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Modeling Bronchial Epithelium in Severe Asthma With Human Induced Pluripotent Stem Cells (iPSC) (MOSAIC)

Primary Purpose

Severe Asthma, Allergic Bronchopulmonary Aspergillosis (ABPA)

Status
Completed
Phase
Not Applicable
Locations
France
Study Type
Interventional
Intervention
Blood sample
Sponsored by
University Hospital, Montpellier
About
Eligibility
Locations
Arms
Outcomes
Full info

About this trial

This is an interventional other trial for Severe Asthma focused on measuring Severe asthma, ABPA, Pluripotent stem cells, Airway epithelium, Eosinophils, Crystal

Eligibility Criteria

18 Years - 85 Years (Adult, Older Adult)All SexesDoes not accept healthy volunteers

Inclusion Criteria: - Smoking < 10 BP and weaned > 1 year. Diagnostic criteria for Severe asthma group T2 : History of severe asthma diagnosed by a physician (according to GINA criteria) Subject on high dose inhaled corticosteroid (ICS 1000 µg beclometasone equivalent) and beta-agonists for at least 6 months prior to inclusion Blood eosinophilia in history (previous years) > 300/mm3 Diagnostic criteria for Allergic bronchopulmonary aspergillosis (ABPA) group - Diagnosis of ABPA defined by the following 3 mandatory criteria: Diagnosis of asthma by the physician for at least 12 months based on the 2019 recommendations of the Global Initiative for Asthma (GINA) group Evidence of hypersensitivity to Aspergillus Fumigatus by skin test (on screening or previous documented positive skin test within the last 12 months), or serum Immunoglobulin E (IgE) specific antibodies to A. Fumigatus (≥ 0.35 kUnit/l) at screening. Elevated total serum IgE (> 1000 IU/ml). If the 3 ancillary criteria for the diagnosis of of ABPA (below) are met, an IgE level ≤ 1,000 IU/ml is acceptable. If the patient is receiving oral corticosteroids (OCs) at screening, a previous documented IgE level >1000 IU/ml within the last 12 months is acceptable. And at least 2 of the following ancillary criteria: Blood eosinophil count >500 cells/μl at screening for patients not receiving OCs at screening. For patients receiving OCs at screening, blood eosinophil count > 500 cells/μl at screening or documented previous eosinophil count > 500 cells/μl in the last 12 months. Presence of precipitating antibodies or serum immunoglobulin G (IgG) to A. Fumigatus at screening. Documented radiological abnormalities consistent with ABPA (such as transient mucoid impaction, hyperdense mucus [high attenuation of mucous plugs], opacities of centro-lobular nodules attenuation of mucous plugs], opacities of centro-lobular nodules, telectasis, bronchiectasis, etc.) by chest X-ray or high-resolution computed tomography (HR-CT) within the last 18 months or at screening. Exclusion Criteria: Other associated respiratory diseases (e.g. chronic obstructive pulmonary disease (COPD), cystic fibrosis) Patient in an exclusion period determined by another protocol Protected populations according to the French public health code: Parturient, nursing or pregnant women; subjects deprived of liberty by judicial or administrative decision; Major protected by law (under any form of guardianship). Lack of informed consent Non-beneficiary of the national health insurance system

Sites / Locations

  • university Hospital of Montpellier

Arms of the Study

Arm 1

Arm Type

Experimental

Arm Label

Eligible patients

Arm Description

Outcomes

Primary Outcome Measures

Obtention of functional bronchial epithelium from iPSC: yes/no
Was a functional bronchial epithelium obtained from the patient's induced pluripotent stem cells from blood? (yes/no) The achievement of functional bronchial epithelium (iALI) from iPSCs of a T2 severe asthma patient and an ABPA patient will be assessed by quantification of differentiation markers by immunofluorescence, integrity of the bronchial epithelium by measurement of trans-epithelial resistance (TEER), secretory function by measurement of mucin concentrations (CCSP, MUC5AC and MUC5B) and analysis of ciliary beat

