Genetic and Epigenetic Strategies for the Acquisition of Telomere Maintenance in Human Cancer Cells

人类癌细胞端粒维持的遗传和表观遗传策略

• 批准号: 10669077
• 负责人: Floris Barthel
• 金额: $ 24.4万
• 依托单位: TRANSLATIONAL GENOMICS RESEARCH INST
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10669077
• 关键词: 3-Dimensional; Adult; Advisory Committees; Affect; Aneuploidy; Automobile Driving; Award; Binding; Biological Models; Bone Marrow; Cancer Biology; Cancer Control; Cancer Model; Cells; Chimeric Proteins; Chromatin; DNA; DNA Methylation; Development; Development Plans; Disease; Elements; Environment; Epigenetic Process; Event; Exhibits; Faculty; Fibroblasts; Genes; Genetic; Genetic Engineering; Genetic Transcription; Genomic medicine; Genomics; Goals; Grant; Growth; Harvest; Human; In Vitro; Laboratories; Literature; Malignant Neoplasms; Measures; Mentors; Mentorship; Messenger RNA; Methylation; Methyltransferase; Molecular; Molecular Biology; Molecular Conformation; Mutagenesis; Mutation; Nature; Pathway interactions; Positioning Attribute; Public Health; RNA-Directed DNA Polymerase; Regulatory Element; Reporting; Research; Retinoblastoma Protein; Role; Site; TP53 gene; Technology; Telomerase; Telomerase inhibition; Telomere Maintenance; Testing; The Jackson Laboratory; Therapeutic; Time; Tissues; Toxic effect; Training; Training Programs; Tumor Suppressor Proteins; Work; Writing; cancer biomarkers; cancer cell; cancer therapy; career; experimental study; improved; induced pluripotent stem cell; innovation; insight; large datasets; multidisciplinary; novel; pluripotency; promoter; skills training; targeted cancer therapy; telomere; therapeutic target; transcription factor

PROJECT SUMMARY/ABSTRACT Telomerase reactivation is a fundamental event in the genesis of nearly every human cancer. Although transcriptionally silent in differentiated adult cells, its catalytic component telomerase reverse transcriptase (TERT) is expressed in over 80% of human cancers. Despite the recent discovery of reactivating TERT promoter mutations, very little is known about mechanisms leading to the reactivation of telomerase in human cancer. This proposal aims to determine the underlying molecular mechanisms regulating TERT expression in cancer. Dr. Barthel has previously shown that methylation of a region in the TERT promoter was associated with increased transcription. Here, he will functionally resolve that DNA methylation is responsible for reactivating telomerase in cancer development (Aim 1). This Aim uses epigenetic editing to measure the impact of manipulating DNA methylation on telomerase activity. Furthermore, he will assess molecular mechanisms that are affected by methylation at the TERT promoter (Aim 2). He will determine whether chromatin conformation and transcription factor (TF) binding are regulated via methylation. Finally, he will identify what changes in DNA methylation, TF binding and chromatin conformation are associated with telomerase reactivation in cancer development (Aim 3). Collectively, the proposed studies will establish the role of methylation in reactivating telomerase in cancer and provide insights into the nature of its spontaneous reactivation. The long-term goal of Dr. Barthel is to identify vulnerabilities of telomerase reactivation during cancer development. Over the course of this award Dr. Barthel will be supported by his primary mentor, Dr. Roel Verhaak, a renowned computational biologist with a remarkable track record studying the molecular biology of cancer. His co-mentor, Dr. Albert Cheng has pioneered innovative genetic engineering technologies. In addition, Dr. Barthel has assembled an advisory committee that includes: Dr. Jerry Shay, a leader on telomeres and telomerase in cancer; Dr. Yijun Ruan, an expert on three-dimensional chromatin conformation; and Dr. Suneet Agarwal, who uses induced pluripotent stem cells to study telomere disease. Together, this multidisciplinary team will enable Dr. Barthel to successfully execute the proposed experiments and advance his professional development plan to facilitate his transition to an independent academic position. Pillars of the proposed research skills training program include advanced training in genetic engineering, chromatin conformation analysis and induced pluripotency. Professional development will feature key elements including mentorship, grant writing and laboratory management. Work towards the proposed project will primarily be conducted at The Jackson Laboratory for Genomic Medicine, which offers all the state-of-the art facilities required for the successful completion of the Aims in addition to a collegial scientific environment. Given his detailed research plan, excellent advisory committee and comprehensive training plan it is expected that Dr. Barthel will quickly transition to an independent faculty position through this award.

项目摘要/摘要 端粒酶重新激活是几乎每个人类癌症的起源中的一个基本事件。虽然 在分化的成年细胞中的转录静音，其催化成分端粒酶逆转录酶 （TERT）在超过80％的人类癌症中表达。尽管最近发现了重新激活的tert 启动子突变对导致人类端粒酶重新激活的机制知之甚少 癌症。该建议旨在确定调节TERT表达的基本分子机制 癌症。 Barthel博士先前表明，TERT启动子中区域的甲基化是相关的 随着转录的增加。在这里，他将在功能上解决DNA甲基化负责 在癌症发展中重新激活端粒酶（AIM 1）。此目标使用表观遗传编辑来测量 操纵DNA甲基化对端粒酶活性的影响。此外，他将评估分子 受TERT启动子甲基化影响的机制（AIM 2）。他将确定是否 染色质构象和转录因子（TF）结合通过甲基化调节。最后，他会的 确定DNA甲基化，TF结合和染色质构象的变化与 癌症发展中的端粒酶重新激活（AIM 3）。拟议的研究共同确定 甲基化在癌症中重新激活端粒酶中的作用，并为其自发的性质提供见解 重新激活。巴瑟博士的长期目标是确定端粒酶重新激活的漏洞 癌症发展。在这个奖项中，巴瑟博士将得到其主要导师博士的支持。 Roel Verhaak，一位著名的计算生物学家，具有出色的记录，研究了分子 癌症的生物学。他的同事艾伯特·郑博士（Albert Cheng）博士开创了创新的基因工程技术。 此外，巴特尔博士还组建了一个咨询委员会，其中包括：杰里·谢伊（Jerry Shay）博士， 癌症中的端粒和端粒酶；三维染色质构象专家Yijun Ruan博士； Suneet Agarwal博士，他使用诱导的多能干细胞研究端粒疾病。一起，这个 多学科团队将使Barthel博士能够成功执行拟议的实验并提前 他的专业发展计划旨在促进他过渡到独立的学术职位。柱子 拟议的研究技能培训计划包括基因工程的高级培训，染色质 构象分析和诱导多能性。专业发展将以关键要素为特征 指导，赠款写作和实验室管理。为拟议项目而进行的工作将主要是 在杰克逊基因组医学实验室进行，该实验室提供所有最先进的设施 除了大学科学环境外，还需要成功完成目标。鉴于他 详细的研究计划，出色的咨询委员会和全面的培训计划，预计博士 巴瑟将通过该奖项迅速过渡到独立教师职位。