Towards defining the functional methylome in acute lymphoblastic leukemia

到目前为止，对候选基因的DNA甲基化或使用低分辨率全基因组技术的研究已经 提供了存在于各种肿瘤类型中的异常甲基化的不完整视图。当前的目标是 建议缩小这一知识差距，并阐明造成这种异常的机制(S) 急性淋巴细胞白血病(ALL)基因甲基化的疾病特异性靶向性研究我们建议追求三个目标 目的：L)在正常和恶性前体B细胞中生成高分辨率的全基因组甲基化图谱 对照和儿童个体都使用全基因组甲基化微阵列；2)阐明序列特异性 通过比较肿瘤中CGI甲基化模式和区域特异性异常甲基化靶点 和正常骨髓；以及3)生成CpG位点特定的CGI甲基化图谱以确定这些关系 使用大规模并行测序技术在甲基化位置和程度与基因表达之间的关系 QRT-PCR法。这项工作将推动对ALL甲基组的破译及其对ALL表型的贡献 开发的分析策略应该为研究其他基因异常甲基化定义一个很好的模板 肿瘤类型。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；以及2)调查宫内妊娠的后果 将继续就表观遗传事件的建立进行曝光。培训计划包括高级分析、癌症病理生物学、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、一对一指导和负责任的研究工作培训。培训将由密苏里大学医学院的M.Sharon Stack博士和Charles Caldwell博士指导。Stack博士将通过帮助候选人融入地方和国家两级的科学界，帮助候选人过渡到独立。考德威尔博士将通过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，帮助候选人过渡到独立。

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:l) 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 sequencespecific 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.