Esrp regulated programs of alternative splicing in skin development and function

Esrp 调控皮肤发育和功能中的选择性剪接程序

• 批准号: 8899793
• 负责人: RUSS Paul CARSTENS
• 金额: $ 36.97万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8899793
• 关键词: Ablation; Accounting; Adult; Affect; Alleles; Alopecia; Alternative Splicing; Bullous Skin Diseases; Cell Proliferation; Cells; Defect; Development; Disease; Electrolyte Balance; Embryo; Epidermis; Epithelial; Epithelial Cell Proliferation; Epithelial Cells; Epithelium; Erinaceidae; Exons; Family member; Fibroblast Growth Factor Receptor 2; Foundations; Gene Expression; Gene Expression Regulation; Genes; Genetic Programming; Growth; Hair; Hair follicle structure; Health; High-Throughput Nucleotide Sequencing; Impaired wound healing; Investigation; Knock-out; Knockout Mice; Lead; Liquid substance; Maintenance; Mediating; Mesenchymal; Mesenchyme; MicroRNAs; Molecular; Morphology; Mus; Natural regeneration; Pathway interactions; Phenotype; Population; Process; Production; Protein Isoforms; Psoriasis; Publications; Qualifying; RNA Splicing; Regulation; Regulator Genes; Role; Signal Pathway; Signal Transduction; Skin; Skin Cancer; Skin graft; Stem cells; Technology; Transcript; Variant; appendage; arm; genome-wide; in vivo; insight; knockout gene; mouse model; novel; programs; receptor; tool; transcriptome sequencing

DESCRIPTION (provided by applicant): The epidermis and skin appendages perform essential barrier functions that protect the body from environmental insults and maintain fluid-electrolyte balance. Defects in the development, differentiation, and proliferation of the cells tht comprise the skin and hair follicles lead to numerous diseases such as skin cancer, psoriasis, bullous skin disease, and alopecia. Transcriptional programs that control skin and hair follicle development have defined processes important for epidermal integrity and hair regeneration. More recently microRNAs have also emerged as important regulators of gene expression in the skin. In contrast, the role of alternative splicing (AS) in skin development and function is essentially unstudied~ there have been no publications focused on alternative splicing factors in the skin or its appendages. Reciprocal epithelial- mesenchymal interactions (EMIs) underlie formation of the epidermis and hair follicles. One important arm involves the interaction of mesenchymal derived Fgf7 and/or Fgf10 with an epithelial-specific splice isoform of fibroblast growth factor receptor 2, Fgfr2-IIIb. Fgf7 and Fgf10 specifically interact with Fgfr2-IIIb, but not mesenchymal Fgfr2-IIIc. Abrogation of this directional signaling pathway results in defects in the epidermis and hair follicles. My lab identified the epithelial-specific splicing factors Esrp1 and Esrp2 that are both necessary and sufficient for the expression of the Fgfr2-IIIb splice variant in epithelial cells. We generated Esrp1 and Esrp2 knockout mice and demonstrate that combined Esrp1/Esrp2 KO is lethal and results in epidermal hypoplasia, reduced follicle numbers, and sparse hair. We hypothesize that the Esrps are required for normal epidermal and follicular development and function by enforcing the expression of epithelial-specific splice isoforms. We will determine the phenotypes of Esrp deletion in specific epithelial cell populations in skin and appendages and identify a comprehensive set of Esrp target transcripts through the following Aims: 1) Determine the phenotypes associated with Esrp ablation in the interfollicular epidermis and in hair follicles. We will define phenotypes associated with conditionally ablation of the Esrps in developing and adult epidermis and hair follicles. 2) Define comprehensive programs of Esrp regulated alternative splicing in the epidermis and hair follicle. We will use high throughput sequencing (RNA-Seq) and splicing sensitive microarrays to define genome-wide programs of alternative splicing in the different cell populations that populate the skin. These technologies will be used in conjunction with conditional deletion strategies to define Esrp regulated targets i the epidermis and hair follicle stem cells. The proposed aims constitute the first comprehensive analysis of alternative splicing in skin development and function, thereby introducing a new paradigm to the field. These studies will reveal novel insights into the processes that impact skin development and function.

描述（由申请人提供）：表皮和皮肤附属物具有基本的屏障功能，可保护身体免受环境侵害并维持液体-电解质平衡。构成皮肤和毛囊的细胞的发育、分化和增殖的缺陷导致许多疾病，如皮肤癌、牛皮癣、大疱性皮肤病和脱发。控制皮肤和毛囊发育的转录程序已经定义了对表皮完整性和毛发再生重要的过程。最近，microRNA也成为皮肤基因表达的重要调节因子。相比之下，选择性剪接（AS）在皮肤发育和功能中的作用基本上是未研究的-没有出版物关注皮肤或其附件中的选择性剪接因子。上皮-间充质相互作用（EMIs）是表皮和毛囊形成的基础.一个重要的分支涉及间充质来源的Fgf 7和/或Fgf 10与成纤维细胞生长因子受体2的上皮特异性剪接同种型Fgfr 2-IIIb的相互作用。Fgf 7和Fgf 10与Fgfr 2-IIIb特异性相互作用，但不与间充质Fgfr 2-IIIc相互作用。这种定向信号通路的消除导致表皮和毛囊的缺陷。我的实验室鉴定了上皮特异性剪接因子Esrp 1和Esrp 2，它们对于Fgfr 2-IIIb剪接变体的表达是必要的，也是足够的。 上皮细胞我们产生了Esrp 1和Esrp 2基因敲除小鼠，并证明联合Esrp 1/Esrp 2 KO是致命的，并导致表皮发育不全，毛囊数量减少和毛发稀疏。我们推测，Esrps是需要正常的表皮和滤泡的发展和功能，通过执行上皮特异性剪接异构体的表达。我们将确定皮肤和附件中特定上皮细胞群体中Esrp缺失的表型，并通过以下目的鉴定一组全面的Esrp靶转录物：1）确定与毛囊间表皮和毛囊中Esrp消融相关的表型。我们将定义与发育和成人表皮和毛囊中的Esrps的条件性消融相关的表型。2)定义表皮和毛囊中Esrp调节的选择性剪接的综合方案。我们将使用高吞吐量 测序（RNA-Seq）和剪接敏感的微阵列，以确定在不同的细胞群体，填充皮肤的选择性剪接的全基因组程序。这些技术将与条件性缺失策略结合使用，以确定表皮和毛囊干细胞中的Esrp调节靶点。所提出的目标构成了第一个全面的分析选择性剪接在皮肤发育和功能，从而引入了一个新的范式领域。这些研究将揭示影响皮肤的过程的新见解 发展和功能。