Study of the involvement of RNA modifications during the Leishmania parasite-host interaction

• Grant number: 23/02341-1
• Support Opportunities: Scholarships in Brazil - Master
• Start date: March 01, 2023
• End date: March 31, 2025
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Nilmar Silvio Moretti
• Grantee: Artur Honorato Reis
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Associated research grant: 22/03075-0 - Unraveling the regulatory mechanisms of host-parasite interaction of Leishmania: focus on post-transcriptional and post-translational changes, AP.PNGP.PI
• Associated scholarship(s): 23/13752-2 - Impact of m6A RNA modification on Leishmania drug resistance to antimonial, BE.EP.MS

Abstract

The life cycle of species of the genus Leishmania is complex and involves flagellated promastigotes present in the sandfly insect vector and amastigotes present within the macrophages of the mammalian host. Leishmania has developed different host cell subversion mechanisms, ranging from changes in the expression of specific genes to changes in metabolism, such as glycolysis, ensuring its survival during infection. Post-transcriptional modifications of mRNA molecules, known as epitranscriptome, such as Nv-Methyladenosine (m6A), have been identified as a new regulator of the inflammatory response during infection by infectious agents. m6A levels are regulated by the writers METTL3, METTL14 and WTAP, which add the modification to the mRNA; the erasers, FTO and ALKBH5, which act as demethylases and the readers YTHDF1-2 and YTHDF1-3, which recognize the modification and can trigger other downstream effects. Considering the role of m6A in the host-pathogen interaction during infection by different agents and the ability of Leishmania to manipulate macrophage gene expression, we intend in this project to investigate the role of RNA modifications during Leishmania infection. For this we will: 1) Evaluate the expression profile of the m6A regulatory machinery in samples from patients diagnosed with cutaneous leishmaniasis; 2) Evaluate the protein expression levels of the m6A regulatory machinery during in vitro macrophage infection assays; 3) Perform direct RNA sequencing analyzes to identify the mRNAs modified by m6A during macrophage infection with Leishmania. Together, the data generated with this project will allow us to advance in understanding the mechanisms Leishmania used to progress during the infection in macrophages.