Use of Plasmodium knowlesi as a model for malaria research in vitro

• Grant number: 18/06219-8
• Support Opportunities: Research Grants - Young Investigators Grants
• Start date: March 01, 2019
• End date: February 29, 2024
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Roberto Rudge de Moraes Barros
• Grantee: Roberto Rudge de Moraes Barros
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Associated researchers: Daniel Youssef Bargieri ; David Serre ; Leonardo José de Moura Carvalho
• Associated research grant(s): 24/13739-9 - 2024 ASTMH Annual Meeting, AR.EXT
• Associated scholarship(s): 21/14319-5 - Antimalarial activity evaluation of new compounds using Plasmodium knowlesi and P. falciparum in vitro, BP.IC; 21/01910-7 - Analysis of the chloroquine activity and pkcrt gene transcription in Plasmodium knowlesi blood stages in vitro, BP.IC; 21/02996-2 - Molecular characterization of transcriptional promoter sequences of Plasmodium spp, BP.DD; 20/02303-4 - Transfection of P. knowlesi in vitro and P. cynomolgi in vivo using the CRISPR/Cas9 technology, BP.MS; 19/07223-1 - In vitro evaluation of antimalarial compounds using transgenic parasites, BP.IC

Abstract

Malaria is a major human parasitic disease, responsible for more than 200 million cases and around 500,000 deaths per year worldwide. Five species of Plasmodium cause human malaria: Plasmodium falciparum, P. vivax, P. ovale, P. malariae and P. knowlesi. P. falciparum, the main cause of deaths for malaria, receives most of the attention and investments in research and control, mainly due the existence of robust long-term in vitro culture techniques. However, P. falciparum is evolutionarily and biologically distant from other species, and control measures developed against P. falciparum do not show the same efficacy when applied against other species. This fact highlights the need of specific research, especially against P. vivax, responsible for ~90% of cases in Brazil. The lack of long-term in vitro culture systems has been a challenge to P. vivax research, maintaining this species neglected worldwide. Recently P. knowlesi, a parasite evolutionary close to P. vivax, was adapted to long-term in vitro culture, offering an alternative for malaria research. This project aims to study different aspects of the biology of Plasmodium using in vitro cultures of P. knowlesi and in vivo infections of P. cynomolgi. To achieve this main goal we will work with four secondary goals: (1) development of transgenic parasites using the CRISPR/Cas9 technology; (2) evaluation of antimalarial activity of drugs in vitro; (3) molecular characterization of transcriptional regulatory sequences; and (4) use of the Single Cell RNAseq technology to identify and characterize P. knowlesi sexual stages. Results obtained will improve the understanding of the biology of P. knowlesi and P. vivax, accelerating the development of control strategies. In Brazil, there is currently no laboratory working with P. knowlesi. The establishment of this experimental model will put the country at the forefront of malaria research. (AU)

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)

BARBOSA, CAMILA S.; AHMAD, ANEES; MALUF, SARAH EL CHAMY; MOURA, IGOR M. R.; SOUZA, GUILHERME E.; GUERRA, GIOVANNA A. H.; BARROS, ROBERTO R. MORAES; GAZARINI, MARCOS L.; AGUIAR, ANNA C. C.; BURTOLOSO, ANTONIO C. B.; et al. Synthesis, Structure-Activity Relationships, and Parasitological Profiling of Brussonol Derivatives as New Plasmodium falciparum Inhibitors. PHARMACEUTICALS, v. 15, n. 7, p. 21-pg., 2022-07-01. (13/07600-3, 21/03977-1, 20/14429-2, 19/17721-9, 19/19708-0, 20/12904-5, 18/06219-8, 15/20084-0, 18/07287-7, 13/18009-4, 21/14319-5)

MORAES BARROS, ROBERTO R.; THAWNASHOM, KITTISAK; GIBSON, TYLER J.; ARMISTEAD, JENNIFER S.; CALEON, RAMONCITO L.; KANEKO, MIHO; KITE, WHITNEY A.; MERSHON, J. PATRICK; BROCKHURST, JACQUELINE K.; ENGELS, THERESA; et al. Activity of Plasmodium vivax promoter elements in Plasmodium knowlesi, and a centromere-containing plasmid that expresses NanoLuc throughout the parasite life cycle. Malaria Journal, v. 20, n. 1, p. 13-pg., 2021-06-05. (18/06219-8)

VIEIRA, TAIS BARUEL; ASTRO, THAFNE PLASTINA; DE MORAES BARROS, ROBERTO RUDGE. Genetic Manipulation of Non-Falciparum Human Malaria Parasites. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 11, p. 5-pg., 2021-08-30. (18/06219-8, 19/07223-1, 20/02303-4)