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Identification of factors related to phagocytosis and survival of L. amazonensis by comparison of proteomes and analysis of mutants by CRISPR-Cas9

Grant number: 21/08915-4
Support Opportunities:Regular Research Grants
Duration: June 01, 2022 - January 31, 2025
Field of knowledge:Biological Sciences - Parasitology - Protozoology of Parasites
Principal Investigator:Beatriz Simonsen Stolf
Grantee:Beatriz Simonsen Stolf
Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil


Leishmaniasis, a group of diseases caused by parasites of the genus Leishmania, is endemic in 98 countries and has an estimated annual incidence of more than 1 million cases. The symptomatic infection has several clinical forms, which are divided into cutaneous, mucocutaneous and visceral. In Brazil, three dermotropic species are predominant, L. (V.) braziliensis, L. (L.) amazonensis, and L. (V.) guyanensis. During its life cycle, the parasite presents itself as an extracellular promastigote transmitted by the insect vector and as an intracellular amastigote in the vertebrate host. The parasites express several virulence factors that influence their survival and infectivity. The amastigotes of PH8 and LV79 strains of L. (L.) amazonensis have differences in virulence in mice and in protein expression. In vitro, PH8 promastigotes have higher adhesion to macrophages and internalization. The comparison of the proteomes of membrane enriched fractions of PH8 and LV79 identified several differences, and the protein profile was able to discriminate samples from the two strains. This project aims to validate the differences observed in the proteomes and to evaluate the functional importance of some proteins. We will generate mutants (knockout and/or tagged) for enolase and LABCG1 by CRISPR-Cas9 and compare phagocytosis and infection between mutant and wild type parasites of the two strains. We also intend to analyze the susceptibility of PH8 and LV79 to complement and drugs (in the case of LABCG1 ko) and compare their LPGs and phlebotomine colonization. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
TANO, FABIA TOMIE; TELLERIA, ERICH LOZA; REGO, FELIPE DUTRA; COELHO, FELIPE SOARES; DE REZENDE, ELOIZA; SOARES, RODRIGO PEDRO; TRAUB-CSEKO, YARA MARIA; STOLF, BEATRIZ SIMONSEN. Phenotypical Differences between Leishmania (Leishmania) amazonensis PH8 and LV79 Strains May Impact Survival in Mammal Host and in Phlebotomine Sand Flies. PATHOGENS, v. 12, n. 2, p. 13-pg., . (18/14972-8, 21/08915-4, 17/26197-6)

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