Manipulating chromosome structure to suppress genome instability

操纵染色体结构以抑制基因组不稳定

• 批准号: 9038333
• 负责人: Amity Manning
• 金额: $ 24.9万
• 依托单位: WORCESTER POLYTECHNIC INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-20 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9038333
• 关键词: Address; Agar; Aneuploid Cells; Aneuploidy; Antimitotic Agents; Area; Award; Bioinformatics; Biological; Biological Assay; Candidate Disease Gene; Cell Death; Cell Survival; Cells; Centromere; Cessation of life; Characteristics; Chromatin; Chromatin Structure; Chromosomal Instability; Chromosome Segregation; Chromosome Structures; Chromosomes; DNA Damage; Data; Defect; Development; Drug resistance; Enhancers; Environment; Enzymes; Epigenetic Process; Evolution; Exhibits; Fostering; Future; Gene-Modified; Generations; Genetic Heterogeneity; Genetic Screening; Genome Stability; Genomic Instability; Genomics; Goals; Grant; Growth; Human; Institutes; Malignant Neoplasms; Mediating; Medical; Mentors; Mentorship; Mitotic; Modification; Molecular; Neoplasm Metastasis; Numerical Chromosomal Abnormality; Pathway interactions; Patient-Focused Outcomes; Patients; Phase; Phenotype; Physical condensation; Positioning Attribute; Publications; Regulation; Relapse; Research; Resistance; Resources; Retinoblastoma Protein; Role; Solid Neoplasm; Structural defect; Techniques; Testing; Therapeutic; Time; Training; Transferase; Tumor Suppressor Proteins; Work; Xenograft procedure; abstracting; acquired drug resistance; cancer cell; career; cell growth; cohesion; drug sensitivity; faculty research; genome integrity; high throughput screening; histone methylation; histone methyltransferase; improved; in vivo; innovation; insight; mouse model; neoplastic cell; novel; novel strategies; outcome forecast; research study; response; screening; segregation; skills; stem; success; therapy resistant; training opportunity; tumor; tumor growth; tumor initiation

Project Summary/Abstract , is one form of genome instability that is a common feature of cancer cells. CIN, by definition, The majority of solid tumors exhibit both structural and numerical chromosome aberrations characteristic of genomic instability. Chromosome instability (CIN), the frequent and persistent gains and losses of whole chromosomes results in the generation of aneuploid cells and has important implications. CIN has been proposed to facilitate the evolution of tumor cells by promoting genetic heterogeneity, thereby enabling changes that promote growth and metastasis. Furthermore, genomic diversity generated by CIN promotes the development of cancer cells that are resistant to therapeutics and are more prone to tumor relapse. Consequently, CIN correlates with poor patient prognosis. Limitations in identifying and treating cancers that exhibit CIN stem from a lack of understanding of the underlying molecular mechanisms that contribute to CIN and the inability to manipulate CIN in a therapeutically relevant manner. I have recently demonstrated that pRB, a tumor suppressor whose function is compromised in many human cancers, promotes genome stability. I have shown that pRB loss leads to defects in chromatin structure, and that these abnormalities promote increased DNA damage and corruption of mitotic fidelity. However, corruption of the pRB pathway is not the only mechanism by which chromatin structure is compromised, and other regulators of chromatin structure are likely to similarly impact genome stability. One manner in which chromatin structure is impacted is through regulation of chromatin and histone methylation status. Indeed, a growing body of evidence has suggested that modification of epigenetic characteristics of cells correlates with aneuploidy. In addition, my preliminary studies have shown that modulation of levels of a subset of methyl-transferases can compromise chromatin structure and genome integrity. Together, these data suggest that general deregulation of chromatin structure is broadly relevant to the genesis of genomic instability. However, this has not been analyzed in a systematic manner, and the mechanisms by which chromatin structure are deregulated and contributes to genome instability remain unknown. Significantly, I have shown that by restoring chromosomal structure, I can suppress both DNA damage and promote accurate chromosome segregation in tumor cells. In doing so, I have identified Wapl, a negative regulator of cohesion as a novel regulator of CIN. In the following proposal, this novel strategy will enable me to test, for the first time, whether suppression of CIN renders tumor cells less capable of developing chemotherapeutic resistance in vivo, and thus less likely to relapse following traditional treatments. In addition, using a combination of bioinformatics and cell biological approaches, I will examine whether structural defects associated with CIN can be exploited to increase sensitivity to DNA damaging agents and decrease cell survival. Finally, I will identify regulators of chromatin structure whose deregulation impacts genome stability using a novel screening approach I have developed. The successful completion of the aims described in this proposal will not only provide new and valuable insights into the genesis of CIN in human cancers, but also reveal the therapeutic ramifications of manipulating this common feature in tumors. My career goal is to obtain a research faculty position at a leading institute where I will define the tumor suppressive mechanisms that maintain normal chromosome structure and genomic integrity. However, my successful transition to independence in this field would be significantly bolstered by augmenting my expertise in cell biological techniques with new training opportunities in mouse models of cancer, bioinformatics and high throughput screening approaches. It is with these acquired skills that I will be able to identify cancer-relevant regulators of chromatin structure that impact genome stability and assess their roles as novel tumor suppressors. The success of this project will be greatly enhanced by the outstanding training committee that I have assembled to mentor and advise me as I progress towards independence. This committee includes my co-mentors Dr. Nicholas Dyson and Dr. Jeffrey Engelman, as well as Dr. Daniel Haber, Dr. Cyril Benes, and Dr. David Pellman, all of whom are experts in their respective fields. In addition, the exceptional research environment available at MGH and the Harvard Medical Area has all of the necessary physical resources required for the completion of the proposed training and research studies. The K99/R00 award would afford me the protected time needed for this advanced training and allow me to continue to foster my growth under the mentorship of Dr. Dyson and Dr. Engelman. I expect that the mentored phase of this proposal, which includes using mouse models to investigate the therapeutic relevance of manipulating CIN in solid tumors, and the initiation of a novel screening approach, will take 1-2 years and result in at least one major publication. The following independent phase of the award will allow me to identify and characterize additional, cancer-relevant, regulators of chromosome structure and genome stability. Together these data will be used to justify future studies proposed in an R01 grant that I plan to submit at the start of the third year of the independent phase.

