Modeling mucus plugging during type 2-high severe asthma using human induced pluripotent stem cells (hiPSC).

Severe asthma patients with persistent airflow obstruction are characterized by functional obstruction due to mucus plugs. Mucus plugs contain eosinophil derived-Charcot-Leyden crystals protein galectin-10. Our aim is to establish an in vitro human model of severe asthma that will recapitulate epithelium cell remodelling and mucus plugs using human induced pluripotent stem cells (hiPSCs) derived airway epithelial cells (i-ALI). We would like to explore how galectin-10 (gal-10) crystals could drive mucus plugs in human airways. We generated three hiPSC lines derived from peripheral blood of two highly characterized Type 2 (T2) high severe asthma (SA) patients and an Allergic Bronchopulmonary Aspergillosis (ABPA) patient (RECHMPL22_0384 – MOSAIC, <ext-link>NCT05616338</ext-link>, Montpellier University Hospital, France), named iPS T2 SA-1; T2 SA-2 and ABPA, respectively. Peripheral blood mononuclear cells (PBMCs) were reprogrammed using integration-free Sendai Virus. i-ALI were exposed in a chronic fashion to interleukin 13 to mimic T2 inflammation and then to acute gal-10 crystals apical exposure. Gal-10 crystals in the presence of IL-13 were able to induce MUC5AC formation in iALI derived from asthmatic patients. Confocal imaging and electron microscopy revealead that Gal-10 crystals were part of the mucin network. Gal-10 + IL-13 stimulation increased elasticity and viscosity of the mucus derived from the cultures. Human iPSCs are a powerful tool that recapitulate severe asthma heterogeneity. To our knowledge, it is the first in vitro severe asthma human model of mucus plugging endotype. Further work is needed to unravel the molecular mechanisms between galectin- 10 crystals and mucus plugging.