Proteomic analysis of the soluble proteomes of miltefosine-sensitive and -resistant Leishmania infantum chagasi isolates obtained from Brazilian patients with different treatment outcomes
Proteomic analysis of the soluble proteomes of miltefosine-sensitive and -resistant Leishmania infantum chagasi isolates obtained from Brazilian patients with different treatment outcomes
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DOI：
10.1016/j.jprot.2014.05.010
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发表时间：
2014-08-28
影响因子：
3.3
通讯作者：
Lemos, Elenice M.
中科院分区：
文献类型：
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作者：
Carnielli, Juliana B. T.;de Andrade, Helida M.;Lemos, Elenice M.
利什曼原虫对米替膦的耐药机制在实验耐药品系中已部分确定;然而，仍需要使用具有不同米替福新敏感性的临床分离株进行研究。在我们的研究中，我们使用蛋白质组学2D-DIGE/MS方法来研究来自具有不同米替福新治疗结果的内脏利什曼病患者的米替福新敏感和耐药的恰加斯婴儿利什曼原虫分离株中的不同蛋白质丰度。从这些菌株获得的高分辨率蛋白质组显示823个匹配的点，46个点表现出不同的菌株之间的丰度。在这些差异表达点中，26个（56.5%）显示抗性分离物的表达高于敏感分离物，20个（43.5%）显示抗性分离物的表达低于敏感分离物。MALDI/TOF-TOF质谱允许鉴定32个点，其具有对应于22个非冗余蛋白的独特蛋白质鉴定。大多数蛋白质组米替膦耐药菌株上调与氧化还原稳态，应激反应，保护细胞凋亡，药物转运。这些差异表达的蛋白质可能参与米替福新的天然耐药性，并表明利什曼原虫的米替福新耐药性机制是多因素的。生物学意义内脏利什曼病（VL）是一种严重的疾病，其治疗计划具有挑战性，需要长期且痛苦地应用耐受性差的有毒药物。因此，米替福新作为一种安全有效的口服药物的发现被认为是治疗利什曼病的一个重大进展。然而，在临床相关种属中观察到利什曼原虫对米替福新的不同敏感性，并且对利什曼原虫临床分离株获得耐药性的生物学机制知之甚少。我们的工作旨在阐明利什曼原虫对米替福新天然耐药的机制，通过研究显示不同米替福新治疗结果的VL患者的分离株。(C)© 2014 Elsevier B. V.保留所有权利。
The mechanism of miltefosine-resistance in Leishmania spp. has been partially determined in experimental resistant lines; however, studies using clinical isolates with different miltefosine susceptibilities are still needed. In our study, we used a proteomic 2D-DIGE/MS approach to study different protein abundances in miltefosine-sensitive and -resistant Leishmania infantum chagasi isolates from visceral leishmaniasis patients with different miltefosine treatment outcomes. The high-resolution proteome obtained from these isolates showed 823 matched spots and 46 spots exhibited different abundances between the isolates. Out of these differentially expressed spots, 26 (56.5%) showed greater and 20 (43.5%) showed lower expression of the resistant isolate compared to the sensitive isolate. MALDI/TOF-TOF mass spectrometry allowed the identification of 32 spots with unique protein identification correspondent to 22 non-redundant proteins. Most of the proteins up-regulated in the proteome miltefosine-resistant isolates were associated with redox homeostasis, stress response, protection to apoptosis, and drug translocation. These differentially expressed proteins are likely involved in miltefosine natural resistance and suggest that the miltefosine-resistance mechanism in Leishmania is multifactorial.Biological significanceVisceral leishmaniasis (VL) is a serious disease with a challenging treatment plan requiring the prolonged and painful applications of poorly tolerated toxic drugs. Therefore, the identification of miltefosine, an effective and safe oral drug, was considered a significant advancement in leishmaniasis therapy. However, different sensitivities to miltefosine in Leishmania have been observed in clinically relevant species, and the biological mechanism by which clinical isolates of Leishmania acquire drug resistance is poorly understood. Our work aims to elucidate the mechanism of natural resistance to miltefosine in Leishmania by studying the isolates from VL patients who displayed different miltefosine treatment outcomes. (C) 2014 Elsevier B.V. All rights reserved.