Using organoid platform to investigate the effect of smoking on congenital cystic lung disease [Oral presentation]

Abstract

<div><p><strong>Purpose:</strong><br>This study aimed to investigate the effects of cigarette smoke extract (CSE) on airway organoids (AOs) derived from lung tissue from patient with congenital cystic lung disease, and to evaluate the potential protective role of DAPT, a γ-secretase inhibitor, in modulating smoke-induced airway changes.</p><p><strong>Methods:</strong><br>AOs were established from both normal and diseased regions of intra-lobar pulmonary sequestration tissue. The organoids were cultured in two conditions: without DAPT for 2 weeks, or with 2.5μM DAPT for 1 week followed by no DAPT for 1 week. Both groups were subsequently exposed to 4% CSE for 24 hours. Analyses included mucus viscosity measurements, MUC5AC protein expression (ELISA), PAS staining for mucin visualization, and qPCR for key airway epithelial markers (MUC5AC, MUC5AB, FOXJ1), tight junction proteins (E-cadherin, ZO-1), and inflammatory cytokines (IL-6, IL-8).</p><p><strong>Results:</strong><br>Disease-derived AOs demonstrated higher baseline mucus secretion compared to normal tissue-derived AOs. CSE exposure significantly increased mucus viscosity and MUC5AC expression in both normal and diseased AOs, with a more pronounced effect in diseased organoids. Gene expression analysis revealed that CSE substantially upregulated mucins (MUC5AC, MUC5AB) and pro-inflammatory cytokines (IL-6, IL-8), particularly in disease-derived organoids without DAPT treatment. DAPT treatment effectively suppressed these CSE-induced changes, maintaining lower levels of mucus secretion and inflammatory markers. The expression of tight junction proteins (E-cadherin and ZO-1) remained relatively stable across all conditions.</p><p>​​​​​​​<strong>Conclusion:</strong><br>This study demonstrates that pediatric pulmonary sequestration-derived organoids show enhanced susceptibility to CSE-induced mucus hypersecretion and inflammatory responses compared to normal tissue-derived organoids. These findings provide valuable insights into the pathophysiology of pulmonary sequestration and confirm the potentially higher risk of chest infections int these patients.</p></div>