Ros3 (Lem3p/CDC50) Gene Dosage Is Implicated in Miltefosine Susceptibility in Leishmania (Viannia) braziliensis Clinical Isolates and in Leishmania (Leishmania) major

Autor(es)

Espada, Caroline R. ^[1] ; Albuquerque-Wendt, Andreia ^{[2, 3, 4]} ; Hornillos, Valentin ^{[5, 6]} ; Gluenz, Eva ^{[2, 4]} ; Coelho, Adriano C. ^[7] ; Uliana, Silvia R. B. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Inst Ciencias Biomed, Dept Parasitol, Sao Paulo - Brazil

^[2] Univ Oxford, Sir William Dunn Sch Pathol, Oxford - England

^[3] Univ Lisbon, Inst Higiene & Med Trop, Global Hlth & Trop Med, Lisbon - Portugal

^[4] Univ Glasgow, Coll Med Vet & Life Sci, Wellcome Ctr Integrat Parasitol, Inst Infect Immun & Inflammat, Glasgow, Lanark - Scotland

^[5] Univ Seville, Dept Quim Organ, Seville - Spain

^[6] Ctr Innovac Quim Avanzada, Seville - Spain

^[7] Univ Estadual Campinas, Inst Biol, Dept Biol Anim, Campinas - Brazil

• Tipo de documento: Artigo Científico
• Fonte: ACS INFECTIOUS DISEASES; v. 7, n. 4, p. 849-858, APR 9 2021.
• Citações Web of Science: 0

Resumo

The Ros3 protein is a component of the MT-Ros3 transporter complex, considered as the main route of miltefosine entry in Leishmania. L. braziliensis clinical isolates presenting differences in miltefosine susceptibility and uptake were previously shown to differentially express ros3. In this work, we showed that the ros3 gene copy number was increased in the isolate presenting the highest rates of miltefosine uptake and, thus, the highest susceptibility to this drug. The role of the ros3 gene dosage in miltefosine susceptibility was then investigated through a modulation of the gene copy number using two distinct approaches: through an overexpression of ros3 in a tolerant L. braziliensis clinical isolate and in L. major and by generating mono- and diallelic knockouts of this gene in L. major using clustered regularly interspaced short palindromic repeats (CRISPR) Cas9 (Cas = CRISPR-associated). Although the levels of ros3 mRNA were increased at least 40-fold in overexpressing clones, no significant reduction in the half-maximal effective concentration (EC50) for miltefosine was observed in these parasites. The partial or complete deletion of ros3 in L. major, in turn, resulted in a significant increase of 3 and 20 times, respectively, in the EC50 to miltefosine. We unequivocally showed that the ros3 copy number is one of the factors involved in the differential susceptibility and uptake of miltefosine. (AU)

• Processo FAPESP: 15/09080-2 - Avaliação de candidatos a fármacos para o tratamento de leishmaniose no Brasil
• Beneficiário: Silvia Reni Bortolin Uliana
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 18/25299-2 - Investigação do papel da proteína Ros3 na suscetibilidade diferencial de Leishmania à miltefosina utilizando uma abordagem de CRISPR/Cas9
• Beneficiário: Caroline Ricce Espada
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado Direto
• Processo FAPESP: 16/23405-4 - Suscetibilidade à miltefosina em isolados clínicos de Leishmania (Viannia) braziliensis: caracterização fenotípica e investigação das bases envolvidas na redução de suscetibilidade ao fármaco
• Beneficiário: Caroline Ricce Espada
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
• Processo FAPESP: 16/21171-6 - Paromomicina no tratamento da Leishmaniose Tegumentar: investigação in vitro, in vivo e na identificação de marcadores moleculares associados à suscetibilidade e resistência
• Beneficiário: Adriano Cappellazzo Coelho
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores