L’insuffisance ovarienne primaire (IOP) est une cause majeure d’anovulation et d’infertilité chez la femme. Cette insuffisance est le résultat d’une déplétion du stock des follicules ou d’un blocage de leur maturation. De ce fait, la stérilité est le plus souvent définitive. Une origine génétique de cette maladie est parfois retrouvée avec des mutations du chromosome X ou des autosomes, mais dans plus de 80 % des cas l’IOP est idiopathique. L’enjeu est donc d’identifier de nouveaux gènes candidats. Dans cette étude nous validons la prévalence des mutations du gène NOBOX faisant de ce facteur un des gènes clés de l’IOP. Par ailleurs, le séquençage de 2e génération multiplex par puces PGM™ ION TORRENT nous a permis de mettre en évidence dans 20 % de la cohorte étudiée des mutations de 10 gènes, dont 4 nouveaux candidats. De façon intéressante, la présence d’au moins deux gènes mutés chez 7 patientes induit une apparition plus précoce de la maladie. Cette étude contribue à une meilleure compréhension de l’origine génétique de l’IOP et met pour la première fois en lumière le phénomène d’oligogénicité chez des patientes en IOP.

L’insuffisance ovarienne primaire (IOP) touchant environ 1 % des femmes, est une pathologie multifactorielle caractérisée par une aménorrhée et une élévation des gonadotrophines avant l’âge de 40 ans. L’IOP peut être expliquée par un défaut du développement folliculaire ou une déplétion en follicules consécutive à une anomalie de la formation ou à un épuisement anormalement rapide du stock folliculaire. L’IOP est associée à un certain nombre de syndromes et d’anomalies génétiques mais dans 80 % des cas la cause reste inconnue. Série de 100 patientes présentant une IOP avec une aménorrhée primaire ou secondaire sans prémutation de FMR1 ou d’anomalie du caryotype. Séquençage multiplex de 19 gènes par la technologie PGM™ ION TORRENT. Nous avons identifié des variations de 12 gènes parmi lesquelles 5 mutations faux-sens de trois nouveaux gènes codant des facteurs de transcription. Deux mutations de SOHLH1 (spermatogenesis and oogenesis bHLH 1) p.S174L et p.P218 T sont identifiées chez une même patiente en aménorrhée primaire et la mutation p.L306 M chez une autre patiente en aménorrhée secondaire. Des variations de FIGLA (Factor in the germline-alpha) p.A41 V et de LHX8 (Lim Homeobox 8) p.A325 V sont retrouvées chez deux patientes en aménorrhée secondaire. L’analyse fonctionnelle des variations de ces gènes montre une perte drastique de leur activité transcriptionnelle confirmant leur implication chez les patientes porteuses. Cette étude contribue à une meilleure compréhension de l’origine génétique de l’IOP et témoigne du rôle critique et précoce de ces facteurs de transcription dans la mise en place du stock folliculaire.

L’insuffisance ovarienne primaire (IOP) est une maladie rare, définie par une aménorrhée de plus de 4 mois avant l’âge de 40 ans. Une origine génétique de cette maladie est parfois retrouvée avec des mutations des autosomes (FSHR, NOBOX…) et/ou du chromosome X, mais dans plus de 80 % des cas l’IOP est idiopathique. De nouveaux gènes issus des études de modèles murins reproduisant la maladie représentent de bons candidats pour mieux comprendre cette pathologie. Les mutations de 19 nouveaux gènes ont été recherchées par puce PGM ION TORRENT au sein d’une cohorte de 109 femmes atteintes d’IOP sporadique primaire ou secondaire. Toutes les variations retrouvées ont été validées par la technique de Sanger. Nous confirmons la prévalence des mutations de NOBOX ou ESR1. De plus, nous mettons en évidence de nouvelles variations d’une kinase impliquée dans une des voies de signalisation essentielle à la croissance folliculaire. De façon intéressante, ces variations sont associées à des mutations perte de fonction de BMP15 ou GDF9, deux acteurs essentiels au dialogue ovocyte-granulosa. Les analyses fonctionnelles de ces variants sont en cours d’analyse afin de confirmer leur participation à la genèse de la maladie. Cette étude contribue à une meilleure compréhension de l’origine génétique de l’IOP et met pour la première fois en évidence le phénomène d’oligogénisme chez des patientes en IOP.

