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Molecular Mycobacterial Pathogenesis

Christophe Guilhot

Group Leader

The overall objectives of the group Guilhot are to understand what make some mycobacterial species, such as Mycobacterium tuberculosis and Mycobacterium leprae, such deadly human pathogens and to use this knowledge to propose new ways to fight them.  The biosynthesis and translocation to the cell surface of pathogen specific lipids and their contribution to pathogenesis has been the main focus of this group in the recent years.

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Bacterial lipids in the context of host/pathogen interaction constitute a poorly explored scientific field.  Mycobacteria, and Mycobacterium tuberculosis (MTB) in particular, are among the best examples of bacterial pathogens using lipids to promote replication and persistence in the human host.

Consistently, a large portion of their genome is devoted to lipid metabolism and to the construction of a very complex cell envelope containing a large variety of unique lipid components. Remarkably several surface lipids, located in an ideal position for interacting with host cells, are restricted to a very limited number of mycobacterial pathogen species. Understanding how these lipids are built and translocated to the cell surface, and how they contribute to the MTB virulence are therefore major issues in the fields of microbiology and bacterial pathogenesis.

The group Guilhot has tackled these questions for more than 10 years using complementary approaches in molecular genetics, biochemistry, cellular microbiology and animal experimentation. This research team has unraveled several lipid biosynthetic pathways and has proposed several roles and molecular mechanisms of action for MTB lipids during the interaction with host cells. More recently, this group has launched several new research programs to decipher the persistence and transmission steps of the MTB infectious cycle.

 

Main Publications

  • Burbaud S et al. (2016) Trehalose polyphleate, are produced by a glycolipid biosynthetic pathway conserved across phylogenetically distant mycobacteria. Cell Chem Biol 23: 278-289
  • Boritsch E et al. (2016) pks5-recombination-mediated cell surface remodelling in Mycobacterium tuberculosis emergence. Nature Microbiol 1:15019. C. Guilhot * co-corresponding author
  • Gonzalo-Asensio J et al. (2014) Evolutionary history of tuberculosis shaped by conserved mutations in the PhoPR virulence regulator. Proc Natl Acad Sci USA 111:11491-11496
  • Passemar C et al. (2014) Multiple deletions in the polyketide synthase gene repertoire of Mycobacterium tuberculosis reveal functional overlap of cell envelope lipids in host-pathogen interactions. Cell Microbiol 16 :195-213
  • Liu CF et al. (2013) Bacterial protein-O-mannosylating enzyme is crucial for virulence of Mycobacterium tuberculosis. Proc Natl Acad Sci USA 110:6560-6565

 

 

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Interaction of human macrophages (labeled in blue) with a model mycobacterial strain (green) genetically engineered to synthesize a phenolglycolipid (red) specific to Mycobacterium leprae.

 

Scientists

Christophe Guilhot

CNRS Research director

Catherine Astarie-Dequeker

CNRS Research associate

Christian Chalut

CNRS Research associate

Kaymeuang Cam

Associate Professor - University of Toulouse

Research Assistants

Celine Berrone

European Union H2020 Engineer

Wladimir Malaga

CNRS Engineer

Gautier Prevot

CNRS Fixed-term contract Engineer

Alice Marchand

ANR-funded Engineer

Post-doctoral Fellow

Delphine Payros

Fellow of the Fondation pour la Recherche Médicale

Aideen Allen

ANR Fellow

PhD Student

Laurie Thouvenel

Fellow of the Ministry of Higher Education & Research

Publications

2017

  • Augenstreich J., Arbues, A., Simeone, R., Haanappel, E., Wegener, A., Sayes, F., Le Chevalier, F., Chalut, C., Malaga, W., Guilhot, C., Brosch*, R. and C. Astarie-Dequeker*. (2017) ESX-1 and phthiocerol dimycocerosates of Mycobacterium tuberculosis act in concert to cause phagosomal rupture and host cell apoptosis. Cell. Microbiol. (in press)

