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1. Molecular Genetics of Cystic Fibrosis

Author

Tsui, L.-C., Rommens, J., Kerem, B., Rozmahel, R., Zielenski, J., Kennedy, D., Markiewicz, D., Plavsic, N., Chou, J.-L., Bozon, D., Dobbs, M., Tsui, Lap-Chee, editor, Romeo, Giovanni, editor, Greger, Rainer, editor, and Gorini, Sergio, editor

Published
1991
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3. Characterization of a novel 21-kb deletion, CFTRdele2,3(21 kb), in the CFTR gene: a cystic fibrosis mutation of Slavic origin common in Central and East Europe

Author

Dörk, T., Macek Jr, M., Mekus, F., Tümmler, B., Tzountzouris, J., Casals, T., Krebsová, A., Koudová, M., Sakmaryová, I., Macek Sr, M., Vávrová, V., Zemková, D., Ginter, E., Petrova, N.V., Ivaschenko, T., Baranov, V., Witt, M., Pogorzelski, A., Bal, J., Zékanowsky, C., Wagner, K., Stuhrmann, M., Bauer, I., Seydewitz, H.H., Neumann, T., Jakubiczka, S., Kraus, C., Thamm, B., Nechiporenko, M., Livshits, L., Mosse, N., Tsukerman, G., Kadási, L., Ravnik-Glavač, M., Glavač, D., Komel, R., Vouk, K., Kučinskas, V., Krumina, A., Teder, M., Kocheva, S., Efremov, G.D., Onay, T., Kirdar, B., Malone, G., Schwarz, M., Zhou, Z., Friedman, K.J., Carles, S., Claustres, M., Bozon, D., Verlingue, C., Férec, C., Tzetis, M., Kanavakis, E., Cuppens, H., Bombieri, C., Pignatti, P.F., Sangiuolo, F., Jordanova, A., Kusic, J., Radojkovič, D., Sertić, J., Richter, D., Stavljenić Rukavina, A., Bjorck, E., Strandvik, B., Cardoso, H., Montgomery, M., Nakielna, B., Hughes, D., Estivill, X., Aznarez, I., Tullis, E., Tsui, L.-C., and Zielenski, J.

Published
2000
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7. Single, short in-del, and copy number variations detection in monogenic dyslipidemia using a next-generation sequencing strategy

Author

Marmontel, O., primary, Charrière, S., additional, Simonet, T., additional, Bonnet, V., additional, Dumont, S., additional, Mahl, M., additional, Jacobs, C., additional, Nony, S., additional, Chabane, K., additional, Bozon, D., additional, Janin, A., additional, Peretti, N., additional, Lachaux, A., additional, Bardel, C., additional, Millat, G., additional, Moulin, P., additional, Marçais, C., additional, and Di Filippo, M., additional

Published
2018
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11. Molecular basis of mucopolysaccharidosis type II in Portugal

Author

Silva, IMoreira da, Froissart, R, Santos, HMarques dos, Caseiro, C, Maire, I, Bozon, D, and Repositório da Universidade de Lisboa

Abstract

Made available in DSpace on 2015-12-30T10:19:30Z (GMT). No. of bitstreams: 0 Previous issue date: 2001

Published
2001

12. Characterization of a novel 21-kb deletion, CFTRdele2,3(21 kb), in the CFTR gene: a cystic fibrosis mutation of Slavic origin common in Central and East Europe

