Genomic stability of human pluripotent stem cells

人类多能干细胞的基因组稳定性

• 批准号: 8261878
• 负责人: Paul Hubert Lerou
• 金额: $ 22.83万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8261878
• 关键词: Accounting; Affect; Aneuploidy; Award; Biological; Biological Assay; CDK2 gene; 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; Imaging Techniques; Imaging technology; Infertility; Institutes; Institution; Instruction; Life; Maintenance; Malignant Neoplasms; Mediating; Mentors; Mentorship; Mitotic; Molecular; Mutation; 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; 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; repaired; response; segregation; self-renewal; senescence; small molecule; stem; stem cell biology; telomere; tool; tumorigenesis

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 ofthe 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 ofthe pathways maintaining genomic stability. Viral-mediated gene over expression / 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.

细胞周期确保重复的染色体忠实地重复和分离为子细胞。 细胞周期的失调导致基因组不稳定性，包括非整倍性，癌症的标志和A 早期胚胎或配子中出现非整倍性时，不育和遗传综合征的重要原因。 ES细胞仍保持基因组稳定性，即使它们改变了RB和p53功能，并且能够 逃脱衰老，无限期地分裂，而无需造成DNA损伤。该研究建议将测试 HES细胞具有保持基因组稳定性的独特和专业机制的假设。 我的职业目标是成为干细胞生物学领域的独立资助的临床医生 HES和诱导多能茎（IPS）细胞操作和细胞周期调节方面的专业知识。 获得独立奖的途径（K99/R00）将极大地帮助我过渡到完全独立。在 在不久的将来，该奖项将使我能够获得多能细胞生物学，实时细胞的深入专业知识 成像和先进的分子生物学技术，用于研究细胞周期调节 Marc W. Kirschner博士的George Q. Daley博士的指导。培训计划 哈佛医学的儿童医院和系统生物学系血液学/肿瘤科 学校汇集了两个机构以及哈佛干细胞研究所的资源， 并为在指导阶段完成培训的培训提供了一个杰出的环境。这会 极大地促进了我有效向独立过渡。 ^ 该研究建议的具体目的是：1）表征RB和p53途径及其在 人多能细胞中的基因组稳定性维持，2）从体细胞中产生IPS细胞 与基因组不稳定性有关的港口突变。高级技术，包括实时和固定细胞 成像，将应用于人多能干细胞，以深入分析该途径 维持基因组稳定性。病毒介导的基因在表达 / RNAi基因敲低技术和 小分子将用于审问这些途径，并将与发展相辅相成 基因组不稳定性疾病特异性IPS细胞系。 这项研究的目的是研究多能细胞中基因组稳定性维持的机制 为了更好地理解非整倍性的分子基础。