DNA repair pathway choice mediates somatic cell reprogramming

DNA修复途径选择介导体细胞重编程

基本信息

批准号：
10413221
负责人：
Dieter Meinrad Egli
金额：
$ 46.71万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10413221
关键词：
Affect Aneuploidy BRCA1 gene Basic Science Biological Assay Cell Cycle Cell Cycle Progression Cell Line Cell Therapy Cells Characteristics Competence DNA DNA Damage DNA Double Strand Break DNA Repair DNA Repair Pathway DNA biosynthesis DNA replication fork Data Development Double Strand Break Repair Frequencies G2 Phase Generations Genetic Transcription Genome Genome Stability Genomic Instability Genomics Impairment Laboratories Lead Location Maps Mediating Mitosis Molecular Mus Mutation Nonhomologous DNA End Joining Optics Pathway interactions Pluripotent Stem Cells Repetitive Sequence Research Role S phase Signal Transduction Site Somatic Cell Structure System TP53 gene Techniques cell type genome integrity homologous recombination improved induced pluripotent stem cell mutant next generation sequencing nuclear transfer overexpression p53-binding protein 1 repaired senescence somatic cell nuclear transfer stem cells whole genome

项目摘要

Project Summary Reprogramming is the conversion of a somatic cell to a pluripotent stem cell. This technique is now routinely used in laboratories around the world, but the vast majority of reprogrammed stem cell lines are not developmentally fully competent, compromising their utility in research and therapy. Reprogramming induces DNA damage, which can have lasting consequences on the quality of the resulting cells. Our studies have shown that DNA damage during reprogramming is induced by abnormalities in DNA replication. However, the cause of DNA damage, the mechanisms of repair, and the developmental consequences of the damage are not well understood. The strength of this proposal is that with the experimental systems used, we are able to identify the specific type of damage induced by reprogramming, and the molecular mechanisms required for repair: we are able to distinguish the role of double strand break HR from the role of stalled replication fork stability. We are also able to distinguish the effect of genome instability on reprogramming efficiency from incomplete transcriptional transitions. We are able to map the sites in the genome with reprogramming-induced damage, and we are able to identify pathways that can be used to increase genome stability and potentially improve developmental competence of reprogrammed stem cells. These studies will provide a mechanistic understanding how genome instability inhibits the induced transition between different cellular states.

项目概要重编程是体细胞向多能干细胞的转化。这种技术现在已成为常规世界各地的实验室都在使用，但绝大多数重编程干细胞系并不是发育上完全有能力，从而损害了他们在研究和治疗中的效用。重编程诱导 DNA 损伤，可能会对所得细胞的质量产生持久的影响。我们的研究有表明重编程过程中的 DNA 损伤是由 DNA 复制异常引起的。然而， DNA 损伤的原因、修复机制以及损伤的发育后果不太理解。该提案的优势在于，通过使用的实验系统，我们能够确定重编程引起的特定类型的损伤，以及重编程所需的分子机制修复：我们能够区分双链断裂 HR 的作用和停滞复制叉的作用稳定。我们还能够区分基因组不稳定性对重编程效率的影响不完整的转录转换。我们能够通过重编程诱导来绘制基因组中的位点损伤，我们能够识别可用于提高基因组稳定性和潜在的途径提高重编程干细胞的发育能力。这些研究将提供一个机制了解基因组不稳定性如何抑制不同细胞状态之间的诱导转变。