Genomic and Proteomic Studies on the Mode of Action of Oxaboroles against the African Trypanosome.
Genomic and Proteomic Studies on the Mode of Action of Oxaboroles against the African Trypanosome.
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DOI：
10.1371/journal.pntd.0004299
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发表时间：
2015-12
影响因子：
3.8
通讯作者：
Fairlamb AH
中科院分区：
文献类型：
--
作者：
Jones DC;Foth BJ;Urbaniak MD;Patterson S;Ong HB;Berriman M;Fairlamb AH
SCYX-7158是一种氧杂硼杂环戊烯，目前正处于治疗人类非洲锥虫病的I期临床试验中。在这里，我们调查可能的模式对布氏锥虫使用正交化学蛋白质组学和基因组学的方法。使用固定在树脂上的氧杂硼杂环戊烯类似物的基于SILAC的蛋白质组学研究用于与可溶性氧杂硼杂环戊烯或固定的无活性对照竞争，以鉴定两种策略共有的13种蛋白质。用亚致死浓度的氧杂硼杂环戊烯孵育的细胞的细胞周期分析鉴定了G2和>G2细胞的细微但显著的积累。考虑到DNA保真度受损的可能性，我们研究了长期接触T。通过体外产生耐药细胞系，事实证明，耐药性比目前在该领域使用的药物更难产生，并且在我们的三个细胞系中有一个是不稳定的。耐药细胞系的全基因组测序显示66个基因的单核苷酸多态性和几个大规模的基因组畸变。耐药细胞系之间缺乏简单一致的机制，蛋白质组学研究中的结合伴侣列表多样，表明一定程度的多药理学应降低该领域出现的此类化合物耐药风险。遗传学和化学生物学相结合的方法提供了候选人的名单进行调查的更详细的信息，这种有前途的新药类的作用方式。目前正在开发用于治疗人类非洲锥虫病的一类新的含硼化学品（氧杂硼杂环戊烯）的作用方式尚不清楚。在这里，我们确定了一些潜在的候选蛋白质，这些蛋白质可能参与这些化合物的作用模式或抗性机制。这些信息可能对保护化合物免受该领域出现的耐药性以及开辟药物发现的新途径至关重要。
SCYX-7158, an oxaborole, is currently in Phase I clinical trials for the treatment of human African trypanosomiasis. Here we investigate possible modes of action against Trypanosoma brucei using orthogonal chemo-proteomic and genomic approaches. SILAC-based proteomic studies using an oxaborole analogue immobilised onto a resin was used either in competition with a soluble oxaborole or an immobilised inactive control to identify thirteen proteins common to both strategies. Cell-cycle analysis of cells incubated with sub-lethal concentrations of an oxaborole identified a subtle but significant accumulation of G2 and >G2 cells. Given the possibility of compromised DNA fidelity, we investigated long-term exposure of T. brucei to oxaboroles by generating resistant cell lines in vitro. Resistance proved more difficult to generate than for drugs currently used in the field, and in one of our three cell lines was unstable. Whole-genome sequencing of the resistant cell lines revealed single nucleotide polymorphisms in 66 genes and several large-scale genomic aberrations. The absence of a simple consistent mechanism among resistant cell lines and the diverse list of binding partners from the proteomic studies suggest a degree of polypharmacology that should reduce the risk of resistance to this compound class emerging in the field. The combined genetic and chemical biology approaches have provided lists of candidates to be investigated for more detailed information on the mode of action of this promising new drug class. The mode of action of a new class of boron-containing chemicals (the oxaboroles), currently under development for the treatment of human African trypanosomiasis, is unknown. Here we identify a number of potential candidate proteins that could be involved either in the mode of action of these compounds or in the mechanism of resistance. This information could prove critical in protecting the compounds against resistance emerging in the field as well as opening up new avenues for drug discovery.