Institute of Pathology, Medizinische Hochschule Hannover, Hannover, Germany.
Institute of Pathology, Medizinische Hochschule Hannover, Hannover, Germany.
J Mol Diagn. 2020 Apr;22(4):591-598. doi: 10.1016/j.jmoldx.2020.01.004. Epub 2020 Feb 7.
Chromosomal translocations resulting in fusion genes represent important oncogenic drivers and potential therapeutic targets in rare leukemia subtypes. Formalin-fixed, paraffin-embedded trephines are frequently used in hematologic diagnostic tests and provide relevant access to leukemic cells for further studies, for example, phenotyping in bone marrow fibrosis. However, high-throughput molecular analysis of nucleic acids obtained from this material is challenging, especially the reliable detection of RNA transcripts. Sixty-three formalin-fixed, paraffin-embedded bone marrow trephines of patients with chronic eosinophilic leukemia, chronic myeloid leukemia, acute myeloid leukemia, and myeloproliferative neoplasms were analyzed for gene mutations and the presence of fusion transcripts with a commercial amplicon-based next-generation sequencing approach. Fusion transcripts relevant for diagnosis and therapy could be detected and validated (by RT-PCR) in 25 patients (39.7%). Retrospectively selected material, up to 10 years old, was used for this purpose, and only one sample failed in the RNA analysis (1.6%). This study concludes that amplicon-based fusion transcript detection in bone marrow trephines is feasible and that bone marrow trephines taken for histologic assessment can also be applied for high-throughput molecular analysis.
染色体易位导致融合基因是罕见白血病亚型中的重要致癌驱动因子和潜在治疗靶点。福尔马林固定、石蜡包埋的环钻常用于血液学诊断测试，并为进一步研究提供了获取白血病细胞的相关途径，例如骨髓纤维化中的表型分析。然而，从这种材料中获得的核酸的高通量分子分析具有挑战性，特别是可靠地检测 RNA 转录本。采用商业的基于扩增子的下一代测序方法，对 63 例慢性嗜酸性粒细胞白血病、慢性髓性白血病、急性髓性白血病和骨髓增生性肿瘤患者的福尔马林固定、石蜡包埋的骨髓环钻进行基因突变和融合转录本的分析。在 25 例患者（39.7%）中可检测到并验证（通过 RT-PCR）与诊断和治疗相关的融合转录本。为此目的使用了回溯性选择的材料，最长可达 10 年，只有一份样本的 RNA 分析失败（1.6%）。本研究得出结论，骨髓环钻中的基于扩增子的融合转录本检测是可行的，并且用于组织学评估的骨髓环钻也可用于高通量分子分析。
