Post-Transcriptional Regulators of Epidermal Homeostasis and Neoplasia

表皮稳态和肿瘤的转录后调节因子

基本信息

批准号：
10161730
负责人：
GEORGE L SEN
金额：
$ 37.02万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-18 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10161730
关键词：
3&apos Untranslated Regions 3-Dimensional Address Atrophic Basal cell carcinoma Basement membrane Binding Cells Complex Dermal Development Differentiation and Growth Disease Epidermis Epithelial Equilibrium Event Foundations Generations Genetic Transcription Genetic Translation Growth Homeostasis Human Immune Lead Malignant Neoplasms Mediating Messenger RNA Modeling Molecular Mus Natural regeneration Neoplasms Organism Pathway interactions Play Population Post-Transcriptional Regulation Process Proteins Psoriasis Publications Publishing RNA RNA Helicase Regulation Research Design Role Skin Squamous cell carcinoma Structure Transcript Translating Translations Tumor stage Water Work cancer site chronic wound clinically relevant crosslinking and immunoprecipitation sequencing design epidermal stem cell experimental study in vivo insight knock-down loss of function mRNA Transcript Degradation member messenger ribonucleoprotein neoplastic novel strategies overexpression premature prevent protein degradation prototype self-renewal skin disorder stem cells therapy development tumor tumor initiation tumor progression tumorigenesis

项目摘要

Project Summary/Abstract Background: Transcriptional mechanisms that regulate epidermal homeostasis have been well established but recently we have discovered that post-transcriptional mechanisms play prominent roles in maintaining epidermal self-renewal. We have shown that the RNA helicase DDX6 is necessary to maintain epidermal self-renewal through the mRNA degradation and translation pathway. By associating with specific members of the translation pathway DDX6 binds to and mediates the translation of self- renewal and proliferation transcripts to maintain self-renewal. DDX6 also associates with mRNA degradation proteins to bind differentiation-inducing transcripts to promote their degradation to prevent premature differentiation. Objective/hypothesis: This proposal seeks to understand the regulation of epidermal homeostasis and tumor initiation through post-transcriptional mechanisms. We previously identified proteins associated with DDX6 and our objective is to characterize the role of each protein in regulating epidermal growth, differentiation, and progression to neoplasia as well as the mechanisms of action. Furthermore we seek to determine the specific transcripts that DDX6 and its associated complexes bind during homeostasis and tumor initiation. Specific Aims: (1) To determine the role of DDX6 associated proteins on epidermal homeostasis and tumor initiation and (2) to identify and characterize the transcripts associated with DDX6 complexes. Study Design: To study epidermal homeostasis in a more clinically relevant setting, we generate 3-dimensionally intact human skin, containing human epidermal cells (that have been permanently knocked down for either DDX6 or its associated proteins) in the context of human dermal stroma and basement membrane, regenerated on immune compromised mice. By using this model, we can perform loss of function experiments on DDX6 and its associated proteins in regenerated human skin to characterize their role in epidermal growth, differentiation, and progression to neoplasia. We will also use CLIP- Seq to determine the RNAs associated with DDX6 complexes during the progression from normal to neoplastic epidermis.

项目摘要/摘要背景：调节表皮稳态的转录机制具有已经建立得很好，但最近我们发现了转录后机制在维持表皮自我更新方面起着重要作用。我们有表明RNA解旋酶DDX6对于维持表皮自我更新是必要的通过mRNA降解和翻译途径。通过与特定翻译途径的成员DDX6与并介导的翻译自我更新和增殖转录本以保持自我更新。 DDX6也是如此与mRNA降解蛋白结合诱导分化的蛋白成绩单以促进其降解以防止过早差异。客观/假设：该提议旨在了解通过转录后机制的表皮稳态和肿瘤起始。我们以前以前确定了与DDX6相关的蛋白质，我们的目标是表征每种蛋白质在调节表皮生长，分化，和肿瘤的进展以及作用机理。此外，我们寻求确定DDX6及其相关复合物的特定成绩单在稳态和肿瘤开始期间结合。具体目的：（1）确定DDX6相关蛋白在表皮上的作用稳态和肿瘤的启动以及（2）识别和表征成绩单与DDX6复合物相关。研究设计：在更临床相关的环境中研究表皮稳态我们产生3维完全完整的人皮肤，包含人表皮细胞（已被永久击倒DDX6或其相关的蛋白质）在人真皮基质和地下膜的背景下，在免疫损害的小鼠上再生。通过使用此模型，我们可以执行 DDX6及其相关蛋白的功能实验损失在再生中人体皮肤以表征其在表皮生长，分化和发展为肿瘤。我们还将使用夹克确定RNA 从正常到肿瘤的进展过程中与DDX6复合物相关表皮。