Identification of Methylated Genes in Chronic Lymphocytic Leukemia

慢性淋巴细胞白血病甲基化基因的鉴定

• 批准号: 6986001
• 负责人: CHRISTOPH PLASS
• 金额: $ 21.76万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6986001
• 关键词: DNA methylation; clinical research; genes; genotype; methylation

Epigenetic changes, including DNA methylation, are a common finding in human malignancies. Research in chronic lymphocytic leukemia (CLL) has focused on genetic deletions that promote impaired apoptosis, but to date very few genes have been identified. In preliminary studies we have utilized Restriction Landmark Genomic Scanning (RLGS), a two-dimensional gel electrophoresis that allows detection of altered DNA methylation patterns, to study methylation in ten CLL patient samples. Our results strongly indicate that DNA methylation, via gene silencing, contributes significantly to the pathogenesis of CLL. Specifically, we demonstrated marked variation in the amount of aberrant methylation in patient samples ranging from 2.5-8.1% as compared to normal B cells. To fully establish the role of methylation in CLL and to exploit these alterations in the clinic, we propose more detailed studies outlined below. Our hypothesis for this Project is that DNA methylation contributes significantly to the development and progression of CLL. In Specific Aim 1, RLGS will be used on 100 CLL patients of different genotypes. RLGS profiles of CLL patients samples will be compared with CD19+ selected normal B-lymphocytes to identify novel CLL methylation sequences specific to CLL. In addition, methylation patterns between favorable and un-favorable genotypes will be compared to identify the most promising gene targets to pursue for investigation in Specific Aim 2. In Specific Aim 2, we plan to clone as many as 200 novel methylated sequences identified from aim 1 using genomic libraries that will guide the search for new tumor suppressor genes in CLL. Furthermore we propose experiments to perform a biostatistical evaluation of DNA methylation patterns in CLL. In aim 3 these genes will then be analyzed in a large cohort of CLL patient samples obtained from the national cooperative oncology groups, and results will be correlated with response to therapy, progression-free survival and overall survival. In Specific Aim 4, novel candidate cancer genes, DAPK1, ID4 and DERMO1, whose expression is altered by DNA methylation, will be studied in more detail in order to determine potential importance to the pathogenesis and progression of CLL.

表观遗传学变化，包括DNA甲基化，是人类恶性肿瘤中常见的发现。研究 慢性淋巴细胞白血病（CLL）的研究重点是促进受损细胞凋亡的基因缺失， 迄今为止，只有很少的基因被发现。在初步研究中，我们使用了限制地标 基因组扫描（RLGS），一种允许检测改变的DNA的二维凝胶电泳 甲基化模式，以研究10个CLL患者样品中的甲基化。我们的研究结果有力地表明， 通过基因沉默，甲基化显著促进CLL的发病机制。我们特别 在患者样品中，异常甲基化的量的显著变化范围为2.5-8.1%， 与正常的B细胞相比。充分确定甲基化在CLL中的作用，并利用这些改变 在临床方面，我们建议进行以下更详细的研究。我们对这个项目的假设是DNA 甲基化显著促进CLL的发生和进展。在具体目标1中，RLGS将 用于100例不同基因型的CLL患者。将比较CLL患者样本的RLGS谱 用CD 19+选择的正常B淋巴细胞鉴定对CLL特异的新的CLL甲基化序列。在 此外，将比较有利和不利基因型之间的甲基化模式，以鉴定 最有希望的基因靶点，以追求在特定目标2的研究。在Specific Aim 2中，我们计划克隆为 使用基因组文库从AIM 1中鉴定出多达200个新的甲基化序列， 寻找新的CLL抑癌基因。此外，我们提出了实验，以执行一个 CLL中DNA甲基化模式的生物统计学评价。在aim 3中，这些基因将被分析， 从国家肿瘤协作组获得的CLL患者样本的大队列，结果将在 与治疗反应、无进展生存期和总生存期相关。在具体目标4中，小说 候选癌症基因，DAPK 1，ID 4和DERMO 1，其表达被DNA甲基化改变，将被 更详细地研究，以确定对CLL的发病机制和进展的潜在重要性。