The endothelial cell (EC) dysfunction is a common characteristic of various pathologies that include atherosclerosis, hypertension, and Fabry's disease. Aware of the role of eNO and ACE in EC dysfunction, we questioned whether polymorphism of eNOS and/or ACE gene may be a common denominator in these pathologies. Patients with CHD (108), HT (109), Fabry's disease (37) and healthy subjects (control, 141) were genotyped for the eNOSG894T by RFLP-PCR technique and for eNOS4b/a, and ACEI/D polymorphisms by PCR amplification. The results of these studies were statistically evaluated. Compared to controls, the frequency of the eNOSG894T (T allele) was higher in CHD (P=0.03) and Fabry (P=0.01), while the eNOS4b/a (a allele) in CHD (P=0.01) and HT patients (P=0.01). The proportion of the ACEI/D was similar in all subjects. In CHD patients at "low risk" of atherogenic factors, the frequency of the T and a alleles of eNOS gene was high (P=0.03 and 0.02, respectively). Carriers of the T allele of eNOSG894T were over-represented (P=0.04) in Fabry subgroup with renal failure. Compared to women, the eNOS894T alleles were more frequent (P=0.03) in men with CHD and HT, whereas ACE I/D in men (P=0.03) with HT. These findings suggest: (i) the frequency of eNOSG894T and/or eNOS4b/a is significantly associated with coronary dysfunction; (ii) eNOS4b/a confers a relatively high risk of hypertension in subjects with atherogenic risk factors; (iii) the frequency of eNOSG894T is high in Fabry hemizygotes with renal complications. Therefore, eNOS gene polymorphism represent a frequent risk factor for vascular abnormalities in CHD, HT and Fabry's disease, afflictions which have in common, the endothelial dysfunction.

Acta PaediatricaVolume 91, Issue s439 p. 119-119 Fabry disease: from clinical manifestations to molecular mechanisms K Azibi, K Azibi Laboratoire de Génétique, Université Paris V, CHU Cochin, Paris, France Institut Cochin, Inserm, CNRS, Paris, FranceSearch for more papers by this authorC Heltianu, C Heltianu Institute of Cellular Biology and Pathology “N. Simionescu”, Bucharest, RumaniaSearch for more papers by this authorC Caillaud, C Caillaud Laboratoire de Génétique, Université Paris V, CHU Cochin, Paris, France Institut Cochin, Inserm, CNRS, Paris, FranceSearch for more papers by this authorJ Manicom, J Manicom Laboratoire de Génétique, Université Paris V, CHU Cochin, Paris, FranceSearch for more papers by this authorJP Puech, JP Puech Laboratoire de Génétique, Université Paris V, CHU Cochin, Paris, FranceSearch for more papers by this authorA Kahn, A Kahn Institut Cochin, Inserm, CNRS, Paris, FranceSearch for more papers by this authorL Poenaru, L Poenaru Laboratoire de Génétique, Université Paris V, CHU Cochin, Paris, France Institut Cochin, Inserm, CNRS, Paris, FranceSearch for more papers by this author K Azibi, K Azibi Laboratoire de Génétique, Université Paris V, CHU Cochin, Paris, France Institut Cochin, Inserm, CNRS, Paris, FranceSearch for more papers by this authorC Heltianu, C Heltianu Institute of Cellular Biology and Pathology “N. Simionescu”, Bucharest, RumaniaSearch for more papers by this authorC Caillaud, C Caillaud Laboratoire de Génétique, Université Paris V, CHU Cochin, Paris, France Institut Cochin, Inserm, CNRS, Paris, FranceSearch for more papers by this authorJ Manicom, J Manicom Laboratoire de Génétique, Université Paris V, CHU Cochin, Paris, FranceSearch for more papers by this authorJP Puech, JP Puech Laboratoire de Génétique, Université Paris V, CHU Cochin, Paris, FranceSearch for more papers by this authorA Kahn, A Kahn Institut Cochin, Inserm, CNRS, Paris, FranceSearch for more papers by this authorL Poenaru, L Poenaru Laboratoire de Génétique, Université Paris V, CHU Cochin, Paris, France Institut Cochin, Inserm, CNRS, Paris, FranceSearch for more papers by this author First published: 02 January 2007 https://doi.org/10.1111/j.1651-2227.2002.tb03129.xAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume91, Issues439November 2002Pages 119-119 RelatedInformation

