Bernard Corvilain – Xavier De Deken – Françoise Miot – Duox Lab

Our research

The DUOXLab – From H2O2 generator DUOX to oxidative stress and cancer development.
Our research group is part of the Institute of Interdisciplinary Research, IRIBHM (Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire). The Institute is localized at the Medical School of the University of Brussels (Université Libre de Bruxelles, ULB).

Hydrogen peroxide (H2O2) is a highly reactive chemical molecule originally considered as an antibacterial agent. In excess, H2O2 induces an oxidative stress responsible for different pathologies including cardiovascular diseases, neurodegeneration, tumorigenesis or ageing. It is now widely accepted that many cell types, other than the phagocytes, produce reactive oxygen species (ROS).

The thyroid and hydrogen peroxide.
The main function of the thyroid is the uptake and concentration of iodide via the Na+/I- symporteur (NIS) for thyroid hormone biosynthesis. Thyroid hormones, tri- and tetra-iodothyronine (T3 and T4), results from thyroperoxidase (TPO)-catalyzed iodination of thyroglobulin (TG) tyrosine residues and coupling of iodotyrosines. The thyroid hormones act on multiple organs in the body and influence critical physiological processes. They increase the metabolic rate and affect protein synthesis modulating the human body thermoregulation. Foremost, they are absolutely required for normal growth and neuronal maturation, preventing cretinism.

 

The thyroid gland generates massive amounts of H2O2 required for the biosynthesis of thyroid hormones. In 2000, we have discovered and characterized the molecular nature of the proteins responsible for this thyroid H2O2-generating system: the DUOX1 and DUOX2 enzymes. These proteins belong to a novel protein family, the NADPH-oxidases, presenting a catalytic domain involved in ROS generation.

 

Our multidisciplinary projects aim to better characterize the function of these new oxidases and to study their physiological roles using human biological samples and transgenic animal models. Our research group tries to elucidate the mechanisms governing the processing and maturation of DUOX enzymes allowing their correct addressing at the cell surface. Moreover, using molecular approaches, the interactions between DUOX and their maturation factors DUOXA that influence the type of ROS generated (H2O2 or O2.-) are investigated.

 

 

In vivo investigation of DUOX/DUOXA physiological roles: Congenital hypothyroidism and transgenic mouse models
Congenital hypothyroidism (CH) is the most common congenital endocrine disorder affecting 1 in every 3000 newborns. In 20% of the patients, permanent CH is due to biochemical defects causing dyshormonogenesis. Transient CH is often associated with a temporally limited exposure to external factors during pregnancy such as a lack or an excess of iodide intake, transplacental antibodies, or antithyroid drug-based treatments. The first genetic basis for transient CH has been proposed with the discovery of affected patients bearing mutations in DUOX2 gene. Several published DUOX2 missense mutations have been characterized in the laboratory. We have demonstrated that they cause DUOX expression and/or maturation defects resulting in the absence or reduced H2O2 generation. Recently, we identified the first inactivating DUOX2 mutation localized in the catalytic core of the oxidase. In collaboration with Pr. G Van Vliet (University of Montreal, Canada) and Pr. DP Carvalho (University of Rio de Janeiro, Brazil), we continue to screen for natural DUOX/DUOXA mutations in CH patients to help for genetic counseling.

In collaboration with several academic groups around the world, we have also access to transgenic mice very useful to analyze the physiological roles of these novel H2O2 generators in the thyroid metabolism, but also in the other DUOX expressing tissues.

 

 

Excess of H2O2: Risk factor in cancer development ?
Up to 50% of the population above 60 years old present thyroid nodules and 5% of these nodules will degenerate into cancers. Irradiation is the only environmental risk factor clearly implicated in thyroid cancer pathogenesis. However, irradiation is certainly not responsible for the majority of thyroid tumors. We hypothesized that the elevated frequency of thyroid tumors could be partially explained by the mutagenic environment present in the thyroid, resulting from the hormonogenic hydrogen peroxide.

We have demonstrated that non-lethal concentrations of H2O2 provoke multiple mutagenic DNA double strand breaks in primo-cultured human thyrocytes. We have also shown that DNA breaks induced by H2O2 were more slowly repaired than those induced by irradiation supporting the hypothesis that generation of H2O2 in thyroid could also play a role in mutagenesis particularly in case of antioxidant defense deficiency .

Characterization of the thyroid-H2O2 generating system in the zebrafish model
Zebrafish has been recognized as an attractive organism to study many biological processes on vertebrate development.