项目概要/摘要 ，是基因组不稳定性的一种形式，是癌细胞的共同特征。 CIN，根据定义， 大多数实体瘤都表现出结构和数量染色体畸变特征 基因组不稳定。染色体不稳定性（CIN），即整个染色体频繁且持续的增减 染色体 导致非整倍体细胞的产生并具有重要意义。 CIN 的提出是为了方便 通过促进遗传异质性来促进肿瘤细胞的进化，从而实现促进生长的变化 和转移。此外，CIN 产生的基因组多样性促进癌细胞的发展 对治疗有抵抗力并且更容易发生肿瘤复发。因此，CIN 与贫困相关 患者预后。识别和治疗表现出 CIN 的癌症的局限性源于缺乏 了解导致 CI​​N 的潜在分子机制并无法进行操作 CIN 以治疗相关的方式。 我最近证明了 pRB，一种肿瘤抑制因子，其功能在许多人类体内受到损害。 癌症，促进基因组稳定性。我已经证明 pRB 丢失会导致染色质结构缺陷，并且 这些异常会促进 DNA 损伤的增加和有丝分裂保真度的破坏。然而，腐败 pRB 途径的改变并不是染色质结构受损的唯一机制，其他机制 染色质结构的调节因子可能会类似地影响基因组稳定性。染色质的一种方式 结构受到影响是通过染色质和组蛋白甲基化状态的调节。确实，身体在成长 大量证据表明细胞表观遗传特征的改变与非整倍性相关。在 此外，我的初步研究表明，调节甲基转移酶子集的水平可以 损害染色质结构和基因组完整性。总之，这些数据表明普遍放松管制 染色质结构的变化与基因组不稳定的起源广泛相关。然而，这并没有被 系统地分析了染色质结构失调和调节的机制 导致基因组不稳定的因素仍然未知。 重要的是，我已经证明，通过恢复染色体结构，我可以抑制 DNA 损伤和 促进肿瘤细胞中染色体的准确分离。在此过程中，我发现了 Wapl，一个负面的 内聚力调节器作为 CIN 的新型调节器。在下面的提案中，这个新颖的策略将使我能够 首次测试抑制 CIN 是否会降低肿瘤细胞的发育能力 体内化疗耐药，因此传统治疗后不太可能复发。此外， 结合生物信息学和细胞生物学方法，我将检查是否存在结构缺陷 与 CIN 相关的 CIN 可用于增加对 DNA 损伤剂的敏感性并减少细胞 生存。最后，我将确定染色质结构的调节因子，其放松调节会影响基因组稳定性 使用我开发的一种新颖的筛选方法。成功完成本文件中描述的目标 该提案不仅将为人类癌症中 CIN 的起源提供新的、有价值的见解，而且 揭示了操纵肿瘤这一共同特征的治疗效果。 我的职业目标是在一家领先的研究所获得研究教职，在那里我将定义肿瘤 维持正常染色体结构和基因组完整性的抑制机制。然而，我的 通过增强我的专业知识，将大大促进在该领域成功过渡到独立 细胞生物学技术，在癌症、生物信息学和高水平小鼠模型方面提供新的培训机会 通量筛选方法。凭借这些获得的技能，我将能够识别与癌症相关的疾病 影响基因组稳定性并评估其作为新型肿瘤的作用的染色质结构调节因子 抑制器。我所领导的优秀培训委员会将极大地促进该项目的成功 在我走向独立的过程中，大家齐聚一堂，为我提供指导和建议。这个委员会包括我的 共同导师 Nicholas Dyson 博士和 Jeffrey Engelman 博士，以及 Daniel Haber 博士、Cyril Benes 博士和 David Pellman 博士，他们都是各自领域的专家。此外，卓越的研究 MGH 和哈佛医学区的环境拥有所有必要的物质资源 完成拟议的培训和研究所需的。 K99/R00 奖足以让我 这种高级培训所需的受保护时间，使我能够在以下条件下继续促进我的成长 戴森博士和恩格曼博士的指导。我预计该提案的指导阶段包括 使用小鼠模型研究操纵 CIN 在实体瘤中的治疗相关性，以及 启动一种新颖的筛选方法将需要 1-2 年时间，并至少发表一篇主要出版物。这 该奖项的独立阶段将使我能够识别和描述其他与癌症相关的、 染色体结构和基因组稳定性的调节因子。这些数据将一起用于证明未来的合理性 我计划在独立阶段第三年开始时提交的 R01 资助中提出的研究。