CHARGE syndrome: a recurrent hotspot of mutations in CHD7 IVS25 analyzed by bioinformatic tools and minigene assays

Legendre, Marine; Rodríguez-Ballesteros, Montserrat; Rossi, Massimiliano; Abadie, Véronique; Amiel, Jeanne; Revençu, Nicole; Blanchet, P.; Brioude, Frédéric; Delrue, Marie‐Ange; Doubaj, Yassamine; Sefiani, Abdelaziz; Francannet, Christine; Holder‐Espinasse, Muriel; Jouk, Pierre‐Simon; Julia, Sophie; Melki, Judith; Mur, Sébastien; Naudion, Sophie; Fabre-Teste, Jennifer; Busa, Tiffany; Stamm, Stephen; Lyonnet, Stanislas; Attié‐Bitach, Tania; Kitzis, Alain; Gilbert‐Dussardier, Brigitte; Bilan, Frédéric

doi:10.1038/s41431-017-0007-0

Cited by 10 publications

(12 citation statements)

References 12 publications

(1 reference statement)

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“…Only one patient had a hot spot of intron variant c.5405 − 7G > A. In our study, it was reported to be pathogenic [ 25 ]; this finding was consistent with the results of two previous studies, which indicated that the CHD7 variant was not the primary cause of KS because only 3–5% of nIHH/KS patients had a CHD7 variant [ 23 , 26 ].…”

Section: Discussionsupporting

confidence: 92%

Clinical, hormonal, and genetic characteristics of 25 Chinese patients with idiopathic hypogonadotropic hypogonadism

2022

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Background Idiopathic hypogonadotropic hypogonadism (IHH) is a type of congenital disease caused by a variety of gene variants leading to dysfunction in the secretion of hypothalamic gonadotropin-releasing hormones (GnRHs). Clinically, IHH can be divided into Kallmann syndrome (KS) with dysosmia and normosmic idiopathic hypogonadotropic hypogonadism (nIHH) according to the presence or absence of an olfactory disorder. Methods We retrospectively evaluated 25 IHH patients (8 KS and 17 nIHH) who were diagnosed at the Department of Endocrinology of Shanghai Children’s Hospital from 2015 to 2021. We analysed the patients’ clinical data, including their hormone levels and gene sequences. Results All male patients exhibited small phalli, and 35% of them exhibited cryptorchidism. A significant difference was observed in the levels of dihydrotestosterone (DHT) after human chorionic gonadotropin (HCG) stimulation (P = 0.028) between the KS group and the nIHH group. Missense variants were the major cause of IHH, and the main pathogenic genes were FGFR1, PROKR2/PROK2, and KAl1. Nine reported and 13 novel variants of six genes were identified. De novo variants were detected in 16 IHH patients; eight patients inherited the variants from their mothers, while only three patients inherited variants from their fathers. One patient had both KAl1 and PROKR2 gene variants, and another patient had two different PROKR2 gene variants. These two patients both had the hot spot variant c.533G > C (p. Trp178Ser) of the PROKR2 gene. Conclusion IHH should be highly suspected in patients with a small phallus and cryptorchidism. Compared with nIHH patients, KS patients exhibited a higher level of DHT after HCG stimulation. Missense variants were the major cause of IHH, and most of the inherited variants were from their mothers who exhibited no obvious clinical symptoms. We identified 9 reported variants and 13 novel variants that led to IHH. A small proportion of patients were at risk of inheriting either the oligogenic variant or the compound heterozygous variant. The hot spot variant c.533G > C (p. Trp178Ser) of PROKR2 might be involved in oligogenic inheritance and compound heterozygous inheritance. These findings provide deeper insight into the diagnosis and classification of IHH and will contribute to its clinical assessment.

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Section: Discussionsupporting

confidence: 92%

Clinical, hormonal, and genetic characteristics of 25 Chinese patients with idiopathic hypogonadotropic hypogonadism

2022

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“…Therefore, whether the gene expression and chromatin structure are correct may be the pathogenesis of CS caused by the CHD7 mutation. The CHD7 gene may be regulated by epigenetic and signaling pathways that function as chromatin remodeling factors and may directly or indirectly affect ectodermal lineage genes (Platt et al, 2017;Legendre et al, 2018;Goodman and Bonni, 2019). Abnormal antenatal examinations were present in ten of the 12 patients.…”

Section: Used the Acronym "Charge" (Ocular Coloboma [C] Heart Malform...mentioning

confidence: 99%

Discovery of Novel Variants on the CHD7 Gene: A Case Series of CHARGE Syndrome

Chen

Lü

et al. 2022

Front. Genet.

