Höllein Alexander, Twardziok Sven O, Walter Wencke, Hutter Stephan, Baer Constance, Hernandez-Sanchez Jesus Maria, Meggendorfer Manja, Haferlach Torsten, Kern Wolfgang, Haferlach Claudia
MLL Munich Leukemia Laboratory, Munich, Germany.
MLL Munich Leukemia Laboratory, Munich, Germany; Centro de Investigación del Cáncer (Universidad de Salamanca-CSIC) Campus Universitario Miguel de Unamuno, Salamanca, Spain.
Cancer Genet. 2020 Apr;242:15-24. doi: 10.1016/j.cancergen.2020.01.001. Epub 2020 Jan 11.
The diagnosis and risk stratification of multiple myeloma (MM) is based on clinical and cytogenetic tests. Magnetic CD138 enrichment followed by interphase FISH (fluorescence in situ hybridisation) is the gold standard to identify prognostic translocations and copy number alterations (CNA). Although clinical implications of gene expression profiling (GEP) or panel based sequencing results are evident, those tests have not yet reached routine clinical application. We set up a single workflow to analyse MM of 211 patients at first diagnosis by whole genome sequencing (WGS) and RNA-Seq and validate the results by FISH analysis. We observed a 96% concordance of FISH and WGS results when assessing translocations involving the IGH locus and an overall concordance of FISH and WGS of 92% when assessing CNA. WGS analysis resulted in the identification of 17 additional MYC-translocations that were missed by FISH analysis. RNA-Seq followed by supervised clustering grouped patients in their expected genetically defined subgroup and prompted the assessment of WGS data in cases that were not congruent with FISH. This allowed the identification of additional IGH-translocations and hyperdiploid cases. We show the reliability of WGS an RNA-Seq in a clinical setting, which is a prerequisite for a novel routine diagnostic test.
多发性骨髓瘤（MM）的诊断和风险分层基于临床和细胞遗传学检测。磁珠富集CD138后进行间期荧光原位杂交（FISH）是识别预后性易位和拷贝数改变（CNA）的金标准。尽管基因表达谱分析（GEP）或基于基因panel的测序结果的临床意义已很明显，但这些检测尚未达到常规临床应用阶段。我们建立了一个单一工作流程，通过全基因组测序（WGS）和RNA测序对211例初诊MM患者进行分析，并通过FISH分析验证结果。在评估涉及IGH基因座的易位时，我们观察到FISH和WGS结果的一致性为96%，在评估CNA时，FISH和WGS的总体一致性为92%。WGS分析发现了17例FISH分析遗漏的额外MYC易位。RNA测序后进行监督聚类，将患者分组到预期的基因定义亚组中，并促使在与FISH结果不一致的病例中评估WGS数据。这使得能够识别出额外的IGH易位和超二倍体病例。我们展示了WGS和RNA测序在临床环境中的可靠性，这是新型常规诊断检测的先决条件。
