Development of advanced in-vitro human bronchial epithelial models and lung cell atlas in COPD enabled by thoracic surgery translational research

Background: Careful analysis of cellular function and differentiation in the human bronchial epithelium is important to understand, treat and prevent disease patterns that characterize chronic lung diseases (CLD, including chronic obstructive lung disease/COPD). To enable the translational research in thoracic surgery, close collaboration between surgical departments and associated laboratories is warranted. A highly standardized long-term culture approach based on primary human bronchial epithelial cells (pHBECs) from non-CLD and end-stage COPD patients was recently developed in our laboratory (Stoleriu et al.,Thorax 2024). To physiologically mimic the native microenvironment of the bronchial epithelium further adaptations of primary cell culture are needed.

Methods: Over 200 human bronchial tissue samples from CLD patients upon lung transplantation at the Thoracic Surgery Division of the LMU Hospital Munich between 01.2018-09.2024 were processed at the CPC-M bioArchive, the LMU Hospital, the Asklepios Clinic Gauting and the Institute for Lung Health and Immunity Munich. Long-term pHBECs cultures isolated from n=4 (each) non-CLD, COPD-II and IV patients at air-liquid-interface were used for comprehensive cell characterization and culture validation on a novel transwell system.

Results: Single cell RNA-seq analysis of pHBECs from non-CLD and COPD-IV patients revealed a novel basal cell population that could drive the hypersecretory phenotype in end-stage COPD derived cultures. The basal cell state was characterized by overrepresentation of Wnt and Notch canonical airway pathways involved in secretory and multi-ciliated cell differentiation. After successful long-term pHBECs culture on conventional silicone-based membrane transwells, a novel culture system based on elastic-porous convex transwell membranes was developed to improve cell differentiation, mimic physiological primary cell cultures and facilitate a reliable measurement of transcellular drug transport.

Conclusion: The collaboration between thoracic surgery and respective scientific laboratories is able to update knowledge about the cellular landscape of the human bronchial epithelium and of the lung cell atlas, thus contributing to a better disease understanding in COPD patients driving therapeutic ideas.

Publikationsverlauf

Artikel online veröffentlicht:
18. März 2025