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Background: CHARGE syndrome (CS) is a single-gene genetic disorder with multiple organ malformations caused by a variant of the chromodomain helicase DNA-binding protein 7 (CHD7) gene on chromosome 8q12.1. In this study, we aimed to investigate new variants that have emerged in these cases compared with typical CS and the relationship between the genes and phenotypes.Methods: Patients with suspected genetic diseases were subjected to Whole Exome Sequencing (WES) at a genetics laboratory in Guangzhou. The average sequencing coverage depth was >200 ×, and 96% was >20 ×. The variant interpretation was manipulated according to the American College of Medical Genetics (ACMG) guidelines. Molecular data on databases for ClinVar and CHD7 were also collected and collated. We reviewed the currently described CHD7 variants and analyzed the genetic variation and phenotypic heterogeneity.Results: Data of 12 patients with CS from four hospitals in China were collected. According to gestational age, most of them (8/12) were near-term babies with a lower birth weight than their peers, averaging 2.62 kg. In this study, the most common phenotypes were respiratory tract malformations (11/12), heart malformations (10/12), and central nervous system malformations (9/12). Two fetuses were confirmed to have brain or heart abnormalities during prenatal testing, while 10/12 were found to have abnormalities during prenatal testing. The maximum Acute Physiology and Chronic Health Evaluation (APACHE II) score at admission was 19, and the average was 11.58. Five variants in the CHD7 gene c.7012C > T (p.Q2338*), c.7868delC (p.P2623Rfs*16), c.5405-3C > G, c.6936 + 2T > C, and c.8077-2A > G) were novel and were located in exons 33, 36, and introns 25, 32, and 37, respectively. There may be a positive correlation between exon location and phenotype.Conclusion: Five novel variants were discovered. These expanded the mutational spectrum of the CHD7 gene and the phenotype of CS. There may be a correlation between the new mutation sites and the phenotype, which has some reference value for the evaluation of mutation sites.

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“…Depending on the objectives of the study, different experimental settings and laboratory protocols can be used. To give an overview of the potential applications of the pDESTsplice and pSpliceExpress vectors, we have compiled 25 published studies [ 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 ], in which these minigene systems were used ( Table 1 and Table 2 ). In most of the works, the usage of pSpliceExpress has been described ( n = 18).…”

Section: Studies Using the Pdestsplice And Pspliceexpress Vectorsmentioning

confidence: 99%

“…However, other procedures were also applied. Legendre et al performed a nested RT-PCR within the region of the lariat structure to determine the branch point [ 51 ]. After PCR, the amplicons were differentiated in size by fluorescent capillary electrophoresis instead of a gel electrophoresis [ 51 ].…”

Section: Studies Using the Pdestsplice And Pspliceexpress Vectorsmentioning

confidence: 99%

“…Legendre et al performed a nested RT-PCR within the region of the lariat structure to determine the branch point [ 51 ]. After PCR, the amplicons were differentiated in size by fluorescent capillary electrophoresis instead of a gel electrophoresis [ 51 ]. In fluorescence capillary electrophoresis, DNA molecules are separated by passing them through a small capillary tube filled with a conductive buffer.…”

Section: Studies Using the Pdestsplice And Pspliceexpress Vectorsmentioning

confidence: 99%

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Principles and Practical Considerations for the Analysis of Disease-Associated Alternative Splicing Events Using the Gateway Cloning-Based Minigene Vectors pDESTsplice and pSpliceExpress

Putscher

Hecker

Fitzner

et al. 2021

IJMS

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Splicing is an important RNA processing step. Genetic variations can alter the splicing process and thereby contribute to the development of various diseases. Alterations of the splicing pattern can be examined by gene expression analyses, by computational tools for predicting the effects of genetic variants on splicing, and by splicing reporter minigene assays for studying alternative splicing events under defined conditions. The minigene assay is based on transient transfection of cells with a vector containing a genomic region of interest cloned between two constitutive exons. Cloning can be accomplished by the use of restriction enzymes or by site-specific recombination using Gateway cloning. The vectors pDESTsplice and pSpliceExpress represent two minigene systems based on Gateway cloning, which are available through the Addgene plasmid repository. In this review, we describe the features of these two splicing reporter minigene systems. Moreover, we provide an overview of studies in which determinants of alternative splicing were investigated by using pDESTsplice or pSpliceExpress. The studies were reviewed with regard to the investigated splicing regulatory events and the experimental strategy to construct and perform a splicing reporter minigene assay. We further elaborate on how analyses on the regulation of RNA splicing offer promising prospects for gaining important insights into disease mechanisms.

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CHARGE syndrome: a recurrent hotspot of mutations in CHD7 IVS25 analyzed by bioinformatic tools and minigene assays

Cited by 10 publications

References 12 publications

Clinical, hormonal, and genetic characteristics of 25 Chinese patients with idiopathic hypogonadotropic hypogonadism

Clinical, hormonal, and genetic characteristics of 25 Chinese patients with idiopathic hypogonadotropic hypogonadism

Discovery of Novel Variants on the CHD7 Gene: A Case Series of CHARGE Syndrome

Principles and Practical Considerations for the Analysis of Disease-Associated Alternative Splicing Events Using the Gateway Cloning-Based Minigene Vectors pDESTsplice and pSpliceExpress

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