2016

  • Arbues A., Malaga, W., Constant, C., Guilhot, C., Prandi*, J. AND C. Astarie-Dequeker*. (2016) The Trisaccharides of Phenolic Glycolipids Confer 1 an Advantage to Major Pathogenic  Mycobacteria through Manipulation of Host-Cell Pattern-Recognition Receptors. ACS Chem. Biol. 11:2865-2875.
  • Burbaud S, Laval F, Lemassu A, Daffé M, Guilhot C and Chalut C (2016). Trehalose polyphleate, are produced by a glycolipid biosynthetic pathway conserved across phylogenetically distant mycobacteria. Cell. Chem. Biol. 23, 1-12.
  • Boritsch E, Frigui W, Cascioferro A, Malaga W, Etienne G, Laval F, Pawlik A, Le Chevalier F, Orgeur M, Ma L, Bouchier C, Stinear TP, Supply P, Majlessi L, Daffé M, Guilhot C and Brosch R (2016). pks5-recombination-mediated cell surface remodelling in Mycobacterium tuberculosis emergence. Nature Microbiol. 1, 15019.

2015

  • Brosch R. and Guilhot C (2015). Les bacilles de la tuberculose bovine: une evolution aux dépens de la transmissibilité chez l’homme. Médecine & Sciences, 31.

2014

  • Arbues A, Lugo-Villarino G, Neyrolles O, Guilhot C and Astarie-Dequeker C (2014). Playing hide-and-seek with host macrophages through the use of mycobacterial cell envelope phthiocerol dimycocerosates and phenol glycolipids. Front. Cell. Infect. Microbiol. 4, 1-7.
  • Gavalda S, Bardou F, Laval F, Bon C, Malaga W, Chalut C, Guilhot C, Mourey L, Daffé M. and Quémard A (2014). One domain may hide another: an acyltransferase function carried by a thioesterase-like domain of polyketide synthase. Chem. Biol. 21, 1660-1669.
  • Gonzalo-Asensio J, Malaga W, Pawlik A, Astarie-Dequeker C, Passemar C, Moreau F, Laval F, Daffé M, Martin C, Brosch R & C Guilhot (2014) Evolutionary history of tuberculosis shaped by conserved mutations in the PhoPR virulence regulator. Proc. Natl. Acad. Sci USA. 111, 11491-11496.
  • Solans L, Gonzalo-Asensio J, Sala C, Benjak A, Uplekar S, Rougemont J, Guilhot C, Malaga W, Martin C and Cole ST (2014). The PhoP-dependent ncRNA Mcr7 modulates the TAT secretion system in Mycobacterium tuberculosis. PLoS Pathog., 10, e1004183. PubMed
  • Caire-Brändli I, Papadopoulos A, Malaga W, Marais D, Canaan S, Thilo L and de Chastellier C (2014). Reversible lipid accumulation and associated division arrest of Mycobacterium avium in lipoprotein (VLDL)-induced foamy macrophages may resemble key events during latency and reactivation of tuberculosis.Infect. Immun., 82, 476-490. PubMed
  • Passemar C, Arbués A, Malaga W, Mercier I, Moreau F, Lepourry L, Neyrolles O, Guilhot C and Astarie-Dequeker C (2014). Multiple deletions in the polyketide synthase gene repertoire of Mycobacterium tuberculosis reveal functional overlap of cell envelope lipids in host-pathogen interactions. Cell. Microbiol., 16, 195-213. PubMed