Author

Dork, T Macek, M Mekus, F Tummler, B Tzountzouris, J and Casals, T Krebsova, A Koudova, M Sakmaryova, I Macek, M and Vavrova, V Zemkova, D Ginter, E Petrova, NV and Ivaschenko, T Baranov, V Witt, M Pogorzelski, A Bal, J and Zekanowsky, C Wagner, K Stuhrmann, M Bauer, I and Seydewitz, HH Neumann, T Jakubiczka, S Kraus, C Thamm, B and Nechiporenko, M Livshits, L Mosse, N Tsukerman, G and Kadasi, L Ravnik-Glavac, M Glavac, D Komel, R Vouk, K and Kucinskas, V Krumina, A Teder, M Kocheva, S Efremov, GD Onay, T Kirdar, B Malone, G Schwarz, M Zhou, ZQ and Friedman, KJ Carles, S Claustres, M Bozon, D and Verlingue, C Ferec, C Tzetis, M Kanavakis, E Cuppens, H and Bombieri, C Pignatti, PF Sangiuolo, F Jordanova, A and Kusic, J Radojkovic, D Sertic, J Richter, D Rukavina, AS and Bjorck, E Strandvik, B Cardoso, H Montgomery, M and Nakielna, B Hughes, D Estivill, X Aznarez, I Tullis, E and Tsui, LC Zielenski, J

Abstract

We report a large genomic deletion of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, viz.. a deletion that is frequently observed in Central and Eastern Europe. The mutation, termed CFTRdele2.3(21 kb), deletes 21,080 bp spanning introns 1-3 of the CFTR gene. Transcript analyses have revealed that this deletion results in the loss of exons 2 and 3 in epithelial CFTR mRNA, thereby producing a premature termination signal within exon 4. In order to develop a simple polymerase chain reaction assay for this allele, we defined the end-points of the deletion at the DNA sequence level. We next screened for this mutation in a representative set of European and European-derived populations. Some 197 CF patients, including seven homozygotes, bearing this mutation have been identified during the course of our study. Clinical evaluation of CFTRdele2.3(21 kb) homozygotes and a comparison of compound heterozygotes for Delta F508/CFTRdele2,3(21 kb) with pairwise-matched Delta F508 homozygotes indicate that this deletion represents a severe mutation associated with pancreatic insufficiency and early age at diagnosis. Current data show that the mutation is particularly common in Czech (6.4% of all CF chromosomes), Russian (5.2%), Belorussian (3.3%). Austrian (2.6%), German (1.5%), Polish (1.5%), Slovenian (1.5%), Ukrainian (1.2%), and Slovak patients (1.1%). It has also been found in Lithuania, Latvia, Macedonia and Greece and has sporadically been observed in Canada, USA, France, Spain, Turkey, and UK, but not in CF patients from Bulgaria, Croatia, Romania or Serbia. Haplotype analysis has identified the same extragenic CF-haplotype: XV-2c/KM. 19 “A” and the same infrequent intragenic microsatellite haplotype 16-33-13 (IVS8CA-IVS17bTA-IVSI7bCA) in all examined CFTRdele2,3(21 kb) chromosomes, suggesting a common origin for this deletion. We conclude that the 21-kb deletion is a frequent and severe CF mutation in populations of Eastern- and Western-Slavic descent.

Published
2000

13. SURF1 deficiency causes demyelinating Charcot-Marie-Tooth disease

Author

Echaniz-Laguna, A., primary, Ghezzi, D., additional, Chassagne, M., additional, Mayencon, M., additional, Padet, S., additional, Melchionda, L., additional, Rouvet, I., additional, Lannes, B., additional, Bozon, D., additional, Latour, P., additional, Zeviani, M., additional, and Mousson de Camaret, B., additional

Published
2013
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14. No Evidence for Segregation Distortion of Cystic Fibrosis Alleles among Sibs of Cystic Fibrosis Patients

