Production of Leishmania amazonensis mutant lines expressing modified luciferases

• Grant number: 15/26121-4
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: March 01, 2016
• End date: November 30, 2018
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Silvia Reni Bortolin Uliana
• Grantee: Victor de Sousa Agostino
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Protozoan parasites of the Leishmania genus are the etiological agents of leishmaniasis, diseases with diverse clinical presentations. Globally, about 2 million cases of tegumentary and visceral leishmaniasis are diagnosed every year. Only a few drugs are available for the treatment of leishmaniasis in Brazil: meglumine antimoniate, amphotericin B and pentamidine. However, these drugs present several limitations such as toxicity, parenteral administration and high cost. Therefore, there is an urgent need for new alternatives for the therapy of this disease. In our laboratory, we have been working to identify new antileishmanial chemotherapeutic agents. To that end, we have been employing experimental models for testing candidate compounds using mutant parasites that express reporter genes. That facilitates parasite detection in vitro and in vivo. Luciferase expressing parasites have been obtained and used successfully for characterizing drug activity. However, some limitations were found on the use of luciferase as a reporter. The aim of this project, therefore, is to obtain Leishmania amazonensis lines expressing variants of the luciferase enzyme named NanoLuc, NanoLuc-PEST and RedLuc. These enzymes are able to catalyze reactions with a higher specific activity or in wavelengths that are distinct from the wild type luciferases. Therefore, the are expected to increase in the sensitivity of parasite detection in vitro and in vivo in drug screening tests. Furthermore, these parasites will be also useful for studies concerning leishmaniasis pathophysiology.