TGF-ß and IL-4+IL-13 induce neuroplasticity in an in vitro model of hPSC-derived sensory neurons
Asthma is a chronic inflammatory disease of the airways characterized by airway hyperresponsiveness. Recent studies using mouse models, bronchial biopsies of asthma patients, and lung tissue of patients with fatal asthma have implicated neuroplasticity as a pathological feature of severe asthma, possibly contributing to airway hyperresponsiveness. Currently, we lack the models to study the mechanisms behind sensory neuroplasticity in vitro. Here, we describe the differentiation of sensory neurons from human pluripotent stem cells. Generation of sensory neurons was validated using immunofluorescence, flow cytometry and RNA-sequencing, as well as functional responsiveness to capsaicin using calcium imaging. We exposed these hPSC-derived sensory neurons to TGF-ß or Th2-cytokines IL-4 together with IL-13, both of which have important roles in asthmatic airway remodelling. Both treatments induced neuroplasticity-related changes such as increased network density and neuronal sensitivity in the sensory neurons, albeit TGF-ß more strongly than IL-4+IL-13. Our results show that hPSC-derived sensory neurons can be generated robustly and reproducibly and provide a good model to investigate the mechanisms behind neuroplasticity. Furthermore, our findings support a role for the asthmatic airway microenvironment, most notably TGF-ß, in the development of neuroplasticity. Overall design: H9WAO9 human Pluripotent Stem cell (hPSC)-derived sensory neurons were used to study neuroplasticity development. Mature hPSC-derived sensory neurons exposed to TGF-ß or IL-4+IL-13 for 5-days were subjected to RNA-sequencing.