Author

Devoto, M, Romeo, G, ten Kate, L P, Chevalier, F, Bozon, D, Estivill, X, Casals, T, Abeliovich, D, Lerer, I, Padoan, R, Seia, M, Hill, A, Liechtigallati, S, Kramer, R, Beards, F, DEAR, S, DALLAPICCOLA, B, SANGIUOLO, F, MACEK, M, MCMAHON, R, CONNARTY, M, HARVEY, JF, CLAUSTRES, M, DESGEORGES, M, de Vries, R, Scheffer, H, CANKIKLAIN, N, AUDREZET, MP, BIENVENU, T, CHOMEL, JC, DZIADEK, [No Value], TUMMLER, B, SCHWARZ, M, HAWORTH, A, BENITEZ, J, MAZURCZAK, T, BAL, J, CREMONESI, L, RONCHETTO, P, CASHMAN, SM, FEREC, C, CUPPENS, H, Bauer, I, ANGELICHEVA, D, WAGNER, K, PACHECO, P, BONIZZATO, A, MCMAHON, CJ, RAVNIKGLAVAC, M, REIS, A, STUHRMANN, M, GARNERONE, S, CURTIS, A, GRUNING, G, KANAVAKIS, E, KLAASSEN, T, GRADE, T, and Witt, Michal

Subjects
Pathology, medicine.medical_specialty, business.industry, Distortion, Genetics, medicine, Allele, medicine.disease, business, Cystic fibrosis, Genetics (clinical)
Published
1995

15. Genetic determination of exocrine pancreatic function in cystic fibrosis

Author

Bozon, D, Markiewicz, D, Rommens, J, Durie, P, Kristidis, P, Tsui, LC, and Corey, M

Subjects
Amino acid sequence, Chromosome mapping, Chromosome deletion, Codon - genetics, Alleles
Abstract

We showed elsewhere that the pancreatic function status of cystic fibrosis (CF) patients could be correlated to mutations in the CF transmembrane conductance regulator (CFTR) gene. Although the majority of CF mutations- including the most common, ΔF508-strongly correlated with pancreatic insufficiency (PI), approximately 10% of the mutant alleles may confer pancreatic sufficiency (PS). To extend this observation, genomic DNA of 538 CF patients with well-documented pancreatic function status were analyzed for a series of known mutations in their CFTR genes. Only 20 of the 25 mutations tested were found in this population. They accounted for 84% of the CF chromosomes, with ΔF508 being the most frequent (71%), and the other mutations accounted for less than 5% each. A total of 30 different, complete genotypes could be determined in 394 (73%) of the patients. The data showed that each genotype was associated only with PI or only with PS, but not with both. This result is thus consistent with the hypothesis that PI and PS in CF are predisposed by the genotype at the CFTR locus; the PS phenotype occurs in patients who have one or two mild CFTR mutations, such as R117H, R334W, R347P, A455E, and P574H, whereas the PI phenotype occurs in patients with two severe alleles, such as ΔF508, ΔI507, Q493X, G542X, R553X, W1282X, 621 + 1G→T, 1717-1G→A, 556delA, 3659delC, I148T, G480C, V520F, G551D, and R560T., published_or_final_version

Published
1992

16. Characterization of a novel 21-kb deletion, CFTRdele2,3(21 kb), in the CFTR gene: a cystic fibrosis mutation of Slavic origin common in Central and East Europe

