DYSREGULATED MUCIN PRODUCTION ASSESSMENT AS A RESPONSE TO ATYPICAL ENTEROPATHOGENIC ESCHERICHIA COLI (aEPEC) STRAINS INFECTIONS

• Grant number: 24/21847-6
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: January 23, 2025
• End date: February 28, 2025
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Principal Investigator: Tânia Aparecida Tardelli Gomes do Amaral
• Grantee: Juan Josue Puño Sarmiento
• Supervisor: Vivek Malhotra
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Institution abroad: Centre For Genomic Regulation, Spain
• Associated to the scholarship: 22/12006-2 - Role of cytokines in modulating the induction of mucus production by atypical enteropathogenic Escherichia coli (aEPEC) strains in intestinal cells in vitro, BP.PD

Abstract

The understanding of the main role of goblet cells was only restricted to the production of mucins; however, this concept has changed drastically because of the discovery of populations of goblet cells exerting metabolic functions and the defense that place them now as the central players in the intestinal homeostasis. Altered goblet cell function could lead to a dysfunctional mucus barrier essential for disease prevention. There is a gap in knowledge about the impact of enteric pathogens on goblet cell function that could contribute to the development of chronic inflammatory diseases. The proposal of this internship project is to explore the mechanisms associated with dysregulated mucin production after infection with aEPEC strains. To this end, we will evaluate gel-forming and cell surface mucin levels in differentiated HT29-N2 cells. In addition, we will examine intracellular calcium levels that correlate with the mucin secretory function. Finally, the key proteins involved in inflammatory processes, cytokine production, and calcium regulators will be detected from cellular contents. These responses would represent a starting point to elucidate the hypersecretion phenomenon induced by some strains in an in vivo model. Recently, through a short research stay funded by the Capes PrInt program, I could conduct a pilot study at the CRG in Barcelona by testing the effect of aEPEC strains on intracellular calcium influx levels from goblet cells culture. In addition, we are expecting results of transcriptome analysis that will provide goblet cells endoplasmic reticulum (ER) stress responses induced by bacterial stimuli. The Malhotra's lab has a large experience identifying potential proteins involved in mucin secretion. Most of their methodologies are already standardized and since a mutual agreement has been signed by both institutions, the aEPEC strains are already stored in the host laboratory.