Correction of Nonsense Mutations in Cystic Fibrosis

• ClinicalTrials.gov Identifier: NCT03670472
• Recruitment Status: Recruiting
• First Posted: September 13, 2018
• Last Update Posted: November 18, 2020
• Sponsor: University Hospital, Lille
• Collaborator: Vaincre la Mucoviscidose
• Information provided by (Responsible Party): University Hospital, Lille

Study Description

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.

Condition or disease	Intervention/treatment
Cystic Fibrosis	Other: smear of nasal fossae

Study Design

• Study Type: Observational
• Estimated Enrollment: 85 participants
• Observational Model: Case-Control
• Time Perspective: Prospective
• Official Title: Optimization of Correcting Molecules of Nonsense Mutations in Epithelial Cells of the Upper Airways of Patients With Cystic Fibrosis With Nonsense Mutations in the CFTR Gene
• Actual Study Start Date: February 3, 2016
• Estimated Primary Completion Date: January 2030
• Estimated Study Completion Date: January 2030

Groups and Cohorts

Intervention Details:

• Other: smear of nasal fossae
  1 smear of nasal fossae during a usual or scheduled visit

Outcome Measures

Primary Outcome Measures :

1. Transport of iodide ions through the CEVAS membrane [ Time Frame: less than 48hrs after the collect. ]
   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

Secondary Outcome Measures :

1. Immortalization of patient cells [ Time Frame: an average 12 months ]
   Immortalization of patient cells will be attempted by transfection of construct expressing the origin-of-replication defective SV40 as described in Gruenert et al.,2004
2. Expression of the CFTR gene at the mRNA and protein level [ Time Frame: less than 1 week. ]

Eligibility Criteria

• Ages Eligible for Study: 8 Years and older (Child, Adult, Older Adult)
• Sexes Eligible for Study: All
• Accepts Healthy Volunteers: No
• 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.

Criteria

Inclusion Criteria:

• 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

Exclusion Criteria:

• 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

Contacts and Locations

Contacts

Contact: Anne Prévotat, MD; 03 20 44 59 48 ext +33; anne.prevotat@chru-lille.fr

Contact: Fabrice Lejeune, PhD; fabrice.lejeune@inserm.fr

Locations

France

Camsp Chu Amiens; Recruiting

Amiens, France

Hopital Femme Mere Enfant - Hcl - Bron; Recruiting

Bron, France

Hôpital Calmette,CHU; Recruiting

Lille, France

Principal Investigator: Anne Prévotat, MD

Aphm Hopital La Timone - Marseille; Recruiting

Marseille, France

Chu Montpellier; Recruiting

Montpellier, France

Cmp Enfants Aphp Robert Debre - Paris; Recruiting

Paris, France

Hu Paris Centre Site Cochin Aphp - Paris 14; Recruiting

Paris, France

Hopitaux Universitaires de Strasbour; Recruiting

Strasbourg, France

Sponsors and Collaborators

University Hospital, Lille

Vaincre la Mucoviscidose

Investigators

• Principal Investigator: Anne Prévotat, MD; University Hospital, Lille

More Information

• Responsible Party: University Hospital, Lille
• ClinicalTrials.gov Identifier: NCT03670472
• Other Study ID Numbers: 2013_59; 2014-A01236-41 ( Other Identifier: ID-RCB number, ANSM )
• First Posted: September 13, 2018
• Last Update Posted: November 18, 2020
• Last Verified: October 2020

Individual Participant Data (IPD) Sharing Statement:

• Plan to Share IPD: Undecided

• Studies a U.S. FDA-regulated Drug Product: No
• Studies a U.S. FDA-regulated Device Product: No

Keywords provided by University Hospital, Lille:

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

Additional relevant MeSH terms:

Cystic Fibrosis; Fibrosis; Pathologic Processes; Pancreatic Diseases; Digestive System Diseases; Lung Diseases; Respiratory Tract Diseases; Genetic Diseases, Inborn; Infant, Newborn, Diseases