Regulation of epidermal growth and differentiation through mRNA export

通过 mRNA 输出调节表皮生长和分化

• 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
• 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

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.

项目总结/摘要 背景：调节表皮稳态的转录机制 但是最近我们发现mRNA的输出机制 在维持表皮自我更新中发挥重要作用。我们已经证明，RBM 15 ZC 3 H18仅在干细胞和祖细胞中与NXF 1输出者相关， 在分化细胞中与NXF 1相关。这种关联允许RBM 15或ZC 3 H18 控制参与表皮生长的关键转录物的mRNA输出， 分化 目的/假设：该提案旨在了解表皮细胞的调节， 干细胞和祖细胞通过转录后自我更新和分化 机制等我们已经确定了出口所需的RNA结合蛋白 来促进表皮的自我更新。同样，我们发现 RNA结合蛋白是将诱导分化的mRNA输出到 促进表皮分化。此外，这些蛋白质的突变可能导致 由于表皮生长调节的改变而引起的皮肤的克隆扩张， 分化 具体目的：（1）研究RBM 15和ZC 3 H18对表皮生长的影响 和差异化。(2)确定RBM 15和ZC 3 H18的分子机制 野生型和突变蛋白影响表皮稳态。 研究设计：为了在更临床相关的环境中研究表皮稳态，我们 生成三维完整的人类皮肤，包含人类表皮细胞（其 在人类环境中，RBM 15或ZC 3 H18已被永久敲除 真皮基质和基底膜，在免疫受损小鼠上再生。 利用该模型，我们可以对RBM 15或ZC 3 H18进行功能丧失实验 以表征它们在表皮生长中的作用， 分化我们还将使用RNA免疫沉淀法， 测序以确定与这些蛋白质相关的RNA。