The gene encoding endothelial nitric oxide synthase (eNOS) is involved in abnormalities in nitric oxide (NO) synthesis that mediates functional damage of vascular cells, especially of endothelial cells (ECs), a common characteristic in cardiovascular diseases. In Fabry's disease, the characteristic mutation in the alpha-galactosidase A (alpha-gal A) gene induces large deposits of glycosphingolipids, particularly concentrated in ECs, a process associated with endothelial dysfunction. To determine whether in addition to alpha-gal A gene mutations, eNOS genetic variations are implicated in this process, we examined the genotypes of the missense Glu298Asp (G894T) variant in exon 7 and 27-bp tandem repeats in intron 4 (4b/a) in 19 patients with Fabry's disease, and 39 normal volunteers. The results showed that both varials have a significant association with Fabry's disease. The frequencies of mutant Glu/Asp + Asp/Asp genotypes and Asp allele are significantly higher in Fabry's disease (68.4%, p = 0.044, and 47.4%, p = 0.022, respectively) than in controls (46.7% and 25%, respectively). The frequencies of eNOS 4b/a polymorphisms are also significantly different in Fabry's disease when compared to controls. The mutant 4b/a + 4a/a genotype frequencies are 55.5% (p = 0.032) and 4a allele 27.8% (p = 0.05) compared with controls (23.1% and 12.8%, respectively). These results indicate that more than half of the patients with Fabry's disease carry the Glu298Asp variant ( approximately 68%) and/or the 4b/a polymorphism ( approximately 55%). To the best of our knowledge, this is the first report showing an influence of eNOS gene polymorphisms in patients with Fabry's disease.

Human MutationVolume 17, Issue 4 p. 353-353 Mutation and Polymorphism ReportFree Access Identification of a novel de novo mutation (G373D) in the α-galactosidase A gene (GLA) in a patient affected with Fabry disease Dominique P. Germain, Dominique P. Germain Département de Génétique, Hôpital Européen Georges Pompidou, Paris, FranceSearch for more papers by this authorDominique Salard, Dominique Salard Service de Dermatologie, Hôpital St Jacques, Besançon, FranceSearch for more papers by this authorFlorence Fellmann, Florence Fellmann Service de Cytogénétique, Hôpital St Jacques, Besançon, FranceSearch for more papers by this authorKemal Azibi, Kemal Azibi Laboratoire de Génétique, CHU Cochin, Paris, FranceSearch for more papers by this authorCatherine Caillaud, Catherine Caillaud Laboratoire de Génétique, CHU Cochin, Paris, FranceSearch for more papers by this authorMarie-Charlotte Bernard, Marie-Charlotte Bernard Département de Génétique, Hôpital Européen Georges Pompidou, Paris, FranceSearch for more papers by this authorLivia Poenaru, Livia Poenaru Laboratoire de Génétique, CHU Cochin, Paris, FranceSearch for more papers by this author Dominique P. Germain, Dominique P. Germain Département de Génétique, Hôpital Européen Georges Pompidou, Paris, FranceSearch for more papers by this authorDominique Salard, Dominique Salard Service de Dermatologie, Hôpital St Jacques, Besançon, FranceSearch for more papers by this authorFlorence Fellmann, Florence Fellmann Service de Cytogénétique, Hôpital St Jacques, Besançon, FranceSearch for more papers by this authorKemal Azibi, Kemal Azibi Laboratoire de Génétique, CHU Cochin, Paris, FranceSearch for more papers by this authorCatherine Caillaud, Catherine Caillaud Laboratoire de Génétique, CHU Cochin, Paris, FranceSearch for more papers by this authorMarie-Charlotte Bernard, Marie-Charlotte Bernard Département de Génétique, Hôpital Européen Georges Pompidou, Paris, FranceSearch for more papers by this authorLivia Poenaru, Livia Poenaru Laboratoire de Génétique, CHU Cochin, Paris, FranceSearch for more papers by this author First published: 02 April 2001 https://doi.org/10.1002/humu.41Citations: 10 Communicated by Mark H. Paalman Online Citation: Human Mutation, Mutation and Polymorphism Report #225 (2001) Online http://journals.wiley.com/1059-7794/pdf/mutation/mpr225.pdf Acknowledgments: We thank the patient and his family for their help during this project. AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article.Citing Literature Volume17, Issue4Special Issue: SNP 2000: Third International Meeting on Single Nucleotide Polymorphism and Complex Genome AnalysisApril 2001Pages 353-353 RelatedInformation

Title : Identification of two polymorphisms (c189G>C; c190T>C) in exon 2 of the human MRP6 gene (ABCC6) by screening of pseudoxanthoma elasticum patients: possible sequence correction?