Secondary Outcome Measures

Comparison of the transcriptomic profile between iALI and airway epithelial cells
the transcriptomic profile of iALI from iPSCs of a T2 severe asthma patient and an ABPA patient will be compare to the one of airway epithelial cells from nasal brushing from the same patients
Comparison of the transcriptomic profile between bronchial epithelia generated from severe asthma patients and from healthy subjects
The transcriptomic profile of iALI from iPSCs of a T2 severe asthma patient and an ABPA patient will be compare to those from healthy subjects
Differentiation of iPSC into mature eosinophils : yes/no
Characterization of eosinophils obtained from iPSCs of a T2 severe asthma patient and an ABPA patient by the presence of specific markers by flow cytometry, immunohistochemistry and immunofluorescence
Evaluation of immune cell/bronchial epithelium dialogue
Co culture of the epithelia with iPSC derived eosinophils and the immune cell/bronchial epithelium dialogue in the co-culture of eosinophils with iALI will be evaluated by the activation of eosinophils in contact with the epithelium and by the characterization and study of the phenotype of the exposed epithelium
Obtention of iPSC from peripheral blood sampling : yes/no
Qualification of blood-derived iPSC lines by morphology, culture behavior, karyotype and expression of specific markers

Full Information

First Posted
November 7, 2022
Last Updated
August 28, 2023
Sponsor
University Hospital, Montpellier
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1. Study Identification

Unique Protocol Identification Number
NCT05616338
Brief Title
Modeling Bronchial Epithelium in Severe Asthma With Human Induced Pluripotent Stem Cells (iPSC)
Acronym
MOSAIC
Official Title
Modeling Bronchial Epithelium in Severe Asthma With Human Induced Pluripotent Stem Cells (iPSC)
Study Type
Interventional

2. Study Status

Record Verification Date
August 2023
Overall Recruitment Status
Completed
Study Start Date
November 29, 2022 (Actual)
Primary Completion Date
March 16, 2023 (Actual)
Study Completion Date
March 16, 2023 (Actual)

3. Sponsor/Collaborators

Responsible Party, by Official Title
Sponsor
Name of the Sponsor
University Hospital, Montpellier

4. Oversight

Studies a U.S. FDA-regulated Drug Product
No
Studies a U.S. FDA-regulated Device Product
No
Data Monitoring Committee
No

5. Study Description

Brief Summary
Asthma is severe when it cannot be controlled with maximum-dose inhaled therapies while management of comorbidities and other precipitating or aggravating factors has been optimized. Allergic bronchopulmonary aspergillosis (ABPA) is a complex bronchopulmonary disease resulting from immunological reactions against Aspergillus Fumigatus. The development of a model of bronchial epithelium generated from patients with chronic lung disease will allow the modeling of bronchial tissue to understand the formation of these mucus plugs. This study aims to validate this model The investigators propose to verify the feasibility of obtaining and comparing two epithelia in two populations based on the following experiments: Differentiation of an Induced Pluripotent Stem cell (iPSC) clone derived from blood sample (Peripheral Blood Mononuclear Cells) of Type 2 inflammation (T2) severe asthma and Allergic Bronchopulmonary Aspergillosis (ABPA) in order to obtain differentiated bronchial epithelia in vitro.
Detailed Description
Asthma is severe when it cannot be controlled with maximum-dose inhaled therapies while management of comorbidities and other precipitating or aggravating factors has been optimized. Allergic bronchopulmonary aspergillosis (ABPA) is a complex bronchopulmonary disease resulting from immunological reactions against Aspergillus Fumigatus. At the cellular and molecular level, severe asthma and ABPA are chronic bronchial diseases characterized by type 2 (T2) airway inflammation, bronchial smooth muscle hyperplasia, excessive mucus production by mucus cell metaplasia and epithelial remodeling. Type 2 inflammation in asthma is predominant and is characterized by the accumulation of immune cells, such as eosinophils, mast cells, T-helper 2 (Th2) cells, innate lymphoid cells type 2 (ILC2s) and dendritic cells. Persistent airway obstruction despite maximal therapy is currently considered the greatest unmet medical need in asthma treatment. Fahy and colleagues published a key paper in 2018 that may explain airway obstruction. Using chest CT scans, they demonstrated the presence of mucus plugs that completely occlude segments of the airways of severe asthmatics. These mucus plugs are associated with a high number of circulating eosinophils in the blood. There is no large-scale model to model severe asthma and ABPA. The investigators propose to develop a bronchial epithelium with pluripotent stem cells (iPSC) in air-liquid interface called iALI generated from these patients in aim to model severe asthma and ABPA and in particular the epithelium. The secondary objectives aims to answer are: Comparison of the iPSC derived airway epithelium with nasal epithelial cells isolated from the same patient (nasal brush). To characterize the phenotype of the iPSC derived airway epithelium from ABPA and T2 severe asthma patient, in comparison to a healthy control. Differentiation of iPSC into mature eosinophils Co culture of the epithelia with iPSC derived eosinophils Culture of iPSC derived airway epithelium with Charcot-Leyden crystal Demonstrate the feasibility of obtaining iPSC from peripheral blood sampling.

