Spanish National Cancer Centre (CNIO), Melchor Fernández Almagro 3, Madrid 28029, Spain.
Haematologica. 2010 Apr;95(4):613-21. doi: 10.3324/haematol.2009.013870. Epub 2010 Feb 4.
Vorinostat (suberoylanilide hydroxamic acid, SAHA), an inhibitor of class I and II histone deacetylases, has been approved for the treatment of cutaneous T-cell lymphoma. In spite of emerging information on the effect of vorinostat in many types of cancer, little is yet known about this drug's mechanism of action, which is essential for its proper use in combination therapy. We investigated alterations in gene expression profile over time in cutaneous T-cell lymphoma cells treated with vorinostat. Subsequently, we evaluated inhibitors of PI3K, PIM and HSP90 as potential combination agents in the treatment of cutaneous T-cell lymphoma.
The genes significantly up- or down-regulated by vorinostat over different time periods (2-fold change, false discovery rate corrected P value<0.05) were selected using the short-time series expression miner. Cell viability was assessed in vitro in cutaneous T-cell lymphoma cells through measuring intracellular ATP content. Drug interactions were analyzed by the combination index method with CalcuSyn software.
The functional analysis suggests that vorinostat modifies signaling of T-cell receptor, MAPK, and JAK-STAT pathways. The phosphorylation studies of ZAP70 (Tyr319, Tyr493) and its downstream target AKT (Ser473) revealed that vorinostat inhibits phosphorylation of these kinases. With regards to effects on cutaneous T-cell lymphoma cells, combining vorinostat with PI3K inhibitors resulted in synergy while cytotoxic antagonism was observed when vorinostat was combined with HSP90 inhibitor.
These results demonstrate the potential targets of vorinostat, underlining the importance of T-cell receptor signaling inhibition following vorinostat treatment. Additionally, we showed that combination therapies involving histone deacetylase inhibitors and inhibitors of PI3K are potentially efficacious for the treatment of cutaneous T-cell lymphoma.
伏立诺他（琥珀酰亚胺基羟肟酸，SAHA），一种 I 类和 II 类组蛋白去乙酰化酶抑制剂，已被批准用于治疗皮肤 T 细胞淋巴瘤。尽管有越来越多的信息表明伏立诺他对多种类型的癌症有效，但对于这种药物的作用机制知之甚少，而这对于其在联合治疗中的正确应用至关重要。我们研究了伏立诺他处理后的皮肤 T 细胞淋巴瘤细胞中基因表达谱随时间的变化。随后，我们评估了 PI3K、PIM 和 HSP90 的抑制剂作为治疗皮肤 T 细胞淋巴瘤的潜在联合药物。
使用短时序列表达挖掘程序，选择伏立诺他在不同时间段（2 倍变化，校正后的错误发现率 P 值<0.05）显著上调或下调的基因。通过测量细胞内 ATP 含量，在皮肤 T 细胞淋巴瘤细胞中体外评估细胞活力。使用 CalcuSyn 软件的组合指数方法分析药物相互作用。
功能分析表明，伏立诺他改变了 T 细胞受体、MAPK 和 JAK-STAT 途径的信号。ZAP70（Tyr319、Tyr493）及其下游靶标 AKT（Ser473）的磷酸化研究表明，伏立诺他抑制这些激酶的磷酸化。关于对皮肤 T 细胞淋巴瘤细胞的影响，伏立诺他与 PI3K 抑制剂联合使用产生协同作用，而当伏立诺他与 HSP90 抑制剂联合使用时则观察到细胞毒性拮抗作用。
这些结果表明了伏立诺他的潜在靶点，强调了伏立诺他治疗后抑制 T 细胞受体信号的重要性。此外，我们表明，组蛋白去乙酰化酶抑制剂与 PI3K 抑制剂的联合治疗可能对皮肤 T 细胞淋巴瘤的治疗有效。
