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Stage proposé par Michel POLAK
Professeur, Responsable du service d’endocrinologie pédiatrique,
Hôpital Necker Enfants Malades, Paris
INSERM U845, Equipe Raphaël Scharfmann
Faculté de médecine Necker, Paris 5
156 rue de Vaugirard
75 015 Paris
Téléphone : 01 44 49 48 03
Mail : [email protected]
Directeur du Laboratoire : Paul Kelly
Titre : Implication d’une mutation de FOXE1 dans les anomalies du développement de la thyroïde
Projet de stage :
L’hypothyroïdie congénitale (HC) est une des plus fréquentes maladies endocriniennes du nouveauné (1/3800). Dans 85% des cas, l’HC est due à une anomalie du développement de la glande thyroïde
(dysgénésies thyroïdiennes, DT) (Polak, 2004). Nous avons récemment montré que cette affection
avait un caractère familial (Castanet, 2000). Les données des études génétiques permettent
d'impliquer trois facteurs de transcription TTF-1, PAX8 et FOXE1 dans le développement de la glande
thyroïde (Castanet, 2001, Trueba, 2005). Au laboratoire, nous avons trouvé une mutation de FOXE1
chez un patient atteint de DT. Nous proposons au stagiaire de faire l’étude fonctionnelle de ce facteur
de transcription muté. Ainsi, à l’aide de transfection cellulaire, la transactivation d’un gène rapporteur
sera étudiée in vitro. La responsabilité de la mutation dans le phénotype du patient pourra être ainsi
établie.
Techniques mises en œuvre par le stagiaire :
Mutagénèse dirigée de FOXE1 inséré dans un plasmide (incluant le séquençage pour vérifier la
présence de la mutation)
Techniques de bactériologie (minipreps et maxipreps)
Culture cellulaire : culture et transfection de cellules
Etude de la transactivation : tests luciférase et Galactosidase
Publications du responsable de stage au cours des 5 dernières années :
En gras publications en liaison avec le thème du stage proposé.
Carre A, Castanet M, Sura-Trueba S, Szinnai G, Van Vliet G, Trochet D, Amiel J, Leger J, Czernichow P, Scotet V, Polak
M. Polymorphic length of FOXE1 alanine stretch: evidence for genetic susceptibility to thyroid dysgenesis. Hum Genet.
2007 Aug 24
Szinnai G, Sarnacki S, Polak M. Hereditary medullary thyroid carcinoma: how molecular genetics made multiple
endocrine neoplasia type 2 a paediatric disease. Endocr Dev. 2007;10:173-87.
Castanet M, Polak M, Leger J. Familial forms of thyroid dysgenesis. Endocr Dev. 2007;10:15-28.
Beltrand J, Beregszaszi M, Chevenne D, Sebag G, De Kerdanet M, Huet F, Polak M, Tubiana-Rufi N, Lacombe D, De Paoli AM,
Levy-Marchal C. Metabolic correction induced by leptin replacement treatment in young children with Berardinelli-Seip
congenital lipoatrophy. Pediatrics. 2007 Aug;120(2):e291-6.
Vaxillaire M, Dechaume A, Busiah K, Cave H, Pereira S, Scharfmann R, de Nanclares GP, Castano L, Froguel P, Polak M;
SUR1-Neonatal Diabetes Study Group. New ABCC8 mutations in relapsing neonatal diabetes and clinical features. Diabetes.
2007 Jun;56(6):1737-41.
Valayannopoulos V, Vaxillaire M, Aigrain Y, Jaubert F, Bellanne-Chantelot C, Ribeiro MJ, Brunelle F, Froguel P, Robert JJ,
Polak M, Nihoul-Fekete C, de Lonlay P. Coexistence in the same family of both focal and diffuse forms of hyperinsulinism.
Diabetes Care. 2007 Jun;30(6):1590-2. Epub 2007 Mar 23
Polak M, Cave H. Neonatal diabetes mellitus: a disease linked to multiple mechanisms. Orphanet J Rare Dis. 2007 Mar 9;2:12.
Delonlay P, Simon A, Galmiche-Rolland L, Giurgea I, Verkarre V, Aigrain Y, Santiago-Ribeiro MJ, Polak M, Robert JJ, BellanneChantelot C, Brunelle F, Nihoul-Fekete C, Jaubert F. Neonatal hyperinsulinism: clinicopathologic correlation.Hum Pathol. 2007
Mar;38(3):387-99.