6. Conditions and Keywords

Primary Disease or Condition Being Studied in the Trial, or the Focus of the Study
Severe Asthma, Allergic Bronchopulmonary Aspergillosis (ABPA)
Keywords
Severe asthma, ABPA, Pluripotent stem cells, Airway epithelium, Eosinophils, Crystal

7. Study Design

Primary Purpose
Other
Study Phase
Not Applicable
Interventional Study Model
Single Group Assignment
Masking
None (Open Label)
Allocation
N/A
Enrollment
4 (Actual)

8. Arms, Groups, and Interventions

Arm Title
Eligible patients
Arm Type
Experimental
Intervention Type
Other
Intervention Name(s)
Blood sample
Other Intervention Name(s)
nasal brush
Intervention Description
A blood sample and a nasal brush for each participant. The nasal brushing will allow the isolation of epithelial cells that will serve as a comparison for the bronchial tissue produced from blood-derived iPS. A blood sample of approximately 14 ml will be taken for isolation and freezing of the blood mononuclear cells allowing the generation of iPS.
Primary Outcome Measure Information:
Title
Obtention of functional bronchial epithelium from iPSC: yes/no
Description
Was a functional bronchial epithelium obtained from the patient's induced pluripotent stem cells from blood? (yes/no) The achievement of functional bronchial epithelium (iALI) from iPSCs of a T2 severe asthma patient and an ABPA patient will be assessed by quantification of differentiation markers by immunofluorescence, integrity of the bronchial epithelium by measurement of trans-epithelial resistance (TEER), secretory function by measurement of mucin concentrations (CCSP, MUC5AC and MUC5B) and analysis of ciliary beat
Time Frame
Day 0 + culture (cross-sectional study)
Secondary Outcome Measure Information:
Title
Comparison of the transcriptomic profile between iALI and airway epithelial cells
Description
the transcriptomic profile of iALI from iPSCs of a T2 severe asthma patient and an ABPA patient will be compare to the one of airway epithelial cells from nasal brushing from the same patients
Time Frame
Day 0 + culture (cross-sectional study)
Title
Comparison of the transcriptomic profile between bronchial epithelia generated from severe asthma patients and from healthy subjects
Description
The transcriptomic profile of iALI from iPSCs of a T2 severe asthma patient and an ABPA patient will be compare to those from healthy subjects
Time Frame
Day 0 + culture (cross-sectional study)
Title
Differentiation of iPSC into mature eosinophils : yes/no
Description
Characterization of eosinophils obtained from iPSCs of a T2 severe asthma patient and an ABPA patient by the presence of specific markers by flow cytometry, immunohistochemistry and immunofluorescence
Time Frame
Day 0 + culture (cross-sectional study)
Title
Evaluation of immune cell/bronchial epithelium dialogue
Description
Co culture of the epithelia with iPSC derived eosinophils and the immune cell/bronchial epithelium dialogue in the co-culture of eosinophils with iALI will be evaluated by the activation of eosinophils in contact with the epithelium and by the characterization and study of the phenotype of the exposed epithelium
Time Frame
Day 0 + culture (cross-sectional study)
Title
Obtention of iPSC from peripheral blood sampling : yes/no
Description
Qualification of blood-derived iPSC lines by morphology, culture behavior, karyotype and expression of specific markers
Time Frame
Day 0 + culture (cross-sectional study)

