Analysis of the role of enolase and LABCG1 in L. (L.) virulence amazonensis using tags and silencing by CRISPR-Cas9

• Grant number: 21/08666-4
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: April 01, 2023
• End date: April 30, 2025
• Field of knowledge: Biological Sciences - Parasitology
• Principal Investigator: Beatriz Simonsen Stolf
• Grantee: Gustavo Rolim Barbosa
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Leishmaniasis is a complex of diseases caused by parasites of the genus Leishmania, endemic in 98 countries and responsible for more than 1 million cases every year. The human symptomatic infection has different clinical forms, which are grouped into tegumentary/cutaneous and visceral. The parasite is transmitted to man (and other vertebrates) by insect vectors, in which it remains in the promastigote form. Inside phagocytic cells of the vertebrate host it differentiates and multiplies in the amastigote form (mandatory intracellular). Recent works from our group demonstrated that parasites from PH8 and LV79 strains of Leishmania (L.) amazonensis have differences in virulence in mice and in infectivity in macrophages (in vitro). Promastigotes from the two strains show differences in Lipophosphoglycan (LPG) and in proteomes of membrane enriched fractions. The proteins enolase and LABCG1 are more abundant in the proteome of PH8 strain, and we believe that they may contribute to the higher virulence of this strain. Therefore, this project aims to determine the role of these proteins on the infectivity and survival of L. (L.) amazonensis. To achieve that, we will follow the abundance and location of these proteins in PH8 and LV79 using a protein tag method and will develop a knockout lineage for LABCG1 in PH8 by CRISPR-Cas9. We intend to evaluate the labeled proteins and the knockout lineage during infection to determine their roles in the host-pathogen relationship.