The parathyroid hormone family member TIP39 interacts with sarco/endoplasmic reticulum Ca2+- ATPase activity by influencing calcium homeostasis

Abstract

Darier disease (DD) is a genetic skin disease that is associated with mutations in the ATP2A2 gene encoding the type 2 sarco/endoplasmic reticulum Ca²⁺-ATPase (SERCA2). Mutations of this gene result in alterations of calcium homeostasis, abnormal epidermal adhesion and dyskeratosis. Silencing of ATP2A2 in monolayer cell culture of keratinocytes reduces desmoplakin expression at the borders of cells and impacts cell adhesion. Here, we report establishment of a three-dimensional (3D) epidermal model of DD, and use this model to evaluate peptide therapy with tuberoinfundibular peptide of 39 residues (TIP39) to normalize calcium transport. Gene silencing of ATP2A2 in keratinocytes grown in a 3D model resulted in dyskeratosis, partial parakeratosis and suprabasal clefts that resembled the histological changes seen in skin biopsies from patients with DD. TIP39, a peptide recently identified as a regulator of keratinocyte calcium transport,, was then applied to this ATP2A2 silenced 3D epidermal model. In normal keratinocytes, TIP39 increased [Ca²⁺]_i through the inositol trisphosphate (IP3) receptor pathway and stimulated differentiation. In monolayer ATP2A2-silenced keratinocytes, although TIP39 increased cytosolic calcium from the endoplasmic reticulum (ER), the response was incomplete compared with its control. TIP39 was observed to reduce intercellular clefts of the gene silenced epidermal model but did not significantly upregulate keratinocyte differentiation genes such as keratin10 and filaggrin. These findings indicate that TIP39 is a modulator of ER calcium signaling and may be used as a potential strategy for improving aspects of DD.

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