Genome-wide RNAi screens in African trypanosomes identify the nifurtimox activator NTR and the eflornithine transporter AAT6
Genome-wide RNAi screens in African trypanosomes identify the nifurtimox activator NTR and the eflornithine transporter AAT6
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DOI：
10.1016/j.molbiopara.2010.11.010
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发表时间：
2011-03-01
影响因子：
1.5
通讯作者：
Horn, David
中科院分区：
文献类型：
--
作者：
Baker, Nicola;Alsford, Sam;Horn, David
为了有效，治疗化合物通常必须进入靶细胞，并且在某些情况下，必须浓缩或修饰。因此，摄取和活化机制通常形成对感染因子的选择性的基础。功能丧失筛选可用于鉴定参与药物摄取和代谢的蛋白质，也可鉴定临床相关的潜在耐药机制。我们使用基因组规模的RNA干扰（RNAi）文库来鉴定血流型布氏锥虫的功能丧失抗性机制。硝呋莫司-依氟鸟氨酸联合治疗（NECT）最近被引入用于人类非洲锥虫病，我们在这里关注这些药物。对硝呋替莫和相关药物苄硝哒唑的耐药性筛查发现了硝基还原酶（NTR）前药激活剂功能的丧失。对氨基酸类似物依氟鸟氨酸的耐药性筛选确定了氨基酸转运蛋白（AAT 6）功能的丧失。我们的研究结果证实了最近的研究结果，并表明NTR或AAT 6功能丧失代表了对这些药物耐药的主要潜在机制。因此，血流型T.布鲁氏菌RNAi文库为选择性遗传筛选和快速鉴定药物活化、摄取和潜在抗性机制提供了一种通用工具。(C)2010 Elsevier B.V.保留所有权利。
To be effective, therapeutic compounds must typically enter target cells and, in some cases, must be concentrated or modified. Thus, uptake and activation mechanisms often form the basis of selectivity against infectious agents. Loss-of-function screens can be used to identify proteins involved in drug uptake and metabolism and may also identify clinically relevant potential resistance mechanisms. We used a genome-scale RNA interference (RNAi) library to identify loss-of-function resistance mechanisms in bloodstream-form Trypanosoma brucei. Nifurtimox-Eflornithine Combination Therapy (NECT) was recently introduced for Human African Trypanosomiasis and we focus on these drugs here. Screens for resistance to nifurtimox and a related drug, benznidazole, identified loss of nitroreductase (NTR) pro-drug activator function. A screen for resistance to the amino-acid analogue, eflornithine, identified loss of amino-acid transporter (AAT6) function. Our results confirm recent findings and suggest that NTR or AAT6 loss-of-function represent major potential mechanisms of resistance to these drugs. Thus, bloodstream-form T. brucei RNAi libraries present a versatile tool for selective genetic screening and for the rapid identification of drug-activation, uptake and potential resistance mechanisms. (C) 2010 Elsevier B.V. All rights reserved.