Characterization of genes potentially involved in the metabolism and transport of iron to the Leishmania glycosome

• Grant number: 22/04551-0
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: December 01, 2022
• Status: Discontinued
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Maria Fernanda Laranjeira da Silva
• Grantee: Romário Lopes Boy
• Host Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated scholarship(s): 24/21947-0 - Exploring Iron Metabolism and Antimonial Resistance in Leishmania amazonensis: A Metabolomic and Genetic Analysis of AQP1 and LIRG1, BE.EP.DR

Abstract

Protozoa of the genus Leishmania are the etiological agents of diseases known as leishmaniasis that affect millions of people. These organisms are parasites that alternate between the invertebrate and vertebrate host where they are able to survive and replicate in macrophages. One of the critical conditions found by Leishmania in the macrophage is the lack of nutrients, including iron. It has already been shown that, in Leishmania, iron deprivation induces the expression of a series of genes not yet characterized. Trypanosomatids have unique organelles called glycosomes that constitute one of the main differences between parasite and host. These organelles compartmentalize enzymes that use iron as a cofactor, however, the transport of iron into the glycossomes has not yet been characterized. From in silico analysis of L. amazonensis transcripts modulated by iron deprivation, we identified 11 genes with predicted glycosome-targeting signals and started the characterization of 5 of these genes with three or more predicted transmembrane domains. By immunofluorescence, we demonstrated the glycosomal compartmentalization of the proteins encoded by 2 of these genes in promastigotes. Therefore, our goal is to determine the role of these 2 genes in Leishmania iron metabolism by obtaining knockout parasites using the CRISPR/Cas9 strategy. The functional characterization of these knockouts will contribute to the elucidation of the mechanisms involved in the metabolism and transport of iron to the Leishmania glycosome and may indicate new targets for the development of chemotherapeutic agents for the treatment of leishmaniasis.