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Barrière de mucus

Workpackage : WP1

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Our team is highly interdisciplinary. It brings together students and researchers from a variety of disciplines, including pharmaceutical sciences, medical sciences, biochemistry, molecular biology and physiology, to work side by side.

Current members of the team are:
Jean-Luc Desseyn, PhD, HDR, head
Valérie Gouyer, PhD, HDR, Eng. CHU
Ségolène Plet, Tech.
Mylène Magnien, Tech.
Frédéric Gottrand, MD, PhD, PUPH
David Séguy, MD, PhD, PUPH (adjunct to the group)
Guillaume Lacroix, Thesis D2

Previous PhD students:
Hélène Valque, PhD (2008-2011), Project leader/Eng., Servier Labs, France
Céline Portal, PhD (2013-2016), Postdoctoral fellow, Johns Hopkins University, Wilmer Eye Institute, Baltimore
Bastien Demouveau, PhD (2016-2019), Project manager at OptimHal-pProtecSom, France
Salah Amini, Postdoc (under supervision of D. Séguy)

[Recent achievements] [Events] [Publications][Gallery]

Research topics

The laboratory uses recombinant molecules, cellular biology, genetically modified murine models and in vivo imaging as part of an integrated approach (i) to study the mechanistic basis of human diseases linked to mucus and mucins, (ii) to identify new biomarkers and (iii) to develop innovative therapeutic treatments. We examine the mechanisms involved in the protection by the mucus hydrogel of the underlying epithelium (nose, trachea, lung, intestine, ear, eye, vagina) and how we can modulate the mucus properties.

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Recent achievements

Our events

2019
Microrheology experiments using bead scattering and tracking of bacteria and sperm cells in mucus from murine colon and from HT29-MTX cells were performed. Comparison between wild-type mice and our transgenic line, secreting in its intestine a recombinant molecule composed of 12 CYS domains (rCYSx12) of gelling mucins, and mucus samples from co-cultured HT29 cells with HT29 cells secreting rCYSx12, demonstrate that the recombinant molecule decreases mucus permeability and impedes movement of motile cells. In addition, the model of emperical obstruction scaling shows that poly-CYS enrichment decreases the size of the mucin mesh in mice. Our data show that the CYS-domain is crucial in the properties of mucus hydrogels.
Bastien_SciRep2019
The full paper in open access here.

2017
The changes in mucus cell density and mucin production are established characteristics of many mucosal conditions. To better understand the role of the gel-forming mucin MUC5B and its value as a biomarker, we have created a genetically modified mouse Muc5b-GFP using homologous recombination. Our model allows the visualization of the mucin production in anesthetized mice as green mucus in the trachea and nose. In the vagina, Muc5b production greatly varies during estrous cycle, with a peak during proestrus and in pregnant mice. We also observed that Muc5b is expressed earlier in embryonic lung than previously thought. We were also surprised by the high number of Muc5b+ goblet cells in the conjunctiva. We then used this tissue to demonstrate that the density of Muc5b+ goblet cells can easily be monitored in living mice using confocal laser endomicroscopy in an experimental dry eye model. Full story in open access here and here.
pCLE Conjunctiva Muc5b
Eye conjunctiva of Muc5b-GFP transgenic mouse observed by probe-based confocal laser endomicroscopy