2013

  • Arbués A, Aguilo JI, Gonzalo-Asensio J, Marinova D, Uranga S, Puentes E, Fernandez C, Parra A, Cardona PJ, Villaplana C, Ausina V, Williams A, Clark S, Malaga W, Guilhot C, Gicquel B and C Martin (2013). Construction, characterization and preclinical evaluation of MTBVAC, the first live-attenuated M. tuberculosis-based vaccine to enter clinical trials. Vaccine, 31, 4867-4873. PubMed
  • Rottier K, Faille A, Prudhomme T, Leblanc C, Chalut C, Cabantous S, Guilhot C, Mourey L, Pedelacq JD (2013). Detection of soluble co-factor dependent protein expression in vivo: application to the 4’-phosphopantetheinyl transferase PptT from Mycobacterium tuberculosis. J. Struct. Biol., 183(3), 320-328.PubMed
  • Martin P, Marcq I, Magistro G, Penary M, Garcie C, Payros D, Boury M, Olier M, Nougayrède J-P, Audebert M, Chalut C, Schubert S and Oswald E (2013). Interplay between siderophores and colibactin genotoxin biosynthetic pathways in Escherichia coli. PLoS Pathog., 9(7), e1003437.PubMed
  • Liu C-F, Tonini L, Malaga W, Beau M, Stella A, Bouyssié D, Jackson MC, Nigou J, Puzo G, Guilhot C, Burlet-Schiltz O and Rivière M (2013). Bacterial protein-O-mannosylating enzyme is crucial for virulence of Mycobacterium tuberculosis. Proc. Natl. Acad. Sci. USA, 110, 6560-6565. PubMed
  • Siméone R, Huet G, Constant P, Malaga W, Lemassu A, Laval F, Daffé M, Guilhot C and Chalut C (2013). Functional characterisation of three O-methyltransferases involved in the biosynthesis of phenolglycolipids in Mycobacterium tuberculosis. PLoS ONE, 8(3), e58954. PubMed

2012

  • Leblanc C, Prudhomme T, Tabouret G, Ray A, Burbaud S, Cabantous S, Mourey L, Guilhot C and Chalut C (2012). 4’-Phosphopantetheinyl transferase PptT, a new drug target required for Mycobacterium tuberculosis growth and persistence in vivo. PLoS Pathog., 8(12), e1003097. PubMed
  • Bergeret F, Gavalda S, Chalut C, Malaga W, Quémard A, Pédelacq J-D, Daffé M, Guilhot C, Mourey L and Bon C (2012). Biochemical and structural study of the atypical acyltransferase domain from the mycobacterial polyketide synthase Pks13. J. Biol. Chem., 287, 33675-33690. PubMed
  • Manca C, Peixoto B, Malaga W, Guilhot C and Kaplan G (2012) Modulation of the cytokine response in human monocytes by Mycobacterium leprae phenolic glycolipid-1 (PGL-1). J. Interferon Cytokines Res., 32(1), 27-33. PubMed

2011

  • Krishna S, Ray A, Dubey SK, Larrouy-Maumus G, Chalut C, Castanier R, Noguera A, Gilleron M, Puzo G, Vercellone A, Nampoothiri KM and Nigou J (2011). Lipoglycans contribute to innate immune detection of mycobacteria. PLoS ONE, 6(12), e28476. PubMed
  • Krishnan N, Malaga W, Constant P, Caws M, Chau TTH, Salmons J, Lan NTN, Bang ND, Daffé M, Young DB, Robertson BD, Guilhot C and Thwaites GE (2011) Mycobacterium tuberculosis lineage influences innate immune response and virulence and is associated with distinct cell envelope lipid profiles. PLoS ONE, 6(9), e23870. PubMed
  • Kirksey MA, Tischler AD, Simeone R, Hisert KB, Uplekar S, Guilhot C, and McKinney JD (2011) Spontaneous loss of phthiocerol dimycocerosates (PDIM) production in Mycobacterium tuberculosis H37Rv.Infect. Immun., 79, 2826-2838. PubMed
  • Neyrolles O and Guilhot C (2011). Recent advances in deciphering the contribution of Mycobacterium tuberculosis lipids to pathogenesis. Tuberculosis, 91, 187-95. PubMed

2010

  • Astarie-Dequeker C, Nigou J, Passemar C, and Guilhot C (2010). The role of mycobacterial lipids in host pathogenesis. Drug Discovery Today: Disease Mechanisms, 7:e33-e41.
  • Tabouret G, Astarie-Dequeker C, Demangel C, Malaga C, Constant P, Ray A, Honoré N, Bello NF, Perez E, Daffé M and Guilhot C (2010). Mycobacterium leprae phenolglycolipid-1 expressed by engineered M. bovis BCG modulates early interaction with human phagocytes. PLoS Pathog., 6:e1001159. PubMed
  • Siméone R, Léger M, Constant P, Malaga W, Marrakchi H, Daffé M, Guilhot C, and Chalut C (2010). Delineation of the roles of FadD22, FadD26 and FadD29 in the biosynthesis of phthiocerol dimycocerosates and related compounds in Mycobacterium tuberculosis. FEBS J., 277:2715-2725. PubMed