Author

Dork, T, Macek, M, Mekus, F, Tummler, B, Tzountzouris, J, Casals, T, Krebsova, A, Koudova, M, Sakmaryova, I, Vavrova, V, Zemkova, D, Ginter, E, Petrova, NV, Ivaschenko, T, Baranov, V, Witt, M, Pogorzelski, A, Bal, J, Zekanowsky, C, Wagner, K, Stuhrmann, M, Bauer, I, Seydewitz, HH, Neumann, T, Jakubiczka, S, Kraus, C, Thamm, B, Nechiporenko, M, Livshits, L, Mosse, N, Tsukerman, G, Kadasi, L, Ravnik-Glavac, M, Glavac, D, Komel, R, Vouk, K, Kucinskas, V, Krumina, A, Teder, M, Kocheva, S, Efremov, GD, Onay, T, Kirdar, B, Malone, G, Schwarz, M, Zhou, ZQ, Friedman, KJ, Carles, S, Claustres, M, Bozon, D, Verlingue, C, Ferec, C, Tzetis, M, Kanavakis, E, Cuppens, H, Bombieri, C, Pignatti, PF, Sangiuolo, F, Jordanova, A, Kušić-Tišma, Jelena, Radojković, Dragica, Sertić, J, Richter, D, Rukavina, AS, Bjorck, E, Strandvik, B, Cardoso, H, Montgomery, M, Nakielna, B, Hughes, D, Estivill, X, Aznarez, I, Tullis, E, Tsui, LC, Zielenski, J, Dork, T, Macek, M, Mekus, F, Tummler, B, Tzountzouris, J, Casals, T, Krebsova, A, Koudova, M, Sakmaryova, I, Vavrova, V, Zemkova, D, Ginter, E, Petrova, NV, Ivaschenko, T, Baranov, V, Witt, M, Pogorzelski, A, Bal, J, Zekanowsky, C, Wagner, K, Stuhrmann, M, Bauer, I, Seydewitz, HH, Neumann, T, Jakubiczka, S, Kraus, C, Thamm, B, Nechiporenko, M, Livshits, L, Mosse, N, Tsukerman, G, Kadasi, L, Ravnik-Glavac, M, Glavac, D, Komel, R, Vouk, K, Kucinskas, V, Krumina, A, Teder, M, Kocheva, S, Efremov, GD, Onay, T, Kirdar, B, Malone, G, Schwarz, M, Zhou, ZQ, Friedman, KJ, Carles, S, Claustres, M, Bozon, D, Verlingue, C, Ferec, C, Tzetis, M, Kanavakis, E, Cuppens, H, Bombieri, C, Pignatti, PF, Sangiuolo, F, Jordanova, A, Kušić-Tišma, Jelena, Radojković, Dragica, Sertić, J, Richter, D, Rukavina, AS, Bjorck, E, Strandvik, B, Cardoso, H, Montgomery, M, Nakielna, B, Hughes, D, Estivill, X, Aznarez, I, Tullis, E, Tsui, LC, and Zielenski, J

Abstract

We report a large genomic deletion of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, viz.. a deletion that is frequently observed in Central and Eastern Europe. The mutation, termed CFTRdele2.3(21 kb), deletes 21,080 bp spanning introns 1-3 of the CFTR gene. Transcript analyses have revealed that this deletion results in the loss of exons 2 and 3 in epithelial CFTR mRNA, thereby producing a premature termination signal within exon 4. In order to develop a simple polymerase chain reaction assay for this allele, we defined the end-points of the deletion at the DNA sequence level. We next screened for this mutation in a representative set of European and European-derived populations. Some 197 CF patients, including seven homozygotes, bearing this mutation have been identified during the course of our study. Clinical evaluation of CFTRdele2.3(21 kb) homozygotes and a comparison of compound heterozygotes for Delta F508/CFTRdele2,3(21 kb) with pairwise-matched Delta F508 homozygotes indicate that this deletion represents a severe mutation associated with pancreatic insufficiency and early age at diagnosis. Current data show that the mutation is particularly common in Czech (6.4% of all CF chromosomes), Russian (5.2%), Belorussian (3.3%). Austrian (2.6%), German (1.5%), Polish (1.5%), Slovenian (1.5%), Ukrainian (1.2%), and Slovak patients (1.1%). It has also been found in Lithuania, Latvia, Macedonia and Greece and has sporadically been observed in Canada, USA, France, Spain, Turkey, and UK, but not in CF patients from Bulgaria, Croatia, Romania or Serbia. Haplotype analysis has identified the same extragenic CF-haplotype: XV-2c/KM. 19 "A" and the same infrequent intragenic microsatellite haplotype 16-33-13 (IVS8CA-IVS17bTA-IVSI7bCA) in all examined CFTRdele2,3(21 kb) chromosomes, suggesting a common origin for this deletion. We conclude that the 21-kb deletion is a frequent and severe CF mutation in populations of Eastern- and Western-Slavic descent