The zebrafish thyroid is not organized in one compact gland encapsulated by connective tissue but consist of follicles of variable shape and diameter that lie individually between the first gill arch and the bulbus arteriosus along the ventral aorta. Compared to the two DUOX and two DUOXA genes present in mammals, only one duox and one single duoxa gene have been identified in the zebrafish genome. Expression of duox mRNA has been demonstrated in various extra-thyroid tissues including the brain, the epidermis, the intestine and the ultimobranchial bodies. In collaboration with the Costagliola Lab  (Pr. S. Costagliola), we explored the role of duox and duoxa in thyroid function during zebrafish development using transgenic fish lines invalidated for the related genes.

Group members

Principal Investigators
Françoise Miot, PhD (fmiot@ulb.ac.be)

phone # +32 (0)2 555 4152

Françoise Miot obtained a Master’s degree in Biochemical Sciences from the Université Libre de
Bruxelles (ULB-1979). After her PhD in Biochemical Sciences “Study of the mechanisms controlling
cAMP accumulation by phosphodiesterase activity” (IRIBHM-ULB-1986), Françoise performed a
postdoctoral training at the Catholic University of Leuven (KUL) in the laboratory of Pr. De Wulf
(1986-1988). Until 1991, she worked in the Unit of Biomedical Magnetic Resonance (ULB-VUB).
She obtained a permanent research position in the IRIBHM research center in 1991, and founded the
DUOX laboratory with Dr. B. Corvilain in 1997.

Bernard Corvilain, MD (bcorvila@ulb.ac.be)

phone # +32 (0)2 555 3407

Bernard Corvilain graduated as a Medical Doctor in 1984 and was board certified in Internal
Medicine in 1992 at ULB. Bernard obtained his PhD in Medical sciences “Regulation of the thyroid
H2O2 generating system: Implications in the control of the thyroid hormone biosynthesis” (IRIBHM ULB-1995). He received a research fellowship from the Erasme Foundation (1987-1988, 1998-1999) and he was a research assistant from the Belgian National Research Scientific Fund (FRS-FNRS – 1988-1990).
Bernard is the clinical director of the endocrinology department at the Erasme Hospital (ULB) and board member of the Belgian Thyroid Club.

Xavier De Deken, PhD (xdedeken@ulb.ac.be)

phone # +32 (0)2 555 4152

Xavier De Deken obtained a Master’s degree in Biomedical Sciences from ULB (1997). After his PhD
in Biomedical Sciences “Cloning and the characterization of the thyroid H2O2 generating system (2002 – ULB), Xavier performed a postdoctoral training at the IRCM, Montreal, Canada (2002 – 2004). Xavier joined back the laboratory in 2004 as a postdoctoral fellow. He obtained a permanent research position in the lab in 2010. His main research interests concern the biochemical characterization of the DUOX oxidases and their role in pathophysiological processes.

PhD Students :

Nicoletta Giusti
Nicoletta Giusti obtained a Master’s degree in Medical Biotechnology in 2010 from the University of
Perugia (Italy) and a Master’s degree in Bioinformatic in 2012 from the University “La Sapienza” of
Roma (Italy). She joined the DUOX Lab in January 2013 starting her PhD in Biomedical Sciences at
ULB. Her main research concerns are the characterization of the thyroid-H2O2 generating system
during zebrafish development.

Louise Poncelet
Louise Poncelet obtained her Master’s degree in Biomedical Sciences from ULB (2013). She started
her PhD in Biomedical Sciences in 2013 at IRIBHM-ULB. Her main research concerns are the study
of the molecular mechanisms involved in the regulation of DUOX activity and maturation mediated by
their maturation factors DUOXA.

KYRILLI Elina, MD
Aglaia Kyrilli was graduated as Medical Doctor in 2007 from the National University of Athens Greece and board certified specialist in Internal Medicine-Endocrinology at ULB in 2014. She joined the DUOX Lab in October 2014 and started her PhD in Medical Sciences. Her main research concerns the characterization of thyrocyte DNA damage response after radioiodine exposure in comparison to γ-radiation and H2O2

Karima Merakchi
Karima Merakchi, after a Master in Biotechnology and Molecular and Cellular Pathology in the University of Science and Technology of Algiers, started and obtained her Master’s degree in Biochemistry and Molecular and Cellular Biology from ULB in 2017. She started her PhD in Biomedical Sciences in 2017 and joined the lab. Her main research concerns are the study of the implication of the interleukin-4 in thyroid auto-immune diseases using a new mouse model overexpressing this cytokine in the thyroid tissue.

