Core facility for molecular cytogenetics, genetic diagnostics and profiling

分子细胞遗传学、遗传诊断和分析的核心设施

• 批准号: 144856273
• 负责人: Professor Dr. Lars Bullinger
• 金额: --
• 依托单位: Klinik für Innere Medizin III
• 依托单位国家: 德国
• 项目类别: Clinical Research Units
• 财政年份: 2009
• 资助国家: 德国
• 起止时间: 2008-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/144856273?language=en
• 关键词: Core; facility; molecular; cytogenetics; genetic

During the last funding period we have established a centralized sample collection within the Clinical Research Unit (CRU). Patient samples from the University of Würzburg as well as from the University of Ulm have been characterized for cytogenetic and molecular genetic changes. The CD138 purified plasma cells are processed for archiving of tumor sample slides and of tumor cell pellets for further analysis. Additionally, corresponding germline material is archived as reference control. High-resolution genome-wide screening for genetic alterations of MM samples was performed using SNP chip analysis (Affymetrix Microarray Human Genome SNP 6.0). During the next funding period we will expand the collection of clinical samples. The incorporation of well characterized patients from clinical trials run by the German Study Group on Multiple Myeloma (DSMM) will strengthen the tumor bank and will serve as an invaluable resource for translational research within the CRU. The recently published characterization of the mutational landscape of 38 MM tumor genomes through massively parallel sequencing highlighted new aspects of the pathogenesis of multiple myeloma including the detection of novel mutated candidate genes.Consequently, we will put a special focus on a refined molecular characterization of the primary MM samples by the use of novel next-generation sequencing (NGS) technologies. From a number of MM samples from very well characterized patients participating in DSMM trials we will perform a mutational screen in a comprehensive set of target genes: NRAS, KRAS, BRAF, TP53, DIS3, FAM46C, KDM6A [also known as UTX], TRAF3, CYLD, CDKN2A/CDKN2B and CDKN2C. This research will be accompanied by a transcriptome sequencing (RNA-seq) approach in order to allow integrative analyses of DNA variation and gene expression changes. Furthermore, RNA-seq data offers the possibility to detect MMassociated fusion genes as well as aberrant/differential splicing patterns which recently have been shown to be also of pathogenetic relevance. Similarly, over the last decade small noncoding RNAs, particularly microRNAs (miRNA) were identified as key regulators of normal and pathological hematopoiesis. However, the role of miRNAs in the development of plasma cell disorders and the oncogenic pathways that lead to the development of MM is not fully understood. Therefore, we will also try to identify differential expression patterns of miRNAs in various MM subgroups investigated by the CRU projects.

在上一个资助期间，我们在临床研究单位 (CRU) 内建立了集中样本收集中心。来自维尔茨堡大学和乌尔姆大学的患者样本已被表征为细胞遗传学和分子遗传学变化。对 CD138 纯化的浆细胞进行处理，用于存档肿瘤样本载玻片和肿瘤细胞沉淀以供进一步分析。此外，相应的种系材料被存档作为参考对照。使用 SNP 芯片分析（Affymetrix 微阵列人类基因组 SNP 6.0）对 MM 样本的遗传改变进行高分辨率全基因组筛查。在下一个资助期内，我们将扩大临床样本的收集。德国多发性骨髓瘤研究组 (DSMM) 开展的临床试验中纳入特征明确的患者，将加强肿瘤库，并将成为 CRU 内转化研究的宝贵资源。最近发表的通过大规模并行测序对 38 MM 肿瘤基因组突变情况进行的表征，突出了多发性骨髓瘤发病机制的新方面，包括检测新型突变候选基因。因此，我们将特别关注通过使用新型下一代测序 (NGS) 技术对初级 MM 样本进行精细的分子表征。从参与 DSMM 试验的大量特征明确的患者的 MM 样本中，我们将对一组全面的目标基因进行突变筛选：NRAS、KRAS、BRAF、TP53、DIS3、FAM46C、KDM6A [也称为 UTX]、TRAF3、CYLD、CDKN2A/CDKN2B 和 CDKN2C。这项研究将伴随转录组测序 (RNA-seq) 方法，以便对 DNA 变异和基因表达变化进行综合分析。此外，RNA-seq 数据提供了检测 MM 相关融合基因以及异常/差异剪接模式的可能性，最近已证明这些剪接模式也与致病相关。同样，在过去十年中，小非编码 RNA，特别是 microRNA (miRNA) 被确定为正常和病理造血的关键调节因子。然而，miRNA 在浆细胞疾病发展中的作用以及导致 MM 发展的致癌途径尚未完全了解。因此，我们还将尝试鉴定 CRU 项目研究的各个 MM 亚组中 miRNA 的差异表达模式。