Mechanisms of chromosome segregation, aneuploidy, and tumorigenesis

染色体分离、非整倍性和肿瘤发生的机制

基本信息

批准号：
10406521
负责人：
Don W Cleveland
金额：
$ 94.14万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-05-01 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10406521
关键词：
ATP phosphohydrolase Aneuploidy Antisense Oligonucleotide Therapy Back Cells Centromere Chromosome Segregation Chromosome abnormality Chromosomes Complex Cyclins Cytokinesis DNA biosynthesis Development Epigenetic Process Event Extrachromosomal Inheritance Frequencies Genome Genomics Glioblastoma Grant Haploidy Human Immune checkpoint inhibitor Link Malignant Neoplasms Malignant neoplasm of brain Mammals Mediating Mitotic Mitotic Checkpoint Molecular Molecular Conformation National Institute of General Medical Sciences Role Testing Y Chromosome chromosome missegregation chromosome number abnormality chromothripsis extrachromosomal DNA genome sequencing micronucleus nervous system disorder nuclease prevent therapy development tumor tumorigenesis ubiquitin-protein ligase whole genome

ATP phosphohydrolase Aneuploidy Antisense Oligonucleotide Therapy Back Cells Centromere Chromosome Segregation Chromosome abnormality Chromosomes Complex Cyclins Cytokinesis DNA biosynthesis Development Epigenetic Process Event Extrachromosomal Inheritance Frequencies Genome Genomics Glioblastoma Grant Haploidy Human Immune checkpoint inhibitor Link Malignant Neoplasms Malignant neoplasm of brain Mammals Mediating Mitotic Mitotic Checkpoint Molecular Molecular Conformation National Institute of General Medical Sciences Role Testing Y Chromosome chromosome missegregation chromosome number abnormality chromothripsis extrachromosomal DNA genome sequencing micronucleus nervous system disorder nuclease prevent therapy development tumor tumorigenesis ubiquitin-protein ligase whole genome

展开

项目摘要

PROJECT SUMMARY Chromosome missegregation or errors in cytokinesis produce aneuploidy, a chromosome content other than a multiple of the haploid number. The linkage of aneuploidy to tumorigenesis has long been recognized. A striking chromosomal abnormality linked to chromosome missegregation is chromothripsis (also known as chromoanagenesis), an event in which one (or two) chromosomes appear to have been shattered into tens to hundreds of small genomic fragments and religated back together in random order. Chromotriptic chromosomes are now recognized to be present in a broad range of cancers. With support from an NIGMS R35 grant, we have identified mechanisms of normal chromosome segregation that act to prevent aneuploidy in the normal situation and have determined that single chromosome missegregation or transient spindle pole amplification is a driver of tumorigenesis. We have identified the epigenetic mark of centromere identity and determined that DNA replication acts as an error correction mechanism to maintain that identity. We have identified key molecular mechanisms underlying the mitotic checkpoint (also known as the spindle assembly checkpoint), the primary guard against chromosome missegregation in mammals. We have identified how both mitotic checkpoint activation and silencing involve the catalytic action of a conformation altering AAA+ ATPase TRIP13. We have also determined that mitotic exit has an absolute requirement for TRIP13-mediated disassembly of the checkpoint inhibitor or the non-essential APC15 subunit of the E3 ubiquitin ligase that targets mitotic cyclin destruction. By exploiting a unique feature of the human Y centromere, we have produced cells in which we can induce selective, transient inactivation of the Y centromere, with the Y chromosome missegregated into micronuclei at high frequency. With these and whole genome sequencing, we determined that simple missegregation into a micronucleus can initiate chromothripsis and drive the complex genome rearrangements frequently found in human cancer. In the upcoming 5 years, we propose to determine mechanisms of fragmentation of a chromosome during chromothripsis, identify and validate nucleases that fragment micronuclear chromosomes, determine how shattered chromosomes are reassembled and produce extrachromosomal DNA (ecDNA), determine mechanisms of inheritance of ecDNA, and determine the role of spatial proximity in the inheritance of centromere identity, including neocentromere formation and other genomic abnormalities. We will also exploit our development over the last 15 years of antisense oligonucleotide (ASO) therapy for nervous system disease to undertake proof of principle therapy development targeting inactivation of the mitotic checkpoint by testing suppression of TRIP13/APC15 for the major brain cancer glioblastoma.

项目摘要染色体的错误分析或细胞因子的错误会产生非整倍性，染色体含量以外的染色体含量单倍体数的倍数。长期以来已经认识到非整倍性与肿瘤发生的联系。一个惊人的与染色体错误分类有关的染色体异常是染色体（也称为染色体），一个事件，其中一个（或两个）染色体似乎已被粉碎成十亿数百个小基因组碎片和宗教以随机的顺序恢复在一起。染色体染色体现在被认为存在于广泛的癌症中。在NIGMS R35赠款的支持下，我们确定了正常染色体分离的机制该作用是在正常情况下防止非整倍性的，并确定了单个染色体错误分析或瞬态主轴电极扩增是肿瘤发生的驱动因素。我们已经确定了 Centromere身份的表观遗传标记，并确定DNA复制是误差校正的保持这种身份的机制。我们已经确定了有丝分裂的关键分子机制检查点（也称为主轴装配检查站），反对染色体的主要防护哺乳动物的错误分析。我们已经确定了有丝分裂检查点的激活和沉默如何涉及构象改变AAA+ ATPase Trip13的催化作用。我们还确定有丝分裂出口具有对检查点抑制剂或非必需的TRIP13介导的拆卸的绝对要求 E3泛素连接酶的APC15亚基靶向有丝分裂细胞周期蛋白的破坏。通过利用独特的功能人类的Y层，我们产生了细胞，在其中我们可以诱导选择性，瞬时失活的细胞 Y Centromere，Y染色体错误地将其分为高频中的微核。与这些和整个基因组测序，我们确定将简单的错误呈现为微核可以启动铬骨并推动人类癌症经常发现的复杂基因组重排。在接下来的5年中，我们建议确定染色体碎片的机制染色体，识别和验证碎屑微核染色体的核酸酶，确定如何重新组装破碎的染色体并产生外染色体DNA（ECDNA），确定 ECDNA继承的机制，并确定空间接近性在Centromere继承中的作用身份，包括新中心粒形成和其他基因组异常。我们还将利用我们的在过去的15年中，反义寡核苷酸（ASO）治疗神经系统疾病承担针对旨在通过测试灭活有丝分裂检查点的原理治疗开发的证明主要脑癌胶质母细胞瘤的TRIP13/APC15抑制。