Using Repopulated Epithelium on Bronchial Scaffolds to Study Epithelial Cell – Extracellular Matrix Interactions in patients with Chronic Obstructive Pulmonary Disease and Healthy Donors

Project: Dissertation

Description

The extracellular matrix (ECM) in each organ/tissue is unique with respect to architecture and composition. It is also a highly dynamic complex, changing continuously to adapt to different physiological and pathophysiological circumstances. Epithelial–mesenchymal interactions play an important role in epithelial differentiation and repair and 3D air liquid interface (ALI) culture models have been used to study the mechanisms of epithelial differentiation and repair. However, these models do not recapitulate the dynamic interactions between epithelial cells and the ECM, not even in the presence of ECM components such as collagens and proteoglycans. As stem cell research progresses, new methodologies become available for repopulating stem cells and primary airway epithelial cells on decellularized tracheal scaffolds and differentiate them into pseudostratified tracheal epithelial cells. This work suggests it may be possible to establish an ex vivo airway model using a human bronchial ECM scaffold repopulated with human bronchial epithelial cells
(HBECs). This approach would allow us to study the interactions between airway epithelial cells and bronchial ECM from patients with airway diseases, e.g. COPD and other chronic diseases such as IPF and CF. This system could potentially be used as a disease related ex vivo model for early screening for adverse airway
effects, such as airway irritant potency of new candidate drugs.

Hypothesis
Alterations in composition of the bronchial ECM modulate the phenotype of epithelial cells and cause pathological remodeling of airway epithelial cells in COPD patients.

Objectives and aims
1. To establish an ex vivo airway model by growing normal human bronchial epithelial (NHBE) cells on decellularized human bronchial scaffolds.

2. To study components including carbohydrate context and glycosaminoglycans of bronchial scaffolds of COPD patients and healthy donors.

3. To study the effects of different scaffolds on epithelial cell phenotype.

4. To investigate epigenetic modifications in NHBE cells grown on scaffolds from COPD patients and healthy donors. Later, the same experiment will be done in HBECs from COPD patients grown on COPD and normal bronchial scaffolds.

5. To use RNA deep sequencing to compare the gene signatures in NHBE cells grown on scaffolds from COPD patients and healthy donors. Later, the same experiment will be done in HBECs from COPD patients (with/without cigarette smoke extract stimulation) grown on COPD and normal bronchial scaffolds
StatusNot started

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