Multiplex PCR combining deltaF508 mutation and intragenic microsatellites of the CFTR gene for pre-implantation genetic diagnosis (PGD) of cystic fibrosis

OBJECTIVE: To evaluate the diagnostic contribution of molecular analysis of the cystic fibrosis transmembrane conductance regulator (CFTR) gene in patients suspected of having mild or atypical cystic fibrosis (CF). METHODS: This was a cross-sectional study involving adolescents and adults aged ≥ 14 years. Volunteers underwent clinical, laboratory, and radiological evaluation, as well as spirometry, sputum microbiology, liver ultrasound, sweat tests, and molecular analysis of the CFTR gene. We then divided the patients into three groups by the number of mutations identified (none, one, and two or more) and compared those groups in terms of their characteristics. RESULTS: We evaluated 37 patients with phenotypic findings of CF, with or without sweat test confirmation. The mean age of the patients was 32.5 ± 13.6 years, and females predominated (75.7%). The molecular analysis contributed to the definitive diagnosis of CF in 3 patients (8.1%), all of whom had at least two mutations. There were 7 patients (18.9%) with only one mutation and 26 patients (70.3%) with no mutations. None of the clinical characteristics evaluated was found to be associated with the genetic diagnosis. The most common mutation was p.F508del, which was found in 5 patients. The combination of p.V232D and p.F508del was found in 2 patients. Other mutations identified were p.A559T, p.D1152H, p.T1057A, p.I148T, p.V754M, p.P1290P, p.R1066H, and p.T351S. CONCLUSIONS: The molecular analysis of the CFTR gene coding region showed a limited contribution to the diagnostic investigation of patients suspected of having mild or atypical CF. In addition, there were no associations between the clinical characteristics and the genetic diagnosis.

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Multiplex sequence variation detection throughout the CFTR gene appropriate for preimplantation genetic diagnosis in populations with heterogeneity of cystic fibrosis mutations

MHR Basic science of reproductive medicine ◽

10.1093/molehr/8.9.880 ◽

2002 ◽

Vol 8 (9) ◽

pp. 880-886 ◽

Cited By ~ 11

Author(s):

C. Vrettou

Keyword(s):

Cystic Fibrosis ◽

Preimplantation Genetic Diagnosis ◽

Sequence Variation ◽

Genetic Diagnosis ◽

Cftr Gene

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Clinical hallmarks and genetic polymorphisms in the CFTR gene contribute to the disclosure of the A1006E mutation

Clinical & Investigative Medicine ◽

10.25011/cim.v33i4.14226 ◽

2010 ◽

Vol 33 (4) ◽

pp. 234

Author(s):

Anna Cristina Tomaiuolo ◽

Federico Alghisi ◽

Stefano Petrocchi ◽

Cecilia Surace ◽

Maria Cristina Roberti ◽

...

Keyword(s):

Cystic Fibrosis ◽

Clinical Presentation ◽

Genetic Diagnosis ◽

Cftr Gene ◽

Transmembrane Conductance Regulator ◽

Transmembrane Conductance ◽

Bilateral Absence ◽

Speed Up ◽

Idiopathic Chronic Pancreatitis ◽

Dna Strand

Since the identification of the Cystic Fibrosis transmembrane conductance regulator (CFTR) gene in 1989, many genetic mutations have been found in cystic fibrosis (CF) patients. Dysfunctions of the CFTR gene are responsible for the highly variable clinical presentation ranging from severe CF, disseminated bronchiectasis, idiopathic chronic pancreatitis and congenital bilateral absence of vas deferens (CBAVD). Linkage disequilibrium studies have shown that some mutations are stringently coupled with polymorphisms in a genetic complex called haplotype. From a familial study of a patient with CBAVD, carrier of the A1006E mutation, we have observed its strict association with the polymorphism 5T-TG11. In order to speed up the genetic diagnosis and to correlate the clinical setting to this genetic feature, we have directly investigated the exon 17a, where the A1006E mutation is located, of five cystic fibrosis patients belonging to two unrelated families. All patients had the 5T-TG11 tract, F508del and one unknown mutation. One more family with two affected individuals carrying the Q220X/A1006E mutations was investigated for the poly-T polymorphism. All the members were found to have the A1006E mutation and the 5T-TG11 in the same DNA strand, demonstrating that this strategy is a reliable and inexpensive method for genotyping the CFTR gene. A detailed description of the clinical presentation and follow-up are provided in order to highlight common phenotypic features useful to improve the management of cystic fibrosis patients.

