Gregorová J, Vrábel D, Radová L, Gablo N A, Almaši M, Štork M, Slabý O, Pour L, Minařík J, Ševčíková S
Klin Onkol. 2018 Spring;31(Supplementum1):148-150.
Multiple myeloma (MM) is the second most common hematooncological disease. Patient survival has been greatly improved by the introduction of new drugs into clinical practice, but survival is negatively affected by the so-called extramedullary relapse (EM), caused by the loss of plasma cell dependence on the bone marrow microenvironment and their migration out of the bone marrow. The nature and causes of this process are currently unclear. MicroRNAs (miRNAs) are short, non-coding RNA molecules involved in many physiological and pathological processes. Their significance in the pathogenesis of MM has been demonstrated by several studies. We assume that they are also involved in the development of the EM. The aim of this study was to analyze different miRNA expression between MM and EM patients.
Using next generation sequencing, we analyzed 39 samples of bone marrow cells from MM patients at diagnosis and 9 bone marrow plasma samples of EM patients.
In total, 2,278 miRNA were sequenced, but only 658 miRNAs were analyzed as they were expressed in all samples and had at least 20 reads. Expression data were generated using the Chimira tool from fastq data. All sequences were mapped using miRBase v20. Further analyses were performed using the R/Bioconductor package. The Bayesian procedure was used for normalization of expression. P values were adjusted using the Benjamini-Hochberg method. Analysis found 10 miRNA (p < 0.0005) that are statistically significantly expressed in EM vs. MM patients - these are miR-26a-5p, miR-26b-5p, miR-30e-5p, miR-424-3p, miR-503-5p, miR-767-5p, miR-105-5p, miR-5695-5p, miR-450b-5p and miR-92b-3p. These miRNAs will be further verified by qPCR method on a larger set of MM and EM patients.
Our pilot study has shown that there are differentially expressed miRNAs between MM and EM patients.Key words: multiple myeloma - microRNA - carcinogenesis - next generation sequencing The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papersThis work was supported by grant MZ ČR AZV 17- 29343A. Submitted: 17. 3. 2018Accepted: 20. 3. 2018.
多发性骨髓瘤（MM）是第二常见的血液肿瘤疾病。新药引入临床实践后患者生存率有了显著提高，但所谓的髓外复发（EM）对生存产生负面影响，髓外复发是由浆细胞对骨髓微环境的依赖性丧失及其从骨髓中迁移所致。目前该过程的性质和原因尚不清楚。微小RNA（miRNA）是参与许多生理和病理过程的短链非编码RNA分子。多项研究已证实它们在MM发病机制中的重要性。我们推测它们也参与了EM的发生发展。本研究旨在分析MM患者与EM患者之间不同的miRNA表达情况。
我们使用下一代测序技术分析了39例MM患者诊断时的骨髓细胞样本以及9例EM患者的骨髓血浆样本。
总共对2278个miRNA进行了测序，但仅分析了658个miRNA，因为它们在所有样本中均有表达且读数至少为20。使用Chimira工具从fastq数据生成表达数据。所有序列均使用miRBase v20进行映射。使用R/Bioconductor软件包进行进一步分析。采用贝叶斯方法对表达进行归一化。使用Benjamini-Hochberg方法调整P值。分析发现10个miRNA（p < 0.0005）在EM患者与MM患者中具有统计学显著差异表达，它们分别是miR-26a-5p、miR-26b-5p、miR-30e-5p、miR-424-3p、miR-503-5p、miR-767-5p、miR-105-5p、miR-5695-5p、miR-450b-5p和miR-92b-3p。这些miRNA将通过qPCR方法在更大规模的MM和EM患者组中进一步验证。
我们的初步研究表明，MM患者与EM患者之间存在差异表达的miRNA。关键词：多发性骨髓瘤 - 微小RNA - 致癌作用 - 下一代测序 作者声明他们在研究中使用的药物、产品或服务不存在潜在利益冲突。编辑委员会声明该手稿符合ICMJE对生物医学论文的推荐标准。本研究得到了捷克卫生部AZV 17 - 29343A项目的支持。提交日期：2018年3月17日。接受日期：2018年3月20日。
