An Optimized And Druggable Human Keratinocyte And Ipsc-Derived Sensory Neuron Co-Culture System For Atopic Itch

Atopic dermatitis (AD) is a highly prevalent, relapse-remitting, inflammatory skin disease, the hallmark symptom of which is chronic itch. Mechanisms underlying AD itch are multifactorial, involving various cells, receptors, and mediators. Developing a physiologically relevant, human model system for AD itch research and drug development is crucial. To this end, human induced pluripotent stem cell-derived sensory neurons (iPSCSNs) were cultured with human primary keratinocytes to form deconstructed skin models. Using Ca2+-imaging in a direct contact, 2.5D co-culturing format, which mimics natural skin innervation and permits both paracrine exchange and juxtacrine signaling, iPSCSNs exhibited functional TRPA1 responses not seen in monotypic iPSCSN cultures or in iPSCSNs conditioned with keratinocyte medium. Different AD-associated cytokines were used to stimulate the co-culture systems to mimic an inflamed lesional skin environment, whereby TNF was found to increase iPSCSN chemosensitivity. Finally, both TRPA1 and JAK1/2 inhibition reduced iPSCSN responses to pruritogens (TSLP, IL-31), thus supporting TRPA1 as a therapeutic target for AD itch in humans. This study demonstrates that human deconstructed skin models can be a useful tool in AD and broader pruritus research. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=181 SRC="FIGDIR/small/724000v1_ufig1.gif" ALT="Figure 1"> View larger version (35K): org.highwire.dtl.DTLVardef@11c0c9borg.highwire.dtl.DTLVardef@7fa518org.highwire.dtl.DTLVardef@2fe7a2org.highwire.dtl.DTLVardef@1105fa7_HPS_FORMAT_FIGEXP M_FIG C_FIG