Modeling Bronchial Epithelium Modifications Associated With COPD Using iPS

Modeling Bronchial Epithelia Modifications Associated With Chronic Obstructive Pulmonary Disease (COPD) Using Induced Pluripotent Stemcells (iPS)

The primary objective of this study is to 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 (iPS) clone derived from cutaneous biopsy in a population of heavy smokers (plus patients with chronic obstructive pulmonary disease) in order to obtain differentiated bronchial epithelia in vitro.
• For each of these same patients, generation of bronchial epithelium in vitro from bronchial biopsy using human bronchial epithelial cells (HBECs) in air-liquid interface (ALI) cultures.

Secondary objectives:

• Confrontation of the two models of in vitro generation of bronchial epithelium by iPS and ALI in order to validate our model of study of the human bronchial epithelium COPD generated by the iPS.
• Inducing a "COPD-like" epithelial phenotype by exposing the bronchial epithelia generated from the iPS to a source of in vitro toxin (tobacco) and studying its effects by transcriptomic analysis via high-throughput sequencing (NGS) at key steps during the model.
• Demonstrate the feasibility of obtaining iPS from peripheral blood sampling.

Inclusion Criteria for COPD patients:

• The patient has given his/her signed, informed consent
• Forced expiratory volume in 1 second / forced vital capacity < 0.7 [FEV1/FVC < 0.7]
• FEV1 < 50% of predicted value
• Exclusion of other respiratory pathologies, based on anamnesis, computed tomography and complementary respiratory function tests and transthoracic ultrasound,
• Smoking >10 pack-years, ongoing or weaned
• Absence of exacerbation or intercurrent infectious episode for at least 4 weeks at time of inclusion
• Medical indication for a bronchial fibroscopy (indication validated by the physician in charge of the patient and the study investigating physician): pulmonary peripheral nodule exploration with computed tomography, interventional endoscopy for emphysema reduction, removal of foreign bodies, other indications.
• Absence of contraindications for bronchial biopsy (left to physician's discretion): not limited to: regular intake of anticoagulants or anti-platelet aggregation, hemostasis abnormalities

Inclusion Criteria for smokers:

• The patient has given his/her signed, informed consent
• Forced expiratory volume in 1 second / forced vital capacity > 0.7 [FEV1/FVC > 0.7]
• FEV1 > 80% of predicted value
• Exclusion of other respiratory pathologies, based on anamnesis, computed tomography (absence of extensive emphysema lesions defined by the presence of less than <5% relative surface area occupied by densities less than 910 UH (measured on millimeter cuts)) and respiratory function tests (Residual Volume <120%, Diffusion Capacity for Carbon Monoxide > 60% and Total Pulmonary Capacity> 120%) and transthoracic ultrasound (normal),
• Smoking >40 pack-years, ongoing or weaned
• Absence of exacerbation or intercurrent infectious episode for at least 4 weeks at time of inclusion
• Medical indication for a bronchial fibroscopy (indication validated by the physician in charge of the patient and the study investigating physician): pulmonary peripheral nodule exploration with computed tomography, interventional endoscopy for emphysema reduction, removal of foreign bodies, other indications.
• Absence of contraindications for bronchial biopsy (left to physician's discretion): not limited to: regular intake of anticoagulants or anti-platelet aggregation, hemostasis abnormalities

Exclusion Criteria:

• Contra-indication for bronchial fibroscopy and cutaneous biopsy
• Deficiency in α-1 antitrypsin
• Significant bronchiectasis on chest computed tomography scan
• Colonization of the airways by Pseudomonas Aeroginosa or Aspergillus
• Chemotherapy or other cytotoxic treatments in the last 12 months
• Systemic corticotherapy in the 4 weeks prior to inclusion
• Long-term antibiotic treatment, notably by macrolides
• Any other treatment that may affect the bronchial epithelium, this being left to the discretion of the investigating physician
• Consumer of illicit drugs or alcohol
• Participant in an exclusion period determined by a previous study
• Subject not affiliated with, or not beneficiary of, a social security plan.
• Pregnant or nursing women according to Article L1121-5 of the Public Health Code.
• Vulnerable persons according to Article L1121-6 of the Public Health Code
• An adult who is protected or incapable of giving consent in accordance with Article L1121-8 of the Public Health Code
• Subject deprived of freedom by judicial or administrative decision.
• Those patients for whom two epithelia cultures (1 from bronchial biopsy ("ALI") and 1 from fibroblast reprogrammation of redifferentiated iPS ("iALI")) were not obtained will be excluded (post-inclusion).