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Validation of Therapeutic Efficacy Targeting the Splicing Variants in Cystic Fibrosis and CFTR Pathologies (ONB-CFTR)

Primary Purpose

Cystic Fibrosis, CFTR Gene Mutation

Status

Recruiting

Phase

Not Applicable

Locations

France

Study Type

Interventional

Intervention

Nasal cells sampling

Rectal biopsy sampling

About this trial

This is an interventional basic science trial for Cystic Fibrosis focused on measuring genetics, cystic fibrosis, personalized therapy, Antisense OligoNucleotide strategy, splicing events, intronic variant

Eligibility Criteria

12 Years - undefined (Child, Adult, Older Adult)All SexesDoes not accept healthy volunteers

Inclusion Criteria:

The subject must have given their free and informed consent and signed the consent
The subject must be affiliated or beneficiary of a health insurance plan Women and men are included
The patient is at least 12 years old.
The patient has cystic fibrosis or a CFTR pathology and therefore carries two mutations (with at least one mutation affecting splicing) in the CFTR gene.
Patients who volunteer for rectal biopsy collection (only from Montpellier University Hospital) must be at least 18 years old.

Exclusion Criteria:

The subject is in a period of exclusion determined by a previous study.
The subject is under judicial protection, under guardianship or under curatorship
The subject does not accept to sign consent
It turns out to be impossible to give informed information to the subject
The subject does not read the French language fluently
The subject is a pregnant or breastfeeding woman
The subject has porphyria, or has hepatic insufficiency, or suffers from epilepsy, or suffers from conduction disorders, or suffers from severe heart failure, has a cons-indication to the use of a local anesthetic spray.

Specific non-inclusion criteria for rectal sampling:

the subject has thrombocytopenia
the subject has a bleeding disorder
The patient has severe inflammation of the rectum.

Sites / Locations

Montpellier University HospitalRecruiting

Arms of the Study

Arm 1

Arm Type

Experimental

Arm Label

Nasal cells sampling and/or rectal biospy

Arm Description

Depending of the patient' genotype, specific ONB-CFTR (50 nM) will be incubated at the apical face of in vitro epithelium, alone and in combination with CFTR modulators. Efficacy of ONB will be compared to a condition with oligonucleotide control incubation. Rectal biopsies from volunteer patients were stored as a bio-bank of organoids.

Outcomes

Primary Outcome Measures

Restoration of the correctly spliced CFTR mRNA (full length) using specific ONB-CFTR (designed for one splicing variant).

The increase will be assessed in comparison to oligonucleotide-control effect by using semi-quantitative fluorescent PCR.

Restoration of the mature CFTR protein using specific ONB-CFTR (designed for one splicing variant).

The increase will be assessed in comparison to oligonucleotide-control effect by using western blot

Restoration of CFTR channel function using specific ONB-CFTR (designed for one splicing variant).

The increase will be assessed in comparison to oligonucleotide-control effect by using electrophysiological assays (Ussing chamber).

Secondary Outcome Measures

Restoration of the correctly spliced CFTR mRNA (full length) and mature CFTR protein and CFTR channel function using a pool of ONB-CFTR (a mix of specific ONB-CFTR).

The increase will be assessed in comparison to oligonucleotide-control effect by using semi-quantitative fluorescent PCR, western blot and electrophysiological assays (Ussing chamber).

Assessment of the amount of CFTR mRNA with normal splicing under the conditions tested.

That parameter will be quantified in comparison to oligonucleotide-control effect by using quantitative PCR assays.

Assessment of the amount of mature CFTR proteins under the conditions tested.

That parameter will be quantified in comparison to oligonucleotide-control effect by using western blot assays.

Assessment of the CFTR channel activity under the conditions tested.

That parameter will be quantified in comparison to oligonucleotide-control effect by using electrophysiological assays (Ussing chamber).

Increase of CFTR channel function using ONB-CFTR and CFTR modulators (correctors and/or potentiators) under the conditions tested.

The increase will be assessed in comparison to oligonucleotide-control effect by using electrophysiological assays (Ussing chamber).

