Aquaporins e connexins: establishment of salivary gland organoid in vitro model as a tool for the analysis of communication channels in salivary gland structure and to unravel structural aspects of Lupus sialadenitis

• Grant number: 21/13517-8
• Support Opportunities: Regular Research Grants
• Start date: April 01, 2022
• End date: September 30, 2024
• Field of knowledge: Health Sciences - Dentistry
• Principal Investigator: Silvia Vanessa Lourenço
• Grantee: Silvia Vanessa Lourenço
• Host Institution: Faculdade de Odontologia (FO). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Kátia Cândido Carvalho ; Victor Elias Arana-Chavez
• Associated research grant(s): 22/00577-5 - Role of high salt diet and transcription factor NFTA5 on Sjögren's syndrome, AP.R SPRINT

Abstract

Lupus erythematosus (LE) is an autoimmune disease that affects multiple organs. It may affect the oral cavity and around 75% of the patients report subjective sensation of dry mouth (xerostomia). Our research group has demonstrated in multiple studies that xerostomia in LE is related to a type of manifestation specific of the disease - the lupus sialadenitis - which includes morphological patterns such as thickening of the ductal basal membrane, apoptosis, among others. The hypothesis of this proposal is that inflammatory factors may affect membrane surface molecules with alteration in communication channels, such as aquaporins and connexins, with loss of homeostasis, alterations in saliva composition, and probably culmination with xerostomia. In order to test this hypothesis we the following question will be addressed: Could inflammatory factors potentially alter the patterns of aquaporins and connexins in salivary glands? Research strategies: 1) establishment of in vitro salivary gland organoid model to be used as a tool for the studies of lupus sialadenitis; 2) tissue analysis: we will analyze the expression patterns of aquaporins and connexins in salivary glands from patients diagnosed with lupus erythematosus (LE), in normal controls and in salivary glands from human fetuses at the various developmental stages; 3) the organoid model will be exposed to inflammatory factors and posteriorly the expression patterns of aquaporins, connexins as well as FGF10 and SOX-9 will be evaluated using immunofluorescence and RT-qPCR. 4) analytic techniques: metabolomic and lipidomic analysis will be performed in the in vitro models to understand the biological lipid and metabolite hubs involved in salivary gland injury exposed to inflammatory factors. All results will be analyzed comparatively with the morphological aspects of salivary glands affected by LE. Expected results: to create a biological model to study salivary gland degenerative alteration to be able to create strategies to address salivary gland regenerative therapies. (AU)