3'Nucleotidase/nuclease is required for Leishmania infantum clinical isolate susceptibility to miltefosine.
3'Nucleotidase/nuclease is required for Leishmania infantum clinical isolate susceptibility to miltefosine.
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DOI：
10.1016/j.ebiom.2022.104378
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发表时间：
2022-12
期刊：
影响因子：
11.1
通讯作者：
Mottram, Jeremy C.
中科院分区：
文献类型：
--
作者：
Carnielli, Juliana B. T.;Dave, Anuja;Romano, Audrey;Forrester, Sarah;de Faria, Pedro R.;Monti-Rocha, Renata;Costa, Carlos H. N.;Dietze, Reynaldo;Graham, Ian A.;Mottram, Jeremy C.
关键词：
巴西内脏利什曼病的米替福新治疗失败与婴儿利什曼原虫的米替福新易感基因座（MSL）缺失有关。MSL包含4个基因，3′-核苷酸酶/核酸酶（NUC 1和NUC 2）、解旋酶样蛋白（HLP）和3，2-反式-烯酰-CoA异构酶（TEI）。在这项研究中，CRISPR-Cas9用于表位标签或删除NUC 1，NUC 2，HLP和TEI，以研究它们在米替福新耐药机制中的作用。此外，在26株婴儿乳杆菌临床分离株中评估了米替福新转运蛋白基因和米替福新介导的活性氧稳态。还进行了比较脂质组学分析以研究米替福新抗性的分子基础。从MSL中删除NUC 1，NUC 2与米替福新敏感性显著降低相关，重新表达后恢复。缺乏MSL或NUC 1和NUC 2的寄生虫的代谢组学分析发现寄生虫脂质含量增加，包括麦角固醇;这些脂质可能通过在膜中结合药物而导致米替福新耐药性。缺乏MSL的寄生虫对米替福新引起的脂质代谢紊乱更有抵抗力，NUC 1和NUC 2参与了这一途径。此外，发现从米替福新治疗后复发的患者中分离的缺乏MSL的婴儿利什曼原虫寄生虫调节宿主巨噬细胞中的一氧化氮积累。总之，这些数据表明，多因素机制参与了婴儿乳杆菌对米替福新的天然耐药性，3 '核苷酸酶/核酸酶基因NUC 1和NUC 2的缺失促成了表型。 和.
Miltefosine treatment failure in visceral leishmaniasis in Brazil has been associated with deletion of the miltefosine susceptibility locus (MSL) in Leishmania infantum. The MSL comprises four genes, 3′-nucleotidase/nucleases (NUC1 and NUC2); helicase-like protein (HLP); and 3,2-trans-enoyl-CoA isomerase (TEI). In this study CRISPR-Cas9 was used to either epitope tag or delete NUC1, NUC2, HLP and TEI, to investigate their role in miltefosine resistance mechanisms. Additionally, miltefosine transporter genes and miltefosine-mediated reactive oxygen species homeostasis were assessed in 26 L. infantum clinical isolates. A comparative lipidomic analysis was also performed to investigate the molecular basis of miltefosine resistance. Deletion of both NUC1, NUC2 from the MSL was associated with a significant decrease in miltefosine susceptibility, which was restored after re-expression. Metabolomic analysis of parasites lacking the MSL or NUC1 and NUC2 identified an increase in the parasite lipid content, including ergosterol; these lipids may contribute to miltefosine resistance by binding the drug in the membrane. Parasites lacking the MSL are more resistant to lipid metabolism perturbation caused by miltefosine and NUC1 and NUC2 are involved in this pathway. Additionally, L. infantum parasites lacking the MSL isolated from patients who relapsed after miltefosine treatment were found to modulate nitric oxide accumulation in host macrophages. Altogether, these data indicate that multifactorial mechanisms are involved in natural resistance to miltefosine in L. infantum and that the absence of the 3'nucleotidase/nuclease genes NUC1 and NUC2 contributes to the phenotype. and .
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影响因子：
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作者：
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通讯作者：
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DOI：
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发表时间：
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期刊：
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