Bachelot A, Plu-Bureau G, Thibaud E, Laborde K, Pinto G, Samara D, Nihoul-Fekete C, Kuttenn F, Polak M, Touraine P. Longterm outcome of patients with congenital adrenal hyperplasia due to 21-hydroxylase deficiency. Horm Res. 2007;67(6):268-76.
Epub 2006 Dec 14.
Cauchi S, Vaxillaire M, Choquet H, Durand E, Duval A, Polak M, Froguel P. No major contribution of TCF7L2 sequence variants
to maturity onset of diabetes of the young (MODY) or neonatal diabetes mellitus in French white subjects.Diabetologia. 2007
Jan;50(1):214-6.
Szinnai G, Lacroix L, Carre A, Guimiot F, Talbot M, Martinovic J, Delezoide AL, Vekemans M, Michiels S, Caillou B,
Schlumberger M, Bidart JM, Polak M. Sodium/iodide symporter (NIS) gene expression is the limiting step for the onset
of thyroid function in the human fetus.
J Clin Endocrinol Metab. 2007 Jan;92(1):70-6.
Flechtner I, de Lonlay P, Polak M. Diabetes and hypoglycaemia in young children and mutations in the Kir6.2 subunit of the
potassium channel: therapeutic consequences. Diabetes Metab. 2006 Dec;32(6):569-80.
Alharbi M, Polak M. Complete congenital anterior pituitary insufficiency and syndrome of inapropriate antidiuretic hormone
secretion: a rare association in children.J Pediatr Endocrinol Metab. 2006 Dec;19(12):1445-9.
Pearson ER, Flechtner I, Njolstad PR, Malecki MT, Flanagan SE, Larkin B, Ashcroft FM, Klimes I, Codner E, Iotova V,
Slingerland AS, Shield J, Robert JJ, Holst JJ, Clark PM, Ellard S, Sovik O, Polak M, Hattersley AT; Neonatal Diabetes
International Collaborative Group. Switching from insulin to oral sulfonylureas in patients with diabetes due to Kir6.2 mutations.
N Engl J Med. 2006 Aug 3;355(5):467-77.
Babenko AP, Polak M, Cave H, Busiah K, Czernichow P, Scharfmann R, Bryan J, Aguilar-Bryan L, Vaxillaire M, Froguel P.
Activating mutations in the ABCC8 gene in neonatal diabetes mellitus. N Engl J Med. 2006 Aug 3;355(5):456-66.
Druet C, Dabbas M, Baltakse V, Payen C, Jouret B, Baud C, Chevenne D, Ricour C, Tauber M, Polak M, Alberti C, LevyMarchal C. Insulin resistance and the metabolic syndrome in obese French children. Clin Endocrinol (Oxf). 2006 Jun;64(6):6728.
Polak M, Legac I, Vuillard E, Guibourdenche J, Castanet M, Luton D. Congenital hyperthyroidism: the fetus as a patient.
Horm Res. 2006;65(5):235-42. Epub 2006 Mar 30.
Banaei-Bouchareb L, Peuchmaur M, Czernichow P, Polak M. A transient microenvironment loaded mainly with macrophages in
the early developing human pancreas. J Endocrinol. 2006 Mar;188(3):467-80.
Massin N, Czernichow C, Thibaud E, Kuttenn F, Polak M, Touraine P. Idiopathic premature ovarian failure in 63 young
women.Horm Res. 2006;65(2):89-95.
Szinnai G, Kosugi S, Derrien C, Lucidarme N, David V, Czernichow P, Polak M. Extending the clinical heterogeneity of
iodide transport defect (ITD): a novel mutation R124H of the sodium/iodide symporter gene and review of genotypephenotype correlations in ITD. J Clin Endocrinol Metab. 2006 Apr;91(4):1199-204.
de Lonlay P, Simon-Carre A, Ribeiro MJ, Boddaert N, Giurgea I, Laborde K, Bellanne-Chantelot C, Verkarre V, Polak M, Rahier
J, Syrota A, Seidenwurm D, Nihoul-Fekete C, Robert JJ, Brunelle F, Jaubert F. Congenital hyperinsulinism: pancreatic
[18F]fluoro-L-dihydroxyphenylalanine (DOPA) positron emission tomography and immunohistochemistry study of DOPA
decarboxylase and insulin secretion. J Clin Endocrinol Metab. 2006 Mar;91(3):933-40.