10. Eligibility

Sex
All
Minimum Age & Unit of Time
18 Years
Maximum Age & Unit of Time
85 Years
Accepts Healthy Volunteers
No
Eligibility Criteria
Inclusion Criteria: - Smoking < 10 BP and weaned > 1 year. Diagnostic criteria for Severe asthma group T2 : History of severe asthma diagnosed by a physician (according to GINA criteria) Subject on high dose inhaled corticosteroid (ICS 1000 µg beclometasone equivalent) and beta-agonists for at least 6 months prior to inclusion Blood eosinophilia in history (previous years) > 300/mm3 Diagnostic criteria for Allergic bronchopulmonary aspergillosis (ABPA) group - Diagnosis of ABPA defined by the following 3 mandatory criteria: Diagnosis of asthma by the physician for at least 12 months based on the 2019 recommendations of the Global Initiative for Asthma (GINA) group Evidence of hypersensitivity to Aspergillus Fumigatus by skin test (on screening or previous documented positive skin test within the last 12 months), or serum Immunoglobulin E (IgE) specific antibodies to A. Fumigatus (≥ 0.35 kUnit/l) at screening. Elevated total serum IgE (> 1000 IU/ml). If the 3 ancillary criteria for the diagnosis of of ABPA (below) are met, an IgE level ≤ 1,000 IU/ml is acceptable. If the patient is receiving oral corticosteroids (OCs) at screening, a previous documented IgE level >1000 IU/ml within the last 12 months is acceptable. And at least 2 of the following ancillary criteria: Blood eosinophil count >500 cells/μl at screening for patients not receiving OCs at screening. For patients receiving OCs at screening, blood eosinophil count > 500 cells/μl at screening or documented previous eosinophil count > 500 cells/μl in the last 12 months. Presence of precipitating antibodies or serum immunoglobulin G (IgG) to A. Fumigatus at screening. Documented radiological abnormalities consistent with ABPA (such as transient mucoid impaction, hyperdense mucus [high attenuation of mucous plugs], opacities of centro-lobular nodules attenuation of mucous plugs], opacities of centro-lobular nodules, telectasis, bronchiectasis, etc.) by chest X-ray or high-resolution computed tomography (HR-CT) within the last 18 months or at screening. Exclusion Criteria: Other associated respiratory diseases (e.g. chronic obstructive pulmonary disease (COPD), cystic fibrosis) Patient in an exclusion period determined by another protocol Protected populations according to the French public health code: Parturient, nursing or pregnant women; subjects deprived of liberty by judicial or administrative decision; Major protected by law (under any form of guardianship). Lack of informed consent Non-beneficiary of the national health insurance system
Overall Study Officials:
First Name & Middle Initial & Last Name & Degree
Anne Sophie GAMEZ, MD
Organizational Affiliation
University Hospital, Montpellier
Official's Role
Principal Investigator
Facility Information:
Facility Name
university Hospital of Montpellier
City
Montpellier
ZIP/Postal Code
34295
Country
France

12. IPD Sharing Statement

Plan to Share IPD
Yes
IPD Sharing Plan Description
The general goal is to make the study data available to interested researchers as well as to provide proof of transparency for the study. Data will be made available to persons who address a reasonable request to the study director. Individual participant data (and an accompanying data dictionary) will be deidentified and potentially further cleaned or aggregated as the investigators deem necessary to protect participant anonymity.
IPD Sharing Time Frame
Datasets that underlie the results reported in the article (text, tables, figures, and appendices) can be requested after the publication process has been completed.
IPD Sharing Access Criteria
The conditions under which members of the public will be granted access to datasets are: The data will be used/examined in a not-for-profit manner; The data will not be used in an attempt to identify a participant or group of participants; The user does not work for a private insurance company; The data will not be used in support of any kind of private insurance policy or health penalties; The data will be used/examined for the advancement of science/ teaching while respecting participant/patient privacy and rights; The user will state why they wish to access the data.

Learn more about this trial

Modeling Bronchial Epithelium in Severe Asthma With Human Induced Pluripotent Stem Cells (iPSC)

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