Full Information

NCT ID

NCT05100823

First Posted

September 6, 2021

Last Updated

March 28, 2023

Sponsor

University Hospital, Montpellier

Collaborators

Foch Hospital, Suresnes, FRANCE, Hôpital Necker-Enfants Malades, Hôpital Cochin

1. Study Identification

Unique Protocol Identification Number

NCT05100823

Brief Title

Validation of Therapeutic Efficacy Targeting the Splicing Variants in Cystic Fibrosis and CFTR Pathologies

Acronym

ONB-CFTR

Official Title

Validation of Therapeutic Efficacy Targeting the Splicing Variants in Cystic Fibrosis and CFTR Pathologies

Study Type

Interventional

2. Study Status

Record Verification Date

March 2023

Overall Recruitment Status

Recruiting

Study Start Date

March 30, 2022 (Actual)

Primary Completion Date

March 30, 2025 (Anticipated)

Study Completion Date

March 30, 2025 (Anticipated)

3. Sponsor/Collaborators

Responsible Party, by Official Title

Sponsor

Name of the Sponsor

University Hospital, Montpellier

Collaborators

Foch Hospital, Suresnes, FRANCE, Hôpital Necker-Enfants Malades, Hôpital Cochin

4. Oversight

Studies a U.S. FDA-regulated Drug Product

Studies a U.S. FDA-regulated Device Product

Data Monitoring Committee

5. Study Description

Brief Summary

Cystic Fibrosis, an inherited autosomal recessive disease, arises from mutations in the CFTR gene. For intronic mutations affecting splicing events, oligonucleotides therapy has the potential to restore the production of the full length CFTR protein. Recent scientific research has demonstrated the potential of this approach to restore full length mRNA CFTR in in vitro human airway cells. The study aims to validate the therapeutic efficacy of oligonucleotide blockers (ONB) that target splicing defects associated to splicing variants in epithelia obtained from patients with Cystic Fibrosis and CFTR-related disorders.

Detailed Description

The study will include patients with various CFTR genotypes. The assessment of ONB (named ONB-CFTR) will be performed using an air-liquid interface model of airway epithelium, developed from nasal cells of patients, without or with a combination of existing CFTR modulators, depending on the patient' genotype. This study will also aim to build a local biobank of rectal organoids from patients (only from Montpellier, France) carrying rare CFTR disease-causing variants.

6. Conditions and Keywords

Primary Disease or Condition Being Studied in the Trial, or the Focus of the Study

Cystic Fibrosis, CFTR Gene Mutation

Keywords

genetics, cystic fibrosis, personalized therapy, Antisense OligoNucleotide strategy, splicing events, intronic variant

7. Study Design

Primary Purpose

Basic Science

Study Phase

Not Applicable

Interventional Study Model

Single Group Assignment

Masking

None (Open Label)

Allocation

N/A

Enrollment

20 (Anticipated)

8. Arms, Groups, and Interventions

Arm Title

Nasal cells sampling and/or rectal biospy

Arm Type

Experimental

Arm Description

Intervention Type

Procedure

Intervention Name(s)

Nasal cells sampling

Intervention Description

Nasal epithelium brushing in intermediate turbinate using a specific curette following a local anesthesia with Xylocaine 5% nebulizer.

Intervention Type

Procedure

Intervention Name(s)

Rectal biopsy sampling

Intervention Description

Forceps Biopsy Procedure (Servidoni et al., 2013) Only for volunteer patients included in the Montpellier center.

Primary Outcome Measure Information:

Title

Restoration of the correctly spliced CFTR mRNA (full length) using specific ONB-CFTR (designed for one splicing variant).

Description

The increase will be assessed in comparison to oligonucleotide-control effect by using semi-quantitative fluorescent PCR.

Time Frame

21 days after the air-liquid switch of epithelia (i.e. full differentiation)

Title

Restoration of the mature CFTR protein using specific ONB-CFTR (designed for one splicing variant).