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INFLUENCE OF GENE POLYMORPHISM OF THE 1st PHASE OF XENOBIOTICS METABOLISM ON ANTIBACTERIAL THERAPY EFFICACY IN PATIENTS WITH CYSTIC FIBROSIS HOMOZYGOUS FOR F508DEL MUTATION OF CFTR GENE

PEDIATRIA Journal named after G N SPERANSKY ◽

10.24110/0031-403x-2018-97-2-94-98 ◽

2018 ◽

Vol 97 (2) ◽

pp. 94-98

Author(s):

O.G. Novoselova ◽

◽

N.V. Petrova ◽

E.I. Kondratyeva ◽

S.A. Krasovskii ◽

...

Keyword(s):

Cystic Fibrosis ◽

Gene Polymorphism ◽

Antibacterial Therapy ◽

Cftr Gene ◽

Therapy Efficacy

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CFTR Cooperative Cis-Regulatory Elements in Intestinal Cells

International Journal of Molecular Sciences ◽

10.3390/ijms22052599 ◽

2021 ◽

Vol 22 (5) ◽

pp. 2599

Author(s):

Mégane Collobert ◽

Ozvan Bocher ◽

Anaïs Le Nabec ◽

Emmanuelle Génin ◽

Claude Férec ◽

...

Keyword(s):

Gene Expression ◽

Cystic Fibrosis ◽

Gene Regulation ◽

Genetic Diseases ◽

Regulatory Elements ◽

Cftr Gene ◽

Intestinal Cells ◽

Cooperative Effects ◽

Cis Acting ◽

Study Techniques

About 8% of the human genome is covered with candidate cis-regulatory elements (cCREs). Disruptions of CREs, described as “cis-ruptions” have been identified as being involved in various genetic diseases. Thanks to the development of chromatin conformation study techniques, several long-range cystic fibrosis transmembrane conductance regulator (CFTR) regulatory elements were identified, but the regulatory mechanisms of the CFTR gene have yet to be fully elucidated. The aim of this work is to improve our knowledge of the CFTR gene regulation, and to identity factors that could impact the CFTR gene expression, and potentially account for the variability of the clinical presentation of cystic fibrosis as well as CFTR-related disorders. Here, we apply the robust GWAS3D score to determine which of the CFTR introns could be involved in gene regulation. This approach highlights four particular CFTR introns of interest. Using reporter gene constructs in intestinal cells, we show that two new introns display strong cooperative effects in intestinal cells. Chromatin immunoprecipitation analyses further demonstrate fixation of transcription factors network. These results provide new insights into our understanding of the CFTR gene regulation and allow us to suggest a 3D CFTR locus structure in intestinal cells. A better understand of regulation mechanisms of the CFTR gene could elucidate cases of patients where the phenotype is not yet explained by the genotype. This would thus help in better diagnosis and therefore better management. These cis-acting regions may be a therapeutic challenge that could lead to the development of specific molecules capable of modulating gene expression in the future.

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A Peptide-Nucleic Acid Targeting miR-335-5p Enhances Expression of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Gene with the Possible Involvement of the CFTR Scaffolding Protein NHERF1

Biomedicines ◽

10.3390/biomedicines9020117 ◽

2021 ◽

Vol 9 (2) ◽

pp. 117

Author(s):

Anna Tamanini ◽

Enrica Fabbri ◽

Tiziana Jakova ◽

Jessica Gasparello ◽

Alex Manicardi ◽

...

Keyword(s):

Cystic Fibrosis ◽

Nucleic Acid ◽

Peptide Nucleic Acid ◽

Stop Codon ◽

Scaffolding Protein ◽

Cftr Gene ◽

Scaffolding Proteins ◽

Transmembrane Conductance Regulator ◽

Transmembrane Conductance ◽

Cystic Fibrosis Transmembrane

(1) Background: Up-regulation of the Cystic Fibrosis Transmembrane Conductance Regulator gene (CFTR) might be of great relevance for the development of therapeutic protocols for cystic fibrosis (CF). MicroRNAs are deeply involved in the regulation of CFTR and scaffolding proteins (such as NHERF1, NHERF2 and Ezrin). (2) Methods: Content of miRNAs and mRNAs was analyzed by RT-qPCR, while the CFTR and NHERF1 production was analyzed by Western blotting. (3) Results: The results here described show that the CFTR scaffolding protein NHERF1 can be up-regulated in bronchial epithelial Calu-3 cells by a peptide-nucleic acid (PNA) targeting miR-335-5p, predicted to bind to the 3′-UTR sequence of the NHERF1 mRNA. Treatment of Calu-3 cells with this PNA (R8-PNA-a335) causes also up-regulation of CFTR. (4) Conclusions: We propose miR-335-5p targeting as a strategy to increase CFTR. While the efficiency of PNA-based targeting of miR-335-5p should be verified as a therapeutic strategy in CF caused by stop-codon mutation of the CFTR gene, this approach might give appreciable results in CF cells carrying other mutations impairing the processing or stability of CFTR protein, supporting its application in personalized therapy for precision medicine.

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