Sami Djerbib

Sami Djerbib obtained his veterinary diploma at the National Veterinary School of Algiers (ENSV) in 2015 and completed a Master of specialization at the University of Liège in 2016. He started his PhD in Biomedical Sciences in 2018 at ULB and joined the lab. His main research concerns the study of the implication of hydrogen peroxide (H2O2) in thyroid cancers using a new mouse model overexpressing the H2O2 generator DUOX/DUOXA in the thyroid tissue.

Technicians :
Chantal Degraef

Sang Van Tran
Sang Van Tran obtained his BS in Biotechnology from the Institut Roger Lambion in 2018. After performing a research training at the IRIBHM institute, Sang Van Tran joins our lab in 2018 to replace our former technician.

Publications

1: Dickinson JD, Sweeter JM, Warren KJ, Ahmad IM, De Deken X, Zimmerman MC, Brody SL. Autophagy regulates DUOX1 localization and superoxide production in airway epithelial cells during chronic IL-13 stimulation. Redox Biol. 2018 Apr;14:272-284. doi: 10.1016/j.redox.2017.09.013. Epub 2017 Sep 22. PubMed PMID: 28982074; PubMed Central PMCID: PMC5635347.

2: Strickaert A, Saiselet M, Dom G, De Deken X, Dumont JE, Feron O, Sonveaux P, Maenhaut C. Cancer heterogeneity is not compatible with one unique cancer cell metabolic map. Oncogene. 2017 May 11;36(19):2637-2642. doi: 10.1038/onc.2016.411. Epub 2016 Oct 31. Review. PubMed PMID: 27797377; PubMed Central PMCID: PMC5442421.

3: Eskalli Z, Achouri Y, Hahn S, Many MC, Craps J, Refetoff S, Liao XH, Dumont JE, Van Sande J, Corvilain B, Miot F, De Deken X. Overexpression of Interleukin-4 in the Thyroid of Transgenic Mice Upregulates the Expression of Duox1 and the Anion Transporter Pendrin. Thyroid. 2016 Oct;26(10):1499-1512. PubMed PMID: 27599561; PubMed Central PMCID: PMC5067804.

4: Ameziane-El-Hassani R, Talbot M, de Souza Dos Santos MC, Al Ghuzlan A, Hartl D, Bidart JM, De Deken X, Miot F, Diallo I, de Vathaire F, Schlumberger M, Dupuy C. NADPH oxidase DUOX1 promotes long-term persistence of oxidative stress after an exposure to irradiation. Proc Natl Acad Sci U S A. 2015 Apr 21;112(16):5051-6. doi: 10.1073/pnas.1420707112. Epub 2015 Apr 6. PubMed PMID: 25848056; PubMed Central PMCID: PMC4413347.

5: De Deken X, Corvilain B, Dumont JE, Miot F. Roles of DUOX-mediated hydrogen peroxide in metabolism, host defense, and signaling. Antioxid Redox Signal. 2014 Jun 10;20(17):2776-93. doi: 10.1089/ars.2013.5602. Epub 2013 Oct 25. Review. PubMed PMID: 24161126.

6: Fink K, Martin L, Mukawera E, Chartier S, De Deken X, Brochiero E, Miot F, Grandvaux N. IFNβ/TNFα synergism induces a non-canonical STAT2/IRF9-dependent pathway triggering a novel DUOX2 NADPH oxidase-mediated airway antiviral response. Cell Res. 2013 May;23(5):673-90. doi: 10.1038/cr.2013.47. Epub 2013 Apr 2. Erratum in: Cell Res. 2014 Apr;24(4):509. PubMed PMID: 23545780; PubMed Central PMCID: PMC3641604.

7: Hadad I, Veithen A, Springael JY, Sotiropoulou PA, Mendes Da Costa A, Miot F, Naeije R, De Deken X, Entee KM. Stroma cell-derived factor-1α signaling enhances calcium transients and beating frequency in rat neonatal cardiomyocytes. PLoS One. 2013;8(2):e56007. doi: 10.1371/journal.pone.0056007. Epub 2013 Feb 27. PubMed PMID: 23460790; PubMed Central PMCID: PMC3584107.

