| Autor: |
Fujii TTS; Laboratório de Imunofarmacologia, Instituto de Biofísica Carlos Chagas Filho (IBCCF), Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-901, Brazil., Gomes PS; Laboratório de Imunobiotecnologia, Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-901, Brazil.; Laboratório Interdisciplinar de Pesquisas Médicas, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz-Fiocruz, Rio de Janeiro 21040-900, Brazil., do Monte-Neto RL; Grupo de Pesquisas em Biotecnologia Aplicada a Patógenos-Instituto René Rachou, Fundação Oswaldo Cruz-Fiocruz Minas, Belo Horizonte 30190-002, Brazil., de Oliveira Gomes DC; Núcleo de Doenças Infecciosas, Universidade Federal do Espírito Santo, Vitória 29075-910, Brazil., Ouellette M; CHU de Quebec Research Center, Division of Infectious Disease and Immunity, Department of Microbiology Infectious Disease and Immunology, Laval University, Quebec, QC G1V 4G2, Canada., Torres-Santos EC; Laboratório de Bioquímica de Tripanosomatídeos, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz-Fiocruz, Rio de Janeiro 21040-900, Brazil., Andrade-Neto VV; Laboratório de Bioquímica de Tripanosomatídeos, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz-Fiocruz, Rio de Janeiro 21040-900, Brazil., de Matos Guedes HL; Laboratório de Imunofarmacologia, Instituto de Biofísica Carlos Chagas Filho (IBCCF), Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-901, Brazil.; Laboratório de Imunobiotecnologia, Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-901, Brazil.; Laboratório Interdisciplinar de Pesquisas Médicas, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz-Fiocruz, Rio de Janeiro 21040-900, Brazil. |
| Abstrakt: |
The sterol biosynthesis pathway of Leishmania spp. is used as a pharmacological target; however, available information about the mechanisms of the regulation and remodeling of sterol-related genes is scarce. The present study investigated compensatory mechanisms of the sterol biosynthesis pathway using an inhibitor of HMG-CoA reductase (simvastatin) and by developing drug-resistant parasites to evaluate the impact on sterol remodeling, cross-resistance, and gene expression. Simvastatin-resistant L. amazonensis parasites ( La SimR) underwent reprogramming of sterol metabolism manifested as an increase in cholestane- and stigmastane-based sterols and a decrease in ergostane-based sterols. The levels of the transcripts of sterol 24-C-methyltransferase (SMT), sterol C14-α-demethylase (C14DM), and protease subtilisin (SUB) were increased in La SimR. La SimR was cross-resistance to ketoconazole (a C14DM inhibitor) and remained sensitive to terbinafine (an inhibitor of squalene monooxygenase). Sensitivity of the La SimR mutant to other antileishmanial drugs unrelated to the sterol biosynthesis pathway, such as trivalent antimony and pentamidine, was similar to that of the wild-type strain; however, La SimR was cross-resistant to miltefosine, general serine protease inhibitor N - p -tosyl-l-phenylalanine chloromethyl ketone (TPCK), subtilisin-specific inhibitor 4-[(diethylamino)methyl]- N -[2-(2-methoxyphenyl)ethyl]- N -(3R)-3-pyrrolidinyl-benzamide dihydrochloride (PF-429242), and tunicamycin. The findings on the regulation of the sterol pathway can support the development of drugs and protease inhibitors targeting this route in parasites. |
