Toward Defining the Functional Methylome in Acute Lymphoblastic Leukemia

描述（由申请人提供）：迄今为止，候选基因中的DNA甲基化研究或使用低分辨率全基因组技术提供了各种肿瘤类型中存在的异常甲基化的不完整视图。该提案的直接目标是缩小这一知识差距，并阐明导致急性淋巴细胞白血病（ALL）中甲基化位点异常疾病特异性靶向的机制。我们提出了三个目标：1）使用全基因组甲基化微阵列在来自对照和儿童ALL个体的正常和恶性前体B细胞中产生高分辨率的全基因组甲基化谱; 2）通过比较肿瘤和正常骨髓中的CGI甲基化模式来阐明异常甲基化的序列特异性和区域特异性靶标;和3）使用大规模平行测序技术和qRT-PCR产生CpG位点特异性CGI甲基化图谱以确定甲基化的位置和程度与基因表达之间的关系。这项工作将进一步破译ALL甲基化组及其对ALL表型的贡献，开发的分析策略应该为研究其他肿瘤类型中的异常甲基化定义一个很好的模板。拟议的调查将为候选人的长期职业目标提供基础，成为一级研究机构的独立研究员和教员，长期目标：1）研究遗传学和表观遗传学之间的关系; 2）调查子宫内暴露对建立表观遗传事件的后果。培训计划包括高级分析、癌症病理学、科学写作和技术培训。还包括职业发展活动，如建立网络、提供科学数据的机会、一对一的指导和负责任地进行研究的培训。培训将在M博士指导下进行。密苏里大学医学院的莎伦·斯塔克和查尔斯·考德威尔博士。斯塔克博士将协助候选人过渡到独立，帮助候选人融入地方和国家两级的科学界。考德威尔博士将协助候选人过渡到独立，向候选人公布所有数据，促进新的合作，并承诺完成培训所需的资金，设施，设备和技术支持。 相关性：阐明表观遗传学改变对肿瘤抑制基因沉默的影响将提供有关癌症病因的基础知识。这将允许鉴定可用作诊断或预后标志物的潜在靶序列。这是最重要的，因为它可能导致使用DNA甲基化抑制剂恢复基因功能的新治疗方法。

DESCRIPTION (provided by applicant): To date, investigations of DNA methylation in candidate genes or using low-resolution genome-wide technologies have provided an incomplete view of the aberrant methylation present in various tumor types. The immediate goal of the proposal is to narrow this knowledge gap and to elucidate the mechanism(s) that are responsible for the aberrant disease-specific targeting of loci methylated in acute lymphoblastic leukemia (ALL). We propose to pursue three aims: 1) Generate high-resolution, genome-wide methylation profiles in normal and malignant precursor B-cells from controls and individuals with childhood ALL using a genome-wide methylation microarray; 2) Elucidate sequence specific and region-specific targets of aberrant methylation by comparing CGI methylation patterns among neoplastic and normal bone marrows; and 3) Generate CpG site-specific CGI methylation maps to determine the relationships between location and extent of methylation and gene expression using a massively parallel sequencing technology and qRT-PCR. This work will progress the deciphering of the ALL methylome and its contribution to the ALL phenotype and the developed analytical strategy should define an excellent template for studying aberrant methylation in other tumor types. The proposed investigations will provide the foundation for the candidate's long-term career goal to become an independent researcher and faculty member at a Tier I research institution where, the long-term objectives: 1) Researching the relationships between genetics and epigenetics; and 2) Investigating the consequences of in utero exposures on the establishment of epigenetic events will be pursued. The training plan includes advanced analytical, cancer pathobiology, scientific writing and technical training. Also included are career development activities such as networking, opportunities to present scientific data, one-on-one mentoring and training in the responsible conduct of research. The training will be under the tutelage of Dr. M. Sharon Stack and Dr. Charles Caldwell at the University of Missouri-School of Medicine. Dr. Stack will assist the candidate in the transition to independence by aiding in the candidate's integration into the scientific community both at the local and national levels. Dr. Caldwell will assist the candidate in the transition to independence by releasing all data to the candidate, facilitating new collaborations, and committing the finances, facilities, equipment and technical support required to complete the training. RELEVANCE: Elucidation of the effects of epigenetic alterations on the silencing of tumor-suppressor genes will provide fundamental knowledge regarding the causes of cancer. This will allow for the identification of potential target sequences that can be used as diagnostic or prognostic markers. This is of the utmost importance because it could lead to novel treatments to restore gene function using DNA methylation inhibitors.