Druet C, Tubiana-Rufi N, Chevenne D, Rigal O, Polak M, Levy-Marchal C. Characterization of insulin secretion and resistance in
type 2 diabetes of adolescents. J Clin Endocrinol Metab. 2006 Feb;91(2):401-4.
Luton D, Le Gac I, Noel M, Guibourdenche J, Polak M. Thyroid function during pregnancy in women with past Graves'
disease.BJOG. 2005 Nov;112(11):1565-7.
Luton D, Le Gac I, Vuillard E, Castanet M, Guibourdenche J, Noel M, Toubert ME, Leger J, Boissinot C, Schlageter MH,
Garel C, Tebeka B, Oury JF, Czernichow P, Polak M. Management of Graves' disease during pregnancy: the key role of
fetal thyroid gland monitoring. J Clin Endocrinol Metab. 2005 Nov;90(11):6093-8.
Castanet M, Leenhardt L, Leger J, Simon-Carre A, Lyonnet S, Pelet A, Czernichow P, Polak M. Thyroid hemiagenesis is
a rare variant of thyroid dysgenesis with a familial component but without Pax8 mutations in a cohort of 22 cases.
Pediatr Res. 2005 Jun;57(6):908-13.
Castanet M, Sura-Trueba S, Chauty A, Carre A, de Roux N, Heath S, Leger J, Lyonnet S, Czernichow P, Polak M.
Linkage and mutational analysis of familial thyroid dysgenesis demonstrate genetic heterogeneity implicating novel
genes. Eur J Hum Genet. 2005 Feb;13(2):232-9.
Trueba SS, Auge J, Mattei G, Etchevers H, Martinovic J, Czernichow P, Vekemans M, Polak M, Attie-Bitach T. PAX8,
TITF1, and FOXE1 gene expression patterns during human development: new insights into human thyroid
development and thyroid dysgenesis-associated malformations.J Clin Endocrinol Metab. 2005 Jan;90(1):455-62
Polak M, Shield J. Neonatal Diabetes Mellitus -- genetic aspects 2004. Pediatr Endocrinol Rev. 2004 Dec;2(2):193-8.
Polak M, Sura-Trueba S, Chauty A, Szinnai G, Carre A, Castanet M. Molecular mechanisms of thyroid dysgenesis. Horm
Res. 2004;62 Suppl 3:14-21.
Vaxillaire M, Populaire C, Busiah K, Cave H, Gloyn AL, Hattersley AT, Czernichow P, Froguel P, Polak M. Kir6.2 mutations are
a common cause of permanent neonatal diabetes in a large cohort of French patients. Diabetes. 2004 Oct;53(10):2719-22.
Shield JP, Temple IK, Sabin M, Mackay D, Robinson DO, Betts PR, Carson DJ, Cave H, Chevenne D, Polak M. An assessment
of pancreatic endocrine function and insulin sensitivity in patients with transient neonatal diabetes in remission.Arch Dis Child
Fetal Neonatal Ed. 2004 Jul;89(4):F341-3.
Banaei-Bouchareb L, Gouon-Evans V, Samara-Boustani D, Castellotti MC, Czernichow P, Pollard JW, Polak M. Insulin cell
mass is altered in Csf1op/Csf1op macrophage-deficient mice. J Leukoc Biol. 2004 Aug;76(2):359-67.
Polak M, Le Gac I, Vuillard E, Guibourdenche J, Leger J, Toubert ME, Madec AM, Oury JF, Czernichow P, Luton D. Fetal
and neonatal thyroid function in relation to maternal Graves' disease. Best Pract Res Clin Endocrinol Metab. 2004
Jun;18(2):289-302.
Polak M, Shield J. Neonatal and very-early-onset diabetes mellitus. Semin Neonatol. 2004 Feb;9(1):59-65.
Donadieu J, Rolon MA, Thomas C, Brugieres L, Plantaz D, Emile JF, Frappaz D, David M, Brauner R, Genereau T, Debray D,
Cabrol S, Barthez MA, Hoang-Xuan K, Polak M; French LCH Study Group. Endocrine involvement in pediatric-onset
Langerhans' cell histiocytosis: a population-based study.J Pediatr. 2004 Mar;144(3):344-50.
