Genomic stability of human pluripotent stem cells

人类多能干细胞的基因组稳定性

• 批准号: 7714486
• 负责人: Paul Hubert Lerou
• 金额: $ 9.72万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7714486
• 关键词: Accounting; Affect; Aneuploidy; Award; Biological; Biological Assay; Cell Culture Techniques; Cell Cycle; Cell Cycle Regulation; Cell Cycle Regulation Pathway; Cell Line; Cell division; Cell model; Cells; Cellular biology; Chromosomes; Complement; Coupled; Cyclin E; Cytogenetic Analysis; DNA; DNA Damage; DNA Repair; DNA biosynthesis; Defect; Development; Disease; Embryo; Ensure; Environment; Fibroblasts; Funding; Future; Generations; Genes; Genetic; Genome Stability; Genomic Instability; Germ Cells; Goals; Hematology; Hospital Departments; Human; Image; Imaging Techniques; Imaging technology; Infertility; Institutes; Institution; Life; Maintenance; Malignant Neoplasms; Mediating; Mentors; Mentorship; Mitotic; Molecular; Mutation; Pathway Analysis; Pathway interactions; Patients; Pediatric Hospitals; Phase; Pluripotent Stem Cells; Production; RNA Interference; Regulation; Reporter; Research; Research Proposals; Resources; Role; Scientist; Somatic Cell; Stem cells; Syndrome; Systems Biology; TP53 gene; Techniques; Telomere Maintenance; Testing; Training; Training Programs; Viral; base; cancer genetics; career; cell fixing; cellular imaging; daughter cell; embryonic stem cell; human embryonic stem cell; improved; induced pluripotent stem cell; insight; knock-down; medical schools; novel; oncology; overexpression; repaired; response; segregation; self-renewal; senescence; small molecule; stem; stem cell biology; telomere; tool; tumorigenesis

DESCRIPTION (provided by applicant): The cell cycle ensures faithful duplication and segregation of duplicated chromosomes into daughter cells. Dysregulation of the cell cycle results in genomic instability, including aneuploidy, a hallmark of cancer and a significant cause of infertility and genetic syndromes when aneuploidy arises in the early embryo or gametes. ES cells maintain genomic stability even though they have altered Rb and p53 function and they are able to escape senescence, dividing indefinitely without the accruing DNA damage. This research proposal will test the hypothesis that hES cells possess unique and specialized mechanisms to maintain genomic stability. My career goal is to become an independently funded clinician-scientist in the field of stem cell biology with expertise in hES and induced pluripotent stem (iPS) cell manipulation and cell cycle regulation. The pathway to Independence Award (K99/R00) will greatly assist my transition to full independence. In the immediate future, this award will allow me to gain in-depth expertise in pluripotent cell biology, live-cell imaging and the advanced molecular biological techniques to study cell cycle regulation under the mentorship of Dr. George Q. Daley and co-mentorship of Dr. Marc W. Kirschner. The training program in the hematology/oncology division at Children's Hospital and department of systems biology at Harvard Medical School brings together resources from both institutions as well as those of the Harvard Stem Cell Institute, and provides an outstanding environment for the completion of training during the mentored phase. This will greatly facilitate my effective transition to independence. The specific aims of this research proposal are to: 1) characterize the Rb and p53 pathways and their role in genomic stability maintenance in human pluripotent cells, and 2) generate IPS cells from somatic cells that harbor mutations implicated in genomic instability. Advanced techniques, including live and fixed cell imaging, will be applied to human pluripotent stem cells to perform in depth analysis of the pathways maintaining genomic stability. Viral-mediated gene overexpression / RNAi gene knock-down techniques and small molecules will be used to interrogate these pathways and will be complemented by the development genomic instability disease specific iPS cell lines. The goal of this research is to study the mechanisms of genomic stability maintenance in pluripotent cells in order to better understand the molecular basis of aneuploidy. PUBLIC RELEVANCE: Cells have specialized molecular pathways to maintain genetic stability. These pathways are disrupted in a wide variety of disease including cancer and genetic syndromes. By studying how human embryonic stem cells maintain genetic stability we will gain an improved understanding of how genetic instability can arise in human cells and how it may result in disease. This research will also provide insight into basic embryonic stem cell biology.

描述（由申请人提供）：细胞周期确保复制染色体的忠实复制和分离到子细胞中。细胞周期失调会导致基因组不稳定，包括非整倍性，这是癌症的标志，也是早期胚胎或配子中出现非整倍性时导致不孕和遗传综合征的重要原因。 ES 细胞即使改变了 Rb 和 p53 功能，也能保持基因组稳定性，并且能够逃避衰老，无限期分裂而不会造成 DNA 损伤。该研究计划将检验 hES 细胞拥有独特且专门的机制来维持基因组稳定性的假设。我的职业目标是成为干细胞生物学领域的一名独立资助的临床医生科学家，拥有 hES 和诱导多能干 (iPS) 细胞操作和细胞周期调控方面的专业知识。 获得独立奖（K99/R00）的途径将极大地帮助我过渡到完全独立。在不久的将来，该奖项将使我能够在多能细胞生物学、活细胞成像和先进分子生物学技术方面获得深入的专业知识，以在 George Q. Daley 博士和共同指导下研究细胞周期调控Marc W. Kirschner 博士。儿童医院血液/肿瘤科和哈佛医学院系统生物学系的培训项目汇集了双方机构以及哈佛干细胞研究所的资源，为培训期间的完成提供了良好的环境。指导阶段。这将极大地促进我向独立的有效过渡。 该研究计划的具体目标是：1) 表征 Rb 和 p53 通路及其在人类多能细胞基因组稳定性维持中的作用，2) 从含有与基因组不稳定相关的突变的体细胞生成 IPS 细胞。包括活细胞和固定细胞成像在内的先进技术将应用于人类多能干细胞，以对维持基因组稳定性的途径进行深入分析。病毒介导的基因过度表达/RNAi基因敲低技术和小分子将用于探究这些途径，并将通过开发基因组不稳定疾病特异性iPS细胞系来补充。本研究的目的是研究多能细胞基因组稳定性维持的机制，以便更好地了解非整倍性的分子基础。 公众相关性：细胞具有专门的分子途径来维持遗传稳定性。这些途径在包括癌症和遗传综合症在内的多种疾病中被破坏。通过研究人类胚胎干细胞如何维持遗传稳定性，我们将更好地了解遗传不稳定性如何在人类细胞中出现以及它如何导致疾病。这项研究还将深入了解基本的胚胎干细胞生物学。