2009

  • Méniche X, Labarre C, de Sousa-D’Auria C, Huc E, Laval F, Tropis M, Bayan N, Portevin D, Guilhot C, Daffé M and Houssin C (2009). Identification of a stress-sensing factor of Corynebacterineae that is essential for the mycobacteria physiology. J. Bacteriol., 191:7323-7332. PubMed
  • Huet G, Constant P, Malaga W, Lanéelle M-A, Kremer K, van Soolingen D, Daffé M, and Guilhot C (2009). A lipid profile typifies the Beijing strains of Mycobacterium tuberculosis: identification of a mutation responsible for a modification of the structures of phthiocerol dimycocerosates and phenolic glycolipids. J. Biol. Chem., 284:27101-27113. PubMed
  • Etienne G, Malaga W, Laval F, Lemassu A, Guilhot C and Daffé M (2009). Identification of the polyketide synthase involved in the biosynthesis of cell surface-exposed lipooligosaccharides in Mycobacteria. J. Bacteriol., 191:2613-2621. PubMed
  • Gavalda S, Léger M, van der Rest B, Stella A, Bardou F, Montrozier H, Chalut C, Burlet-Schiltz O, Marrakchi H, Daffé M and Quémard A (2009). The Pks13/FadD32 crosstalk for the biosynthesis of mycolic acids in Mycobacterium tuberculosis. J. Biol. Chem., 284:19255-19264. PubMed
  • Astarie-Dequeker C, Le Guyader L, Malaga W, Seaphanh F-K, Chalut C, Lopez A and Guilhot C (2009). Phthiocerol dimycocerosates of Mycobacterium tuberculosis participate in the invasion of human macrophages by inducing changes in the lipid organization of the plasma membrane. PLoS Pathog., 5:e1000289. PubMed

2008

  • Sinsimer D, Huet G, Manca C, Tsenova L, Koo M-S, Kurepina N, Kana B, Mathema B, Marras SAE, Kreiswirth BN, Guilhot C and Kaplan G (2008). The phenolic glycolipid of Mycobacterium tuberculosis differentially modulates the early host cytokine response but does not in itself confer hypervirulence. Infect. Immun., 76:3027-3036. PubMed
  • Malaga W, Constant P, Euphrasie D, Cataldi A, Daffé M, Reyrat J-M and Guilhot C (2008). Deciphering the genetic bases of the structural diversity of phenolic glycolipids in strains of the Mycobacterium tuberculosis complex. J. Biol. Chem., 283:15177-15184. PubMed

2007

  • Simeone R, Constant P, Malaga W, Guilhot C, Daffé M and Chalut C (2007). Identification of the missing trans-acting enoyl reductase required for the phthiocerol dimycocerosates and phenolglycolipids biosynthesis in Mycobacterium tuberculosis. J. Bacteriol., 189:4597-4602. PubMed
  • Simeone R, Constant P, Malaga W, Guilhot C, Daffé M and Chalut C (2007). Molecular dissection of the biosynthetic relationship between phthiocerol and phthiodiolone dimycocerosates and their critical role in the virulence and permeability of Mycobacterium tuberculosis. FEBS J., 274:1957-1969. PubMed

2006

  • Sacco E, Legendre V, Laval F, Zerbib D, Montrozier H, Eynard N, Guilhot C, Daffé M and Quemard A (2006). Rv3389c from Mycobacterium tuberculosis, a member of the (R)-specific hydratase/dehydratase family. Biochem. Biophys. Act., 1774:303-311. PubMed
  • Chalut C, Botella L, de Sousa-D’Auria C, Houssin C and Guilhot C (2006). The non redundant roles of two 4’-phosphopantheinyl transferases in vital processes of Mycobacteria. Proc. Natl. Acad. Sci. USA, 103:8511-8516. PubMed

