Development of 3D in vitro systems for studies of intestinal infections

• Grant number: 17/50158-0
• Support Opportunities: Regular Research Grants
• Start date: March 01, 2018
• End date: August 31, 2019
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Agreement: Uppsala University
• Mobility Program: SPRINT - Projetos de pesquisa - Mobilidade
• Principal Investigator: Renata Rosito Tonelli
• Grantee: Renata Rosito Tonelli
• Principal researcher abroad: Staffan Svärd
• Institution abroad: Uppsala University (UU), Sweden
• Host Institution: Instituto de Ciências Ambientais, Químicas e Farmacêuticas (ICAQF). Universidade Federal de São Paulo (UNIFESP). Campus Diadema. Diadema , SP, Brazil
• Associated research grant: 16/19429-5 - Protein post-translational modification in Giardia lamblia: study of SUMOyloma and SUMOylation of cytoskeketal proteins, AP.R

Abstract

Giardia lamblia and Cryptosporidium spp. are important enteric protozoa parasite responsible for Giardiasis and Cryptosporidiosis, respectively. Both are diarrheic diseases affecting humans and animais worldwide. Giardia causes disease without invading adjacent tissues and secreting toxins. Several mechanisms have been proposed to be importanf for induction of symptoms during a Giardia infection but there is no consensus. Svärd's group have identified several new putative virulence factors in Giardia that need to be evaluated. A major challenge in Cryptosporidium research has been the lack of an in vitro culture system, which supports continuous infection and completion of the life cycle. This has limited our understanding of the mechanisms involved in pathogenesis. Considering these, the aim of this proposal is to establish a human 3D intestinal co-culture model for long-term infection with Giardia and Cryptosporidium. This system will be used to study earlier virulence mechanisrn like mucus deqradatlon, apoptosis and changes in intestinal permeability upon Giardia and Cryptosprodium infections. RNA sequencing using illumina next generation sequencing will be used to identify genes in the host cells and Giardia that are influenced by the interaction at different time-points. For Cryptosporidium we wilt do single cell RNA sequencing (RNAseq) on parasites of different morphologies at different times. (AU)