TGF-β and IL-4+IL-13 induce neuroplasticity in an in vitro model of hPSC-derived sensory neurons

Carli Sanna Koster^1,2*I. Sophie T. Bos^1,2Chiara Lavitola^1,2Mihály Balogh¹Barbro N Melgert^1,2Reinoud Gosens^1,2*

• ¹Department of Molecular Pharmacology, Groningen Institute of Pharmacy, University of Groningen, Groningen, Netherlands
• ²Universitair Medisch Centrum Groningen Groningen Research Institute for Asthma and COPD, Groningen, Netherlands

Chronic type 2 inflammation is known to drive neuroplasticity of both afferent and efferent vagal nerves innervating many organs. This results in increased neuronal density and sensitivity, possibly contributing to pathologies such as eczema and asthma. However, the mechanisms driving these neuronal changes, particularly in sensory pathways, remain poorly understood, and appropriate in vitro models to study them are lacking. Here, we describe the differentiation of sensory neurons from human pluripotent stem cells. Generation of sensory neurons was validated by verifying expression of sensory neuron markers such as β3-tubulin, PGP9.5, TRPV1, Nav1.8 and Piezo1/2 using immunofluorescence, flow cytometry and RNA-sequencing, as well as functional responsiveness to capsaicin using calcium imaging and spontaneous firing usuing multi-electrode array. We exposed these hPSC-derived sensory neurons to TGF-β or the type 2 cytokines IL-4 and 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-β more strongly than IL-4+IL-13. Our results show robust a reproducible generation of functional hPSC-derived sensory neurons and their usability as a model to investigate mechanisms behind neuroplasticity. Furthermore, our findings support a role for TGF-β and type 2 cytokines in the development of neuroplasticity.