Team Projects


Camille Locht

Team Leaders:

Nathalie Mielcarek & Camille Locht

 

Group members:

 

Sophie Lecher, Technician

Emmanuelle Petit, Technician

Stephanie Slupek, Technician

Anne-Sophie Debrie, engineer

Dominique Raze, engineer

Anaïs Thiriard, Post-Doc

Romain Veyron-Churlet, CRCN CNRS

Stéphane Cauchi, CRCN CNRS

Loïc Coutte, CRCN Inserm

Olivier Gaillot, MCU-PH

Carine Rouanet, CR IPL

Camille Locht, Research Director Inserm

Nathalie Mielcarek, Research Director Inserm

The group headed by Nathalie Mielcarek and Camille Locht focuses on the development of a live vaccine against pertussis and the characterization of the protective and diagnostic antigen HBHA from Mycobacterium tuberculosis.

The live attenuated pertussis vaccine BPZE1 has been developed by the group and was shown to be protective against Bordetella pertussis infection. It has successfully undergone a phase I trial in humans and is now in further clinical development. The role of mucosal immunity in BPZE1-mediated protection is being investigated, as well as the role of the RisA regulon, involved in survival of B. pertussis in the environment. Interesting anti-inflammatory properties of the vaccine have been discovered in several models and are now the focus of mechanistic investigations.

The HBHA adhesion of M. tuberculosis, discovered by the group, is a surface-associated methylated protein involved in extra-pulmonary dissemination. The group has also established its protective potential in murine models, as well as its value for the diagnostics of latent M. tuberculosis


Selected publications

Schnöller C. et al. (2014) Attenuated Bordetella pertussis vaccine protects against respiratory syncytial virus disease via an IL-17-dependent mechanism. Am J Respir Crit Care Med 189:194-202.

Thorstensson R. et al. (2014) A phase I clinical study of a live attenuated Bordetella pertussis vaccine-BPZE1; a single centre, double-blind, placebo-controlled, dose-escalating study of BPZE1 given intranasally to healthy adult male volunteers. PLoS One 9:e83449.

Verwaerde C. et al. (2014) HBHA vaccination may require both Th1 and Th17 immune responses to protect mice against tuberculosis. Vaccine 32:6240-50.

Mascart F. & Locht C. (2015) Integrating knowledge of Mycobacterium tuberculosis pathogenesis for the design of better vaccines. Expert Rev Vaccines 14:1573-85.

Coutte L. et al. (2016) The multifaceted RisA regulon of Bordetella pertussis. Sci Rep 6:32774.

Zimmermann N. et al. (2016) Human isotype-dependent inhibitory antibody responses against Mycobacterium tuberculosis. EMBO Mol Med 8:1325-1339.


Team Leader:

Françoise Jacob-Dubuisson

 

Group members:

 

Sophie Lecher, Technician

Emmanuelle Petit, Technician

Stephanie Slupek, Technician

Anne-Sophie Debrie, engineer

Alex Rivera-Millot, PhD Student

Loïc Coutte, CRCN Inserm

Rudy Antoine, Senior Scientist, CRCN Inserm

Françoise Jacob-Dubuisson, Research Director CNRS

Our group studies the molecular mechanisms of Bordetella pertussis pathogenesis. We are interested in deciphering processes by which the bacterium interacts with its environment, such as transport and signaling, and in understanding specific genetic regulations in B. pertussis. We are using a combination of approaches including molecular biology, transcriptomics, biochemistry and biophysics to address those questions.

A long-standing project in the group is the secretion of a major adhesin of Bordetella, FHA, across the outer membrane via a specific transporter, FhaC.  The FHA/FhaC pair is a model for the Two-Partner Secretion (‘TPS’) pathway in Gram-negative bacteria. We are interested in the relationship between the structure, the dynamics and the function of FhaC.

We are also interested in two-component sensory transduction. BvgAS is the system that controls the virulence regulon of B. pertussis. We particularly focus on the structure and the function of the sensor-kinase BvgS, which represents a prototype for a large family of bacterial sensor kinases.