Published
2000

18. Identification of mutations in regions corresponding to the two putative nucleotide (ATP)-binding folds of the cystic fibrosis gene

Author

Markiewicz, D, Tsui, LC, Kerem, B, Zielenski, J, Bozon, D, Gazit, E, Yahav, J, Kennedy, D, Riordan, JR, Collins, FS, and Rommens, JM

Subjects
mutational hot spot, Nonsense mutation, genetic disease, missense mutation, pancreatic function
Abstract

Additional mutations in the cystic fibrosis (CF) gene were identified in the regions corresponding to the two putative nucleotide (ATP)-binding folds (NBFs) of the predicted polypeptide. The patient cohort included 46 Canadian CF families with well-characterized DNA marker haplotypes spanning the disease locus and several other families from Israel. Eleven mutations were found in the first NBF, 2 were found in the second NBF, but none was found in the R-domain. Seven of the mutations were of the missense type affecting some of the highly conserved amino acid residues in the first NBF; 3 were nonsense mutations; 2 would probably affect mRNA splicing; 2 corresponded to small deletions, including another 3-base-pair deletion different from the major mutation (ΔF508), which could account for 70% of the CF chromosomes in the population. Nine of these mutations accounted for 12 of the 31 non-ΔF508 CF chromosomes in the Canadian families. The highly heterogeneous nature of the remaining CF mutations provides important insights into the structure and function of the protein, but is also suggests that DNA-based genetic screening for CF carrier status will not be straightforward., published_or_final_version

Published
1990

36. δ°-Thalassemia in cisof β Knossos-globin gene Normal structure and normal transient expression of the δ-globin gene

Author

Ouazana, R., Bozon, D., Baklouti, F., Gonnet, C., Delaunay, J., and Godet, J.

Abstract

We have previously described the first homozygous cases of Hb Knossos in an Algerian family. The Hb A 2was completely absent, ascertaining the presence of a δ°-thalassemia determinant in cisof the β Knossoss gene. Here, we investigate the affected δ-globin gene. The complete DNA sequence of the gene and its 5′ and 3′ flanking regions was determined. Only two nucleotide changes were recorded: a C → T substitution at −199 and an AT insertion at −448 upstream from the cap site. To examine the involvement of these changes in gene function, the δ-gene was subcloned in an expression vector and introduced into COS cells. Analysis of RNA derived from these cells, using an S 1protection assay and dot-blot hybridization, revealed qualitatively and quantitatively normal transcription. The loss of δ-globin gene activity in vivo may be due to the alteration of a tissue-specific control.

Published
1989
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37. IDS Transfer from Overexpressing Cells to IDS-Deficient Cells

Author

Millat, G., Froissart, R., Maire, I., and Bozon, D.

Abstract

Iduronate sulfatase (IDS) is responsible for mucopolysaccharidosis type II, a rare recessive X-linked lysosomal storage disease. The aim of this work was to test the ability of overexpressing cells to transfer IDS to deficient cells. In the first part of our work, IDS processing steps were compared in fibroblasts, COS cells, and lymphoblastoid cell lines and shown to be identical: the two precursor forms (76 and 90 kDa) were processed by a series of intermediate forms to the 55- and 45-kDa mature polypeptides. Then IDS transfer to IDS-deficient cells was tested either by incubation with cell-free medium of overexpressing cells or by coculture. Endocytosis and coculture experiments between transfected Lβ and deleted fibroblasts showed that IDS transfer occurred preferentially by cell-to-cell contact as IDS precursors are poorly secreted by transfected Lβ. The 76- and 62-kDa IDS polypeptides transferred to deleted fibroblasts were correctly processed to the mature 55- and 45-kDa forms. Lβ were not able to internalize the 90-kDa phosphorylated precursor forms excreted in large amounts in the medium of overexpressing fibroblasts. Enzyme transfer occurred only by cell-to-cell contact, but the precursor forms transferred in Lβ after cell-to-cell contact were not processed. This absence of maturation was probably due to a mistargeting of IDS precursors in these cells.

