Nasal epithelial cells are a reliable source to study splicing variants in Usher syndrome

Vaché, Christel; Besnard, Thomas; Blanchet, C.; Baux, David; Larrieu, Lise; Faugère, Valérie; Mondain, M.; Hamel, Christian; Malcolm, Sue; Claustres, Mireille; Roux, Anne‐Françoise

doi:10.1002/humu.21255

Cited by 29 publications

(27 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Together with published studies from our laboratory, [5][6][7][8]10,11 we provide evidence of a powerful service for patients. A total of 92 patients have received diagnoses, with no ambiguous USH1 genotypes.…”

Section: Diagnostic Approachsupporting

confidence: 54%

“…To these, an additional 18 private variants can be added that were published separately by our group in the course of the creation of USHbases, ex vivo assays and nasal cell transcript studies. 7,8,11 Only a few founder effects have been observed-for example the CDH23 p.Arg1502X mutation in Swedes, 26 PCDH15 p.Arg245X in Ashkenazi Jews, and USH1C c.216GϾA in Acadians 29,30 and Quebecois. 33 As a consequence the Asper chip based on known mutations would have been inefficient and would have solved only 52% of the cases.…”

Section: Diagnostic Approachmentioning

confidence: 95%

“…Some remove the canonical AG/GT sites, but some also lie in the introns, outside the invariant sites, or correspond to exonic nucleotide substitutions. Ex vivo assays and transcript analyses from nasal cells for CDH23 c.6050 -9GϾA and ex vivo assays for USH1C c.1210ϩ6TϾC provided evidence for deleterious outcomes with the creation of PTC, either by use of a created acceptor site 8,25 in the first case or by activation of an upstream exonic cryptic splice site in the …”

Section: Summary Of Mutationsmentioning

confidence: 98%

“…7,8 The multistep analysis for determining the predicted effect of alteration of a variant on protein structure has been described. 9 The National Center for Biotechnology (NCBI) RefSeq IDs were MYO7A, 4647; CDH23, 64072 (with the initiation codon located in exon 1); PCDH15, 65217; USH1C, 10083; and USH1G, 124590 (available at www.ncbi.nlm.nih.gov/locuslink/refseq/ NCBI, Bethesda MD).…”

Section: In Silico Studiesmentioning

confidence: 99%

See 3 more Smart Citations

Four-Year Follow-up of Diagnostic Service in USH1 Patients

Roux

Faugère

Vaché

et al. 2011

Invest. Ophthalmol. Vis. Sci.

Self Cite

View full text Add to dashboard Cite

show abstract

Section: Diagnostic Approachsupporting

confidence: 54%

Section: Diagnostic Approachmentioning

confidence: 95%

Section: Summary Of Mutationsmentioning

confidence: 98%

Section: In Silico Studiesmentioning

confidence: 99%

See 2 more Smart Citations

Four-Year Follow-up of Diagnostic Service in USH1 Patients

Roux

Faugère

Vaché

et al. 2011

Invest. Ophthalmol. Vis. Sci.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Subsequently, Vaché et al [11] provided evidence that splicing mutations occurring in most USH genes can be identified through ex vivo analysis of mRNA from nasal epithelial cells.…”

Section: Introductionmentioning

confidence: 99%

Study of USH1 Splicing Variants through Minigenes and Transcript Analysis from Nasal Epithelial Cells

et al. 2013

View full text Add to dashboard Cite

Usher syndrome type I (USH1) is an autosomal recessive disorder characterized by congenital profound deafness, vestibular areflexia and prepubertal retinitis pigmentosa. The first purpose of this study was to determine the pathologic nature of eighteen USH1 putative splicing variants found in our series and their effect in the splicing process by minigene assays. These variants were selected according to bioinformatic analysis. The second aim was to analyze the USH1 transcripts, obtained from nasal epithelial cells samples of our patients, in order to corroborate the observed effect of mutations by minigenes in patient’s tissues. The last objective was to evaluate the nasal ciliary beat frequency in patients with USH1 and compare it with control subjects. In silico analysis were performed using four bioinformatic programs: NNSplice, Human Splicing Finder, NetGene2 and Spliceview. Afterward, minigenes based on the pSPL3 vector were used to investigate the implication of selected changes in the mRNA processing. To observe the effect of mutations in the patient’s tissues, RNA was extracted from nasal epithelial cells and RT-PCR analyses were performed. Four MYO7A (c.470G>A, c.1342_1343delAG, c.5856G>A and c.3652G>A), three CDH23 (c.2289+1G>A, c.6049G>A and c.8722+1delG) and one PCDH15 (c.3717+2dupTT) variants were observed to affect the splicing process by minigene assays and/or transcripts analysis obtained from nasal cells. Based on our results, minigenes are a good approach to determine the implication of identified variants in the mRNA processing, and the analysis of RNA obtained from nasal epithelial cells is an alternative method to discriminate neutral Usher variants from those with a pathogenic effect on the splicing process. In addition, we could observe that the nasal ciliated epithelium of USH1 patients shows a lower ciliary beat frequency than control subjects.

show abstract

Molecular Genetics of U sher Syndrome: Current State of Understanding

Slijkerman

Kremer

Wijk

2017

Encyclopedia of Life Sciences

View full text Add to dashboard Cite

Usher syndrome affects hearing, vision and balance. The syndrome is genetically heterogeneous and mutations in ten genes have been identified to be disease causing. Proteins encoded by these genes cofunction with each other and with other proteins, composing an Usher interactome with diverse functions including structural support, transport and potentially signalling at different subcellular locations in hair cells of the inner ear and in photoreceptors of the retina. In hair cells, Usher proteins mainly contribute to the formation of fibrous links, including the tip link, that connect stereocilia and stereocilia to the kinocilium. In addition, Usher proteins are found in the synaptic region where they might contribute to signal transduction by regulating the cell surface levels of Ca v 1.3 Ca 2+ channels and exocytosis. In photoreceptor cells, the Usher proteins are seen in the region of the connecting cilium including the periciliary region, the basal body, accessory centriole and calyceal processes. Also here, Usher proteins are present in the synaptic region. Although the function of Usher proteins in photoreceptors and hair cells is not yet fully understood, promising therapeutic developments are ongoing to alleviate the phenotypic burden resulting from mutations in any of the Usher genes. Key Concepts Usher syndrome is clinically and genetically heterogeneous. Usher syndrome type I (USH1) and type II (USH2) proteins interact and form highly dynamic protein networks. Usher protein networks fulfil a role in providing structural support, play a role in intracellular transport mechanisms and are suggested to be involved in cellular signalling cascades. A range of cellular and animal models are used to unravel the molecular pathogenesis of Usher syndrome and to evaluate promising therapeutic strategies. Promising progress in the development of (pre)clinical genetic (i.e. gene augmentation, splice modulation, translational read‐through and gene editing) and nongenetic (i.e. small compounds, neurotrophic factors and cell replacement) therapeutic strategies have been published for Usher syndrome.

show abstract

Nasal epithelial cells are a reliable source to study splicing variants in Usher syndrome

Cited by 29 publications

References 37 publications

Four-Year Follow-up of Diagnostic Service in USH1 Patients

Four-Year Follow-up of Diagnostic Service in USH1 Patients

Study of USH1 Splicing Variants through Minigenes and Transcript Analysis from Nasal Epithelial Cells

Molecular Genetics of U sher Syndrome: Current State of Understanding

Contact Info

Product

Resources

About