June 2020: The next Gordon Research Conference on Cilia, Mucus and Mucociliary Interactions has been postponed to the week of May 2-7 and relocated to Switzerland (Les Diablerets).
Feb. 2020: J-L Desseyn will moderate a session on Mucoactive agents to alter mucus properties in cystic fibrosis and will contribute as a speaker at the next CF Conference, to be held in Lyon, France from 3-6 June 2020. Cancelled
Jan. 2020: Jean-Luc Desseyn will be the keynote speaker at the annual meeting of the AE2BM (French association of professors and teachers in Biochemistry and Molecular Biology from the faculties of pharmacy), which will be held on 20 March 2020 in the Salle des Actes of the Facuté de Pharmacie de Paris. Cancelled
Dec. 2019: Salah Amini defended his thesis with flying colors. Congratulations to him. The post-defence reception was an opportunity to appreciate the culinary specialties of the region of origin of Salah. Thank you to his loved ones for this wonderful shared moment.
Oct. 2019: Congratulations to Bastien Demouveaux and Thomas Hueso for successfully defending their PhD thesis. Bastien is now Project Manager in a Biotech and Thomas joined the Gustave Roussy Cancer Centre. We wish them best luck.
March 2019: Congrat. to Bastien who received a financial support to present his thesis work at the 19th FEBS Young Scientists’ Forum and the 44th FEBS Congress that wiil be held in Krakow, 3-11 July.
Jan. 2019: Congrat. to Bastien who has been selected for an oral presentation at the Gordon Research Seminar next month.
Feb. 2019: The conference program of the 2019-Gordon Research Conference (Feb. 17-22) in Italy on Cilia, Mucus and Mucociliary Interactions is known. See the exciting program here. We hope to meet many people from the field.
June 2018: Congratulation to Guillaume Lacroix for his 3-yr PhD competitive fellowship starting next October.
Mar. 2018: After the exciting meeting on CF in Greece, we’ll be delighted to present our data at the 41^st European Cystic Fibrosis Conference in Belgrade (6-9 June).
Jan. 2018: It’s a pleasure to welcome Salah Amini, a new PhD student, who will work on the mucus layer reinforcement during chemotherapy.
Dec. 2017: We will present our most recent results at the 15^th ECFS (European Cystic Fibrosis) Basic Science Conference that will take place in Loutraki, Greece in March 2018.
Nov. 2017: Next January (2018), we will be at the 33^rd Transatlantic Airway conference (Lucerne, Switzerland) on lung biology.
Jul. 2017: We were at the International Workshop on Carcinoma-Associated Mucins in Cambridge in July to present our most recent results.
Jan. 2017: Céline Portal, PhD, joined the Iomini laboratory as a post-doctoral fellow. Good luck to her in New York!
Dec. 2016: Céline Portal successfully defended her PhD thesis with flying colors.
Oct. 2016: Bastien Demouveaux stays in the group for a 3-year PhD thesis on gel-forming mucin and mucus properties. Congratulations to Bastien for earning the Région Hauts-de-France/ hospital Fellowship!
Aug. 2016: Congratulations to our PhD student Céline Portal for winning one of the 15^th TFOS awards to attend the upcoming 8^th International Conference (Sept. 2016) on the Tear Film & Ocular Surface: Basic Science and Clinical Relevance (Montpellier, France). She will present her recent data on mucins and dry eye, and she will also co-chair the session ‘Are there good animal models for human dry eye disease?’.

[Top][Recent achievements] [Events] [Gallery]

Publications

Articles

Impact and consequences of intensive chemotherapy on intestinal barrier and microbiota in acute myeloid leukemia: The role of mucosal strengthening
Hueso T, Ekpe K, Mayeur C, Gatse A, Joncquel-Chevallier Curt M, Gricourt G, Rodriguez C, Burdet C, Ulmann G, Neut C, Amini SE, Lepage P, Raynard B, Willekens C, Micol JB, De Botton S, Yakoub-Agha I, Gottrand F, Desseyn JL, Thomas M, Woerther PL, Seguy D
Gut Microbes 2020;12(1):e1800897. doi:1080/19490976.2020.1800897

Importance of the Phospholipid Core for Mucin Hydrogel Penetration and Mucosal Cell Uptake of Maltodextrin Nanoparticles
Fasquelle F, Carpentier R, Demouveaux B, Desseyn JL, Betbeder D
ACS Appl Bio Mater 2020;3(9):5741-5749. doi:10.1021/acsabm.0c00521