Description

The increase will be assessed in comparison to oligonucleotide-control effect by using western blot

Time Frame

21 days after the air-liquid switch of epithelia (i.e. full differentiation)

Title

Restoration of CFTR channel function using specific ONB-CFTR (designed for one splicing variant).

Description

The increase will be assessed in comparison to oligonucleotide-control effect by using electrophysiological assays (Ussing chamber).

Time Frame

21 days after the air-liquid switch of epithelia (i.e. full differentiation)

Secondary Outcome Measure Information:

Title

Restoration of the correctly spliced CFTR mRNA (full length) and mature CFTR protein and CFTR channel function using a pool of ONB-CFTR (a mix of specific ONB-CFTR).

Description

The increase will be assessed in comparison to oligonucleotide-control effect by using semi-quantitative fluorescent PCR, western blot and electrophysiological assays (Ussing chamber).

Time Frame

21 days after the air-liquid switch of epithelia (i.e. full differentiation)

Title

Assessment of the amount of CFTR mRNA with normal splicing under the conditions tested.

Description

That parameter will be quantified in comparison to oligonucleotide-control effect by using quantitative PCR assays.

Time Frame

21 days after the air-liquid switch of epithelia (i.e. full differentiation)

Title

Assessment of the amount of mature CFTR proteins under the conditions tested.

Description

That parameter will be quantified in comparison to oligonucleotide-control effect by using western blot assays.

Time Frame

21 days after the air-liquid switch of epithelia (i.e. full differentiation)

Title

Assessment of the CFTR channel activity under the conditions tested.

Description

That parameter will be quantified in comparison to oligonucleotide-control effect by using electrophysiological assays (Ussing chamber).

Time Frame

21 days after the air-liquid switch of epithelia (i.e. full differentiation)

Title

Increase of CFTR channel function using ONB-CFTR and CFTR modulators (correctors and/or potentiators) under the conditions tested.

Description

The increase will be assessed in comparison to oligonucleotide-control effect by using electrophysiological assays (Ussing chamber).

Time Frame

21 days after the air-liquid switch of epithelia (i.e. full differentiation)

10. Eligibility

Sex

All

Minimum Age & Unit of Time

12 Years

Accepts Healthy Volunteers

Eligibility Criteria

Inclusion Criteria: The subject must have given their free and informed consent and signed the consent The subject must be affiliated or beneficiary of a health insurance plan Women and men are included The patient is at least 12 years old. The patient has cystic fibrosis or a CFTR pathology and therefore carries two mutations (with at least one mutation affecting splicing) in the CFTR gene. Patients who volunteer for rectal biopsy collection (only from Montpellier University Hospital) must be at least 18 years old. Exclusion Criteria: The subject is in a period of exclusion determined by a previous study. The subject is under judicial protection, under guardianship or under curatorship The subject does not accept to sign consent It turns out to be impossible to give informed information to the subject The subject does not read the French language fluently The subject is a pregnant or breastfeeding woman The subject has porphyria, or has hepatic insufficiency, or suffers from epilepsy, or suffers from conduction disorders, or suffers from severe heart failure, has a cons-indication to the use of a local anesthetic spray. Specific non-inclusion criteria for rectal sampling: the subject has thrombocytopenia the subject has a bleeding disorder The patient has severe inflammation of the rectum.

Central Contact Person:

First Name & Middle Initial & Last Name or Official Title & Degree

Anne Bergougnoux, PhD, PharmD

Phone

+33 411759879

anne.bergougnoux@inserm.fr

First Name & Middle Initial & Last Name or Official Title & Degree

Magali Taulan-Cadars, PhD

Phone

+33 411759846

magali.taulan@inserm.fr

Facility Information:

Facility Name

Montpellier University Hospital

City

Montpellier

ZIP/Postal Code

34090

Country

France

Individual Site Status

Recruiting

Facility Contact:

First Name & Middle Initial & Last Name & Degree

Anne Bergougnoux, phD, PharmD

12. IPD Sharing Statement

Learn more about this trial

Validation of Therapeutic Efficacy Targeting the Splicing Variants in Cystic Fibrosis and CFTR Pathologies

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