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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要端粒酶重新激活是几乎所有人类癌症发生过程中的一个基本事件。虽然在分化的成体细胞中转录沉默，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。尽管最近发现重新激活TERT 启动子突变，对于导致人类端粒酶重新激活的机制知之甚少癌症。该提案旨在确定调节 TERT 表达的潜在分子机制癌症。 Barthel 博士此前已证明 TERT 启动子中某个区域的甲基化与随着转录的增加。在这里，他将在功能上解析 DNA 甲基化是造成重新激活癌症发展中的端粒酶（目标 1）。该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，他还将评估分子受 TERT 启动子甲基化影响的机制（目标 2）。他将决定是否染色质构象和转录因子 (TF) 结合通过甲基化进行调节。最后，他将确定 DNA 甲基化、TF 结合和染色质构象的哪些变化与癌症发展中的端粒酶重新激活（目标 3）。总的来说，拟议的研究将建立甲基化在癌症中端粒酶重新激活中的作用，并提供对其自发性的本质的见解重新激活。 Barthel 博士的长期目标是确定端粒酶重新激活的脆弱性癌症的发展。在获得该奖项的过程中，Barthel 博士将得到他的主要导师 Dr. Barthel 的支持。 Roel Verhaak 是一位著名的计算生物学家，在分子生物学研究方面有着出色的记录癌症生物学。他的合作导师郑良豪博士开创了创新基因工程技术。此外，Barthel 博士还组建了一个咨询委员会，其中包括： Jerry Shay 博士，该领域的领导者癌症中的端粒和端粒酶；阮一军博士，三维染色质构象专家； Suneet Agarwal 博士利用诱导多能干细胞研究端粒疾病。在一起，这个多学科团队将使 Barthel 博士能够成功执行拟议的实验并推进他的专业发展计划，以促进他过渡到独立的学术职位。的支柱拟议的研究技能培训计划包括基因工程、染色质方面的高级培训构象分析和诱导多能性。专业发展的关键要素包括指导、资助写作和实验室管理。拟议项目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之外，还需要成功完成目标。鉴于他的详细的研究计划、优秀的顾问委员会和全面的培训计划，期待博士的到来。巴塞尔将通过该奖项迅速过渡到独立教职职位。