Mucin CYS domain stiffens the mucus gel hindering bacteria and spermatozoa
Demouveaux B, Gouyer V, Robbe-Masselot C, Gottrand F, Narita T, Desseyn JL
Sci Rep 2019;9(1):16993. doi:10.1038/s41598-019-53547-x

Early life nutrition influences susceptibility to chronic inflammatory colitis in later life
Ley D, Desseyn JL, Gouyer V, Plet S, Tims S, Renes I, Mischke m, Gottrand F
Sci Rep 2019;9(1):18111. doi:10.1038/s41598-019-54308-6

Muc5b-deficient mice develop early histological lung abnormalities
Valque H, Gouyer V, Duez C, Leboeuf C, Marquillies P, Le Bert M, Plet S, Ryffel B, Janin A, Gottrand F, Desseyn JL
Biol Open 2019;8(11):bio046359. doi:10.1242/bio.046359

Muc5b is mainly expressed and sialylated in the nasal olfactory epithelium whereas Muc5ac is exclusively expressed and fucosylated in the nasal respiratory epithelium
Amini S-E, Gouyer V, Portal C, Gottrand F, Desseyn JL
Histochem Cell Biol 2019;152:167-174. doi:10.1007/s00418-019-01785-5

Non-C-mannosylable mucin CYS domains hindered proper folding and secretion of mucin
Gouyer V, Demouveaux B, Lacroix G, Valque H, Gottrand F, Desseyn JL
BBRC 2018;506(4):812-818. doi:10.1016/j.bbrc.2018.10.138
Open-access HAL here

Long-term dietary (n-3) polyunsaturated fatty acids show benefits to the lungs of Cftr F508del mice
Portal C, Gouyer V, Léonard R, Husson MO, Gottrand F, Desseyn JL
PLoS One 2018;13(6):e0197808. doi:10.1371/journal.pone.0197808

Systematic gene overexpression in Candida albicans identifies a regulator of early adaptation to the mammalian gut
Znaidi S, van Wijlick L, Hernández‐Cervantes A, Sertour N, Desseyn JL, Vincent F, Atanassova R, Gouyer V, Munro CA, Bachellier‐Bassi S, Dalle F, Jouault T, Bougnoux ME, d'Enfert C
Cell Microbiol 2018;20(11):e12890. doi:10.1111/cmi.12890

In vivo imaging of the Muc5b gel-forming mucin
Portal C, Gouyer V, Magnien M, Plet S, Gottrand F, Desseyn JL
Sci Rep 2017;7:44591. doi:10.1038/srep44591

Preclinical mouse model to monitor live Muc5b-producing conjunctival goblet cell density under pharmacological treatments
Portal C, Gouyer V, Gottrand F, Desseyn JL
PLoS One 2017;12(3):e0174764. doi:10.1371/journal.pone.0174764

Delivery of a mucin domain enriched in cysteine residues strengthens the intestinal mucous barrier
Gouyer V, Dubuquoy L, Robbe-Masselot C, Neut C, Singer E, Plet S, Geboes K, Desreumaux P, Gottrand F, Desseyn JL
Sci Rep 2015;5:9577. doi:10.1038/srep09577

Dietary pectin-derived acidic oligosaccharides improve the pulmonary bacterial clearance of Pseudomonas aeruginosa lung infection in mice by modulating intestinal microbiota and immunity
Bernard H, Desseyn JL, Bartke N, Kleinjans L, Stahl B, Belzer C, Knol J, Gottrand F, Husson MO
J Infect Dis 2015;211(1):156-165. doi: 10.1093/infdis/jiu391

Impact of fish oils on the outcomes of a mouse model of acute Pseudomonas aeruginosa pulmonary infection
Caron E, Desseyn JL, Sergent L, Bartke N, Husson MO, Duhamel A, Gottrand F
Br J Nutr 2015;113(2):191-199. doi: 10.1017/S0007114514003705