Published
1997
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38. δ°‐Thalassemia in cisof βKnossos‐globin gene Normal structure and normal transient expression of the δ‐globin gene

Author

Ouazana, R., Bozon, D., Baklouti, F., Gonnet, C., Delaunay, J., and Godet, J.

Abstract

We have previously described the first homozygous cases of Hb Knossos in an Algerian family. The Hb A2was completely absent, ascertaining the presence of a δ°‐thalassemia determinant in cisof the βKnossoss gene. Here, we investigate the affected δ‐globin gene. The complete DNA sequence of the gene and its 5′ and 3′ flanking regions was determined. Only two nucleotide changes were recorded: a C → T substitution at −199 and an AT insertion at −448 upstream from the cap site. To examine the involvement of these changes in gene function, the δ‐gene was subcloned in an expression vector and introduced into COS cells. Analysis of RNA derived from these cells, using an S1protection assay and dot‐blot hybridization, revealed qualitatively and quantitatively normal transcription. The loss of δ‐globin gene activity in vivo may be due to the alteration of a tissue‐specific control.

Published
1989
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39. Unusual segregation of cystic fibrosis alleles [6]

Author

Kitzis, A., Chomel, J. C., Haliassos, A., Tesson, L., Kaplan, J. C., Feingold, J., Giraud, G., Labbe, A., Dastugue, B., Dumur, V., Farriaux, J. P., Roussel, P., Ferrec, C., Vidaud, M., Goossens, M., Bozon, D., Auvinet, M., Chambon, V., Andre, J., Lissens, W., Bonduelle, M., Liebaers, I., Cochaux, P., Vassart, G., Willems, P., Duckworth-Raysiecki, G., Kerem, B. S., Tsui, L. C., Ray, P. N., Krawczak, M., Schmidtke, J., Novelli, G., Bruno Dallapiccola, Gasparni, G., Pignatti, P. P., Seia, M., Ferrari, M., Devoto, M., Romeo, G., Schwarz, M., Super, M., Ivinson, A., Read, A. P., Meredith, L., Curtis, A., Williamson, R., Beaudet, A. L., Feldman, G. L., O Brien, W. E., Bowcock, A. M., Cavalli-Sforza, L. L., Gilbert, F., Braman, J., and King, M. C.

47. Normal serum ApoB48 and red cells vitamin E concentrations after supplementation in a novel compound heterozygous case of abetalipoproteinemia.

Author

Di Filippo M, Collardeau Frachon S, Janin A, Rajan S, Marmontel O, Decourt C, Rubio A, Nony S, Dumont S, Cuerq C, Charrière S, Moulin P, Lachaux A, Hussain MM, Bozon D, and Peretti N

Subjects
Abetalipoproteinemia blood, Abetalipoproteinemia genetics, Carrier Proteins genetics, Child, Female, Follow-Up Studies, Heterozygote, Humans, Infant, Newborn, Mutation, Abetalipoproteinemia metabolism, Apolipoprotein B-48 blood, Erythrocytes chemistry, Vitamin E analysis
Abstract