8: Raad H, Eskalli Z, Corvilain B, Miot F, De Deken X. Thyroid hydrogen peroxide production is enhanced by the Th2 cytokines, IL-4 and IL-13, through increased expression of the dual oxidase 2 and its maturation factor DUOXA2. Free Radic Biol Med. 2013 Mar;56:216-25. doi: 10.1016/j.freeradbiomed.2012.09.003. Epub 2012 Sep 23. PubMed PMID: 23010498.

9: Hoste C, Dumont JE, Miot F, De Deken X. The type of DUOX-dependent ROS production is dictated by defined sequences in DUOXA. Exp Cell Res. 2012 Nov 1;318(18):2353-64. doi: 10.1016/j.yexcr.2012.07.007. Epub 2012 Jul 16. PubMed PMID: 22814254.

10: Grasberger H, De Deken X, Mayo OB, Raad H, Weiss M, Liao XH, Refetoff S. Mice deficient in dual oxidase maturation factors are severely hypothyroid. Mol Endocrinol. 2012 Mar;26(3):481-92. doi: 10.1210/me.2011-1320. Epub 2012 Feb 2. PubMed PMID: 22301785; PubMed Central PMCID: PMC3286189.

11: Donkó A, Ruisanchez E, Orient A, Enyedi B, Kapui R, Péterfi Z, de Deken X, Benyó Z, Geiszt M. Urothelial cells produce hydrogen peroxide through the activation of Duox1. Free Radic Biol Med. 2010 Dec 15;49(12):2040-8. doi: 10.1016/j.freeradbiomed.2010.09.027. PubMed PMID: 21146788.

12: Kwon J, Shatynski KE, Chen H, Morand S, de Deken X, Miot F, Leto TL, Williams MS. The nonphagocytic NADPH oxidase Duox1 mediates a positive feedback loop during T cell receptor signaling. Sci Signal. 2010 Aug 3;3(133):ra59. doi: 10.1126/scisignal.2000976. PubMed PMID: 20682913; PubMed Central PMCID: PMC2941205.

13: Hoste C, Rigutto S, Van Vliet G, Miot F, De Deken X. Compound heterozygosity for a novel hemizygous missense mutation and a partial deletion affecting the catalytic core of the H2O2-generating enzyme DUOX2 associated with transient congenital hypothyroidism. Hum Mutat. 2010 Apr;31(4):E1304-19. doi: 10.1002/humu.21227. PubMed PMID: 20187165.

14: Dumont JE, De Deken X, Miot F, Corvilain V, Contempré B, Goyens R, Massart C, Van Sande J, Allaoui A, Botteaux A. H2O2, signal, substrate, mutagen and chemorepellent from physiology to biochemistry and disease. Bull Mem Acad R Med Belg. 2010;165(5-6):231-4; discussion 235. PubMed PMID: 21510483.

15: Song Y, Ruf J, Lothaire P, Dequanter D, Andry G, Willemse E, Dumont JE, Van Sande J, De Deken X. Association of duoxes with thyroid peroxidase and its regulation in thyrocytes. J Clin Endocrinol Metab. 2010 Jan;95(1):375-82. doi: 10.1210/jc.2009-1727. Epub 2009 Dec 1. PubMed PMID: 19952225.

16: Allaoui A, Botteaux A, Dumont JE, Hoste C, De Deken X. Dual oxidases and hydrogen peroxide in a complex dialogue between host mucosae and bacteria. Trends Mol Med. 2009 Dec;15(12):571-9. doi: 10.1016/j.molmed.2009.10.003. Epub 2009 Nov 11. PubMed PMID: 19913458.
17: Driessens N, Versteyhe S, Ghaddhab C, Burniat A, De Deken X, Van Sande J, Dumont JE, Miot F, Corvilain B. Hydrogen peroxide induces DNA single- and double-strand breaks in thyroid cells and is therefore a potential mutagen for this organ. Endocr Relat Cancer. 2009 Sep;16(3):845-56. doi: 10.1677/ERC-09-0020. Epub 2009 Jun 9. PubMed PMID: 19509065.

18: Rigutto S, Hoste C, Grasberger H, Milenkovic M, Communi D, Dumont JE, Corvilain B, Miot F, De Deken X. Activation of dual oxidases Duox1 and Duox2: differential regulation mediated by camp-dependent protein kinase and protein kinase C-dependent phosphorylation. J Biol Chem. 2009 Mar 13;284(11):6725-34. doi: 10.1074/jbc.M806893200. Epub 2009 Jan 14. PubMed PMID: 19144650; PubMed Central PMCID: PMC2652333.