Donadieu J, Rolon MA, Pion I, Thomas C, Doz F, Barkaoui M, Robert A, Deville A, Mazingue F, David M, Brauner R, Cabrol S,
Garel C, Polak M; French LCH Study Group. Incidence of growth hormone deficiency in pediatric-onset Langerhans cell
histiocytosis: efficacy and safety of growth hormone treatment. J Clin Endocrinol Metab. 2004 Feb;89(2):604-9.
Beregszaszi M, Jaquet D, Levine M, Ortega-Rodriguez E, Baltakse V, Polak M, Levy-Marchal C. Severe insulin resistance
contrasting with mild anthropometric changes in the adipose tissue of HIV-infected children with lipohypertrophy.Int J Obes
Relat Metab Disord. 2003 Jan;27(1):25-30.
Martin D, Licha-Muntz G, Grasset E, Greneche MO, Nouet D, Francois L, Legrand C, Polak M, Augendre-Ferrante B, TubianaRufi N, Robert JJ. Efficacy of Humalog injections before an afternoon meal and their acceptance by children and adolescents
with type 1 diabetes. Diabet Med. 2002 Dec;19(12):1026-31.
Van Maldergem L, Magre J, Khallouf TE, Gedde-Dahl T Jr, Delepine M, Trygstad O, Seemanova E, Stephenson T, Albott CS,
Bonnici F, Panz VR, Medina JL, Bogalho P, Huet F, Savasta S, Verloes A, Robert JJ, Loret H, De Kerdanet M, Tubiana-Rufi N,
Megarbane A, Maassen J, Polak M, Lacombe D, Kahn CR, Silveira EL, D'Abronzo FH, Grigorescu F, Lathrop M, Capeau J,
O'Rahilly S. Genotype-phenotype relationships in Berardinelli-Seip congenital lipodystrophy. J Med Genet. 2002
Oct;39(10):722-33. Erratum in: J Med Genet. 2003 Feb;40(2):150.
Metz C, Cave H, Bertrand AM, Deffert C, Gueguen-Giroux B, Czernichow P, Polak M; NDM French Study Group. Neonatal
diabetes mellitus. Neonatal diabetes mellitus: chromosomal analysis in transient and permanent cases.J Pediatr. 2002
Oct;141(4):483-9.
Castanet M, Park SM, Smith A, Bost M, Leger J, Lyonnet S, Pelet A, Czernichow P, Chatterjee K, Polak M. A novel lossof-function mutation in TTF-2 is associated with congenital hypothyroidism, thyroid agenesis and cleft palate. Hum Mol
Genet. 2002 Aug 15;11(17):2051-9.
Gloyn AL, Ellard S, Shield JP, Temple IK, Mackay DJ, Polak M, Barrett T, Hattersley AT. Complete glucokinase deficiency is not
a common cause of permanent neonatal diabetes. Diabetologia. 2002 Feb;45(2):290.
Vaxillaire M, Samson C, Cave H, Metz C, Froguel P, Polak M. Glucokinase gene mutations are not a common cause of
permanent neonatal diabetes in France. Diabetologia. 2002 Mar;45(3):454-5.
Leger J, Marinovic D, Garel C, Bonaiti-Pellie C, Polak M, Czernichow P. Thyroid developmental anomalies in first
degree relatives of children with congenital hypothyroidism.J Clin Endocrinol Metab. 2002 Feb;87(2):575-80.
Beringue F, Blondeau B, Castellotti MC, Breant B, Czernichow P, Polak M. Endocrine pancreas development in growth-retarded
human fetuses. Diabetes. 2002 Feb;51(2):385-91.
Castanet M, Polak M, Bonaiti-Pellie C, Lyonnet S, Czernichow P, Leger J. Nineteen years of national screening for
congenital hypothyroidism: familial cases with thyroid dysgenesis suggest the involvement of genetic factors. J Clin
Endocrinol Metab 2001; 86(5):2009-2014.
Castanet M, Lyonnet S, Bonaiti-Pellie C, Polak M, Czernichow P, Leger J. Familial forms of thyroid dysgenesis among
infants with congenital hypothyroidism. N Engl J Med 2000; 343(6):441-442.