Pectin-Derived Acidic Oligosaccharides Improve the Outcome of Pseudomonas aeruginosa Lung Infection in C57BL/6 Mice
Bernard H, Desseyn JL, Gottrand F, Stahl B, Bartke N, Husson MO
PLoS One 2015;10(11):e0139686. doi:10.1371/journal.pone.0139686

Assembly of the respiratory mucin MUC5B: a new model for a gel-forming mucin
Ridley C, Kouvatsos N, Raynal BD, Howard M, Collins RF, Desseyn JL, Jowitt TA, Baldock C, Davis CW, Hardingham TE, Thornton DJ
J Biol Chem 2014;289(23):16409-16420. doi: 10.1074/jbc.M114.566679

MUC5B leads to aggressive behavior of breast cancer MCF7 cells
Valque H, Gouyer V, Gottrand F, Desseyn JL
PLoS One 2012;7(10):e46699. doi:10.1371/journal.pone.0046699

The mucin MUC4 and its membrane partner ErbB2 regulate biological properties of human CAPAN-2 pancreatic cancer cells via different signalling pathways
Jonckheere N, Skrypek N, Merlin J, Dessein AF, Dumont P, Leteurtre E, Harris A, Desseyn JL, Susini C, Frénois F, Van Seuningen I
PLoS One 2012;7(2):e32232. doi:10.1371/journal.pone.0032232

Abnormal expression of Muc5b in Cftr-null mice and in mammary tumors of MMTV-ras mice
Valque H, Gouyer V, Husson MO, Gottrand F, Desseyn JL
Histochem Cell Biol 2011;136(6):699-708. doi:10.1007/s00418-011-0872-5

(n-3) long-chain PUFA differentially affect resistance to Pseudomonas aeruginosa infection of male and female cftr–/– mice
Tiesset H, Bernard H, Bartke N, Beermann C, Flachaire E, Desseyn JL, Gottrand F, Husson MO
J Nutr 2011;141(6):1101-1107. doi:10.3945/jn.110.134585

The extraordinarily complex but highly structured organization of intestinal mucus-gel unveiled in multicolor images
Gouyer V, Gottrand F, Desseyn JL
PLoS One 2011;6(4):e18761. doi:10.1371/journal.pone.0018761

The characterization of the first anti-mouse Muc6 antibody shows an increased expression of the mucin in pancreatic tissue of Cftr-knockout mice
Gouyer V, Leir SH, Tetaert D, Liu Y, Gottrand F, Harris A, Desseyn JL
Histochem Cell Biol 2010;133(5):517-525. doi:10.1007/s00418-010-0688-8

Dietary (n-3) polyunsaturated fatty acids affect the kinetics of pro- and antiinflammatory responses in mice with Pseudomonas aeruginosa lung infection
Tiesset H, Pierre M, Desseyn JL, Guéry B, Beermann C, Galabert C, Gottrand F, Husson MO
J Nutr 2009;139(1):82-89. doi: 10.3945/jn.108.096115

Mucin CYS domains are ancient and highly conserved modules that evolved in concert
Desseyn JL
Mol Phylogenet Evol 2009;52(2):284-292. doi:10.1016/j.ympev.2009.03.035

Autocrine induction of invasion and metastasis by tumor-associated trypsin inhibitor in human colon cancer cells
Gouyer V, Fontaine D, Dumont P, de Wever O, Fontayne-Devaud H, Leteurtre E, Truant S, Delacour D, Drobecq H, Kerckaert JP, de Launoit Y, Bracke M, Gespach C, Desseyn JL, Huet G
Oncogene 2008;27(29):4024-4033. doi: 10.1038/onc.2008.42

Dietary n-3 fatty acids have suppressive effects on mucin upregulation in mice infected with Pseudomonas aeruginosa
Tetaert D, Pierre M, Demeyer D, Husson MO, Béghin L, Galabert C, Gottrand F, Beermann C, Guery B, Desseyn JL
Respir Res 2007;8:39. doi:10.1186/1465-9921-8-39