2005

  • Portevin D, de Sousa-D’Auria C, Montrozier H, Houssin C, Stella A, Lanéelle, M-A, Bardou F, Guilhot C and Daffé M (2005). The Acyl-AMP ligase FadD32 and AccD4-containing Acyl-CoA carboxylase are required for the synthesis of mycolic acids and essential for the mycobacterial growth. Identification of the carboxylation product and determination of the Acyl-CoA carboxylase components. J. Biol. Chem., 280:8862-8874.PubMed
  • Veyron-Churlet R, Bigot S, Guerrini O, Verdoux S, Malaga W, Daffé M and Zerbib D (2005). The biosynthesis of mycolic acids in Mycobacterium tuberculosis relies on multiple specialized elongation complexes interconnected by specific protein-protein interactions. J. Mol. Biol., 3534:847-58. PubMed

2004

  • Perez E, Constant P, Lemassu A, Laval F, Daffé M and Guilhot C (2004). Characterization of three glycosyltransferases involved in the biosynthesis of the phenolic glycolipid antigens from the Mycobacterium tuberculosis complex. J. Biol. Chem., 279:42574-42583. PubMed
  • Perez E, Constant P, Laval F, Lemassu A, Lanéelle M-A, Daffé M and Guilhot C (2004). Molecular dissection of the role of two methyltransferases in the biosynthesis of phenolglycolipids and phthiocerol dimycoserosate in the Mycobacterium tuberculosis complex. J. Biol. Chem., 279:42584-42592. PubMed
  • Portevin D, de Sousa-D’Auria C, Houssin C, Grimaldi C, Chami M, Daffé M and Guilhot C (2004). A polyketide synthase catalyzes the last condensation step of mycolic acid biosynthesis in mycobacteria and related organisms. Proc. Natl. Acad. Sci. USA, 101:314-319. PubMed

2003

  • Malaga W, Perez E and Guilhot C (2003). Production of unmarked mutations in mycobacteria using site specific recombination. FEMS Microbiol. Lett., 219:261-268. PubMed

2002

  • Constant P, Perez E, Malaga W, Lanéelle M-A, Saurel O, Daffé M and Guilhot C (2002). Role of the pks15/1 gene in the biosynthesis of phenolglycolipids in the Mycobacterium tuberculosis complex: evidence that all strains synthesize glycosylated p-hydroxybenzoic methyl esters and that strains devoid of phenolglycolipids harbor a frameshift mutation in the pks15/1 gene. J. Biol. Chem., 277:38148-38158. PubMed
  • Raynaud C, Guilhot C, Rauzier J, Bordat Y, Pelicic V, Manganelli R, Smith I, Gicquel B and Jackson M (2002). Phospholipase C are involved in the virulence of Mycobacterium tuberculosis. Mol. Microbiol., 45:203-217. PubMed
  • Puech V, Guilhot C, Perez E, Tropis M, Armitige LY, Gicquel B and Daffé M (2002). Evidence for a partial redundancy of the fibronectin-binding proteins for the transfer of mycoloyl residues onto the cell wall arabinogalactan termini of Mycobacterium tuberculosis. Mol. Microbiol., 44:1109-1122. PubMed

2001

  • Camacho LR, Constant P, Raynaud C, Lanéelle M-A, Gicquel B, Daffé M and Guilhot C (2001). Analysis of the phthiocerol dimycoserosate locus of Mycobacterium tuberculosis: evidence that this lipid is involved in the cell wall permeability barrier. J. Biol. Chem., 23:19845-19854. PubMed
  • Perez E, Samper S, Bordat Y, Guilhot C, Gicquel B and Martin C (2001). Essential role for phoP in Mycobacterium tuberculosis virulence. Mol. Microbiol., 41:179-187. PubMed

