Histone isomerization and pluripotency

组蛋白异构化和多能性

基本信息

批准号：
8678949
负责人：
Nobuaki Kikyo
金额：
$ 28.88万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8678949
关键词：
Acetylation Affect Amino Acids Autologous Transplantation Binding Biological Biology Cell Differentiation process Cell Nucleus Cell Therapy Cell Transplantation Cells Chromatin Cloning Complement Complex Cytoplasm DNA DNA Microarray Chip Development Egg Proteins Employee Strikes Enzymes Epigenetic Process FOS gene Fibroblasts Gene Activation Gene Expression Gene Expression Profile Gene Expression Regulation Gene Targeting Generations Genes Goals Histone H3 Histones In Vitro Incubated Isomerase JUN gene Laboratories Lead Life Lysine Mammalian Cell Mediating Methylation Microarray Analysis Mitogens Modification Molecular Molecular Chaperones Molecular Conformation Mus Nuclear Patients Peptide Conformation Peptide Fragments Peptidylprolyl Isomerase Phosphorylation Physiological Play Polycomb Preparation Process Production Proline Proteins Recording of previous events Regenerative Medicine Relaxation Reporting Repression Research Research Personnel Role Series Site Somatic Cell Stem cells Tail Testing Tissues Xenopus Xenopus oocyte base chromatin immunoprecipitation chromatin protein design egg embryonic stem cell genome-wide histone modification human disease improved in vivo Model induced pluripotent stem cell innovation insight interest knock-down novel novel strategies nuclear reprogramming nucleoplasmin pluripotency stem cell biology stem cell technology tool

项目摘要

DESCRIPTION (provided by applicant): Project Summary Successful somatic cell nuclear cloning has demonstrated that egg cytoplasm contains powerful nuclear reprogramming activities. These activities can induce dedifferentiation of the terminally differentiated donor nuclei, leading to their acquisition of a totipotent state. Despite the long history of the cloning study and particularly intensive recent research, however, little is known about the molecular mechanisms that reprogram donor nuclei during cloning. The long-term goal of this project is to identify these egg-derived nuclear reprogramming factors and apply the factors to reprogram the differentiation status of living mammalian cells. This project i expected to lead to efficient preparation of patients-derived new tissues for autologous transplantation, complementing the induced pluripotent stem (iPS) cell technology. Global chromatin decondensation is one of the most striking cell biological changes observed in the donor nuclei. The investigator's group has recently found that the egg protein nucleoplasmin and the peptidyl isomerase p16 are responsible for the chromatin decondensation. These two proteins trigger acetylation, phosphorylation and methylation on specific histones and promote gene activation. Transduction of p16 and nucleoplasmin significantly facilitate the production of iPS cells from mouse fibroblasts. Based on these observations, it was hypothesized that nucleoplasmin and p16 organize a specific set of epigenetic modifications on the target genes, which leads to chromatin relaxation and nuclear reprogramming. To test this hypothesis, following specific aims are proposed. Specific Aim 1 is designed to understand how nucleoplasmin and p16 contributes to epigenetic reprogramming of key pluripotency genes during iPS cell formation. In Specific Aim 2, the role of p16 in pluripotency of ES cells will be studied by gene knockdown, DNA microarray analysis and chromatin immunoprecipitation. In Specific Aim 3, the relationship among phosphorylation, methylation and isomerization of histones will be investigated by using peptide fragments, chromatin and living cells. Together, these studies are expected to provide a novel insight into the reprogramming of cell differentiation through epigenetic remodeling.

描述（由申请人提供）：项目概要成功的体细胞核克隆证明卵细胞质含有强大的核重编程活性。这些活性可以诱导终末分化供体核的去分化，导致它们获得全能状态。尽管克隆研究有着悠久的历史，而且最近的研究特别深入，但人们对克隆过程中重新编程供体核的分子机制知之甚少。该项目的长期目标是鉴定这些卵源性核重编程因子，并应用这些因子来重编程活哺乳动物细胞的分化状态。我预计该项目将有效地制备来自患者的新组织以用于自体移植，从而补充诱导多能干（iPS）细胞技术。整体染色质解凝是在供体细胞核中观察到的最显着的细胞生物学变化之一。研究小组最近发现，鸡蛋蛋白核蓝蛋白和肽基异构酶 p16 负责染色质解缩。这两种蛋白触发特定组蛋白的乙酰化、磷酸化和甲基化，并促进基因激活。 p16 和核蓝蛋白的转导显着促进小鼠成纤维细胞产生 iPS 细胞。基于这些观察结果，推测核蓝蛋白和 p16 在靶基因上组织了一组特定的表观遗传修饰，从而导致染色质松弛和核重编程。为了检验这一假设，提出了以下具体目标。具体目标 1 旨在了解核蓝蛋白和 p16 在 iPS 细胞形成过程中如何促进关键多能性基因的表观遗传重编程。在具体目标 2 中，将通过基因敲除、DNA 微阵列分析和染色质免疫沉淀研究 p16 在 ES 细胞多能性中的作用。在具体目标3中，将利用肽片段、染色质和活细胞研究组蛋白磷酸化、甲基化和异构化之间的关系。总之，这些研究预计将为通过表观遗传重塑来重新编程细胞分化提供新的见解。