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Construction and characterization of a genetically modified Leishmania expressing the fluorescent protein GCaMP6: a new tool for antileishmanial compounds screening that might interfer in the parasite calcium homeostasis

Grant number: 18/26416-2
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): April 01, 2019
Effective date (End): December 31, 2021
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal researcher:Marcia Aparecida Silva Graminha
Grantee:Eduarda de Carvalho Le Sénéchal Horta
Home Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

The leishmaniasis correspond to a set of clinical manifestations caused by different species of Leishmania, transmitted to the mammal host through the bite of infected female phlebotomines during the blood repast. This parasitic disease is labeled as neglected and therefore presents few therapeutic options for its control, which are toxic and present very difficult routes of administration. The increase of resistance to antimonials, the limited amount of available drugs, as well as the toxic effects associated to them and, recently, the increase in the number of visceral leishmaniasis cases in patients with HIV virus, make the search for new leishmanicidal agents indispensable. Due to the importance of Ca2+ as a second messenger in trypanosomatids for controlling parasite life and death, the main goal of this proposal is the construction of a genetically modified Leishmania amazonensis strain bearing an episome expressing the genetic encoded calcium indicator protein GCAMP6, allowing monitoring calcium fluctuations in the parasite cytosol. The data herein obtained will be useful not just for comprehension of the Ca2+ role in parasite survival but mostly the application of this tool in drug discovery since prolonged high intracellular Ca2+ level lead cell death, thus helping us in discover new bioactive compounds that interfere with Ca2+ homeostasis.Keywords: Leishmaniasis, signaling pathways, homeostasis.

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