Regulation of epidermal growth and differentiation through mRNA export

通过 mRNA 输出调节表皮生长和分化

基本信息

批准号：
10675700
负责人：
GEORGE L SEN
金额：
$ 44.69万
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：
2022
资助国家：
美国
起止时间：
2022-08-02 至 2027-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10675700
关键词：
3-Dimensional Adaptor Signaling Protein Address Basal Cell Basement membrane Binding Binding Proteins Cell Differentiation process Cell Proliferation Cells Clonal Expansion Code Complex Cytoplasm DNA sequencing Defect Dermal Differentiated Gene Differentiation and Growth Disease Equilibrium Future Gene Expression Generations Genes Genetic Transcription Growth Homeostasis Human Immune Immunoprecipitation Impairment Lead Malignant Neoplasms Mediating Messenger RNA Mitosis Modeling Molecular Mus Mutate Mutation NOTCH1 gene Natural regeneration Nuclear Pore Complex Pathway interactions Phenotype Play Process Proliferating Protein Export Pathway Proteins Psoriasis RNA RNA-Binding Proteins Regulation Research Design Role Skin Skin Carcinoma Specificity Squamous cell carcinoma Stratum Basale Tissues Transcript Translating Water chronic wound clinically relevant crosslinking and immunoprecipitation sequencing design embryonic stem cell epidermal stem cell experimental study knock-down loss of function loss of function mutation mRNA Export mutant next generation sequencing pluripotency posttranscriptional protein complex self-renewal skin disorder stem stem cell growth stem cells transcription factor transcriptome sequencing

项目摘要

Project Summary/Abstract Background: Transcriptional mechanisms that regulate epidermal homeostasis have been well established but recently we have discovered that mRNA export mechanisms play prominent roles in maintaining epidermal self-renewal. We have shown that RBM15 associates with the NXF1 exporter only in stem and progenitor cells while ZC3H18 associates with NXF1 in differentiated cells. This association allows RBM15 or ZC3H18 to control the mRNA export of key transcripts involved in epidermal growth and differentiation. Objective/hypothesis: This proposal seeks to understand the regulation of epidermal stem and progenitor cell self-renewal and differentiation through post-transcriptional mechanisms. We have identified RNA binding proteins that are necessary for the export of self-renewal mRNAs to promote epidermal self-renewal. Similarly we have identified RNA binding proteins that are necessary to export differentiation inducing mRNAs to promote epidermal differentiation. Furthermore mutations in these proteins can lead to clonal expansion of the skin due to altered regulation of epidermal growth and differentiation. Specific Aims: (1) To determine the role of RBM15 and ZC3H18 on epidermal growth and differentiation. (2) To determine the molecular mechanisms of RBM15 and ZC3H18 wildtype and mutant proteins impact on epidermal homeostasis. 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 RBM15 or ZC3H18) 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 RBM15 or ZC3H18 in regenerated human skin to characterize their role in epidermal growth and differentiation. We will also use RNA immunoprecipitations followed by next generation sequencing to determine the RNAs associated with these proteins.

项目概要/摘要背景：调节表皮稳态的转录机制已经很成熟，但最近我们发现 mRNA 输出机制在维持表皮自我更新方面发挥着重要作用。我们已经证明 RBM15 仅在干细胞和祖细胞中与 NXF1 输出蛋白相关，而 ZC3H18 与分化细胞中的 NXF1 相关。该关联允许 RBM15 或 ZC3H18 控制参与表皮生长的关键转录物的 mRNA 输出差异化。目标/假设：本提案旨在了解表皮的调节干细胞和祖细胞通过转录后自我更新和分化机制。我们已经鉴定出出口所需的 RNA 结合蛋白自我更新 mRNA 促进表皮自我更新。同样我们也确定了 RNA 结合蛋白是输出分化诱导 mRNA 所必需的促进表皮分化。此外，这些蛋白质的突变可能导致由于表皮生长调节改变而导致的皮肤克隆性扩张差异化。具体目的：（1）确定RBM15和ZC3H18对表皮生长的作用和差异化。 (2) 确定RBM15和ZC3H18的分子机制野生型和突变蛋白对表皮稳态的影响。研究设计：为了在更具临床相关性的环境中研究表皮稳态，我们生成 3 维完整的人类皮肤，包含人类表皮细胞（即在人类环境中，RBM15 或 ZC3H18 已被永久敲除真皮基质和基底膜在免疫受损的小鼠身上再生。通过使用该模型，我们可以在RBM15或ZC3H18上进行功能损失实验在再生人类皮肤中表征其在表皮生长中的作用和差异化。我们还将使用 RNA 免疫沉淀，然后是下一代测序以确定与这些蛋白质相关的 RNA。