Molecular Oncology Laboratory, Murdoch Children's Research Institute, Parkville, Australia.
Epigenetics. 2012 Aug;7(8):875-82. doi: 10.4161/epi.20983. Epub 2012 Jul 9.
LBH589 is one of the many histone deacetylase inhibitors (HDACi) that are currently in clinical trial. Despite their wide-spread use, there is little literature available describing the typical levels of histone acetylation in untreated peripheral blood, the treatment and storage of samples to retain optimal measurement of histone acetylation nor methods by which histone acetylation analysis may be monitored and measured during the course of a patient's treatment. In this study, we have used cord or peripheral blood as a source of human leukocytes, performed a comparative analysis of sample processing methods and developed a flow cytometric method suitable for monitoring histone acetylation in isolated lymphocytes and liquid tumors. Western blotting and immunohistochemistry techniques have also been addressed. We have tested these methods on blood samples collected from four patients treated with LBH589 as part of an Australian Children's Cancer Clinical Trial (CLBH589AAU03T) and show comparable results when comparing in vitro and in vivo data. This paper does not seek to correlate histone acetylation levels in peripheral blood with clinical outcome but describes methods of analysis that will be of interest to clinicians and scientists monitoring the effects of HDACi on histone acetylation in blood samples in clinical trials or in related research studies.
LBH589 是目前正在临床试验中的众多组蛋白去乙酰化酶抑制剂 (HDACi) 之一。尽管它们被广泛使用，但可用的关于未经处理的外周血中组蛋白乙酰化的典型水平、为保留组蛋白乙酰化的最佳测量值而对样本进行处理和储存的描述，以及在患者治疗过程中监测和测量组蛋白乙酰化分析的方法的文献却很少。在这项研究中，我们使用脐带或外周血作为人类白细胞的来源，对样本处理方法进行了比较分析，并开发了一种适用于监测分离的淋巴细胞和液体肿瘤中组蛋白乙酰化的流式细胞术方法。还解决了 Western blot 和免疫组织化学技术。我们已经在来自四名接受 LBH589 治疗的患者的血液样本上测试了这些方法，这些患者是澳大利亚儿童癌症临床研究 (CLBH589AAU03T) 的一部分，并展示了体外和体内数据的可比结果。本文不寻求将外周血中的组蛋白乙酰化水平与临床结果相关联，而是描述了分析方法，这些方法将引起监测临床试验或相关研究中 HDACi 对血液样本中组蛋白乙酰化影响的临床医生和科学家的兴趣。
