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.

在最后一个资金期间，我们在临床研究部门（CRU）中建立了集中式样本收集。来自温尔茨堡大学以及乌尔姆大学的患者样本的特征是细胞遗传学和分子遗传变化。处理CD138纯化的浆细胞以存档肿瘤样品载玻片和肿瘤细胞颗粒，以进一步分析。另外，将相应的种系材料存档为参考控制。使用SNP芯片分析（Affymetrix微阵列人类基因组SNP 6.0），对MM样品的遗传改变进行了高分辨率全基因组筛查。在下一个资金期间，我们将扩大临床样本的收集。由德国研究小组对多发性骨髓瘤（DSMM）进行的临床试验的良好特征患者纳入肿瘤库，并将成为CRU内翻译研究的宝贵资源。最近发表的通过大规模平行测序对38 mM肿瘤基因组突变景观的表征突出了多发性骨髓瘤的发病机理的新方面，包括检测新型突变的候选基因的检测，我们将特别关注新的MM样品的精制分子表征，该样本通过新颖的下一种序列（NG）进行了序列（NG）。从参加DSMM试验的非常有特征的患者的许多MM样品中，我们将在一组全面的靶基因中执行突变筛选：NRAS，KRAS，BRAF，TP53，DIS3，DIS3，FAM46C，KDM6C，KDM6A [也称为UTX]，TRAF3，TRAF3，CYLD，CYLD，CDKN2A/CDKN2A/CDKN2B和CDKN2B和CDKN2C。这项研究将伴随着转录组测序（RNA-SEQ）方法，以便允许对DNA变异和基因表达变化的综合分析。此外，RNA-seq数据提供了检测MMASSASSECIAD FUSION基因以及异常/差异剪接模式的可能性，这些模式最近已被证明也具有致病性。同样，在过去的十年中，小型非编码RNA，尤其是microRNA（miRNA）被确定为正常和病理造血的关键调节剂。但是，miRNA在浆细胞疾病发展和导致MM发展的致癌途径中的作用尚不完全了解。因此，我们还将尝试在CRU项目研究的各种MM亚组中确定miRNA的差异表达模式。