Book Chapter

  • Guilhot, C. and Daffé, M. (2008) Polyketides and polyketides-containing glycolipids of M. tuberculosis: structure, biosynthesis and biological activities. In S.H.E. Kaufmann & E. J. Rubin (eds), Tuberculosis: Molecular Biology and Biochemistry, Vol. 1 Wiley-Vch Verlag GmBH & Co. KGaA, Weinheim. pp. 21-51.
  • Guilhot, C., Chalut, C. and Daffé M. (2008) Biosynthesis and roles of phenolic glycolipids and related molecules in M. tuberculosis. In M. Daffé & J.-M. Reyrat (eds), The mycobacterial cell envelope. ASM press. Washington, D.C. pp. 273-290.
  • Portevin, D., Malaga, W. and Guilhot C. (2008) The use of temperature sensitive plasmid in mycobacteria. In T. Parish & A. Brown (eds.), Methods in Molecular Biology, Vol. : Mycobacteria Protocols. Humana Press Inc., Totowa, N.J. pp 229-242.
  • Jackson, M., Camacho, L. R., Gicquel, B. and Guilhot C. (2001) Gene replacement and transposon delivery using the negative selection marker sacB. In: « Methods in Molecular Biology: Mycobacterium tuberculosis protocols ». Editors: T. Parish & N. Stocker. Humana Press Inc., Totowa, N.J. pp.1-17.
  • McAdam, R., Quan, S. and Guilhot C. (2000) Mycobacterial transposons and their applications. In: « Molecular Genetics of Mycobacteria ». Editors: G. F. Hatfull & W.R. Jacobs Jr. American Society for Microbiology, Washington, D.C. pp. 69-84. 

Patents & Extensions

Patent title: Use of a 4’-Phopshopantetheinyl Transferases as a target for identifying antibiotic molecules 
Deposit N° EP05292610.2 
Deposit date: 08/12/2005 
Publication N° 1795608 
Proprietors : CNRS, Université P. Sabatier 
Inventors : Christian Chalut, Christophe Guilhot

Extension: 
Patent title: Use of a 4’-Phopshopantetheinyl Transferases as a target for identifying antibiotic molecules 
Priority deposit N° EP05292610.2 
Deposit N° PCT/IB2006004075 
Deposit date: 08/12/2006 
Publication N° WO2007069089 
Proprietors : CNRS, Université P. Sabatier 
Inventors : Christian Chalut, Christophe Guilhot

Patent title: Use of pks 13 protein coding for condensase of mycolic acids of mycobacteria and related strains as an antibiotics target 
Priority deposit N° FR20030010470 
Deposit N° US12/240469 
Deposit date: 29/09/2008 
Publication N° 
Proprietors : CNRS, Université Paris Sud 
Inventors : Christophe Guilhot, Mamadou Daffé, Christine Houssin, Damien Portevin, Célia De Sousa 

Collaborations

Collaborations.jpg

National

  • Roland Brosch, Caroline Demangel, Institut Pasteur (Paris)
  • Priscille Brodin, Franck Lafont, Philip Supply, Institut Pasteur (Lille)
  • Maria-Laura Boschiroli, ANSES (Maison-Alfort)
  • Jean-Louis Herrmann, Université de Versailles St Quentin (Montigny le Bretonneux)
  • Nathalie Winter, INRA (Tours)
  • Thierry Wirth, Museum d’Histoire Naturelle (Paris)

 

International

  • Maria-Cristina Vidal Pessolani, Instituto Oswaldo Cruz (Rio de Janeiro, Brazil)
  • Carlos Martin, Universidad de Zaragoza (Zaragoza, Spain)

Regional

  • Mamadou Daffé, Alain Milon, Lionel Mourey, Olivier Neyrolles, Jérome Nigou, Laurence Salomé, Institut de Pharmacologie et de Biologie Structurale (Toulouse) 
  • Eric Oswald, Institut de Recherche en Santé Digestive (Toulouse)  
  • Yves Génisson, Michel Baltas, Laboratoire de Synthèse et Physiochimie de Molécules d’Intérêt Biologique (Toulouse) 
  • Gwennaële Fichant, Laboratoire de Microbiologie et Génétique Moléculaires (Toulouse)
  • Laurent Kremer, Centre d’Etudes d’Agents Pathogènes et Biotechnologies pour la Santé (Montpellier) 
  • Sylvain Godreuil, Centre Hospitalo-Universitaire (Montpellier)
  • Jean-Yves Bouet, Centre de Biologie Intégrative (Toulouse)

Funding

  • Agence National de la Recherche
  • Fondation pour la Recherche Médical
  • Vaincre la Mucoviscidose
  • Centre National de la Recherche Scientifique
  • Université de Toulouse
  • European Union (H2020)