Deletion of the lipid droplet protein kinase gene affects lipid droplets biogenesis, parasite infectivity, and resistance to trivalent antimony in < i > Leishmania infantum < /i >

by Juliana Martins Ribeiro, Paula Alves Silva, H éllida Marina Costa-Silva, Ana Maria Murta Santi, Silvane Maria Fonseca Murta The Lipid Droplet Protein Kinase (LDK) facilitates lipid droplet (LD) biogenesis, organelles involved in various metabolic and signaling functions in trypanosomatids. As LDK ’s function has not been previously explored inLeishmania spp., we utilized CRISPR/Cas9 technology to generateLDK-knockout lines ofLeishmania infantum to investigate its role in this parasite. Our findings demonstrate thatLDK is not an essential gene forL.infantum, as its deletion did not impede parasite survival. Furthermore, removingLDK did not impact the growth of promastigote forms ofL.infantum lackingLDK. However, a noticeable reduction in LDs occurred during the stationary phase of parasite growth following LDK deletion. In the presence of myriocin, a LD inducer,LDK-knockout parasites displayed reduced LD abundance during both logarithmic and stationary growth phases compared to control parasites. Moreover, an infection analysis involving THP-1 cells revealed that 72 h post-infection,LDK-knockoutL.infantum lines exhibited fewer infected macrophages and intracellular amastigotes than control parasites.LDK-knockoutL.infantum lines also displayed 1.7 to 1.8 -fold greater resistance to trivalent antimony than control parasites. There were no observed alterations in susceptibility to amphotericin B, miltefosine, or menadione inLDK-knockoutL.infantum lines. Our results suggest...
Source: PLoS Neglected Tropical Diseases - Category: Tropical Medicine Authors: Source Type: research