Background and Aims: Abetalipoproteinemia (ABL) is a rare recessive monogenic disease due to MTTP (microsomal triglyceride transfer protein) mutations leading to the absence of plasma apoB-containing lipoproteins. Here we characterize a new ABL case with usual clinical phenotype, hypocholesterolemia, hypotriglyceridemia but normal serum apolipoprotein B48 (apoB48) and red blood cell vitamin E concentrations., Methods: Histology and MTP activity measurements were performed on intestinal biopsies. Mutations in MTTP were identified by Sanger sequencing, quantitative digital droplet and long-range PCR. Functional consequences of the variants were studied in vitro using a minigene splicing assay, measurement of MTP activity and apoB48 secretion., Results: Intestinal steatosis and the absence of measurable lipid transfer activity in intestinal protein extract supported the diagnosis of ABL. A novel MTTP c.1868G>T variant inherited from the patient's father was identified. This variant gives rise to three mRNA transcripts: one normally spliced, found at a low frequency in intestinal biopsy, carrying the p.(Arg623Leu) missense variant, producing in vitro 65% of normal MTP activity and apoB48 secretion, and two abnormally spliced transcripts resulting in a non-functional MTP protein. Digital droplet PCR and long-range sequencing revealed a previously described c.1067+1217_1141del allele inherited from the mother, removing exon 10. Thus, the patient is compound heterozygous for two dysfunctional MTTP alleles. The p.(Arg623Leu) variant may maintain residual secretion of apoB48., Conclusions: Complex cases of primary dyslipidemia require the use of a cascade of different methodologies to establish the diagnosis in patients with non-classical biological phenotypes and provide better knowledge on the regulation of lipid metabolism., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Published
2019
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48. Splicing analysis of 26 F8 nucleotide variations using a minigene assay.

Author

Jourdy Y, Fretigny M, Nougier C, Négrier C, Bozon D, and Vinciguerra C

Subjects
Adolescent, Adult, Child, Child, Preschool, Computational Biology methods, Exons, Genes, Reporter, Hemophilia A pathology, Humans, Infant, Male, Mutation, Missense, Polymorphism, Single Nucleotide, RNA Splice Sites, Registries, Severity of Illness Index, Young Adult, Factor VIII genetics, Hemophilia A genetics, RNA Splicing
Abstract

Background: Classically, the study of splicing impact of variation located near the splice site is performed by both in silico and mRNA analysis. However, RNA sample was rarely available., Objective: To characterize a panel of putative haemophilia A splicing variations., Materials and Methods: Twenty-six F8 variations identified from a cohort of 2075 haemophilia A families were studied using both bioinformatic tools and in vitro minigene assays in HeLa and Huh7 cells., Results: An aberrant splicing was demonstrated for 21/26 tested sequence variations. A good correlation between in silico and in vitro analysis was obtained for variations affecting donor splice site (12/14) and for the synonymous variations located inside an exon (6/6). Conversely, no concordant results were observed for the six variations affecting acceptor splice sites. The variations resulted more frequently in exon skipping (n = 13) than in activation of nearby cryptic splice sites (n = 5), in use of a de novo splice site (n = 2) or in insertion of large intronic sequences (n = 1). This study allowed to reclassify 5 synonymous substitutions c.1167A>G (p.Gln389Gln), c.1569G>T (p.Leu523Leu), c.1752G>A (p.Gln584Gln), c.5586G>A (p.Leu1862Leu) and c.6066C>T (p.Gly2022Gly) as splicing variations. The pathological significance of five variations remained unclear (c.222G>A [p.Thr74Thr], c.237C>T [p.Asn79Asn], c.240C>T [p.Ile80Ile], c.2113+5_2113+8del and c.2113+5G>A)., Discussion: The minigene assay herein gave additional evidences for the clinical significance of 21/26 F8 putative splice site mutations. Such investigation should be performed for each F8 putative splice site variation for which no mRNA sample is available, notably to greatly improve the genetic counselling given to female carriers., (© 2019 John Wiley & Sons Ltd.)

Published
2019
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49. Reccurrent F8 Intronic Deletion Found in Mild Hemophilia A Causes Alu Exonization.