Epigenetic regulation (DNA methylation, histone modifications) of the 11p15 mucin genes (MUC2, MUC5AC, MUC5B, MUC6) in epithelial cancer cells
Vincent A, Perrais M, Desseyn JL, Aubert JP, Pigny P, Van Seuningen I
Oncogene 2007;26(5):6566-6576. doi:MUC5B10.1038/sj.onc.1210479

Reviews

Ocular mucins in dry eye disease
Portal C, Gouyer V, Gottrand F, Desseyn JL
Exp Eye Res 2019;186:107724. doi:10.1016/j.exer.2019.107724

Transgenic mouse reporter to study Muc5b in vivo
Desseyn JL, Portal C, Gottrand F, Gouyer V
Ann Am Thorac Soc 2018;15(Suppl 3):S149-S153. doi:10.1513/AnnalsATS.201802-085AW

Gel-forming mucin interactome drives mucus viscoelasticity
Demouveaux B, Gouyer V, Gottrand F, Narita T, Desseyn JL
Adv Colloid Interface Sci 2018;252:69-82. doi:10.1016/j.cis.2017.12.005

Early-life origin of intestinal inflammatory disorders
Ley D, Desseyn JL, Mischke M, Knol J, Turck D, Gottrand F
Nutr Rev 2017;75(3):175-187. doi:10.1093/nutrit/nuw061

Biological modeling of mucus to modulate mucus barriers
Desseyn JL, Gouyer V, Gottrand F
Am J Physiol Gastrointest Liver Physiol 2016;310(4):G225-G227. doi:10.1152/ajpgi.00274.2015

Modulation of host defence against bacterial and viral infections by omega-3 polyunsaturated fatty acids
Husson MO, Ley D, Portal C, Gottrand M, Hueso T, Desseyn JL, Gottrand F
J Infect 2016;73(6):523-535. doi:10.1016/j.jinf.2016.10.001

Long chain polyunsaturated fatty acids: immunomodulators in disease
Desseyn JL, Gottrand F
Book Dietary Components and Immune Function 2010;chapter 9:155-172.

Architecture of the large membrane-bound mucins
Desseyn JL, Tetaert D, Gouyer V
Gene 2008;410(2):215-222. doi: 10.1016/j.gene.2007.12.014

General audience

La structure des mucines conditionne les propriétés viscoélastiques des gels de mucus
Demouveaux B, Gouyer V, Magnien M, Plet S, Gottrand F, Narita T, Desseyn JL
Med Sci (Paris) 2018;34(10):806-812. doi:10.1051/medsci/20182063

Différenciation des cellules à mucus et régulation de la mucine gélifiante Muc5b : un nouvel outil pour des études ex vivo et précliniques in vivo
Desseyn JL, Portal C, Gottrand F, Gouyer V
Med Sci (Paris) 2017;33(5):478-480. doi:10.1051/medsci/20173305006

Le défaut de mucus intestinal, nouvelle cible thérapeutique
Desseyn JL
Biofutur 2016;372(1):42-47.

Modification à façon des propriétés physiques du mucus. Preuve de concept et applications potentielles
Desseyn JL, Gouyer V, Gottrand F
Med Sci (Paris) 2015;31(12):1063-1066. doi:10.1051/medsci/20153112005

Maladie de l'intestin - Renforcer le mucus pour mieux protéger
Scala B
Science et Santé (Paris) 2015;26(juillet-août):8.

Organisation du gène de mucine humaine MUC5B et bases moléculaires d’une nouvelle classification des gènes de mucines
Desseyn JL, Laine A
Regard sur la Biochimie 1999;3:33-43.

Biological tools

We have developed many tools in recent years to study mucins. Among them, here are a few that might be useful for your research.

Antibodies

Anti mouse-Muc6 antibody (rabbit polyclonal). Directed against the peptide TSKIPVTQTTTHRVPSRCIT, which is N-terminal to the tandem repeats of Muc6 (amino acids 1,387–1,406, GenBank: AY184388.1). For WB and IHC, available upon request.

