Chronic itch affects about to 1 in 5 people worldwide (Hay et al., 2014; Weisshaar, 2016). Current anti-itch drugs and treatment options are highly nonspecific and largely ineffective. Thus, for most patients, chronic itch is intractable. Mounting evidence suggests that dysregulation or overactivation of transient receptor potential vanilloid 3 (TRPV3) could drive chronic itch; however, basic TRPV3 research is lacking. This project investigated how the TRPV3 channel functions in the epidermal layer of human skin and asks what role it plays in dermatitis and skin derived itch. Specifically, this work examines TRPV3 signalling, functional activity, sensitisation, and trafficking in normal human epidermal keratinocytes – a key culture model for human epidermis. A variety of methods including phospho-kinase arrays, cytokine arrays, live calcium imaging, and immunocytochemistry were employed. This work also utilised unsupervised clustering analyses to assess calcium imaging data. The data described herein confirm TRPV3 as a potent inducer of epidermal inflammation, with collaborators revealing a clear link between TRPV3-evoked PAI-1 release and skin-derived itch. Unsupervised clustering analyses revealed the phasic nature of TRPV3- induced calcium flux and highlighted the role of SNARE-mediated receptor cycling in normal TRPV3 activity. Lastly, this work exposed a novel neuro-epidermal link: B-type natriuretic peptide (BNP) – a key neuropeptide driving human itch – sensitised TRPV3 activity and promoted kinase-dependent TRPV3 trafficking. This basic work demonstrated the impact of TRPV3 in human skin and, together with in vivo data from collaborators, identified several novel targets for anti-dermatitis and anti-itch therapeutics. Overall, we believe that interruption of the TRPV3-associated pathways outlined herein would alleviate both inflammation and itch in human patients with atopic dermatitis, psoriasis, and other pruritic dermatoses.