We have also started to investigate copper homeostasis in B. pertussis. This transition metal is both essential and toxic, and it is notably used as weapon by phagocytic cells to fight infections. Copper thus plays an important role at the host-pathogen interface. We have identified original systems used by B. pertussis to defend against excess copper and to import it when needed. We are characterizing their function and their regulation 

To address those questions, we are collaborating with experts in crystallography (V. Villeret and B. Clantin, U. Lille; R. Wintjens, U. Brussels), NMR (R. Schneider, U. Lille), mass spectrometry (S. Cianferani and O. Alba, U. Strasbourg, F. Sobott, U. Leeds), chemistry (O. Melnyk, CIIL; G. Billon, U. Lille), and with platforms of the CILL (JM Saliou, D. Hot, N. Baroi).


Selected publications

Dupré et al. (2015) Extracytoplasmic domain of the Bordetella BvgS sensor-kinase: a new signaling paradigm. PLoS Pathogens 11(3): e1004700.

Guérin et al. (2017) Two-Partner Secretion: combining efficiency and simplicity in the secretion of large proteins involved in bacteria-host and bacteria-bacteria interactions. Frontiers Cell Infect Microbiol 7:148.

Lesne et al. (2018) Coiled coils antagonism regulates activity of Venus flytrap-domain-contaning senor-kinases of the BvgS family. mBIO 9:e02052-17.

Jacob-Dubuisson et al., (2018) Structural insights into the signaling mechanisms of two-component systems. Nature Rev. Microbiol 2018 16(10):585-593.

Antoine et al. (2019) Relationships between copper-related proteomes and lifestyles in beta proteobacteria. Frontiers in Microbiology, 10:2217.


Team Leader:

Philip Supply

 

Group members:

 

Stephanie Slupek, Technician

Carine Rouanet, Research Scientist Institut Pasteur de Lille

Christine Demanche, MCU, Université Lille 2

Philip Supply, Research Director CNRS

Our group focuses on the genomics and evolution of M. tuberculosis, and on translational research to develop new diagnostic tools that are urgently needed to better combat drug resistant TB. Based on genomic screening and identification of genetic markers called MIRU-VNTRs, we developed a powerful genotyping approach for molecular tracing of M. tuberculosis strains, which is used as an international standard e.g. by the US CDC and ECDC for molecular-guided epidemiological surveillance of tuberculosis (TB). We complement this approach with whole genome sequencing (WGS) for studying TB strain population structures and lineages with high or low epidemiological success from worldwide. We identified early branching lineages of non-clonal TB bacilli called M. canettii/M. prototuberculosis with exceptional properties, including a lower virulence/persistence relative to M. tuberculosis, linked to a gene recombination-mediated change in bacterial surface that likely played a role in the emergence of the pathogen. We also reconstituted the evolutionary history of a major M. tuberculosis lineage called Beijing, comprising two major, epidemic multi-drug resistant clones in Eurasia.

Based on results from analysis of >3,600 strain genomes, a new all-in-one next-generation sequencing-based diagnostics directly applicable on clinical isolates, called Deeplex®-MycTB, was developed with Genoscreen for predicting M. tuberculosis drug resistance. We exploit this deep sequencing-based assay for studying heteroresistant TB and the prevalence and transmission of (drug resistant) TB in different world regions. P. Supply is among the nine lead authors having two or more papers among the 100 top-cited TB research studies over the last 15 years (Chen et al., 2015).

This research involves multiple collaborations with the industry (Genoscreen), and with national (R. Brosch, Institut Pasteur, Paris; C. Guilhot, IPBS, Toulouse; T. Wirth, Museum d’Histoire Naturelle, Paris) and international academic teams and reference centers (S. Niemann, Borstel Research Center, Germany; D. Crook, Univ. Oxford, UK; B. De Jong, Institute of Tropical Medicine/WHO Supranational TB Reference Center, Antwerp, and V. Mathys, Public Health Institute, Belgium).


Selected publications

Boritsch EC, et al. 2016. pks5-recombination-mediated surface remodelling in Mycobacterium tuberculosis emergence. Nat Microbiol 1:15019.

Merker M, et al. 2015. Evolutionary history and global spread of the Mycobacterium tuberculosis Beijing lineage. Nat Genet 47:242-249.

