TRP ion channels are non-specific cationic channels with 6 transmembrane domains, most of them are permeable to Ca 2+ ions. Primarily, the multimodal thermosensitive members of the TRP family, TRPV1-4, TRPA1, and TRPM8, are proven to play role in skin sensory functions. In addition to sensory neurons in the skin, some of our previous work has also described the presence of sensory TRP channels on a number of non-neural cells and has been implicated in the physiological and pathological biological processes of keratinocytes and skin appendages (eg sebaceous gland, hair follicles). In this project, we investigate the role of keratinocytes and their heat-sensitive TRP channels in the development of itch and related pruriceptive signaling. Primary normal human epidermal keratinocytes (NHEK) isolated from surgical specimens and We studied the expression (RNASeq, Q-PCR, ELISA) and function (intracellular Ca2+ measurements) of TRP channels and other molecules involved in pruritic signaling in normal human epidermal keratinocytes (NHEKs). Expression of several sensory TRP channels was detected on NHEKs, among which TRPV3 and TRPV4 wee expressed at highest level. Their functionality was confirmed by Ca2+ measurements. In addition to TRP channels, keratinocytes also expressed a number of receptors and mediators that may play a role in the development of pruritus or pruritus-related dermatological disorders (eg, proteinase-activated receptor 2, TLR3, histamine H1 receptor, etc.). Subsequently, we investigated the role of TRP channels in the signaling pathways related to these potentially pruritogenic molecules in NHEKs. According to our results, activation of TLR3 upregulated TRPV3 and enhanced subsequent Ca2+ signals, which may contribute to other TLR3 evoked responses. Based on our results, it appears that keratinocytes may play a role in the transduction of certain itch-inducing signals. Supported by: NKFIH 120187; GINOP-2.3.2-15-2016-00050;ÚNKP-19-4-DE-285; ÚNKP-19-3-I-DE-140; János Bolyai Research Scholarship (TIB)