Secreted Regulators of Epidermal Homeostasis

表皮稳态的分泌调节剂

• 批准号: 9313177
• 负责人: Bryan Sun
• 金额: $ 18.13万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-08 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9313177
• 关键词: Biological; Biological Assay; CRISPR/Cas technology; Calcium; Calcium Binding; Calmodulin; Catalogs; Cells; Cervical; Critical Pathways; Data; Development; EF-Hand Domain; Epidermis; Epithelial; Esophageal Tissue; Evaluation; Extracellular Matrix Proteins; Extracellular Protein; Genes; Genetic; Genetic Techniques; Genomics; Glycine; Health; Homeostasis; Human; Impairment; Individual; Knowledge; Mass Spectrum Analysis; Mechanics; Mediating; Molecular; Mutagenesis; Mutation; Mutation Analysis; Peptide Hydrolases; Play; Process; Production; