencing of Reptin blocks the progression of human
hepatocellular carcinoma in xenografts and is associated
22 │CANCER
with replicative senescence. J Hepatol 2010, 52: 681-9
4.Haurie V, Ménard L, Nicou A, Touriol C, Metzler P,
Fernandez J, Taras D, Lestienne P Balabaud C, BioulacSage P, Prats H, Zucman-Rossi J, Rosenbaum J. The
ATPase Pontin is overexpressed in hepatocellular
carcinoma and co-regulated with Reptin through a new
post-translational mechanism. Hepatology 2009, 50 :
1871-1883.
5.Rousseau B, Ménard L, Haurie V, Taras D, Blanc JF,
Moreau-Gaudry F, Metzler P, Hugues M, Boyault S, Lemière
S, Canron X, Costet P, Cole M, Balabaud C, Bioulac-Sage
P, Zucman-Rossi J, Rosenbaum J. Overexpression and
role of the ATPase and putative DNA helicase RuvB-like
2 in human hepatocellular carcinoma. Hepatology 2007,
46 : 1108-18.
immunology
microbiology
Intracellular dynamic of
subviral structures
(CNRS UMR 5234)
www.mfp.cnrs.fr/mfp/team_dissv_en.php
équipe FRM
Michael Kann and Harald Wodrich
● HBV: the human hepatitis B virus is a major human
pathogen responsible for circa 1 Mio death per year.
Our research is focused on 1) the intracytosolic
trafficking of the capsid, which surrounds the viral
genome towards the nucleus, 2) the still unknown
passage – for all viruses – from the microtubules
to the nuclear pore and 3) the subsequent steps.
These comprise the crossing of the viral genome
inside the capsid through the nuclear pore (Fig. 1),
the liberation of the viral genome from the capsid
and the intranuclear genome conversion/repair.
The investigations are based on microscopical Fig. 1. HBV capsids at the techniques for localization and on biochemical cyto-plasmic and nuclear
methods for adjacent investigations. The project was side of the nuclear pore.
electron
recently expanded to the naturally polymorphism of Transmission
HBV in collaboration with the Research Centre for microscopy. From Kann &
Epidemiology and Biostatistics, Bordeaux (ISPED). Panté, MBC, 2002.
● Parvoviruses: recent investigations showed that parvoviruses enter the
nucleus of resting cells but circumvent the passage through the nuclear
pore. Our recent investigations using time lapse microscopy showed that
parvoviruses dock to the nuclear pore and induce disintegration of the
nuclear membrane. This process occurred even in the absence of cytosol and
requires some key enzymes of mitosis. The underlying mechanism of how
parvoviruses cross the nuclear membrane are thus of high interest in basic
science in order to understand the activating intranuclear cascades. However
the impact of this topic is not restricted to the gain of basic knowledge but has
applied consequences as these viruses are frequently used candidates in
cancer therapy and are moreover the vector for the first licensed drug in gene
therapy world-wide.
● Adenoviruses: the research focuses on the mechanism of endosomal
escape, cytosolic trafficking and nuclear import. These investigations are
linked to adenoviral application in gene therapy, vaccination and cancer
therapy. Our research has shown that these pathways require the interaction
of the capsid protein VI with the cellular ubiquitin/proteasome system. Protein
VI also mediates endosomal escape, which triggers innate immunity, thus
causing Ad mediated inflammation. Work in progress investigates protein VI
function in pathogenesis, intracellular trafficking and cell activation related to
post-translational modifications with ubiquitin and SUMO. The project includes
biochemical assays and cell based systems as well as advanced microscopic
techniques. For a better understanding of adenoviral pathogenesis, protein VI
sequences from patients with adenovirus infections are investigated for their
role in disease development. Further analysis on adenoviruses is performed
in collaboration with the hospital analysing the impact of adenoviral variability
on different clinical parameters.
Fig. 2. Top: Ad capsids (green
signal) separate from protein
VI (red signal) and accumulate
at the microtubule organizing
centre 45 min after infection.
Bottom panel: Ad capsids with
mutated protein VI incapable
of binding to cellular ubiquitin
ligases remain with the
capsid (yellow signal) and
show altered intracellular
trafficking (Wodrich et al.
PLoS Pathogens 2010)
• Research team
Marie-Edith LAFON (MD-PhD)
Christian Cazenave (scientist - CR1)
Fabienne Rayné (scientist – MCU)
Aurélia Cassany (postdoc)
Tetsuro Komatsu (postdoc)
Jessica Raque (technician)
Cindy Aknin (technician)
Muriel Faure della Corte (technician)
Quentin Osseman (PhD student)
Sisley Austin (PhD student)
Remi Vaillant (PhD student in
collaboration with the Heinrich-Pette
Institut, Hamburg, Gemany)
Kenza Snoussi (PhD student in
collaboration with the University of
Tsukuba, Japan)
Somar Kassab (MD-PhD student)
Schreiner S,Martinez R, Groitl P, Rayne
F, Vaillant R, Wimmer P, Bossis G,
Sternsdorf T, Marcinowski L, Ruzsics
Z, Dobner T, Wodrich H. Transcriptional
activation of the adenoviral genome is
mediated by capsid protein VI.. PLoS
Pathog. 2012 Feb;8(2):e1002549
Schmitz A, Schwarz A, Foss M, Zhou
L, Rabe B, Hoellenriegel J, Stoeber
M, Panté N, Kann M. Nucleoporin
153 arrests the nuclear import of
hepatitis B virus capsids in the nuclear
basket. PLoS Pathog. 2010 Jan
29;6(1):e1000741
Wodrich H, Henaff D, Jammart B,
Segura-Morales C, Seelmeir S, Coux
O, Ruzsics Z, Wiethoff CM, Kremer
EJ. A capsid-encoded PPxY-motif
facilitates adenovirus entry. PLoS
Pathog. 2010 Mar 19;6(3):e1000808
(must read Faculty of 1000)
Rabe B, Delaleau M, Bischof A, Foss M,
Sominskaya I, Pumpens P, Cazenave
C, Castroviejo M,Kann M. Nuclear entry
of hepatitis B virus capsids involves
disintegration to protein dimers followed
by nuclear reassociation to capsids.
PLoS Pathog. 2009 Aug;5(8):e1000563
immunology-microbiology │ 25
African Trypanosoma
virulence factor and
pathogenicity
(CNRS UMR 5234)
www.mcmp.aquitaine.cnrs.fr/mfp/
Théo Baltz and Virginie Coustou
• Research team
Zeinab AMMAR (PhD student)
Corinne ASENCIO (AI)
Nicolas BITEAU (IR)
Virginie COUSTOU-LINARES (CR1)
Julien IZOTTE (AI)
Davita PILLAY (Post-doctorante)
Nicolas PLAZOLLES (Tcn)
Loïc RIVIERE (MCU)
Magali THONNUS (Tcn)
Guegan F, Plazolles N, Baltz T, Coustou
V. Erythrophagocytosis of desialylated
red blood cells is responsible for
anaemia during Trypanosoma vivax
infection. Cell Microbiol. 2013 Feb 20.
doi: 10.1111/cmi.12123. [Epub ahead
of print]
Coustou V, Plazolles N, Guegan F,
Baltz T. Sialidases play a key role in
infection and anaemia in Trypanosoma
congolense animal trypanosomiasis.
Cell Microbiol. 2012 Mar;14(3):431-45.
Epub 2012 Feb 13.
Coustou V, Guegan F, Plazolles
N, Baltz T. Complete in vitro life
cycle of Trypanosoma congolense:
development of genetic tools. PLoS
Negl Trop Dis. 2010 Mar 2;4(3):e618.
Giroud C, Ottones F, Coustou V,
Dacheux D, Biteau N, Miezan B, Van
Reet N, Carrington M, Doua F, Baltz T.
Murine Models for Trypanosoma brucei
gambiense disease progression--from
silent to chronic infections and early
brain tropism. PLoS Negl Trop Dis.
2009 Sep 1;3(9):e509
Virulence and pathogenesis factors of African Trypanosomes. T.b. gambiense
is responsible for more than 90% of reported cases of human sleeping
sickness in West and Central Africa. Isolation by in vitro culture of different T.
b. gambiense field isolates directly from the CSF of patients allowed the group
to develop murine models with chronic, sub-chronic and silent infections,
mimicking the situation in the field. Engineering of a T. b. gambiense isolate
to express Renilla Luciferase revealed an unexpected early tropism not only
for the brain but also for other organs such as the spleen and lungs. These
murine models, combined with bioluminescence and immune-histochemistry,
provide new insights into disease progression and could possibly reveal new
mechanisms involved in host-parasite interactions (Fig 6, Giroud et al., PLoS
Negl Trop Dis 2009).
T. congolense is the main causative agent of animal trypanosomiasis in Africa.
Pathogenesis is mainly due to anaemia The group succeeded in producing
long-term cultures of all the developmental stages and to finally complete the
whole lifecycle in vitro. This improved model offers the opportunity to conduct phenotype analyses of genetically modified strains throughout the in
vitro lifecycle and also during experimental infections.(Fig 7, Coustou et al.,
PLoS Negl Trop Dis 2010). Ongoing research focuses on the identification
and characterisation of novel pathogenic factors, in terms of their protective
potential, specifically in the context of an “anti-anaemia” vaccine. Several
factors will be expressed as recombinant proteins and their purifications done
in collaboration with the platform of the unit.
Fig. 1: Bioluminescence analysis
of mice infected with the silent T.
b. gambiense isolate and of their
organs (1, uninfected; 2, infected).
Fig. 2: In vitro culture system for T.
congolense. Insect stages (PCF,
EMF, MCF) are represented on
the left as individual or adherent
(colonies) cells. The mammalian
stage (BSF) on the right is
represented in blood or on BAE
26 │Immunology-microbiology
Trypanosome cytoskeleton
biogenesis (CNRS UMR 5234)
http://www.mfp.cnrs.fr/mfp/team_bct_en.php
Derrick Robinson
Trypanosoma brucei is a flagellated protozoan that is the causal agent for
African sleeping sickness. It is used in studies of basic parasite biology
for health perspectives and also as a model organism to study numerous
biological phenomena such as characterization of gene expression,
cell cycle analysis, protein trafficking, glycobiology and cytoskeleton
biogenesis.
In order to obtain a better understanding of flagella and cell cytoskeleton
biogenesis in general, our lab’s research have focused on immunological
and proteomic analysis of minor flagellar proteins of this model organism.
Subsequently, our group has made a panel of polyclonal / monoclonal
antibodies against minor cytoskeleton proteins. We have also used
bioinformatics approaches to identify the biochemical means for basal
body segregation in T. brucei.
We also focus on novel proteins and structures including uncharacterized
basal body, spindle proteins, and flagellar proteins. We have recently
identified a microtubule-associated protein, TbSAXO, involved in flagellum
motility in the parasite. This protein belongs to a family of proteins involved
in the stabilization of microtubules from in ciliated and/or flagellated
organisms from protozoa to mammals. Their role in the cilia/flagella
function is under study.
Currently, our main focus has centered on a component of a previously
unknown collar-like structure localized at the point of emergence the
flagellum from the cell. Using western blots from two-dimensional gels
and mass spectrometry we identified a 67kDa protein. The gene encoding
the 67kDa protein was cloned, histidine-tagged and the protein affinity
purified. Finally mouse polyclonals/monoclonal were raised against this
protein.
Using GFP and Immunofluorescence studies show that this protein is a
component of a collar-like structure located at the site where the flagellum
exits the cell. This collar is also a component of the flagellar pocket (FP).
The FP is the sole organelle responsible for endo and exocytosis in this
these cells (figure 1). We have named the 67kDa protein BILBO1. BILBO1
is essential for parasite survival.
Because BILBO1 is essential for cell survival we have an excellent
subject for further study of flagellar pocket biogenesis.
• Research team
Derrick ROBINSON DR2 CNRS.
Mélanie BONHIVERS CR1 CNRS.
Denis DACHEUX-DESCHAMPS
(MCU) BX2.
Benoit ROGER (MCU BX1).
Nicolas LANDREIN (TCN CNRS)
Elodie BERDANCE (PhD Student).
Dacheux D, Landrein N, Thonnus M, Gilbert G, Sahin A,
Wodrich H, Robinson DR, Bonhivers M. A MAP6-related
protein is present in protozoa and is involved in flagellum
motility. PLoS One. 2012;7(2):e31344
Joice AC, Lyda TL, Sayce AC, Verplaetse E, Morris MT,
Michels PA, Robinson DR, Morris JC. Extra-glycosomal
localisation of Trypanosoma brucei hexokinase 2. Int J
Parasitol. 2012 Apr;42(4):401-9.
May SF, Peacock L, Almeida Costa CI, Gibson WC, Tetley
L, Robinson DR, Hammarton TC. The Trypanosoma
brucei AIR9-like protein is cytoskeleton-associated and
is required for nucleus positioning and accurate cleavage
furrow placement. Mol Microbiol. 2012 Apr;84(1):77-92
Giroud C, Ottones F, Coustou V, Dacheux D, Biteau N,
Miezan B, Van Reet N, Carrington M, Doua F, Baltz T.
Murine Models for Trypanosoma brucei gambiense disease
progression from silent to chronic infections and early brain
tropism. PLoS Negl Trop Dis. 2009 Sep 1;3(9):e509
Absalon S, Blisnick T, Bonhivers M, Kohl L, Cayet N,
Toutirais G, Buisson J, Robinson D, Bastin P. Flagellum
elongation is required for correct structure, orientation and
function of the flagellar pocket in Trypanosoma brucei. J
Cell Sci. 2008 Nov 15;121(Pt 22):3704-16
Bonhivers, M., Nowacki, S., Landrein, N. and Robinson, D.
R. Biogenesis of the trypanosome endo-exocytotic organelle
is cytoskeleton mediated. PLoS PLoS Biol 6, e105.
Bonhivers M, Landrein N, Decossas M, Robinson DR.
A monoclonal antibody marker for the exclusion-zone
filaments of Trypanosoma brucei.Parasit Vectors. 2008 Jul
10;1(1):21.
Candida and Pathogenicity
(CNRS UMR 5234)
www.mcmp.aquitaine.cnrs.fr/mfp/team_pc_en.php
Thierry Noël
• Research team
Accoceberry Isabelle, MCU-PH
Albert Olivier, AHU
Dementhon Karine, MCF
Fitton-Ouhabi valérie, ADT
Gabriel Frédéric, PHC
Noël Thierry, PR, group leader
Sabra Ayman, Doctorant
Dementhon, K., El-Kirat-Chatel, S., and
Noel, T. (2012). Development of an in vitro
model for the multi-parametric quantification
of the cellular interactions between Candida
yeasts and phagocytes. PLoS ONE,
7(3):e32621. PMID 22479332
Noel, T. (2012). The cellular and molecular
defense mechanisms of the Candida yeasts
against aztole antifungal drugs. Journal of
Medical Mycology, 22: 173-178. ISSN 11565233, 10.1016/j.mycmed.2012.04.004
Gabriel, F., Noel, T., and Accoceberry, I.
(2011). Lindnera (Pichia) fabianii blood
infection after mesenteric ischemia. Medical
Mycology 50, 310-314. PMID 21671831.
El-Kirat-Chatel, S., Dementhon, K., and
Noel, T. (2011). A two-step cloning-free
PCR-based method for the deletion of
genes in the opportunistic pathogenic yeast
Candida lusitaniae. Yeast 28, 321-330.
PMID 21456057
Gabriel, F., Noel, T., and Accoceberry, I.
(2011). Fatal invasive trichosporonosis
due to Trichosporon loubieri in a patient
with T-lymphoblastic lymphoma. Medical
Mycology 49, 306-310. PMID 20950223
Patent
Gabriel F, Accoceberry I, Bessoule J-J,
Manon S, Noël T inventors; Université de
Bordeaux, Centre National de la Recherche
Scientifique (CNRS), Centre Hospitalier
Universitaire
(CHU)
de
Bordeaux,
assignee. Novel Beta-oxidation pathway
and hemiascomycetes yeast mutants.
European Patent EP11159920.5, (2011,
March 25).
28 │immunologY-microbiologY
Fungi of medical interest have become the first cause of opportunistic
parasitic diseases in industrialized countries, mainly because of the expanding
immunocompromised population. Our main research topic focuses on the role
of fungal lipid signalling and fungal lipid metabolism during the first stage of the
infectious process, when fungal cells meet the innate immune cells of the host,
macrophages and neutrophils. We are also interested by specific lipid biosynthetic
pathways, which are important targets for antifungals, and by lipid membrane
transporters which allow fungal cells to resist to antifungal therapy. Our team works
preferentially with non-albicans Candida species, notably Candida lusitaniae that
we developed as a model organism for several years, and which is now accessible
to both formal and reverse genetics.
BASIC RESEARCH
A – Cellular and molecular characterization of the interactions between
Candida and phagocytic cells
We developed a new in vitro model for a multi-parameter characterization of
interaction of Candida fungal cells with murine macrophages and human neutrophils
(Dementhon et al, 2012). This model is based on the use of combined microscopy,
fluorometry, flow cytometry and viability assays, and that of fluorochromes specific
to phagocytes and yeasts, and is amenable to the high throughput screening of
mutant strains.
This model allows us to address the molecular mechanisms involved in yeast
response to macrophages versus neutrophils, either by the screening of libraries
of random tagged mutants of Candida, or by inactivating target genes involved in
the biosynthesis of oxygenated lipids.
B – Role of the fungal lipid metabolism during the interaction Candida /
macrophage
We discovered novel pathways of fatty acid oxidation in the opportunistic pathogenic
ascomycetous yeast C. lusitaniae (Gabriel et al, 2011, European Patent). Two of
them are dependent on Fox2p, the multifunctional protein of the β-oxidation. The
third one is peroxisomal, fox2p-independent, and is unknown so far in Fungi. Our
aims are to characterize the peroxisomal fox2p-independent pathway of fatty acids
catabolism, and to define the role of the fatty acids catabolism in the virulence of
the Candida yeasts, notably when they survive to macrophages.
C - Molecular characterization of antifungal resistance in C. lusitaniae.
We demonstrated cross-resistance between flucytosine (5FC) and fluconazole
(FLC) in several genetically unrelated clinical isolates of C. lusitaniae. Our
data support the idea that 5FC behaves as a competitive inhibitor of the entry
of fluconazole, which has never been characterized so far in fungi. We are
investigating the membrane transporters involved in the cross-resistance between
5FC and FLC.
CLINICAL INVESTIGATIONS – TRANSFER OF TECHNOLOGY
Other domains of expertise in clinical and applied research :
• Molecular identification and description of new emergent and rare opportunistic
fungal species
• Suceptibility testing of yeast and moulds to antifungals and to natural and
synthetic molecules under development.
Macrophage infected with Candida cells.
Left: fluorescence panel showing a macrophage, with its membrane stained with APC-antiCD16 antibody (red) and its cyttoplasm with calcein (green), containing several calcofluorlabeled yeast cells (blue). Right: bright field.
Variability, replication and mobility
of viral and bacterial genomes
(CNRS UMR 5234)
www.mcmp.aquitaine.cnrs.fr/mfp/team_vrmg.php
Marie-Line ANDREOLA, Michel VENTURA, Hervé FLEURY
Our group is interested in the replication of human pathogens. Understanding of
such mechanisms must lead to the development of new therapeutic approaches.
Four principal projects are developed. The first project concerns the study of the
Human Immunodeficiency Virus (HIV) DNA synthesis and integration mediated by
the viral reverse transcriptase (RT) and integrase (IN). More recently, the study
of the RNA dependent RNA polymerase (RdRp) of the hepatitis C virus (HCV)
was developed as a new project of the laboratory. These studies allowed us to
better understand the mechanisms involved in replication of the viral genomes.
Furthermore the clinical studies performed in all around the world, mainly in South
East of Asia and Africa, brought numerous informations on the HIV-1 variability,
the impact on viral replication, and the resistance to therapeutic drugs. We
designed biochemical and cellular models for the study of the interactions between
enzymes involved in stabilization of the nucleic acids and/or interactions of the
viral pathogens with potential cellular partners. The recent data that we obtained
about the mobility of viral nucleic acids allowed us to analyze the mechanism of
enzymes mediating the mobility of bacterial genes involved in antibiotic resistance
(integrase/transposase). Our expertise in DNA polymerase characterization led us
to original contributions to the following fields:
• Reverse transcription initiation and HIV RT interactions with its substrates.
• RNA synthesis initiation by HCV RdRp.
• Processing and integration of HIV-1 DNA by viral integrase.
• Involvement of a bacterial integrase (Int1) of class 1 integron and of a transposase
(IS91 family) in the mobility of genes inducing resistance to antibiotics.
A better knowledge of these recombination/transposition mechanisms should lead
to design new therapeutic approaches against highly resistant bacterial strains.
Also we developed in collaboration with chemists and microbiologists new
specific inhibitors of HIV-1 RT and of HCV RdRp. A new antiviral strategy against
RNA viruses has also been established. Three international patents have been
approved related to these projects
HCV replication cycle.
• Research team
Chercheurs, Enseignants-chercheurs
Marie Line ANDREOLA (DR2)
Corinne ARPIN (MCU-HC)
Pantxika BELLECAVE (Post Doc)
Mohamed BENLEULMI (Post Doc)
Jean Luc BRUN (PU-PH)
Véronique DUBOIS (MCU-PH)
Michel DUPON (PU-PH)
Hervé FLEURY (PU-PH)
Isabelle GARRIGUE (MCU-PH)
Cyril MASANTE (Post Doc)
Didier NEAU (PU-PH)
Vincent PARISSI (CR1)
Claudine QUENTIN-NOURY ( PR1)
Jean Marie RAGNAUD ( PU)
Sandrine REIGADAS ( MEB)
Ludivine SINZELLE (Post Doc)
Françoise TESSIER (MCU)
Jean François VELY (PU-PH)
Michel VENTURA (DR2)
ITA :
Catherine ANDRE (ADT2P)
Lucie BESNARD (IE CDD)
Christina CALMELS (AI)
Laure COULANGE-MAYONNOVE (ADT1)
Jennifer PAPUCHON (Tce)
Patricia PINSON (IE)
Etudiants :
Anais JASPART (Doctorante)
Haytham YASSINE (Doctorant)
HIV-1 replication cycle (from Schaker, 2010)
Cosnefroy O, Tocco A, Lesbats P, Thierry S, Calmels C,
Wiktorowicz T, Reigadas S, Kwon Y, De Cian A, Desfarges S,
Bonot P, San Filippo J, Litvak S, Le Cam E, Rethwilm A, Fleury H,
Connell PP, Sung P, Delelis O, Andréola ML, Parissi V. Stimulation
of hRAD51 nucleofilament restricts HIV-1 integration in vitro and
in infected cells. J Virol. (2012) 86(1):513-26. Epub 2011 Oct 19
Reigadas S, Masquelier B, Calmels C, Laguerre M, Lazaro E,
Vandenhende M, Neau D, Fleury H, Andréola ML. Structure
analysis of HIV-1 IN Y143C/R raltegravir resistance mutation in
association with the secondary mutation T97. Antimicrob Agents
Chemother (2011) 55, 3187-94.
Lesbats P, Botbol Y, Chevereau G, Vaillant C, Calmels C, Arneodo
A, Andréola ML, Lavigne M and Parissi V (2011) Functional
coupling between HIV-1 integrase and the SWI/SNF chromatin
remodeling complex for efficient in vitro integration into stable
nucleosomes PLoS Pathogens, 7(2):e1001280.PMID:21347347
Reigadas S, G Anies, B Masquelier, C Calmels, LJ. Stuyver,
V Parissi, H Fleury, ML Andréola. (2010) The HIV-1 integrase
mutations Y143C/R are an alternative pathway for resistance
to raltegravir and impact the enzyme functions. PLoS ONE, Apr
26;5(4):e10311PMID: 20436677
Mahias K, Ahmed-El-Sayed N, Masante C, Bitard J, Staedel C,
Darfeuille F, Ventura M, Astier-Gin T. Therese Identification of
a structural element of the hepatitis C virus minus strand RNA
involved in the initiation of RNA synthesis. Nucleic Acids Res,
2010, Jul 1;38(12):4079-91..
Bitard J, Chognard G, Dumas E, Rumi J, Masante C, Mahias K,
Astier-Gin T, Ventura M. Hijacking hepatitis C viral replication with
a non-coding replicative RNA. Antiviral Res. 2010 Jul;87(1):9-15.
Pluripotency and early
steps of differentiation
(CNRS UMR 5164)
www.umr5164.u-bordeaux2.fr/
Hélène Bœuf
• Research team
Permanent members (alphabetical order)
Group leader : Bœuf Hélène (DR2, CNRS)
Boiron J. M. (PH-EFS)
Brunet de la Grange Philippe (CR-EFS)
Chevaleyre Jean (IR-EFS)
Conrad Véronique (MCF, UBS)
Faucheux Corinne (MCF, UBS)
Fédou Sandrine (Tech, UBS)
Fontenay Bijou (PH-EFS)
Gauthereau Xavier (AI, CNRS)
Ivanovic Zoran (PH-EFS)
Lafarge Xavier (PH-EFS)
Massé Karine (MCF, UBS)
Praloran Vincent (PU – PH)
Thézé Nadine (PU – UBS)
Thiébaud Pierre (CR1, CNRS)
Veschambre Philippe (MCF, UBS)
Villacreces Arnaud (IE, UBS)
Wlaski Marija (IR-EFS)
Non permanent members for the 20102012 period
Fabre Annabelle : ½ ATER position : 20112012
Mathieu ME: PhD student: 2009-2011
Peytour Yann: PhD student : 2009-2011
Peytour Yann (IR-Contractuel, UBS ; 20112012)
Tocco Alice: PhD student (2012-2014)
Our research program is focused on the study of the alternative mechanisms of
self-renewal and of early cell commitment in three models: mouse embryonic
stem cells (ES), mouse and human adult hematopoietic stem cells (HSC) and
Xenopus embryo.
In Mouse ES cell model (maintained pluripotent in vitro, in the presence of
LIF (Leukemia Inhibitory Factor)], we try to understand the mechanisms of
action of LIF and its targets on cell plasticity.
- We have set up a “reversion/ reprogrammation functional test” allowing
the characterization of genes and signaling pathways involved in LIFdependent plasticity of murine ES cells.
- We have investigated the conservation of function of new pluripotencylinked proteins in murine and Xenopus models and found conservation of
function of the Mras gene, a LIF-dependent Pluri gene that we previously
identified in mES cells (Trouillas 2009).
- We have initiated collaborative works to study the impact of “LIF
signatures” in self renewal properties of mES cells in parallel with cancer
stem cells (in glioma model) and iPSC (induced pluripotent Stem Cells).
Xenopus team’s work is centered on the study of cellular and molecular
mechanisms of commitment and differentiation of embryonic cells in Xenopus
laevis model. We are following several leads: the TEAD/ VESTIGIAL protein
complexes during development; the LIF signaling pathway in Xenopus
embryos; the expression profiles and functions of ligands and receptors of the
purinergic pathway during embryonic development of Xenopus.
Adult hematopoetic stem cells (HSC), are used in cellular therapy to cure
hematological are used in cellular therapy to cure hematological malignancies.
We showed that hypoxia favors the maintenance and quiescence of HSC.
We will pursue on the role of physiological hypoxia on hematopoiesis by
investigating quiescence and self renewal mechanisms in normal and
leukemic HSC and will develop a novel strategy to establish iPS (induced
Pluripotent stem cells) starting with human CD34+ hematopoietic stem cells.
Duchez, P., Chevaleyre, J, Brunet de la Grange, P., Vlaski, M.,
Boiron, J.M. and Ivanovic, Z.(2012): Functional stability (at +4°c)
of hematopoietic stem and progenitor cells amplified ex vivo from
cord blood CD34+ cells. Cell Transplant, In press.
Duchez P, Chevaleyre J, Vlaski M, Dazey B, Milpied N, Boiron JM,
Ivanovic Z. (2012): Definitive set-up of clinical-scale procedure
for ex-vivo expansion of cord blood hematopoietic cells for
transplantation. Cell Transplant. 21(11):2517-21
Massé K, Dale N. (2012): Purines as potential morphogens
during embryonic development. Purinergic Signal. 8(3):503-21.
Gan Q, Thiébaud P, Thézé N, Jin L, Xu G, Grant P, Owens GK.
(2011). WD repeat-containing protein 5, a ubiquitously expressed
histone methyltransferase adaptor protein, regulates smooth
muscle cell-selective gene activation through interaction with
pituitary homeobox 2. J Biol Chem. 286(24):21853-64.
Guitart AV, Debeissat C, Hermitte F, Villacreces A, Ivanovic Z,
Boeuf H, Praloran V. (2011) Very low oxygen concentration
(0.1%) reveals two FDCP-Mix cell subpopulations that differ by
their cell cycling, differentiation and p27KIP1 expression. Cell
Death Differ. Jan;18(1):174-8.
30 │Immunology-microbiology
Ivanovic Z, Duchez P, Chevaleyre J, Vlaski M, Lafarge X, Dazey
B, Robert-Richard E, Mazurier F, Boiron JM. (2011) : Clinicalscale cultures of cord blood CD34(+) cells to amplify committed
progenitors and maintain stem cell activity. Cell Transplant.
20(9):1453-63.
Faucheux C, Naye F, Tréguer K, Fédou S, Thiébaud P, Théze
N. (2010). Vestigial like gene family expression in Xenopus:
common and divergent features with other vertebrates. Int J Dev
Biol. 54(8-9):1375-82.
Mathieu M.E., Saucourt C., Mournetas V., Gauthereau X., Thézé
N., Praloran V., Thiébaud P and H. Bœuf (2012) : LIF-dependent
signaling: new pieces in the Lego. Stem cell Reviews and
Reports, Mar;8(1):1-15.
Tréguer K, Faucheux C, Veschambre P, Fédou S, Thézé N,
Thiébaud P (2013) : Comparative functional analysis of ZFP36
genes during Xenopus development. PLos One, 8(1):e54550.
Mathieu, ME, Saucourt, C., Soulet, F., Gauthereau, X., Fedou,
S., Thézé, N., Thiébaud, P. and H. Boeuf (2013): Mras GTPase
is a novel stemness marker with impact on mouse embryonic
stem cell plasticity and on Xenopus embryonic cell fate. In press,
Development.
Activation of human T
lymphocytes (CNRS UMR 5164)
www.umr5164.u-bordeaux2.fr/pages/jdm-presentation.html
Julie Déchanet-Merville
Modulation of T cell activation is a general concern in the prevention or
cure of various diseases including cancer, infections and autoimmune
disorders. The main objective of the team is to understand the basic
mechanisms leading to T lymphocyte activation, regulation and effector
functions, with specific regards to gamma-delta T cells. Three main topics
are investigated: (i) identifying the antigens recognized by gamma-delta
TCRs, (ii) understanding the role of gamma-delta T cells in the response
to infectious pathogens (plasmodium falciparum and cytomegalovirus)
and (iii) characterizing the mechanisms leading to the activation of T
lymphocytes in auto-immune diseases. We combine in vitro and in vivo
approaches using both patient cohorts and animal models to progress in
the understanding of basic biology of T lymphocytes and provide proof of
concept that manipulation of T lymphocytes may impact disease outcome.
This is made possible through the tight links that have been established
between the basic research developed by our unit and the clinical activities
in several Bordeaux University hospital departments, which represent one
hallmark of our research.
• Research team
Team leader
Déchanet-Merville Julie
Heads of groups
Behr Charlotte
Blanco Patrick
Scientists
Capone Myriam
Contin-Bordes Cécile
Couzi Lionel
Duffau Pierre
Mamani-Matsuda Maria
Merville Pierre
Richez Christophe
Lazaro Estibaliz
Taupin Jean-Luc
Research assistants
Daburon Sophie
Douchet Isabelle
Gonzales Santi
Loizon Séverine
Netzer Sonia
Pitard Vincent
Post-doctorants
Faustin Benjamin
Furman David
Marlin Romain
PhD Students
Bachelet Thomas
Howard Jenny
Jacquemin Clément
Khairallah Camille
Khoryati Liliane
Willcox C*, Pitard V*, Netzer S, Couzi L, Salim M, Silberzahn
T, Moreau JF, Hayday A, Willcox B*, and Déchanet-Merville J*.
CMV and tumor stress-surveillance by human gamma-delta T
cell receptor binding to Endothelial Protein C Receptor. *equal
contributors. Nature Immunology (2012) 13: 872-879.
Saint-Basile, V Pitard, J Déchanet-Merville, JF Moreau, M
Troye-Blomberg, O Mercereau-Puijalon, C Behr. Control of
Plasmodium falciparum erythrocytic cycle: gamma-delta T cells
target the red blood cell-invasive merozoites. Blood (2011)
118:6952-6962
Couzi L, Pitard V, Sicard, X, Garrigue I, Hawchar O, Merville
P, JF Moreau and J Déchanet-Merville. Antibody-dependent
anti-cytomegalovirus activity of human gamma-delta T cells
expressing CD16 (FcgammaRIIIa). Blood (2012) 119:1418-27
Duffau P, Seneschal J, Nicco C, Richez C, Lazaro E, Douchet
I, Bordes C, Viallard JF, Goulvestre C, Pellegrin JL, Weill B,
Moreau JF, Batteux F and Blanco P. Platelet CD154 potentiates
interferon-alpha secretion by plamacytoid dendritic cells
in systemic lupus erythematosus. Science Translational
Medicine. (2010) 2:47-56.
G Costa, S Loizon, M Guenot, I Mocan, F Halary, G de
Biochemistry
genetics
Energy transduction
systems and mitochondrial
morphology (SysTEMM)
(IGBC CNRS UMR 5095)
www.ibgc.cnrs.fr/?page=equipe&eq=systemm
Daniel Brèthes
ATP is the universal fuel molecule of any cell. The F1Fo-ATP synthase is a
key enzyme of the energetic metabolism since it is responsible for most of the
cellular synthesis of ATP. This 600 kDa complex is composed in yeast by 17
distinct subunits. It uses the energy of an electrochemical proton gradient to
synthesize ATP. When protons are conducted across its membrane region,
this conduction drives the rotation of a rotor part. This rotor part protrudes in
the hydrophilic catalytic sector and induces conformational changes that lead
to ATP synthesis.
The main project of the team concerns the structural
characterisation of the yeast F1-Fo ATP synthase. We
have recently obtained the first refined structure of the
yeast F1-c10 sub-complex at 3,4 Å resolution (figure). This
work brought new information on the catalytic head, on the
central stalk an on the structure of the membrane rotor.
However, whatever the crystallization conditions or the
inhibited state of the enzyme, the peripheral stalk is always
lost during the crystallization process. We are developing
new techniques of crystallisation in lipidic sponge phase
in order to obtain the structure of the entire complex and
get information on the proton channel domain. We have
also settled collaboration with a team from the CBMN in
Bordeaux to determine the structure and the dynamics of interaction of the
small membrane subunits involved in the dimerization/oligomerization process
by NMR. By cryo-electron microscopy and single particles analysis, we have
obtained the structure at low resolution (30 Å) of the dimer of the ATP synthase
solubilised in mild detergent. The localisation of the interfacial subunits at the
dimerization interface will be done by cryo-EM using functionalized nanogold
particles. The membrane deformation properties of this complex due to its
oligomerization are also studied by reconstitution experiments and electron
microscopy tomography.
The yeast enzyme, like mammalian ATP synthases, is not only involved in ATP
synthesis but also in the organization of the inner mitochondrial membrane:
ATP synthase dimers constitute the building blocks of large oligomers that are
involved in mitochondrial cristae morphology. A second project of the team is
focused on the study of the dynamics and the regulation of the oligomerization
The mitochondrial ultrastructure is normal in control HeLa cells (A) and altered in cells
depleted in dimerization subunits (B, C). bar = 0.5 µm
of this complex in relation with other mitochondrial processes such as fusion,
fission or metabolic modifications in yeast and in human cells. In yeast the
two dimerization subunits are not required for ATP synthase assembly and
activity but are strictly necessary for the enzyme oligomerization. Recently, we
demonstrate that the down-regulation of the mammalian homologues of these
subunits by a shRNA strategy affects the stability of ATP synthase with a 50%
decrease of the available functional enzyme and a destabilization of the ATP
synthase oligomers, correlated with the fission of the mitochondrial network
and the disorganization of mitochondrial ultrastructure.
• Research team
Daniel Brèthes, DR2 CNRS
Alain Dautant, CR1 CNRS
Marie-France Giraud, CR1 CNRS
Isabelle Larrieu, IE2 CNRS
Guy Lauquin, PR0 UBS
Patrick Paumard, MCU UBS
Corinne Sanchez, MCU UBS
Nadir Seddiki, CDD ANR
Collaboration within IBGC
Bénédicte Salin, IR2 CNRS
Habersetzer J, Ziani W, Larrieu I, StinesChaumeil C, Giraud M-F, Brèthes D,
Dautant A and Paumard P. (2013) ATP
synthase oligomerization: From the
enzyme models to the mitochondrial
morphology. Int. J. Biochem. Cell. Biol.
45, 99-105.
Giraud MF, Paumard P, Sanchez C,
Brèthes D, Velours J and Dautant A.
(2012) Rotor architecture in the yeast
and bovine F1-c-ring complexes of
F-ATP synthase. J. Struct. Biol. 177,
490-497.
Velours
J,
Stines-Chaumeil
C,
Habersetzer J, Chaignepain S, Dautant
A and Brèthes D. (2011) Evidence
of the proximity of ATP synthase
subunits 6 (a) in the inner mitochondrial
membrane and in the supramolecular
forms of Saccharomyces cerevisiae
ATP synthase. J. Biol. Chem. 286,
35477-35484.
Dautant A, Velours J, Giraud MF.
(2010) Crystal structure of the Mg·ADPinhibited state of the yeast F1c10-ATP
synthase. J. Biol. Chem. 285, 2950229510.
Talbot J-C, Dautant A, Polidori A, Pucci
B, Cohen-Bouhacina T, Maali A, Salin
B, Brèthes D, Velours J and Giraud M-F.
(2009) Hydrogenated and fluorinated
surfactants derived from Tris(hydroxymethyl)-acrylamidomethane allow the
purification of a highly active yeast F1F0 ATP-synthase with an enhanced
stability. J. Bioenerg. Biomembr. 41,
349-360.
Biochemistry - Genetics │35
Functional analysis
of amyloids (IGBC CNRS UMR 5095)
www.ibgc.cnrs.fr/?page=equipe&eq=hsp
Christophe Cullin
• Research team
Christophe Cullin, Professeur UB2
Loan Lascu, Professeur
Christelle Marchal, MdC UB2
Claude Bobo, TCS CNRS
Hélène Vignaud , Doctorante MRT
Our goal is to understand the molecular characteristics of toxic amyloids.
To answer this challenging question, we have taken advantage of
toxic mutants heterologously expressed in yeast and combine various
approaches from genetics to biophysics. In previous studies, we have
isolated and characterized at a cellular and molecular level the structural
particularities of a toxic amyloid mutant of Het-s, a prion protein from
P.anserina. We are currently working with another amyloid protein related
to Alzheimer disease (AD).
We characterize at the molecular level different Aß variants responsible
for familial Aß and/or selected for their high toxicity in the yeast model. We
will use this model to isolate drugs active against Aß toxicity and analyze
in vitro their interaction with Aß. We have put in place an efficient platform
allowing the production of pure Aß and propose to study both the aggregates
formed but also to characterize the intermediates of amyloidogenesis.
These studies will include “classical” approaches such as Tht binding,
but also various biophysical techniques including FTIR, CryoTEM and
NMR. The toxicity of amyloid-forming proteins will be correlated with their
interactions with lipid membrane during the aggregation. We will follow
this interaction by different approaches (plasmon waveguide resonance,
polarization-modulated infrared reflection absorption spectroscopy
(PMIRRAS), ellipsometry, cryoTEM…) in a frame of a collaboration with
the team of S. Lecomte (CBMN, Pessac).
The comparison of biophysical characteristics of the different Aß variants
should help us to highlight the molecular properties of Aß toxicity.
1- Ta, H. P., Berthelot, K., Coulary-Salin, B., Castano, S.,
Desbat, B., Bonnafous, P., Lambert, O., Alves, I., Cullin,
C. and Lecomte, S. (2012) A yeast toxic mutant of HET-s
amyloid disrupts membrane integrity. Biochim Biophys
Acta
2- D’Angelo, F., Vignaud, H., Di Martino, K., Salin, B.,
Devin, A., Cullin, C. and Marchal, C. (2012) A yeast model
for Aß aggregation exemplifies the role of membrane
trafficking and PICALM in cytotoxicity. Disease Models &
Mechanisms In press
3- Millot, G. A., Berger, A., Lejour, V., Boulé, J. B., Bobo,
C., Cullin, C., Lopes, J., Stoppa-Lyonnet, D. and Nicolas,
A. (2011) Assessment of human Nter and Cter BRCA1
mutations using growth and localization assays in yeast.
Hum Mutat
36 │Biochemistry - Genetics
4-Berthelot, K., Ta, H. P., Géan, J., Lecomte, S. and Cullin,
C. (2011) InVivo and In Vitro Analyses of Toxic Mutants of
HET-s: FTIR Antiparallel Signature Correlates with Amyloid
Toxicity. J Mol Biol 412, 137-152
5- Ta, H. P., Berthelot, K., Coulary-Salin, B., Desbat, B.,
Géan, J., Servant, L., Cullin, C. and Lecomte, S. (2011)
Comparative studies of nontoxic and toxic amyloids
interacting with membrane models at the air-water
interface. Langmuir 27, 4797-4807
6- Berthelot, K., Lecomte, S., Géan, J., Immel, F. and Cullin,
C. (2010) A yeast toxic mutant of HET-s((218-289)) prion
displays alternative intermediates of amyloidogenesis.
Biophys J 99, 1239-1246
7- Couthouis, J., Marchal, C., D’Angelo, F., Berthelot, K.
and Cullin, C. (2010) The toxicity of an «artificial» amyloid
is related to how it interacts with membranes. Prion 4, 283-291
Genetics of Metabolic
Networks (IGBC CNRS UMR 5095)
www.ibgc.cnrs.fr/?page=equipe&eq=grm
• Research team
Cell size, a complex trait at the crossroad between metabolism and
the cell cycle
Cell size can vary more than fifty folds among cell types and organisms,
however for a given cell type, cell size is noticeably constant. Cell
size homeostasis implies that specific mechanisms are devoted to
estimating cell size and coordinating growth (increase in cell volume) and
proliferation (increase in cell number). Although studied for more than 50
years, the mechanisms that regulate cell size in eukaryotes remain largely
mysterious.
Our project is to decipher the genetic control of cell size in yeast.Our aim
is to identify the different pathways involved in cell size control. Cell size
is a highly complex trait affected by hundreds of genes. Therefore, the
global understanding of cell size control cannot be achieved by studying
individual mutations. We use a systems biology approach to get an
integrated picture of the phenomenon. The first pathway that we have
identified is the Sir2-60S pathway (Figure below) that includes more than
30 mutants
Identification of the targets of AICAR, a compound that selectively
kills aneuploid cells
This project is aimed at understanding how the highly promising drug AICAR
is acting in vivo specifically on aneuploid cells. Using yeast, nematode
and mammalian cells, we perform target deconvolution and validation by
combining genetics and affinity chromatography approaches. We have
identified several proteins that are affected by AICAR in vivo in yeast. We
found a strong conservation among the yeast and mouse AICAR-binders,
thus establishing the broad impact of this approach to predict relevant
pharmacological networks.
Benoît Pinson - CR1 CNRS
Chloé Philippe – Master 2
Christelle Saint-Marc - AI CNRS
Delphine Albrecht – Master 2
Jérémy Tissot-Dupont –AI ANR
Johanna Ceschin – ASI Bdx2
José E. Gomes _ MCF Bdx2
Michel Moenner – PR1 Bdx1
Stéphane Puyo – Post-Doc Ligue
Hurlimann HC, Laloo B, Simon-Kayser
B, Saint-Marc C, Coulpier F, Lemoine
S, Daignan-Fornier B, Pinson B.
(2011) Physiological and toxic effects
of the purine intermediate 5-Amino-4ImidazoleCarboxAmide Ribonucleotide
(AICAR) in yeast. J Biol Chem.. 286:
30994-31002.
Laporte D, Lebaudy A, Sahin A, Pinson
B, Ceschin J, Daignan-Fornier B, Sagot
I. (2011) Metabolic status rather than
cell cycle signals control quiescence
entry and exit. J Cell Biol. 192(6):949-57.
Saint-Marc C, Pinson B, Coulpier F,
Jourdren L, Lisova O, Daignan-Fornier
B. (2009) Phenotypic consequences
of purine nucleotide imbalance in
Saccharomyces cerevisiae. Genetics.
183(2):529-38,
Pinson B, Vaur S, Sagot I, Coulpier
F, Lemoine S, Daignan-Fornier B.
(2009)
Metabolic
intermediates
selectively stimulate transcription factor
interaction and modulate phosphate
and purine pathways. Genes Dev. 2009
23(12):1399-407.
Cellular energy metabolism
http://www.ibgc.u-bordeaux2.fr/?page=equipe&eq=mec
Anne Devin
The aim of our studies is to better define the role of mitochondria in the
structuration and regulation of the cellular energy metabolism as well as in a
number of vital process, either normal or pathological: stress adaptation, cell
proliferation.
• Research team
Nicole Avéret - MCU UBS
Anne Devin - CR1 CNRS
Audrey Ladonne - CDD AI INSERM
Jean-Pierre Mazat - Professeur émérite UBS
Stéphane Ransac - MCU UBS
Michel Rigoulet - Professeur UBS
Christophe Rocher - IGE UBS
Edgar Yoboue - Post-Doc
1. Physiological adaptations, energy metabolism and mitochondrial
biogenesis in the yeast levure Saccharomyces cerevisiae
a – Control and regulation of mitochondrial biogenesis
Mitochondrial biogenesis requires both the nuclear and mitochondrial genomes,
and several major enzymatic complexes (respiratory chain, ATPsynthase)
are made of proteins coded by one or the other genome. We aim at better
understanding the relationship between the number, the composition and the
activity of mitochondria and the cellular energy requirements. We specifically
try to better understand the role of the Ras/AMPc pathway and of reactive
oxygen species.
b - Crabtree and Warburg effects
The fate and proliferation of cells are closely linked to their metabolic state and
especially to their energy metabolism. More than 50 years ago, Otto Warburg
showed that the ability of cancer cells to proliferate was directly linked to the
degree of repression of the mitochondrial metabolism associated with and
increase in the glycolytic flux. The more the mitochondrial metabolism is
repressed and glycolysis activated, the more cancer cells can proliferate. We
are working on the molecular mechanisms involved in this process and on
their reversibility.
2. Mechanisms of regulation of phosphorylative oxidations: organization
and regulation of the pathways of reoxydation of cytosolic NADH
The oxidative metabolism in S. cerevisiae is an example of integrated system:
NADH produced during glycolysis or cytosolic oxidation of ethanol is brought
to NADH dehydrogenase located on the external side of the internal de la
membrane. NADH dehydrogenase activity leads to the inhibition of glycerol3-P-déshydrogénase and thus of the glycerol-3-P shuttle. There is evidence
that the 2 NADH dehydrogenases (Nde1p, Nde2p) do not regulate glycerol3-Pdehydrogenase similarly. In addition, variation in the supramolecular
organization of the respiratory chain, that may itself depend on the provided
substrates, may affect its ability to reoxidize either NADH or FADH2. We are
investigating the relationship between the provided substrates, organization
of the respiratory chain, and ROS production.
Baret P, Septembre-Malaterre A, Rigoulet M, Lefebvre
d’Hellencourt C, Priault M, Gonthier MP, Devin A. Dietary
polyphenols preconditioning protects 3T3-L1 preadipocytes
from mitochondrial alterations induced by oxidative stress.
Int J Biochem Cell Biol. 2013 Jan;45(1):167-74
Yoboue ED, Devin A. Reactive oxygen species-mediated
control of mitochondrial biogenesis. Int J Cell Biol.
2012;2012:403870
Casteilla L, Devin A, Salin B, Averet N, Rigoulet M. UCP1
as a water/proton co-transporter. Mitochondrion. 2012
Jul;12(4):480-1
Yoboue ED, Augier E, Galinier A, Blancard C, Pinson
B, Casteilla L, Rigoulet M, Devin A. cAMP-induced
mitochondrial compartment biogenesis: role of glutathione
redox state. J Biol Chem. 2012 Apr 27;287(18):14569-78
Carneiro L, Allard C, Guissard C, Fioramonti X, TourrelCuzin C, Bailbé D, Barreau C, Offer G, Nédelec E, Salin
B, Rigoulet M, Belenguer P, Pénicaud L, Leloup C.
Importance of mitochondrial dynamin-related protein 1 in
hypothalamic glucose sensitivity in rats. Antioxid Redox
Signal. 2012 Aug 1;17(3):433-44
Casteilla L, Devin A, Carriere A, Salin B, Schaeffer
J, Rigoulet M. Control of mitochondrial volume by
mitochondrial metabolic water. Mitochondrion. 2011
Nov;11(6):862-6
Diaz-Ruiz R, Rigoulet M, Devin A. The Warburg and
Crabtree effects: On the origin of cancer cell energy
metabolism and of yeast glucose repression. Biochim
Biophys Acta. 2011 Jun;1807(6):568-76
Rigoulet M, Yoboue ED, Devin A. Mitochondrial ROS
generation and its regulation: mechanisms involved in
H(2)O(2) signaling. Antioxid Redox Signal. 2011 Feb
1;14(3):459-68
Molecular genetics of
mitochondrial systems
http://www.ibgc.u-bordeaux2.fr/?page=equipe&eq=gmsm
Jean-Paul di Rago
ATP synthase is a major enzymatic system of mitochondria, responsible for
the last steps in oxidative phosphorylation. It is made of about 20 polypeptide
subunits with a double genetic origin, nuclear and mitochondrial. We are
interested in the mechanisms involved in the formation of this complex in
the baker yeast Saccharomyces cerevisiae and in Human. To this aim, we
have set up genetic and biochemical strategies in order to identify new
factors involved in the biogenesis of this system.
The cell mitochondrial content is clearly tightly regulated, but the regulatory
mechanisms are poorly understood. This is an important issue, not only
for basic research, but also in diseases since a change in the number of
mitochondria is observed in many diseases (mitochondrial dysfunctions
and cancer). Our approach is based on a collection of yeast ATP synthase
mutants that show a huge increase in the number of mitochondrias.
• Research team
E. Tetaud, CR1 CNRS
S. Duvezin-Caubet, CR1 CNRS
J.-P. Lasserre, MCU Bxd2
J.-P. di Rago, DR2 CNRS
F. Godard, TCE CNRS
A. Martos, Doctorant ANR
A. Kabala, Thèse co-tutelle
F. Courtin, AI FRM
Mitochondrial genomes are supposed to be derived from an ancestral
prokaryotic genome which size was considerably reduced during
evolution, notably because of a massive gene transfer towards the
nucleus. Why a mitochondrial genome is maintained after close to two
billion years of evolution is ill understood. We use the yeast model to
understand the adaptations that are required for a nuclear relocalization
of the mitochondrial genes of ATP synthase.
Finally, we have generated yeast models for human diseases associated
with a defect in ATP synthase, and in collaboration with Marc Blondel
(Brest), we have used these models to screen for drugs that could help
the research for treatments against these diseases. We apply the same
strategy to other diseases that can be modelized in yeast.
Kucharczyk R., Giraud M.-F., Brèthes D., WysockaKapcinska M., Ezkurdia N., Salin B., Velours J.,
Camougrand N, Haraux F. and Jean-Paul di Rago.
Defining the pathogenesis of human mtDNA mutations
using a yeast model : the case of T8851C. Int J Biochem
Cell Biology, 2013 Jan;45(1):130-40
Bietenhader M., Martos A., Tetaud E., Aiyar RS, Sellem
C.H., Kucharczyk R., Clauder-Münster S., Giraud M.F., Godard F., Salin B., Sagot I., Gagneur J., DéquardChablat M., Contamine V., Hermann-Le Denmat S.,
Sainsard-Chanet A., Steinmetz L.M. and di Rago J.-P.
Experimental Relocation of the Mitochondrial ATP9 Gene
to the Nucleus Reveals Forces Underlying Mitochondrial
Genome Evolution. Plos Genetics, 2012;8(8):e1002876
Couplan E, Aiyar RS, Kucharczyk R, Kabala A, Ezkurdia
N, Gagneur J, St Onge RP, Salin B, Soubigou F, Le
Cann M, Steinmetz LM, di Rago JP, Blondel M. A yeastbased assay identifies drugs active against human
mitochondrial disorders. Proc Natl Acad Sci U S A. 2011
Jul 19;108(29):11989-94
Godard F, Tetaud E, Duvezin-Caubet S, di Rago JP.
A genetic screen targeted on the FO component of
mitochondrial ATP synthase in Saccharomyces cerevisiae.
J Biol Chem. 2011 May 20;286(20):18181-9
Déquard-Chablat M, Sellem CH, Golik P, Bidard F,
Martos A, Bietenhader M, di Rago JP, Sainsard-Chanet
A, Hermann-Le Denmat S, Contamine V. Two nuclear life
cycle-regulated genes encode interchangeable subunits c
of mitochondrial ATP synthase in Podospora anserina. Mol
Biol Evol. 2011 Jul;28(7):2063-75
Chromosome Segregation
www.ibgc.cnrs.fr/?page=equipe&eq=mmtc
Jean-Paul Javerzat
team
• Research
Sonia Dheur CR1 CNRS
Sabine Vaur CR2 CNRS
Stéphanie Vazquez T CNRS
Jean-Paul Javerzat DR CNRS
Vaur S, Feytout A, Vazquez S, Javerzat
JP. Pds5 promotes cohesin acetylation
and
stable
cohesin-chromosome
interaction. EMBO Rep. 2012 Jun
29;13(7):645-52.
Feytout A, Vaur S, Genier S, Vazquez
S, Javerzat JP. Psm3 acetylation
on conserved lysine residues is
dispensable for viability in fission
yeast but contributes to Eso1mediated sister chromatid cohesion by
antagonizing Wpl1. Mol Cell Biol. 2011
Apr;31(8):1771-86.
Dheur S, Saupe SJ, Genier S, Vazquez
S, Javerzat JP. Role for cohesin in the
formation of a heterochromatic domain
at fission yeast subtelomeres. Mol Cell
Biol. 2011 Mar;31(5):1088-97.
Javerzat JP. Molecular biology.
Directing the centromere guardian.
Science. 2010 Jan 8;327(5962):150-1.
Bernard P, Drogat J, Dheur S, Genier
S, Javerzat JP. Splicing factor Spf30
assists
exosome-mediated
gene
silencing in fission yeast. Mol Cell Biol.
2010 Mar;30(5):1145-57
Cell division is a common feature of all live forms by which new individuals
are produced from their progenitors. During this process each cell
duplicates its genome and transmits a copy to daughter cells during
mitosis. At that time, the mitotic spindle must “know” which chromosomes
are duplicates so as to segregate them from each other. The linkage of
duplicated chromosomes by the protein complex cohesin enables this to
occur by creating physically associated pairs. These chromosome pairs
(called sister-chromatids) attach to microtubules of the mitotic spindle
through a kinetochore protein complex that forms on the centromeric
region of each chromatid. When all chromosome pairs are attached,
cohesin is cleaved by the enzyme separase, triggering the sudden
disjunction of sister chromatids at the metaphase to anaphase transition.
and their movement to opposite spindle poles.
Cohesin is an essential for chromosome segregation. Loss of function is
lethal and subtle alterations lead to errors in chromosome segregation,
generating cells with an abnormal number of chromosomes (aneuploids)
which have been linked to cancer progression, infertility, and debilitating
genetic diseases such as Down syndrome.
Our main focus is to understand how sister-chromatid cohesion is created
during S phase and maintained until the time of chromosome segregation.
Cohesion maintenance is particularly striking as it can last from a few
hours in actively dividing cells and up to many years as in human oocytes !
Besides its mitotic functions cohesin is involved in the control of gene
expression, presumably by promoting higher order chromosome
structures. In this regard, we investigate the relationship between cohesin,
the heterochromatic domains of chromosomes and gene regulation.
Experiments are carried out in the fission yeast Schizosaccharomyces
pombe, a user-friendly model organism for which all genetic, biochemical
and cytological techniques are available.
Mitochondria, stress
and cell death
www.ibgc.cnrs.fr/?page=equipe&eq=msmc
Stéphen Manon
• Research team
Our research aims at understanding the molecular mechanisms underlying
the role of mitochondria and the Bcl-2 family in apoptosis and mitophagy.
This research is organized along three main themes:
- Identification of the molecular mechanisms regulating the interaction
of pro-apoptotic protein Bax with the outer mitochondrial membrane;
- Regulation of autophagy by the anti-apoptotic protein Bcl-xL;
- Regulation of mitophagy by alterations of the mitochondrial functions.
Recent important results include:
- We have demonstrated that anti-apoptotic protein are able to “prime”
pro-apoptotic proteins, a phenomenon that maybe of crucial importance
in many cancer cells.
- We have provided evidences that functionnal alterations of the
mitochondrial respiratory chain are a signal that triggers autophagy, in
a way dependent on mitochondrial components and mitophagy actors.
- We have unveiled how partial deamidation of Bcl-xL bestows cancer
cells with additional survival advantage by stimulating their autophagic
activity. We have also shown that Bcl-xL controls cancer cells ability to
obtain food from their environment by interfering with the degradation
of plasma membrane nutrient transporters.
Axel Athané, AI-CDD UBS
Nadine Camougrand, DR2 CNRS
David Garenne, M2 student
Cécile Gonzalez, MCf UBS
Stéphen Manon, CR1 CNRS
Bruno Manesse, M2 student
Muriel Priault, CR1 CNRS
Lilit Simonyan, IE-CDD ANR
Dario Trindade, PhD student,
Fundaçao Gulbenkian
Deffieu M, Bhatia-Kissová I, Salin B, Klionsky DJ, Pinson
B, Manon S, Camougrand N (2013) Increased cytochrome
b reduction and mitophagy components are required
to trigger nonspecific autophagy following induced
mitochondrial dysfunction. J Cell Sci., in press
Drullion C, Lagarde V, Gioia R, Legembre P, Priault M,
Cardinaud B, Lippert E, Mahon FX, Pasquet JM (2013)
Mycophenolic Acid overcomes imatinib and nilotinib
resistance of chronic myeloid leukemia cells by apoptosis
or a senescent-like cell cycle arrest. Leuk Res Treatment,
in press
Bhatia-Kiššová I, Camougrand N (2013) Mitophagy: a
process that adapts to the cell physiology. Int J Biochem
Cell Biol 45, 30-33
Renault TT, Teijido O, Antonsson B, Dejean LM, Manon S
(2013) Regulation of Bax mitochondrial localization by Bcl2 and Bcl-x(L): keep your friends close but your enemies
closer. Int J Biochem Cell Biol. 45, 64-67
Renault TT, Grandier-Vazeille X, Arokium H, Velours G,
Camougrand N, Priault M, Teijido O, Dejean LM, Manon S
(2012) The cytosolic domain of human Tom22 modulates
human Bax mitochondrial translocation and conformation
in yeast. FEBS Lett. 586, 116-121
Trocoli A, Mathieu J, Priault M, Reiffers J, Souquère S,
Pierron G, Besançon F, Djavaheri-Mergny M (2011)
ATRA-induced upregulation of Beclin 1 prolongs the life
span of differentiated acute promyelocytic leukemia cells.
Autophagy 7, 1108-1114
Cell Growth and Division
http://www.iecb.u-bordeaux.fr/teams/MCCUSKER/McCuskerlab/
Welcome.html
Derek McCusker
• Research team
Derek McCusker - CR1 CNRS
Mini Jose-Deepak - FRM Fellow
Aurelie Massoni-Laporte - AI CNRS
Romain Mitteau - UB2
Sylvain Tollis - ARC Fellow
Cells grow and divide in a controlled fashion via a regulated series of
events termed the cell cycle. Extensive structural rearrangements occur
during the cell cycle that facilitate the efficient partitioning of the duplicated
genome into a daughter cell. We study how cell cycle cues elicit these
structural transitions at the molecular and cell biological level using high
resolution imaging, biochemistry and mathematical modelling. In doing so,
our aim is to understand how cell growth and cell division are coordinated,
and how failure of these mechanisms contributes to cancer.
Jose M, Tollis S, Nair D, Sibarita
JB and McCusker D. 2013. Robust
polarity establishment occurs via an
endocytosis-based cortical corralling
mechanism. J Cell Biol. 200(4):407418.
Kellogg D*, Royou A, Velours C,
McCusker D*. 2012. Cdk1-dependent
control of membrane trafficking
dynamics. Mol Biol Cell 3336-3347. (*
equal contribution).
McCusker D and Kellogg D. 2012.
Membrane growth during the cell
cycle: Unsolved mysteries and recent
progress. Current Opinion in Cell
Biology. 24: 845-851.
Royou A, McCusker D, Kellogg DR,
Sullivan W. 2008. Distinct pools of
Cdk1 independently drive cytoplasmic
and nuclear mitotic cycles. J Cell Biol.
183(1):63-75.
McCusker D, Denison C, Anderson S,
Egelhofer TA, Yates JR 3rd, Gygi SP,
Kellogg DR. 2007. Cdk1 Coordinates
cell surface growth with the cell cycle.
Nat Cell Biol. 9(5):506-15.
Figure 1. Mathematical modelling and evanescent-field imaging illustrating
the establishment of cell polarity. The cells on the left are non-polarized,
wheras those on the right have established a polarity axis that is used for
the ensuing cell cycle. Results of an in silico mathematical model show
Rho GTPase activity in the top scheme while the bottom scheme shows in
vivo data. Endocytic vesicles are shown in red while exoctic vesicles are
shown in cyan.
Figure 2. Individual Rho GTPase GAP domains (left) display a similar
structure that is revealed when models of their structures are overlaid
(right). The catalytic residue of the GAPs are shown in red while conserved
phosphorylation sites are shown in grey. Phosphorylation of these residues
contributes to the establishment of cell polarity shown in Figure 1.
Mitochondrial organization
and dynamics
www.ibgc.u-bordeaux2.fr/?page=equipe&eq=odm
Manuel Rojo
Mitochondria fulfill several functions that are essential for cell life and
death. They continuously move, fuse and divide and these dynamics are
required for mitochondrial biogenesis, function and maintenance. Fusion
and fission are governed by large GTPases of the dynamin-superfamily
(Mfn1, Mfn2, OPA1, Drp1) that are conserved in all eukaryotes. Mutations
of fusion factors Mfn2 or OPA1 are associated to Charcot-Marie-Tooth
disease type 2A (CMT2A) or to autosomal dominant optic atrophy (DOA)
and defects in dynamics have been also associated to degenerative
neuropathies (Parkinson, Alzheimer).
The dynamics and machineries of mitochondrial membranes differ in
several aspects from those secretory and endocytic compartments.
From the fundamental point of view, their characterization is expected
to reveal mitochondria-specific features as well as mechanisms that are
common to all endomembranes. From the biomedical perspective, the
characterization of mitochondrial dynamics, and of the proteins involved,
will allow the identification of genetic determinants and targets and open
the way for the development of therapeutic strategies. Our team seeks to
characterize the mechanisms of mitochondrial fusion, the role(s) of the
dynamins involved, the functional consequences of pathogenic MFN2 and
OPA1 mutations and the physiological consequences of fusion defects.
The team develops two main projects: (1) the characterization of
the fusion process (ex vivo and in vitro) and (2) the molecular and
functional characterization of fusion factors (Mfn1, Mfn2 and OPA1). We
develop novel assays for the precise and quantitative characterization
of mitochondrial fusion. This will enable the study of fusion in different
physiopathological situations and the search for pharmacological
modulators. We have established strategies for the expression of fusion
factors and investigate their capacity to deform and/or fuse membranes
as well as the consequences of disease-associated mutations. Our
experimental approaches include protein expression, purification and
reconstitution, subcellular fractionation and fluorescence microscopy.
Our team welcomes new researchers and is open to novel projects and
collaborations.
Fusion of differentially labeled mitochondria (mtEGFP, mtDsRed) in a
HeLa cell polykaryon.
• Research team
Manuel Rojo (DR2 CNRS)
Claudine David (IE1 CNRS)
Marine Bouhier (IE CDD FRM)
Paul Charron (Master 2)
Sauvanet C, Duvezin-Caubet S, Salin
B, David C, Massoni-Laporte A, di
Rago JP, Rojo M. Mitochondrial DNA
mutations provoke dominant inhibition
of mitochondrial inner membrane
fusion. PLoS One. 2012;7(11):e49639
Sauvanet, C., S. Duvezin-Caubet, J. P.
di Rago, and M. Rojo. (2010), Energetic
requirements
and
bioenergetic
modulation
of
mitochondrial
morphology and dynamics. Semin Cell
Dev Biol. 21:558-565.
Sauvanet C. , Arnauné-Pelloquin L.
, David C. , Belenguer P., Rojo M.
(2010) Dynamique et morphologie
mitochondriales : acteurs, mécanismes
et pertinence fonctionnelle, Médecine
Sciences 26, 823-829.
Gegg ME, Cooper JM, Chau KY,
Rojo M, Schapira AH, Taanman JW.
(2010) Mitofusin-1 and Mitofusin-2
are ubiquitinated in a PINK1/parkin
dependent manner upon induction
of mitophagy. Hum Mol Genet.
19(24):4861-70.
Landes T, Emorine LJ, Courilleau
D, Rojo M, Belenguer P, ArnaunéPelloquin L. (2010), The BH3-only
Bnip3 binds to the dynamin Opa1 to
promote mitochondrial fragmentation
and apoptosis by distinct mechanisms.
EMBO Rep. 11(6):459-65.
Guillery O, Malka F, Landes T,
Guillou E., Blackstone C, Lombès
A, Belenguer P., Arnoult D., Rojo M
(2008)
Metalloprotease-mediated
OPA1 processing is modulated by the
mitochondrial membrane potential.
Biol. Cell 100(5), 315-325.
Control and Dynamics
of Cell Division
www.iecb.u-bordeaux.fr/index.php/fr/equipes/53-control-and-dynamics-of-cell-division
Anne Royou
• Research team
Anne Royou – CR1 CNRS
Marie-Charlotte Claverie – AI UB2
Nicolas Derive – Doctorant UB2
Nabia Founounou – Post-doc (ATIPAVENIR, Région Aquitaine)
Damien Goutte-Gattat – Post-doc
(ERC)
Emilie Jamet – AI (ERC)
Cédric Landmann –AI (ANR)
Emilie Montembault – CR2 CNRS
Cédric Soler – MCU Clermont-Ferrand
The mechanisms that safeguard cells against aneuploidy are of great
interest as aneuploidy contributes to tumorigenesis. Using live imaging
approaches, we have identified two novel mechanisms that permit the
accurate transmission of chromosomes during cell division of Drosophila
neuronal stem cells. The first mechanism involves the faithful segregation
of damaged chromosomes. We monitor cells entering mitosis with broken
chromosomes. Our studies reveal that chromosome fragments segregate
properly to opposite poles. This poleward motion is mediated through DNA
tethers that connect the chromosome fragments. The second mechanism
involves the coordination of chromosome segregation with cell cleavage.
We found that cells can adapt to a four-fold increase in chromatid length
by elongating transiently during anaphase. This mechanism ensures the
clearance of chromosomes from the cleavage plane prior to completion of
cell division.
Kotadia, S.*, Montembault, E.*,
Sullivan, W. Royou, A. (2012) Cell
elongation is an adaptive response for
clearing long chromatid arms from the
cleavage plane. J. Cell Biol. 199(5):
745-53 *equal contribution
McCusker D., Royou, A. Velours C.,
Kellogg D. (2012) Cdk1-dependent
control of membrane trafficking
dynamics. Mol Biol Cell. 23(17): 333647
Royou, A., Gagou, M., Karess, R.,
D., Sullivan, W. (2010) BubR1 and
Polo-coated DNA tethers facilitate the
segregation of acentric chromatids.
Cell 140(2): 235-45.
Royou, A., McCusker, D., Kellogg,
D., Sullivan, W. (2008) Grapes(Chk1)
prevents nuclear Cdk1 activation by
delaying Cyclin B nuclear accumulation.
J. Cell Biol. 183(1): 63-75
The research team was recently awarded an ERC starting Grant
Cell Biology of Quiescence
www.ibgc.u-bordeaux2.fr/?page=equipe&eq=sagot
Isabelle Sagot
Quiescence, a cellular state defined as a reversible absence of proliferation,
concerns the vast majority of cells on earth, from microbes to stem cells.
Quiescent cells not only have to survive and face aging, but they must
also preserve their ability to re-enter the cell cycle in a tightly regulated
manner and to give rise to a healthy and rejuvenated progeny. Therefore
quiescence is central for two major biological issues: the control of cell
proliferation and cell aging. Remarkably little is known about this specific
cellular state and the mechanisms governing quiescence/proliferation
transitions remain obscure. In fact, even the most recent genomic,
transcriptomic and metabolomic approaches did not reveal any clear
specific quiescence molecular signature, challenging thus the existence
of a «program» that commit cell to the quiescent state.
Few years ago, we have started to characterize quiescent yeast cells
at the cellular level. We have shown that upon entry into quiescence
two evolutionary distant yeast species, S. cerevisiae and S. pombe, reorganize drastically some of their intracellular machineries. Indeed, upon
carbon starvation cells assemble two specific structures Actin Bodies (AB)
and Proteasome Storage Granules (PSGs) that respectively result from
the specific reorganization the actin cytoskeleton and the proteasome.
Importantly, these structures are mobilized within seconds when cells exit
quiescence upon re-feeding. More recently, using AB and PSGs as new
specific markers of the quiescent state at the single cell level, we have
demonstrated that quiescence entry and exit can occur not only in the G1
phase of the cell cycle, but also in other cell cycle stages. Additionally,
we have established that the cell’s metabolic status rather than cell cycle
regulators is critical for the control of quiescence/proliferation transitions.
Our current project is to dress the cartography of all the cellular remodeling
that occur upon transition from proliferation to quiescence and to use
these specific cellular organization as tools to address central questions
in the quiescence field such as: how do quiescent cells survive, what
are the cascade of molecular switches that tightly control quiescence
exit, and how can quiescent cell give rise to a rejuvenated progeny upon
quiescence exit.
• Research team
Isabelle Sagot – CR1 CNRS
Fabien Courtout - Technicien
Damien Laporte – Post-Doc ANR/
Région Aquitaine
Laure Jimenez – Doctorante MESR
UBS
Laporte D, Lebaudy A, Sahin A, Pinson
B, Ceschin J, Daignan-Fornier B, Sagot
I. 2011 Metabolic status rather than cell
cycle signals control quiescence entry
and exit. The Journal of Cell Biology.
192(6):949-57.
Daignan-Fornier
B,
Sagot
I.
2011Proliferation/quiescence : the
controversial
«aller-retour».
Cell
Division. 6(1):10.
Daignan-Fornier B, Sagot I. 2011
Proliferation / Quiescence : When to
start ? Where to stop? What to stock?
Cell Division.6(1) :20.
Laporte D, Salin B, Daignan-Fornier B,
Sagot I. 2008 Reversible cytoplasmic
localization of the proteasome in
quiescent yeast cells. The Journal of
Cell Biology. 181(5): 737-745
Sahin A, Daignan-Fornier B, Sagot I.
2008 Polarized growth in the absence
of F-actin in Saccharomyces cerevisiae
exiting quiescence. PLoS ONE. 3(7)
e2556.
Yeast cells can enter quiescence from all cell cycle stages.
Quiescent cell’s contour is stained with concanavaline A – FITC (Green) ;
Actin Bodies, actin structures that are specific of quiescent cells are stained
with Alexa 548 phalloidine (Red). Round cells have entered quiescence
from G1, budded cells from other cell cycle stages.
Non-self recognition in fungi
http://www.iecb.u-bordeaux.fr/index.php/fr/equipes/53-control-anddynamics-of-cell-division
Sven J. Saupe
• Research team
Annick Breton - CR1 CNRS
Corinne Clavé - Professeur UBS
Asen Daskalov - Doctorant MESR
Marina Lamacchia - Post-Doc ANR
Frederique Ness - MCU UBS
Mathieu Paoletti - CR1 CNRS
Khalid Salamat - Post-Doc ANR
Sven J. Saupe - DR2 CNRS
Martine Sicault - TCE UBS
Debets AJ, Dalstra HJ, Slakhorst M,
Koopmanschap B, Hoekstra RF, Saupe
SJ (2012) High natural prevalence of a
fungal prion. Proc Natl Acad Sci U S A
109: 10432-10437
Daskalov A, Paoletti M, Ness F, Saupe
SJ (2012) Genomic clustering and
homology between HET-S and the
NWD2 STAND protein in various fungal
genomes. Plos One 7: e34854
Mathur V, Seuring C, Riek R, Saupe
SJ, Liebman SW (2012) Localization
of HET-S to the cell periphery, not to
[Het-s] aggregates, is associated with
[Het-s]-HET-S toxicity. Mol Cell Biol 32:
139-153
Saupe SJ, Daskalov A (2012) The
[Het-s] Prion, an Amyloid Fold as a Cell
Death Activation Trigger. PLoS Pathog
8: e1002687
Benkemoun L, Ness F, Sabate R,
Ceschin J, Breton A, Clave C, Saupe
SJ (2011) Two structurally similar
fungal prions efficiently cross-seed in
vivo but form distinct polymers when
coexpressed. Mol Microbiol 82: 13921405
Saupe SJ (2011) The [Het-s] prion of
Podospora anserina and its role in
heterokaryon incompatibility. Semin
Cell Dev Biol 22: 460-468
Our activity is centered on the study of non-self recognition in fungi. We
want to improve our understanding of how fungi recognized and respond
to non-self. We are focused on the description of a non-self recognition
process termed heterokaryon incompatibility which takes place when two
unlike individuals belonging to the same species fuse. But gradually we
expand our studies to include other types of biotic interactions (between
fungi belonging to different species, between fungi and bacteria). Our
main model species is Podospora anserina. We are characterizing the
genes controlling this process at different steps from the initial recognition
event to the ensuing cellular response. In particular, we are interested in
a large gene family encoding STAND proteins which show homology to
animal NOD-like receptors and plant NBS-LRR proteins. We propose that
these genes constitute the fungal counterpart of the pathogen recognition
receptors controlling innate immunity in animals and plants. In addition,
one of the proteins involved in this process is the HET-s prion protein.
Recent studies have shown that the prion fold of HET-s serves as a trigger
to induce the toxicity of the partner protein termed HET-S and that toxicity
of HET-S can also be activated by a STAND protein termed NWD2.
Rare diseases: Genetics and
Metabolism (MRGM, EA4576)
www.univ-bordeauxsegalen.fr/fr/recherche/acteurs-de-la-recherche/biologie-fondamentale-et-appliquee-a-la-medecine/
mrgm.html
Didier Lacombe
Our basic research concerns physiopathological studies of different orphan
genetic diseases affecting developmental programs, energy metabolism
(regulation of energy transducing processes at the level of proteins,
organelle, genes and epigenetics, mitochondrial dynamics, mitophagy
and biogenesis and metabolic flexibility) and lipid metabolism (implications
in nervous system morphogenesis and function, syndromic obesity),
using cellular and animals (e.g. zebrafish) models. Our group drives
translational studies on rare monogenic disorders leading to developmental
abnormalities including malformations, neurogenetic diseases, and
inborn errors of lipid and mitochondrial metabolisms. Pharmacological
research and preclinical drug screening is also in progress for different
diseases (Rubinstein-Taybi syndrome, mitochondria dynamic disorders,
hereditary spastic paraplegias). Our team includes complementary
scientific (genetics, biochemistry and cell biology), technological (live
microscopy, bioenergetics, molecular biology, genomics, zebrafish
models, mice models, cell models) and clinical expertises (rare diseases,
neurodegenerative disorders, heart dysfunction, anesthesia..). Our
laboratory is tightly linked with the clinical department of clinical genetics
and the reference center of mitochondrial diseases, so that an effective
translational research can occur, based on a close proximity between
scientists and clinicians working together in the same group. Our common
objective is to understand the physiopathological mechanisms underlying
new syndromes with multiple congenital abnormalities identified by the
clinical departments associated to our research project at the university
hospital.
• Research team
Didier Lacombe
Patrick Babin
Benoît Arveiler
Rodrigue Rossignol
Cyril Goizet
Anja Knoll-Gellida
Caroline Rooryck-Thambo
Isabelle Coupry
Karine Nouette-Gaulain
Isabelle Vernhet
Rheb regulates mitophagy induced by mitochondrial energetic
status. Melser S, Chatelain EH, Lavie J, Mahfouf W, Jose C, Obre
E, Goorden S, Priault M, Elgersma Y, Rezvani HR, Rossignol R,
Bénard G. Cell Metab. 2013 May 7;17(5):719-30. doi: 10.1016/j.
cmet.2013.03.014. Epub 2013 Apr 18.
spectrum, and potential association with mitochondrial morphofunctional dysfunction. Hum Mutat. 2011; 32(10):1118-27.
Alteration
of
fatty-acid-metabolizing
enzymes
affects
mitochondrial form and function in hereditary spastic paraplegia.
Rossignol R and Tesson C and Nawara M and Salih MA, Zaki
MS, Al Balwi M, Schule R, Mignot C, Obre E, Bouhouche A,
Santorelli FM, Durand CM, Oteyza AC, El-Hachimi KH, Al Drees
A, Bouslam N, Lamari F, Elmalik SA, Kabiraj MM, Seidahmed
MZ, Esteves T, Gaussen M, Monin ML, Gyapay G, Lechner D,
Gonzalez M, Depienne C, Mochel F, Lavie J, Schols L, Lacombe
D, Yahyaoui M, Al Abdulkareem I, Zuchner S, Yamashita A,
Benomar A, Goizet C, Durr A, Gleeson JG, Darios F, Brice A,
Stevanin G. Am J Hum Genet. 2012 Dec 7;91(6):1051-64.
Nurden P et al. Thrombocytopenia resulting from mutations in
filamin A can be expressed as an isolated syndrome. Blood 2011;
118(22):5928-37.
Goizet C, et al. REEP1 mutations in SPG31: frequency, mutational
Tingaud-Sequeira A, Ouadah N, Babin PJ. Zebrafish obesogenic
test: a tool for screening molecules that target adiposity. J Lipid
Res. 2011; 52(9):1765-72.
Tingaud-Sequeira A et al. Vitellogenin expression in white adipose
tissue in female teleost fish. Biol Reprod. 2012; 86(2):38.
Benard G et al. Adaptative Capacity of Mitochondrial Biogenesis
and of Mitochondrial Dynamics in Response to Pathogenic
Respiratory Chain Dysfunction. Antioxid. Redox. Signal. 2012
Apr 19.
Bénard G, et al. Mitochondrial CB1 receptors regulate neuronal
energy metabolism. Nat Neurosci. 2012;15(4):558-64.
cardio
thoracic
Bronchial remodelling
(INserm U1045)
www.univ-bordeauxsegalen.fr/fr/recherche/acteurs-dela-recherche/biologie-fondamentale-et-appliquee-a-lamedecine/centre-de-recherche-cardio-thoracique-debordeaux-crctb.html
Patrick Berger
The project of the team associates human and animal studies in a
multidisciplinary approach (physiologists, chest physicians, radiologists,
pharmacologists, and paediatricians) with strong interconnection between the
team and the clinical investigation center in the hospital
Research brief
Asthma and chronic obstructive pulmonary disease (COPD) are very frequent
inflammatory diseases that are characterized by different patterns of bronchial
remodelling. However, characteristics and localization of the increased in
Bronchial Smooth Muscle (BSM) mass are different. In COPD, there is a BSM
cell hypertrophy which is only present in distal bronchi whereas in asthma,
there are both BSM cell hypertrophy and hyperplasia within the entire bronchial
tree. Anyhow, BSM remodelling has been associated with a poor prognosis,
high morbidity, and deterioration of lung function. As a consequence BSM
remodelling should be a target of innovative treatments.
Mechanisms of BSM remodeling remain largely unknown even if we previously
demonstrated the role of mitochondrial BSM biogenesis (T. Trian. J Exp Med
2007).
Recently, we evaluated the relationship between bronchial epithelium and
BSM. In particular, the epithelial chitinase YKL-40 induces the proliferation
of BSM cells, through a PAR-2 dependant mechanism (Figure A). Moreover,
YKL-40 induces BSM cell migration and pseudopods formation (Figure B).
• Research team
Heading: P Berger
Lecturers: Choukroun, Dupin , Fayon,
Girodet, Hilbert, Lederlin, Laurent,
Marthan, Montaudon, Ozier, Trian,
Vargas
Post-doc: Carvallo
PhD student: Allard, Dournes, Thumerel
Eng : Cattiaux, Ousova
Tech: Maurat
Bara et al. AJRCCM 2012, 185 : 715-22
Girodet PO, Ozier A, Carvalho G, Ilina O, Ousova O,
Gadeau AP, Begueret H, Wulff H, Marthan R, Bradding
P, Berger P. Ca(2+)-activated K(+) channel-3.1 blocker
TRAM-34 attenuates airway remodeling and eosinophilia
in a murine asthma model. Am J Respir Cell Mol Biol. 2013
Feb;48(2):212-9
Lederlin M, Ozier A, Dournes G, Ousova O, Girodet PO,
Begueret H, Marthan R, Montaudon M, Laurent F, Berger
P.In vivo micro-CT assessment of airway remodeling in a
flexible OVA-sensitized murine model of asthma. PLoS
One. 2012;7(10):e48493.
Bara I, Ozier A, Girodet PO, Carvalho G, Cattiaux J,
Begueret H, Thumerel M, Ousova O, Kolbeck R, Coyle AJ,
Woods J, Tunon de Lara JM, Marthan R, Berger P. Role of
YKL-40 in bronchial smooth muscle remodeling in asthma.
Am J Respir Crit Care Med. 2012 Apr 1;185(7):715-22
Lederlin M, Laurent F, Portron Y, Ozier A, Cochet
H, Berger P, Montaudon M. CT attenuation of the
bronchial wall in patients with asthma: comparison with
geometric parameters and correlation with function and
histologic characteristics. AJR Am J Roentgenol. 2012
Dec;199(6):1226-33.
Lederlin M, Laurent F, Dromer C, Cochet H, Berger P,
Montaudon M. Mean bronchial wall attenuation value
in chronic obstructive pulmonary disease: comparison
with standard bronchial parameters and correlation with
function. AJR Am J Roentgenol. 2012 Apr;198(4):800-8
Girodet PO, Ozier A, Trian T, Begueret H, Ousova O,
Vernejoux JM, Chanez P, Marthan R, Berger P, Tunon de
Lara JM. Mast cell adhesion to bronchial smooth muscle in
asthma specifically depends on CD51 and CD44 variant 6.
Allergy. 2010 Aug;65(8):1004-12
CARDIO THORACIC │51
Endothelial cell Biology
(INserm U1045)
www.iecb.u-bordeaux.fr/teams/GENOT/
Elisabeth Génot
• Research team
Elisabeth Génot, DR2 INSERM
IJsbrand Kramer, Pr Bordeaux 1
Thomas Daubon, post-doc
Anne Leclercq, post-doc
Véronique Veillat, post-doc
Pirjo SPuul, post-doc
Isabel Egaña, PhD student
Filipa Curado, PhD student
Paolo Ciufici, PhD student
Génot E. et al., Fgd1 as a central
regulator of extracellular matrix
remodelling: Lessons from Faciogenital
dysplasia J Cell Sci. 2012
Daubon T. et al,. Invadopodia and
rolling-type motility are specific features
of highly invasive p190(bcr-abl)
leukemic cells. Eur J Cell Biol. 2012
Le Roux-Goglin E. et al., Helicobacter
infection induces podosome assembly
in primary hepatocytes in vitro. Eur J
Cell Biol. 2012;91(3):161.
Juin A. et al., Physiological type I collagen
organization induces the formation of a
novel class of linear invadosomes. Mol
Biol Cell. 2012;23:297.
Daubon T., et al., The Aarskog-Scott
syndrome protein Fgd1 regulates
podosome formation and extra-cellular
matrix remodeling in transforming
growth factor β-stimulated aortic
endothelial cells. Mol Cell Biol.
2011;31:4430.
Quideau S. et al., Binding of filamentous
actin and winding into fibrillar aggregates
by the polyphenolic C-glucosidic
ellagitannin vescalagin. Angew Chem
Int Ed Engl. 2011;50:5099.
Saltel F., et al. Invadosomes: intriguing
structures with promise. Eur J Cell Biol.
2011;90:100.
Kremerskothen, J., et al., ZO-1
modulate podosome formation. FASEB
J. 2011, 25(2):505-14.
52 │CARDIO THORACIC
TGFß plays an important role in the homeostasis of the vascular system.
Analysing the effects of TGFß on the cytoskeleton organization of
aortic endothelial cells led us to discover actin-rich structures named
podosomes. These are dynamic actin-rich adhesion plasma membrane
microdomains endowed with extracellular matrix-degrading activities.
Ongoing projects aim at demonstrating the occurrence of podosomes in
tissues and determine their role in (patho)physiology. We are also exploring
the conditions (cytokine, matrix…), which favor podosome formation. We
are working on the elucidation of the molecular mechanisms involved in
podosome formation. The RhoGTPase Cdc42 plays a key role in this
Figure 1 : TGFβ signaling
pathways for podosome
formation in endothelial cells. Both canonical
(smad) and non canonical
(Fgd1) pathways contribute to podosome assembly.
process.
We have now identified Fgd1 as the guanine exchange factor regulating
Cdc42 activation downstream of TGFβ receptors : Fgd1 undergoes Srcdependent tyrosine phosphorylation, translocates to the membrane and
works in a complex with cortactin to activate Cdc42 and induce actin
polymerization for podosome formation (Fig.1). These findings reveal the
involvement of Fgd1 in endothelial cell biology and open up new avenues
to study its role in vascular pathophysiology.
TGFß is also an inflammatory mediator. In collaboration with C. Varon’s
team at U853, we showed that TGFß may induce podosomes in other
cells. We demonstrated that Helicobacter pylori promotes podosome
formation in murine primary hepatocytes in vitro, and this occurs through
the release of TGFβ. Liver cells with podosomes have reduced selfhealing capacities. Helicobacter pylori which colonizes the stomach in
about 50% of all humans is well known as a key risk factor in gastric
diseases, it may also damage liver, causing cirrhosis and liver cancer.
Although it is not yet clear which role podosomes play in the response to
bacterial infection, one may expect that in vivo, podosomes in liver cells
infected with Helicobacter pylori contribute to the pathological state.
Figure 2: A ring of podosomes (at the
center of the image) induced by Helicobacter pylori infection in Hepatocellular
carcinoma cells. F-actin (red), cortactin (green), vinculin (white), and nuclei
(bue). (image, Pirjo Spuul).
Pathophysiology of pulmonary
circulation (INserm U1045)
www.univ-bordeauxsegalen.fr/fr/recherche/acteurs-dela-recherche/biologie-fondamentale-et-appliquee-a-lamedecine/centre-de-recherche-cardio-thoracique-debordeaux-crctb.html
Jean Pierre Savineau
The main scope of the research of the team relates to pulmonary arterial
wall biology, pulmonary arterial hypertension (cellular mechanisms
and pharmacological treatment) and the impact of the environment (air
pollution) on the pulmonary circulation. Focused on the pathophysiology
of the pulmonary circulation, the team works on a real interface between
cardiac and pulmonary diseases. It is multidisciplinary constituted by
colleagues of diverse trainings (basic scientists, physicians, pharmacists)
and of varied disciplines (cellular biology, physiology, pharmacology,
toxicology).
• Research team
C. Guibert, CR1 INSERM
B. Muller, JF. Quignard (Prof.)
I. Baudrimont, A. Courtois, M. Dubois,
T. Ducret, F. Delom, V. Freund-Michel
(Lecturers)
E Dumas de la Roque (PH)
D. Fessart (Post doctoral researcher)
G. Gilbert, N. Khoyrattee, T. Papaite
(PhD students)
J. Cattiau (engineer), H. Crevel
(technician)
The specific objectives of the team are the followings:
1. To characterize the calcium signalling (TRP, RyR, IP3R channels) in
endothelial and smooth muscle cells in physiological conditions and
during pulmonary arterial hypertension.
2. To study the cross talk between endothelial and smooth muscle
cells in the pulmonary arterial wall through myoendothelial junctions
(connexins).
3. To assess innovative treatment in pulmonary arterial hypertension.
4. To study the impact of nanoparticles from air pollution on the pulmonary
circulation
Billaud M, Marthan R, Savineau JP, Guibert
C. Vascular smooth muscle modulates
endothelial control of vasoreactivity via
reactive oxygen species production through
myoendothelial communications. PLoS
One. 2009 4(7):e6432.
Methodologies used
Experiments are conducted (i) on freshly isolated or cultured vascular cells
and tissues (fragments of PA) for molecular biology (PCR, real-time PCR),
cellular biology (electron microscopy, Western blotting, flow cytometry,
siRNA, proliferation, migration and apoptosis assays), electrophysiology
(patch clamp), calcium imaging (fura2, fluo3), immunochemistry,
quantitative proteomic (label-Free); (ii) on isolated and more integrated
vascular preparations (arterial rings, cannulated and pressurized micro
vessels) from both animal and human tissues for reactivity experiments
and electron paramagnetic resonance measurements for NO and
superoxyde anion; (iii) in whole animal for in vivo investigations (Doppler
echocardiography) both wild type and transgenic animals. The team
has developed animal models for pulmonary arterial hypertension
(chronically hypoxic rat and mice, monocrotaline-treated rat) and also
performs experiments on human tissues (pulmonary vessels) through a
collaborative project with the university hospital of Bordeaux.
Delannoy E, Courtois A, Freund-Michel V,
Leblais V, Marthan R, Muller B. Hypoxiainduced hyperreactivity of pulmonary
arteries:
role
of
cyclooxygenase-2,
isoprostanes, and thromboxane receptors.
Cardiovasc Res. 2010, 85(3):582-92.
Ducret T, El Arrouchi j, Courtois A,
Quignard JF, Marthan R, Savineau JP.
Characterization
of
stretch-activated
channels in rat intrapulmonary arterial
smooth muscle cells. Cell Calcium, 2010,
2251-259.
Chevalier M, Gilbert G, Lory P,
Marthan R, Quignard JF, Savineau JP.
Dehydroepiandrosterone (DHEA) inhibits
voltage-gated T-type calcium channels.
Biochemical
Pharmacology,
2012,
83(11):1530-1539.
Below: diffusion of a green fluorescent dye from the endothelium to the
smooth muscle via cell-to-cell communications in basal conditions (left)
and such diffusion was blocked following one hour incubation with a
specific connexion peptide blocker 37-43Gap 27 (right). Scale bars are 15
µm. (from Billaud et al, 2009, Plos one)
Dumas de La Roque E, Bellance N,
Rossignol R, Begueret H, Billaud M, dos
Santos P, Ducret T, Marthan R, Dahan D,
Ramos-Barbón D, Amor-Carro Ó, Savineau
JP, Fayon M. Dehydroepiandrosterone
reverses chronic hypoxia/reoxygenationinduced right ventricular dysfunction
in rats. European Respiratory Journal.
2012;40(6):1420-9
CARDIO THORACIC │53
Cardiovascular adaptation
to ischemia (INserm U1034)
www.u1034.bordeaux.inserm.fr/
Thierry Couffinhal
Our team combines tools of genetics and of in vivo and in vitro imagery
to characterize and follow in time and space the role of target genes in the
vascular morphogenesis.
• Research team
Researchers: JM Daniel-Lamazière, C.
Duplaa, A. Gadeau, MA Renault
ATIP-AVENIR : C. James
University Bordeaux Segalen :
L. Barandon, D. Breilh, T. Couffinhal,
P. Dufourcq, K. Latry, A. Ouattara, E.
Roux, MC Saux, F. Sztarck, JF Viallard
Postdoc fellows:
F. Larrieu, A. Gourgues, J. Vieira-Dias
Technicians: 5
PhD students: 6
Normal tissue functioning depends on adequate supply of oxygen through blood
vessels. Atherosclerosis (i.e. lesion of large vessels or macroangiopathy) or
diabetes, and hypertension (i.e. lesions of small vessels or microangiopathy)
reduce vessel lumen diameter and blood perfusion, leading to tissue ischemia.
There are major causes of mortality by myocardial angina, infarction, or stroke as
well as morbidity by hindlimb arteriopathy, nephropathy, blindness, pre-eclampsia,
neurodegeneration and osteoporosis.
Our project focuses on 4 complementary aspects of vessel formation:
• Understand the mechanisms that maintain or favor formation of vessels in
post-ischemic tissue. In response to ischemia, new blood vessels form and
a collateral circulation are established in order to compensate for the lack of
perfusion. During the regeneration and renewal of adult tissue, the formation
of the vessels is regulated by a combination of signaling factors, of guidance
proteins and by local mechanical interactions between cells. A new concept
is proposed that the need for regeneration reactivates embryonic processes,
redeploying many of the same molecular regulators. Over the past few years,
our team showed that the Hedgehog (Hh) and the Wnt/Frizzled pathways take
a center stage position. We showed that Wnt and Hh signaling pathways,
reactivated in adult ischemic tissues, play a key role in coordinating vessel
formation, maturation, remodeling and 3D organization of the vascular network
and in controlling cardio protective response after ischemic events. Our aim is
to characterize how these two pathways modulate vascular network formation
and organization and to identify how to modulate them to improve ischemic
tissue repair.
• Platelets and endothelium are important players in the regulation of hemostasis
and angiogenesis and it has been reported that platelets can influence the
properties of endothelial cells. We hypothesize that endothelial cells can
be abnormal in patients with platelet disorders, i.e.hyperactivated, leaky
or hyperproliferative. They would be responsible for pathological plateletendothelium interactions, thus giving rise to inappropriate thrombosis, bleeding
or neoangiogenesis.
Using pharmacological and experimental approaches, we explore general
vascular side effects of drugs that may impair angiogenic pathways. Using an
epidemiological approach, we propose to study the cardiovascular side effects
of these newly delivered treatments.
Pre-clinical experiments of stem cell angiogenic therapy are developed in large
animal, using pre-conditioned stem cells and human clinical tools of evaluation.
Effective organ blood flow, perfusion and bio-energetic status in situation of
ischemic heart hemodynamic failure are studied in experimental model and
human clinical trial.
Renault MA, Chapouly C, Yao Q, Larrieu-Lahargue F,
Vandierdonck S, Reynaud A, Petit M, Jaspard-Vinassa B, Belloc
I, Traiffort E, Ruat M, Duplàa C, Couffinhal T, Desgranges C,
Gadeau AP. Desert hedgehog promotes ischemia-induced
angiogenesis by ensuring peripheral nerve survival. Circ Res.
2013 Mar 1;112(5):762-70
Descamps, B., R. Sewduth, N. Ferreira Tojais, B. Jaspard, A
Reynaud, F. Sohet, P. Lacolley, C. Allieres, J.M. Lamaziere, C.
Moreau, P. Dufourcq, T. Couffinhal, and C. Duplaa Frizzled 4
Regulates Arterial Network Organization Through Noncanonical
Wnt/Planar Cell Polarity Signaling. Circulation Research, 2012
;110, 47-58
Barandon L, Casassus F, Leroux L, Moreau C, Allières C, Daniel
Lamazière JM, Dufourcq P, Couffinhal T, Duplàa C. Secreted
Frizzled-Related Protein-1 Improves Postinfarction Scar
54 │CARDIO THORACIC
Formation Through a Modulation of Inflammatory Response.
Arterioscler Thromb Vasc Biol, 2011;31, e80-e87
Josefsson EC, James C, Henley KJ, Debrincat MA, Rogers KL,
Dowling MR, White MJ, Kruse EA, Lane RM, Ellis S, Nurden P,
Mason KD, O’Reilly LA, Roberts AW, Metcalf D, Huang DC, Kile
BT. Megakaryocytes possess a functional intrinsic apoptosis
pathway that must be restrained to survive and produce platelets.
J Exp Med. 2011. 208:2017-31.
Renault MA, Robbesyn F, Réant P, Douin V, le Daret D, Allières
C, Belloc I, Couffinhal T, Arnal JF, Klingel K, Desgrange C, Dos
Santos P, Charpentier F and Gadeau AP. Osteopontin Expression
in Cardiomyocytes Induces Dilated Cardiomyopathy. Circ Heart
Failure, 2010.14(3), 431-439
Technology
Platforms
cytometry
• Research team
Flow cytometry core facility
with high speed cell sorter
www.transbiomed.u-bordeaux2.fr/pages/cytometrieplateforme.
html
The flow cytometry core facility provides three multicolor flow cytometer
analysers with high throughput samplers and a four way high speed cell
sorter with an automated cell deposit unit. Available lasers are 375, 405,
488, 560 and 640nm (see table) giving acces to all the flow cytometry
applications. A dedicated engineer and a technician operate the service.
Scientific manager
Julie Déchanet-Merville DR
Services
High speed four way sorting and cloning on BD FACSAria
Multicolor flow cytometer analysis with high throughput samplers on
Canto, CantoII and Fortessa
Individual training for the use of the analysers and Diva software
Advice on the preparation of samples for analysis or sorting
for all applications : surface or intracellular immunophenotyping, cell cycle,
apotosis, viability, calcium flux, cytokine mutiplexing, GFP and reporter
genes, phosphoflow...
Daylong scientific conferences focused on topics involving flow cytometry
techniques.
Equipment
Technical officer
Vincent Pitard IR
Technician
Santiago Gonzalez
Laboratory
• Contact :
Vincent Pitard
Santiago Gonzalez :
cytometry@cirid.org
Main lab with 3 cytometers and 4 computers for data analysis. All the
computers are connected on a server for data storage and file sharing.
Tel : 05 57 57 57 05
Cell sorter in a biological safety laboratory adapted for sterile cell culture.
58 │cytometry
histopathology
The histopathology platform
www.transbiomed.u-bordeaux2.fr/pages/Histologie.html
• Contact :
Nathalie Dugot-Senant
nathalie.senant@inserm.fr
• Location
The platform is hosted by the INSERM
U1053 “Physiopathologie du cancer du
foie”
60 │histopathology
The purpose of this platform is to provide the scientists with an access to
the equipment and know-how for processing tissue samples for histology
and immunohistochemistry.
Available equipment includes notably a Shandon Citadel 1000 tissue
processor, a TBS 88-MEDITE paraffin inclusion system, several
microtomes and a DAKO Autostainer.
The offer includes counseling, training of the users as well as the realization
of a whole range of services: help with sampling the tissues, freezing,
fixation, inclusion, sectioning, histology and immunohistochemistry.
Laboratory P3
Level 3 safety laboratory
www.transbiomed.u-bordeaux2.fr/pages/P3/labop3.html
• Research team
Michel Ventura
Directeur de recherche CNRS
• Contact
Michel Ventura
Michel.ventura@u-bordeaux2.fr
62 │Laboratory P3
The level 3 safety laboratory offers the possibility to work with class 3
pathogens under adequate safe conditions. University scientific research
groups but also external users are welcomed to use this facility after a
specific training. Analysis and screening of inhibitors, development of
lentiviral vectors as well as fundamental studies on the mechanisms of
replication of some of these pathogens can be performed in this L3 lab.
Four boxes are located around a central room containing 3 freezers, two
at -80°C and one at -135°C freezer. They ensure the conservation of the
samples and the different infected cell lines. An ultracentrifuge is also
available.
Each one of the boxes contains the necessary equipment for pathogens
manipulation (class II PSM, microscope and incubators). The working areas,
separated by a double SAS which ensures the pathogens containment,
are classified under ZRR (Zone à Régime Restrictif) conditions for safety
purposes. Safe processing of the waste is ensured by a double entries
autoclave.
Vectorology
Vectorology platform
www.transbiomed.u-bordeaux2.fr/pages/vectorologie.html
• Research team
Technical officer :
Véronique GUYONNET DUPERAT
(IE1 INSERM- US005)
Scientific manager :
François MOREAU-GAUDRY
(PU-PH-U1035)
Technician :
Alice Bibeyran (AI INSERM – US005)
• Contact
Veronique.guyonnet-duperat@inserm.fr
Alice.bibeyran@inserm.fr
Tel 0557571602
• Location
The platform is hosted by the INSERM
U1035 “Biothérapies des maladies
génétiques et du cancer”
64 │Vectorology
The vectorology platform is an academic structure for the production of
Lentiviral particles. Lentiviral vectors are tools of choice for gene transfer.
They have the particularity to transduce efficiently a large type of cells
including primary stem cells (neurons, retina, HSC). They allow stable
and efficient integration of DNA sequences (reporter genes, cDNA, Sh/
miRNA). The power of this approach and its plasticity explain the success
of this platform.
The main activities of the vectorology platform are the production of
lentiviral vectors for over-expression of gene or knock-down of gene
expression. The platform offer a large choice of lentiviral vectors for
constitutive or inducible expression.
Offered Services
• Advice and assistance in gene transfer (choice of vector, technical
support for cloning and lentiviral infection)
• Production of defective lentiviral particules (batch of 107 to 109
particules/ml).
• Titration of lentiviral particules. The infectious titer is determined
by FACS for various fluorescent genes (GFP, CFP, dsRed).The
quantification of physical particles was made using P24 ELISA.
• Bio-safety test. A PCR-test is achieved to verify the absence of
replicative virus on each batch of virus encoding for an insert of type B
to scale down the vector preparation to safety level 2, according to the
HCB guidelines.
• Development of new lentiviral vectors to propose innovative tools.
• Quantitative PCR technique for estimation of provirus integrated copies
number after gene transfer.
• LAM (linear amplification–mediated) - PCR technique for sequencing
proviral integration sites.
Equipment
L2, L3 laboratory
SFR TransBioMed
www.transbiomed.u-bordeaux2.fr

SFR TransBioMed - Université de Bordeaux

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