ABSTRACT We determined the nucleotide sequence of the bla gene for the Acinetobacter calcoaceticus β-lactamase previously described as CARB-5. Alignment of the deduced amino acid sequence with those of known β-lactamases revealed that CARB-5 possesses an RTG triad in box VII, as described for the Proteus mirabilis GN79 enzyme, instead of the RSG consensus characteristic of the other carbenicillinases. Phylogenetic studies showed that these RTG enzymes constitute a new, separate group, possibly ancestors of the carbenicillinase family.

ABSTRACT A clinical strain of Vibrio cholerae non-O1 non-O139 isolated in France produced a new β-lactamase with a pI of 5.35. The purified enzyme, with a molecular mass of 33,000 Da, was characterized. Its kinetic constants show it to be a carbenicillin-hydrolyzing enzyme comparable to the five previously reported CARB β-lactamases and to SAR-1, another carbenicillin-hydrolyzing β-lactamase that has a pI of 4.9 and that is produced by a V. cholerae strain from Tanzania. This β-lactamase is designated CARB-6, and the gene for CARB-6 could not be transferred to Escherichia coli K-12 by conjugation. The nucleotide sequence of the structural gene was determined by direct sequencing of PCR-generated fragments from plasmid DNA with four pairs of primers covering the whole sequence of the reference CARB-3 gene. The gene encodes a 288-amino-acid protein that shares 94% homology with the CARB-1, CARB-2, and CARB-3 enzymes, 93% homology with the Proteus mirabilis N29 enzyme, and 86.5% homology with the CARB-4 enzyme. The sequence of CARB-6 differs from those of CARB-3, CARB-2, CARB-1, N29, and CARB-4 at 15, 16, 17, 19, and 37 amino acid positions, respectively. All these mutations are located in the C-terminal region of the sequence and at the surface of the molecule, according to the crystal structure of the Staphylococcus aureus PC-1 β-lactamase.

Sarcoglycanopathies are a genetically heterogeneous group of autosomal recessive muscular dystrophies in which the primary defect may reside in any of the genes coding for the different partners of the sarcolemmal sarcoglycan (SG) complex: the alpha-SG (LGMD2D at 17q21.2), the beta-SG (LGMD2E at 4q12), the gamma-SG (LGMD2C at 13q12), and the delta-SG (LGMD2F at 5q33). We report a series of 20 new unrelated families with 14 different mutations in the alpha-SG gene. Along with the mutations that we previously reported this brings our cohort of patients with alpha-sarcoglycanopathy to a total of 31 unrelated patients, carrying 25 different mutations. The missense mutations reside in the extracellular domain of the protein. Five of 15 missense mutations, carried by unrelated subjects on different haplotype backgrounds and of widespread geographical origins, account for 58% of the mutated chromosomes, with a striking prevalence of the R77C substitution (32%). The severity of the disease varies strikingly and correlates at least in part with the amount of residual protein and the type of mutation. The recurrent R284C substitution is associated with a benign disease course.

Primary adhalin (or alpha-sarcoglycan) deficiency due to a defect of the adhalin gene localized on chromosome 17q21 causes an autosomal recessive myopathy. We evaluated 20 patients from 15 families (12 from Europe and three from North Africa) with a primary adhalin deficiency with two objectives: characterization of the clinical phenotype and analysis of the correlation with the level of adhalin expression and the type of gene mutation. Age at onset and severity of the myopathy were heterogeneous: six patients were wheel-chair bound before 15 years of age, whereas five other patients had mild disease with preserved ambulation in adulthood. The clinical pattern was similar in all the patients with symmetric characteristic involvement of trunk and limb muscles, calf hypertrophy, and absence of cardiac dysfunction. Immunofluorescence and immunoblot studies of muscle biopsy specimens showed a large variation in the expression of adhalin. The degree of adhalin deficiency was fairly correlated with the clinical severity. There were 15 different mutations (10 missense, five null). Double null mutations (three patients) were associated with severe myopathy, but in the other cases (null/missense and double missense) there was a large variation in the severity of the disease.