A high-throughput 3D cantilever array to model airway smooth muscle hypercontractility in asthma

Asthma is often characterized by tissue-level mechanical phenotypes that include remodeling of the airway and an increase in airway tightening driven by the underlying smooth muscle. Existing therapies only provide symptom relief and do not improve the baseline narrowing of the airway or halt progression of the disease. To investigate such targeted therapeutics, there is a need for models that can recapitulate the 3D environment present in this tissue, provide phenotypic readouts of contractility, and be easily integrated into existing assay plate designs and laboratory automation used in drug discovery campaigns. To address this, we have developed DEFLCT, a high-throughput plate insert that can be paired with standard labware to easily generate high volumes of microscale tissues in vitro for screening applications. Using this platform, we exposed primary human airway smooth muscle cell-derived microtissues to a panel of six of inflammatory cytokines present in the asthmatic niche, identifying TGF-β1 and IL-13 as strong contractile modulators. RNAseq analysis further demonstrated enrichment of contractile and remodeling-relevant pathways in TGF-β1 and IL-13 treated tissues as well as pathways generally associated with asthma. Taken together, these data establish a disease relevant, 3D tissue model for the asthmatic airway which combines niche specific inflammatory cues and complex mechanical readouts that can be utilized in drug discovery efforts. ### Competing Interest Statement All authors are current or prior employees (were employees at the time of their contributions to the paper) of Novartis.