Intestinal organoid as a tool to investigate the Annexin A1 functionalized multi wall lipid core nanocapsule

• Grant number: 22/05777-2
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Effective date (Start): November 15, 2022
• Effective date (End): November 14, 2023
• Field of knowledge: Biological Sciences - Immunology - Cellular Immunology
• Principal Investigator: Sandra Helena Poliselli Farsky
• Grantee: Milena Fronza Broering
• Supervisor: Soumita Das
• Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Research place: University of Massachusetts, Lowell (UMass Lowell), United States
• Associated to the scholarship: 18/26383-7 - Effects of lipidic nucleus nanocapsule containing Annexin A1 in Ulcerative Colitis model induced in mice, BP.DR

Abstract

The evolution of different types of organoids allowed a better understanding of organogenesis and also of the pathophysiology of different diseases. It is clear that the genesis and progression of Intestinal Bowel Disases (IBDs) share complex mechanisms and an intense multicellular interaction, that is often impossible to reproduce and observe them in 2D in vitro platforms. AnxA1, an endogenous 37KDa protein, is consistently described for its anti-inflammatory functions. In this context, AnxA1 plays the pivotal role on IBDs course, being associated with the protection of gastrointestinal mucosa during conditions of inflammation and epithelial injury. AnxA1 is a pivotal mediator of IBDs resolution. Moreover, AnxA1 seems to be an important mediator to organoids development. This project aims a BEPE fellow in the Laboratory of Pathology, coordinated by Dr. Soumita Das, University of California in San Diego, which has expertise on production and manipulation of murine and human intestinal organoids. After set up the conditions of murine gut organoids, it will investigated the role of recombinant AnxA1 (rAnxA1) or the rAnxA1 functionalized to multi-wall nanocapsule (MLNC-AnxA1) on the development of organoids and on tissue repair evoked by inflammatory agents. The gut organoid will be developed isolating intestinal crypts containing adult-stem cells, and employed in three set of experiments to investigate: 1) the ability of AnxA1, MLNC or MNLCAnxA1 alter the effects of macrophages on enteroid-derived monolayer bacterial injuried; 2) the effects of AnxA1, MLNC or MNLCAnxA1 on bacterial injuried enteroids; 3) the role of AnxA1, LMNC, or LMNCAnxA1 on the ability enteroid-derived monolayer to reorganize into enterocytes. As end-points will be evaluated transepithelial/transendothelial electrical resistance (TEER), expressions of adhesion molecules, tight junctions, AnxA1 expression, goblet cells presence and sequencing of mRNA in samples. Altogether, the BEPE fellow will allow the better understanding of the pathophysiologic mechanisms of IBD also to explore the potential of novel therapeutic approaches. (AU)