THU0223 CHRONIC ADRENERGIC STIMULATION OF MINOR SALIVARY GLANDS OF PATIENTSWITH PRIMARY SJÖGREN’S DRIVES ER STRESS AND ACTIVATION OF THE UNFOLDED PROTEIN RESPONSE
Authors: Kalliopi Moustaka, Stergios Katsiougiannis, Roxane Tenta, Sofia Havaki, Pari Koutsoudaki, Haralampos M. Moutsopoulos, Fotini Skopouli
Journal: Annals of the Rheumatic Diseases (2019)

<h3>Background</h3> Sjögren's syndrome (SS) is a common autoimmune disease in which the main targets of immune injury are specific secretory epithelia, such as salivary and lacrimal glands. Stress appears to play a significant role in the initiation of this entity. <h3>Objectives</h3> The aim of the present study was to investigate whether chronic stress plays a role in triggering endoplasmic reticulum (ER) stress in salivary gland epithelial cells from SS patients. <h3>Methods</h3> Minor salivary gland biopsy specimens were obtained from six SS patients and six control patients with sicca symptoms not fulfilling AECG criteria [1]. The expression and cellular localization of β1-, β2- and α1-adrenoceptors and the levels of cAMP were measured by immunofluorescence. The morphology of the ER was evaluated <i>in situ</i> by Transmission Electron Microscopy (TEM). Primary salivary gland epithelial cell lines (SGEC) derived from minor salivary gland biopsies, were established by the explant out-growth technique [2] and were treated with epinephrine and norepinephrine. The protein levels of the ER stress markers GRP78/Bip and C/EBP homologous protein (CHOP) were determined by immunoblot analysis. <h3>Results</h3> <i>In situ</i> immunofluorescence staining revealed increased expression of β1-, β2- and α1 adrenoceptors as well as cAMP levels in tissues derived from SS patients compared to controls. TEM evaluation of salivary tissues from SS subjects revealed extensive dilation of the ER lumen compared to controls. Treatment of SGEC with epinephrine and norepinephrine did not influence cell survival (cell viability assay). To mimic chronic stress <i>in vitro</i>, epinephrine was applied for 10 days on SGEC. It was found that 20μM Epinephrine induced severe ER stress on SGEC, as attested by increased expression of GRP78/Bip and CHOP, after 3 and 6 days of treatment. The expression of ER stress markers returned to basal levels after 10 days of treatment. <h3>Conclusion</h3> Our data revealed that adrenergic receptors are differentially expressed by salivary epithelium of SS patients compared to control tissues. This finding was further substantiated by the increased levels of cAMP, indicating a profound sympathetic stimulation of this secretory tissue. The dilatated ER lumen of the salivary gland epithelial cells from SS patients confirmed the occurrence of ER stress. Moreover, the finding that catecholamines induce severe ER stress <i>in vitro</i> in SGEC under conditions resembling chronic stress suggests a causal relationship between sympathetic tone and ER stress. Deciphering the role of chronic stress in ER machinery will provide important insights that may lead to novel therapeutic targets for SS. <h3>References</h3> [1] Vitali C, et al. Ann Rheum Dis2002; 61:554–8. [2] Kapsogeorgou EK, et al. J Immunol2001; 166:3107–13. <h3>Acknowledgement</h3> The research has been funded in part by the European Union's Horizon 2020 multicentric protocol HARMONICSS; H2020-SC1-2016; grant agreement no: 731944. <h3>Disclosure of Interests</h3> None declared