19: Znaidi S, Weber S, Al-Abdin OZ, Bomme P, Saidane S, Drouin S, Lemieux S, De Deken X, Robert F, Raymond M. Genomewide location analysis of Candida albicans Upc2p, a regulator of sterol metabolism and azole drug resistance. Eukaryot Cell. 2008 May;7(5):836-47. doi: 10.1128/EC.00070-08. Epub 2008 Apr 4. PubMed PMID: 18390649; PubMed Central PMCID: PMC2394978.

20: Znaidi S, De Deken X, Weber S, Rigby T, Nantel A, Raymond M. The zinc cluster transcription factor Tac1p regulates PDR16 expression in Candida albicans. Mol Microbiol. 2007 Oct;66(2):440-52. PubMed PMID: 17897373.

21: Song Y, Driessens N, Costa M, De Deken X, Detours V, Corvilain B, Maenhaut C, Miot F, Van Sande J, Many MC, Dumont JE. Roles of hydrogen peroxide in thyroid physiology and disease. J Clin Endocrinol Metab. 2007 Oct;92(10):3764-73. Epub 2007 Jul 31. Review. PubMed PMID: 17666482.

22: Rigutto S, Hoste C, Dumont JE, Corvilain B, Miot F, De Deken X. Duox1 is the main source of hydrogen peroxide in the rat thyroid cell line PCCl3. Exp Cell Res. 2007 Nov 1;313(18):3892-901. Epub 2007 Jun 29. PubMed PMID: 17643428.

23: Grasberger H, De Deken X, Miot F, Pohlenz J, Refetoff S. Missense mutations of dual oxidase 2 (DUOX2) implicated in congenital hypothyroidism have impaired trafficking in cells reconstituted with DUOX2 maturation factor. Mol Endocrinol. 2007 Jun;21(6):1408-21. Epub 2007 Mar 20. PubMed PMID: 17374849.

24: Milenkovic M, De Deken X, Jin L, De Felice M, Di Lauro R, Dumont JE, Corvilain B, Miot F. Duox expression and related H2O2 measurement in mouse thyroid: onset in embryonic development and regulation by TSH in adult. J Endocrinol. 2007 Mar;192(3):615-26. PubMed PMID: 17332529.

25: Saidane S, Weber S, De Deken X, St-Germain G, Raymond M. PDR16-mediated azole resistance in Candida albicans. Mol Microbiol. 2006 Jun;60(6):1546-62. PubMed PMID: 16796687.

26: MacPherson S, Akache B, Weber S, De Deken X, Raymond M, Turcotte B. Candida albicans zinc cluster protein Upc2p confers resistance to antifungal drugs and is an activator of ergosterol biosynthetic genes. Antimicrob Agents Chemother. 2005 May;49(5):1745-52. PubMed PMID: 15855491; PubMed Central PMCID: PMC1087678.

27: Wang D, De Deken X, Milenkovic M, Song Y, Pirson I, Dumont JE, Miot F. Identification of a novel partner of duox: EFP1, a thioredoxin-related protein. J Biol Chem. 2005 Jan 28;280(4):3096-103. Epub 2004 Nov 22. PubMed PMID: 15561711.

28: De Deken X, Raymond M. Constitutive activation of the PDR16 promoter in a Candida albicans azole-resistant clinical isolate overexpressing CDR1 and CDR2. Antimicrob Agents Chemother. 2004 Jul;48(7):2700-3. PubMed PMID: 15215129; PubMed Central PMCID: PMC434214.

29: De Deken X, Wang D, Dumont JE, Miot F. Characterization of ThOX proteins as components of the thyroid H(2)O(2)-generating system. Exp Cell Res. 2002 Feb 15;273(2):187-96. PubMed PMID: 11822874.

30: De Deken X, Wang D, Many MC, Costagliola S, Libert F, Vassart G, Dumont JE, Miot F. Cloning of two human thyroid cDNAs encoding new members of the NADPH oxidase family. J Biol Chem. 2000 Jul 28;275(30):23227-33. PubMed PMID: 10806195.

31: De Deken X, Vilain C, Van Sande J, Dumont JE, Miot F. Decrease of telomere length in thyroid adenomas without telomerase activity. J Clin Endocrinol Metab. 1998 Dec;83(12):4368-72. PubMed PMID: 9851779.