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Autophagic process and host cell lysosome mobilization in Trypanosoma cruzi invasion

Grant number: 17/05145-8
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): October 01, 2017
Effective date (End): September 30, 2019
Field of knowledge:Biological Sciences - Parasitology - Protozoology of Parasites
Principal Investigator:Nobuko Yoshida
Grantee:João Paulo Ferreira Rodrigues
Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil


Diverse cellular and molecular aspects of cell invasion by Trypanosoma cruzi, a crucial process for the establishment of infection, have been clarified. However, many questions still remain to be investigated. Among the parasite factors involved in T. cruzi invasion, the surface molecule gp82, specific of metacyclic trypomastigotes, mediates invasion through its signaling activity that induces the host cell lysosome spreading and exocytosis. One of the questions to be addressed is whether the lysosomal glycoproteins LAMP1/2 play a role in invasion by metacyclic forms. This issue is pertinent, provided that conditions that induce lysosome biogenesis/spreading lead to increased parasite internalization. It is also relevant to determine whether gp82 binds to LAMP1/2 proteins. Another aspect to be analyzed is whether the autophagic process is induced by T. cruzi metacyclic forms with reduced invasion capacity expressing the surface molecule gp90 that functions as a negative regulator of invasion. Our main objectives are to: i) determine the effect of anti- LAMP1/2 antibodies on the internalization of metacyclic forms; ii) obtain HeLa cells with reduced or null expression of LAMP1/2, ou both proteins; iii) examine the susceptibility of cells silenced in a LAMP1 and/or LAMP2 toward parasite invasion; iv) determine the ability of gp82 to bind to LAMP1/2; v) verify whether poorly invasive parasites, as well as purified gp90, induce the autophagic process. (AU)

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