← serial sections showing strong labelling of the deep gland cells with the anti-Muc6 antibody (CP4), and Muc6 within the gel layers, while Muc5ac was synthesized in the superficial/foveolar epithelium in agreement with the human distribution.
This antibody has been used in the following studies:
♦ Histochem Cell Biol. 2010; doi: 10.1007/s00418-010-0688-8.
♦ Histochem Cell Biol. 2019; doi: 10.1007/s00418-019-01785-5.

Anti mouse-Muc5b antibody (rabbit polyclonal). Directed against the 19 aa peptide DVSSGLVCYNKDQGGTFQM found in CYS domains #2/3/5 of Muc5b. For IHC, available upon request.
← Immunofluorescence showing Muc5b (green) production in bronchi. This antibody has been used in the following studies:
♦ Histochem Cell Biol. 2011; doi: 10.1007/s00418-011-0872-5.
♦ PLoS One 2012; doi: 10.1371/journal.pone.0046699.
♦ Sci Rep. 2017; doi: 10.1038/srep44591.
♦ Histochem Cell Biol. 2019; doi: 10.1007/s00418-019-01785-5.
♦ Biol Open 2019; doi: 10.1242/bio.046359.

Construct

Mini5B expression vector

Mini5B expression plasmid: Eucaryotic hybrid promoter + intron + Mini5B + Ires-Luc. The plasmid contains a Neo cassette for stable transfection. The mini5B is made of two MUC5B CYS domains flanking a full TR array of MUC5B with a functional N-glycosylation site at the C-term.
This construct with or without the Ires-Luc cassette has been used in the following studies:
♦ PLoS One 2012; doi: 10.1371/journal.pone.0046699.
♦ Biochem Biophys Res Commun. 2018; doi: 10.1016/j.bbrc.2018.10.138.

Transgenic mouse lines

Tff3-rCYSx12-eGFP

Tff3-rCYSx12-eGFP. Mouse Tg^{(Tff3/MUC5B-meGFP)1Jlcd:} Tff3 promoter driving the production/secretion of a meGFP-tagged CYSx12 (human MUC5B) recombinant molecule with Muc2 in the mouse intestine. With its large Tff3 (mItf) promoter, this transgenic mouse can be used as a Tff3 eGFP-reporter mouse. Available under a collaborative MTA.
← Immunofluorescence showing rCYSx12-eGFP (green) secreted with Muc2 (red) in the colonic mucus gel layers. This transgenic mouse has been used in the following studies:
♦ Sci Rep. 2015; doi: 10.1038/srep09577.
♦ Sci Rep. 2019; doi:10.1038/s41598-019-53547-x.
♦ Gut Microbes 2020; doi: 10.1080/19490976.2020.1800897.

Tff3-rCYSx12. The variant without the eGFP tag (named Tg208 or K208/Tg^{(Tff3/MUC5B)1Jlcd}) has been deposited and is available at EMMA. Homozygotes are viable. For details, see MGI/Jax.

Muc5b-meGFP/KO

Muc5b-meGFP/Muc5b knockout Mouse eGFP reporter for Muc5b obtained by homologous recombination (knockin). The full Muc5b glycoprotein is labelled with a monomeric eGFP tag. The last two exons of Muc5b are floxed, giving the opportunity to generate any conditional Muc5b knockout. Available under a collaborative MTA.
These transgenic mice have been used in the following studies:
♦ Sci Rep. 2017; doi: 10.1038/srep44591.
♦ PLoS One 2017; doi: 10.1371/journal.pone.0174764.
♦ Histochem Cell Biol. 2019; doi: 10.1007/s00418-019-01785-5.

Muc4-KO

Muc4 knockout. Mouse Muc4 knockout was obtained by gene targetting. The first exon was removed. This transgenic model has not been published. Available under a collaborative MTA.

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Gallery

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