Regulatory Role of Transcription Factor Slug in Aging

转录因子 Slug 在衰老中的调节作用

基本信息

批准号：
8735056
负责人：
WEN-SHU WU
金额：
$ 32.7万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8735056
关键词：
Adipose tissue Adult Affect Aging Aging-Related Process American Animal Model Attenuated Behavioral Research Binding Binding Sites Biological Assay Body Weight decreased Boxing Bromodeoxyuridine CDKN2A gene Cell Aging Cell Cycle Cell Cycle Arrest Cell Cycle Progression Cell Survival Cells Cellular Stress Consensus Cyclin-Dependent Kinase Inhibitor 2A Cyclin-Dependent Kinases DNA biosynthesis Data Development E-Box Elements Electrophoretic Mobility Shift Assay Embryo Epithelial Event Fibroblasts Foundations Genetic Genetic Transcription Genetically Modified Animals Goals Growth HMGA2 gene Health Hematopoietic Hematopoietic stem cells Human In Vitro Intervention Knock-out Kyphosis deformity of spine Laboratories Learning Link Luciferases Mesenchymal Methods Mission Modeling Molecular Mus Muscle satellite cell Oncogene Activation Osteoporosis Outcome Pathway interactions Personal Satisfaction Phenotype Premature aging syndrome Process Promoter Regions Public Health Regulation Reporter Research Research Project Grants Role Series Site Skeletal Muscle Stem cells Technology Testing Therapeutic Therapeutic Intervention Tissues United States National Institutes of Health Work Zinc Fingers adult stem cell anti aging chromatin immunoprecipitation combat derepression improved in vivo inhibitor/antagonist innovation insight method development novel overexpression oxidative damage p19ARF prevent promoter regenerative research study self-renewal senescence slug small hairpin RNA social stem tissue regeneration transcription factor

项目摘要

DESCRIPTION (provided by applicant): Recent evidence suggests that the cellular senescence machinery and the decline of the regenerative capacity of certain self-renewing stem cell compartments may contribute importantly to mammalian aging. Although derepression of INK4A/ ARF locus is thought to be an intrinsically linked event, very little is known about the specific mechanisms responsible for modulation of p16INK4 and p19ARF expression. Preliminary work from the PI's laboratory suggests that Slug, a zinc-finger transcription factor known to affect cell survival and the epithelial-mesenchymal transition in a variety of tissues, directly inhibits p16INK4 transcription, to the extent that Slug deficiency leads to cellular senescence and premature aging in vitro and in animal models. We therefore hypothesize that Slug influences aging by restraining cellular senescence and sustaining the regenerative potential of adult stem and progenitor cells, primarily through direct negative regulation of the p16INK4 gene. These developments promote our long-term objective -- to discover novel molecular pathways underlying the senescence of both differentiated and stem and progenitor cells -- and have led to three immediate research aims to determine the precise contribution of Slug to organismal aging. Aim 1 will establish the extent to which p16INK4a contributes to accelerated senescence in Slug-deficient MEFs using both knockdown and knockout technologies. Aim 2 will attempt to elucidate the molecular mechanisms by which Slug negatively regulates expression of p16INK4a, by relying on chromatin immunoprecipitation and electrophoretic mobility shift assays. Aim 3 will define the functional role of Slug in tissue senescence and the regenerative potential of adult stem and progenitor cells. These experiments will use a series of genetically modified animal models as well as hematopoietic and skeletal muscle stem cells from Slug and p16INK4a- deficient mice. This proposal is innovative because it exploits a transcription factor, Slug, with unique regulatory activity at the INK4a/ ARF locus. The likelihood that a better understanding of the in vivo regulation of this locus, and hence of mammalian aging, will emerge from our project appears high.

描述（由申请人提供）：最近的证据表明，某些自我更新干细胞区室的细胞衰老机制和再生能力的下降可能对哺乳动物的衰老有重要贡献。尽管认为Ink4a/ ARF基因座的压抑被认为是本质上链接的事件，但对负责调节P16INK4和P19ARF表达的特定机制知之甚少。 PI实验室的初步工作表明，SLUG是一种已知会影响细胞存活和各种组织中上皮间质转变的锌指转录因子，直接抑制p16ink4转录，以至于SLUG缺乏导致细胞衰老和动物模型中的细胞衰变和过度的衰老。因此，我们假设SLUG通过限制细胞衰老并维持成人茎和祖细胞的再生潜力来影响衰老，这主要是通过对P16INK4基因的直接负调控。这些发展促进了我们的长期目标 - 发现分化和茎和祖细胞衰老的基本的新分子途径 - 并导致三个直接研究旨在确定SLUG对生物衰老的确切贡献。 AIM 1将确定P16INK4A在多大程度上使用敲除技术和淘汰技术促进了slug不足的MEF加速衰老的程度。 AIM 2将尝试通过依靠染色质免疫沉淀和电泳迁移率转移测定法来阐明SLUG负面调节p16Ink4a表达的分子机制。 AIM 3将定义SLUG在组织衰老中的功能作用以及成年茎和祖细胞的再生潜力。这些实验将使用一系列转基因的动物模型以及来自slug和p16ink4a缺乏小鼠的造血和骨骼肌干细胞。该提案具有创新性，因为它利用了在Ink4a/ ARF基因座的独特调节活动的转录因子SLUG。更好地了解该基因座的体内调节以及哺乳动物衰老的可能性很高。