Author

Jourdy Y, Janin A, Fretigny M, Lienhart A, Négrier C, Bozon D, and Vinciguerra C

Subjects
Base Sequence, Child, Female, Genes, Reporter, Haplotypes genetics, HeLa Cells, Heterogeneous-Nuclear Ribonucleoprotein Group C genetics, Humans, Male, Middle Aged, Alu Elements genetics, Exons genetics, Factor VIII genetics, Hemophilia A genetics, Introns genetics, Sequence Deletion genetics
Abstract

Incorporation of distant intronic sequences in mature mRNA is an underappreciated cause of genetic disease. Several disease-causing pseudoexons have been found to contain repetitive elements such as Alu elements. This study describes an original pathological mechanism by which a small intronic deletion leads to Alu exonization. We identified an intronic deletion, c.2113+461_2113+473del, in the F8 intron 13, in two individuals with mild hemophilia A. In vivo and in vitro transcript analysis found an aberrant transcript, with an insertion of a 122-bp intronic fragment (c.2113_2114ins2113+477_2113+598) at the exon 13-14 junction. This out-of-frame insertion is predicted to lead to truncated protein (p.Gly705Aspfs ∗ 37). DNA sequencing analysis found that the pseudoexon corresponds to antisense AluY element and the deletion removed a part of the poly(T)-tail from the right arm of these AluY. The heterogenous nuclear riboprotein C1/C2 (hnRNP C) is an important antisense Alu-derived cryptic exon silencer and binds to poly(T)-tracts. Disruption of the hnRNP C binding site in AluY T-tract by mutagenesis or hnRNP C knockdown using siRNA in HeLa cells reproduced the effect of c.2113+461_2113+473del. The screening of 114 unrelated families with mild hemophilia A in whom no genetic event was previously identified found a deletion in the poly(T)-tail of AluY in intron 13 in 54% of case subjects (n = 61/114). In conclusion, this study describes a deletion leading to Alu exonization found in 6.1% of families with mild hemophila A in France., (Copyright © 2017 American Society of Human Genetics. Published by Elsevier Inc. All rights reserved.)

Published
2018
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50. Global molecular analysis and APOE mutations in a cohort of autosomal dominant hypercholesterolemia patients in France.

Author

Wintjens R, Bozon D, Belabbas K, MBou F, Girardet JP, Tounian P, Jolly M, Boccara F, Cohen A, Karsenty A, Dubern B, Carel JC, Azar-Kolakez A, Feillet F, Labarthe F, Gorsky AM, Horovitz A, Tamarindi C, Kieffer P, Lienhardt A, Lascols O, Di Filippo M, and Dufernez F

Subjects
Adult, Apolipoproteins B chemistry, Apolipoproteins E genetics, Child, Cohort Studies, Exons genetics, Female, France, Genotyping Techniques, Humans, Hyperlipoproteinemia Type II diagnosis, Male, Models, Molecular, Phenotype, Proprotein Convertase 9 genetics, Protein Conformation, alpha-Helical, Receptors, LDL genetics, Young Adult, Apolipoproteins B genetics, Hyperlipoproteinemia Type II genetics, Mutation
Abstract

Autosomal dominant hypercholesterolemia (ADH) is a human disorder characterized phenotypically by isolated high-cholesterol levels. Mutations in the low density lipoprotein receptor (LDLR), APOB, and proprotein convertase subtilisin/kexin type 9 (PCSK9) genes are well known to be associated with the disease. To characterize the genetic background associated with ADH in France, the three ADH-associated genes were sequenced in a cohort of 120 children and 109 adult patients. Fifty-one percent of the cohort had a possible deleterious variant in LDLR, 3.1% in APOB, and 1.7% in PCSK9. We identified 18 new variants in LDLR and 2 in PCSK9. Three LDLR variants, including two newly identified, were studied by minigene reporter assay confirming the predicted effects on splicing. Additionally, as recently an in-frame deletion in the APOE gene was found to be linked to ADH, the sequencing of this latter gene was performed in patients without a deleterious variant in the three former genes. An APOE variant was identified in three patients with isolated severe hypercholesterolemia giving a frequency of 1.3% in the cohort. Therefore, even though LDLR mutations are the major cause of ADH with a large mutation spectrum, APOE variants were found to be significantly associated with the disease. Furthermore, using structural analysis and modeling, the identified APOE sequence changes were predicted to impact protein function., (Copyright © 2016 by the American Society for Biochemistry and Molecular Biology, Inc.)

Published
2016
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