Correction of Nonsense Mutations in Cystic Fibrosis

NCT03670472 | Recruiting

• 2 other identifiers: 2013_592014-A01236-41
• Study Type: observational
• Target: 85
• Locations: 1 country
• Sites: 8

Brief Summary

The presence of a nonsense mutation leads to the rapid degradation of the carrier mRNA mutation by a mechanism called NMD (nonsense-mediated mRNA decay) \[6, 13\]. There are currently 3 main strategies at least for correcting nonsense mutations: exon skipping, inhibition of NMD and nonsense mutation readthrough. In the laboratory, we developed a strategy for correcting nonsense mutations combining inhibition of NMD and activation of translecture. For this purpose, we have constructed screening systems to identify NMD-inhibiting and/or readthrough enhancers. The molecules thus identified are then tested on cell lines and in murine models carrying a nonsense mutation. One of our goals is to select a set of molecules that can correct effectively nonsense mutations. For this we have to test these molecules on a great diversity of nonsense mutations. This work will:

• determine if we can correct all the nonsense mutations tested with at least one of our molecules
• determine what is common within a group of mutations corrected by a given molecule
• be able to assign the parameters that make one mutation is corrected by one molecule and not or little by another. This study will therefore improve our theoretical knowledge on the recognition of premature stop codons but also to propose therapeutic approaches for the correction of nonsense mutations of the CFTR gene in cystic fibrosis in a targeted way for a patient.

Conditions

Cystic Fibrosis

Keywords

Cystic fibrosis; nonsense mutations; CFTR gene; nasal epithelial cells; nonsense mutation readthrough

Outcome Measures

Primary Outcomes (1)

• Transport of iodide ions through the CEVAS membrane

  Patient cells with be cultured in BEGM (Lonza) medium and incubated with corrector of nonsense mutations for 20 hours and with a fluorescent molecule called SPQ (for 6-methoxy-N-3'-sulfopropylquinolinium). Iodine can bind SPQ and will quench the SPQ fluorescence. Nitrates bind SPQ without quenching SPQ fluorescence. By placing patient cells first into an iodine-rich medium to quench the SPQ fluorescence and second into a nitrate-rich medium, we will be able to measure the level of functional CFTR protein present in these cells by measuring the re-apparition of fluorescence using fluorimeter. Indeed, nitrate will be able to replace iodine on SPQ without quenching SPQ fluorescence only if iodine exits cells through CFTR channels. This assay allows determining whether a corrector of nonsense mutation is able to lead to the synthesis of functional CFTR protein

  less than 48hrs after the collect.

Secondary Outcomes (2)

• Immortalization of patient cells

  an average 12 months

• Expression of the CFTR gene at the mRNA and protein level

  less than 1 week.

Interventions

smear of nasal fossae OTHER

1 smear of nasal fossae during a usual or scheduled visit

Eligibility Criteria

• Age: 8 Years+
• Sex: all
• Healthy Volunteers: No
• Age Groups: Child (0-17), Adult (18-64), Older Adult (65+)
• Sampling Method: Non-Probability Sample

Study Population

Patients with cystic fibrosis and carry a nonsense mutation on the 2 alleles of the gene coding for the CFTR channel.

You may qualify if:

• Male / female adults and minors aged 8 years and over
• Patients with cystic fibrosis and carry a nonsense mutation on the 2 alleles of the gene coding for the CFTR channel.
• Patients whose genotype of patients concerning the CFTR gene is known.
• Patients with social security
• Major patients who have given their consent
• Minor patients with parental authorization

You may not qualify if:

• Patients who have a mutation other than nonsense in the CFTR gene
• Patients whose CFTR gene was not sequenced on the 2 alleles
• Patients not wishing to participate in this study or persons not giving or not able to give consent.
• Pregnant or lactating women
• Patients under curatorship or guardianship

Sponsors & Collaborators

• University Hospital, Lille: lead
• Vaincre la Mucoviscidose: collaborator

Study Sites (8)

Camsp Chu Amiens

Amiens, France

RECRUITING

Hopital Femme Mere Enfant - Hcl - Bron

Bron, France

RECRUITING

Hôpital Calmette,CHU

Lille, France

RECRUITING

Aphm Hopital La Timone - Marseille

Marseille, France

RECRUITING

Chu Montpellier

Montpellier, France

RECRUITING

Cmp Enfants Aphp Robert Debre - Paris

Paris, France

RECRUITING

Hu Paris Centre Site Cochin Aphp - Paris 14

Paris, France

RECRUITING

Hopitaux Universitaires de Strasbour

Strasbourg, France

RECRUITING

MeSH Terms

Conditions

Cystic Fibrosis

Condition Hierarchy (Ancestors)

Pancreatic Diseases; Digestive System Diseases; Lung Diseases; Respiratory Tract Diseases; Genetic Diseases, Inborn; Congenital, Hereditary, and Neonatal Diseases and Abnormalities; Infant, Newborn, Diseases

Study Officials

• Anne Prévotat, MD

  University Hospital, Lille

  PRINCIPAL INVESTIGATOR

Central Study Contacts

Anne Prévotat, MD

CONTACT

03 20 44 59 48; anne.prevotat@chru-lille.fr

Fabrice Lejeune, PhD

CONTACT

fabrice.lejeune@inserm.fr

Study Design

• Study Type: observational
• Observational Model: CASE CONTROL
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Study Record Dates

• First Submitted: August 1, 2018
• First Posted: September 13, 2018
• Study Start: February 3, 2016
• Primary Completion (Estimated): January 1, 2030
• Study Completion (Estimated): January 1, 2030
• Last Updated: November 18, 2020

Record last verified: 2020-10

Locations

FR (8)