Walker TM, et al. 2015. Whole-genome sequencing for prediction of Mycobacterium tuberculosis drug susceptibility and resistance: a retrospective cohort study. Lancet Infect Dis 15:1193-1202.

Niemann S, Supply P. 2014. Diversity and evolution of Mycobacterium tuberculosis: moving to whole-genome-based approaches. Cold Spring Harb Perspect Med 4:a021188.

Supply, P., et al. 2013. Genome analysis of smooth tubercle bacilli provides insights into ancestry and pathoadaptation of Mycobacterium tuberculosis. Nat Genet 45 : 172-9.


Team Leader:

Alain Baulard

 

Group Members:

 

Sophie Lecher, Technician

Stephanie Slupek, Technician

Kamel Djaout, PostDoc

Rosangela Frita, PostDoc

Rudy Antoine, Senior Scientist, CR Inserm

Alain Baulard, Research Director Inserm

Antibiotic resistance in pathogenic bacteria is now insistently highlighted by scientists, medical doctors, and recently by important decision-makers as one of the most frightening threat that humanity has to combat. This situation is particularly worrisome for tuberculosis (TB) as it now undermines efforts to control the global epidemic.

Our group has been involved in the understanding of the mode of action of various anti-TB drugs called prodrugs, which require bio-activation by Mycobacterium tuberculosis enzymes to acquire their antibacterial effect. We have shown that bioactivation of prodrugs such as ethionamide, isoxyl or thiacetazone, is naturally fine-tuned in M. tuberculosis. In this context, we are developing new therapeutic approaches to by-pass this bacterial control. Using small synthetic molecules we have shown that the antibiotic activity of prodrugs can be drastically boosts, both in vitro and in vivo.

Whereas boosting prodrug-bioactivation is very effective against sensitive M. tuberculosis strains, this strategy is ineffective against bacteria mutated in the corresponding pathways, as observed for isoniazid-, pyrazinamide-, and ethionamide-resistant clinical isolates. Thus, our group has expanded his know-how to reprogram some enzymatic path of M. tuberculosis in order to re-route prodrug bioactivation through unexploited pathways. This concept of pharmacological reversion of resistance using small molecules was successfully applied to treat mice infected with M. tuberculosis otherwise resistant to ethionamide treatment. We are now applying this concept to other important prodrugs such as pyrazinamide, pretomanid and delamanid.

Finally, our team addresses some basic questions in regards to allosteric conformational changes of transcription regulators upon ligand interaction.

This multidisciplinary approach is supported by our local state of the art facilities, in particular our fully automatized screening facility in BioSafety Laboratories level 2 and 3. It involves strong interactions with our close partners of the Faculty of Pharmacy of Lille (N. Willand for Medicinal Chemistry, team U1177), Brussels Universities (R. Wintjens & A. Wohlkönig, for crystallography, ULB & VUB-VIB), Public Health Institute - Belgium (V. Mathys for Animal models). Our small molecules aborting antibiotic resistance (SMARt) are now on preclinical development in collaboration with Bioversys AG (Basel) & GSK (Madrid).


Selected publications

N. Willand et al., Synthetic EthR inhibitors boost antituberculous activity of ethionamide. Nat. Med. 15, 537-544 (2009).

X. Carette et al., Structural activation of the transcriptional repressor EthR from Mycobacterium tuberculosis by single amino acid change mimicking natural and synthetic ligands. Nucleic Acids Res 40, 3018-3030 (2012).

B. Villemagne et al., Ligand efficiency driven design of new inhibitors of Mycobacterium tuberculosis transcriptional repressor EthR using fragment growing, merging, and linking approaches. J. Med. Chem. 57, 4876-4888 (2014).

C. J. Queval et al., STAT3 Represses Nitric Oxide Synthesis in Human Macrophages upon Mycobacterium tuberculosis Infection. Sci Rep 6, 29297 (2016).

N. Blondiaux et al., Reversion of antibiotic resistance in Mycobacterium tuberculosis by spiroisoxazoline SMARt-420. Science. 355, 1206 (2017).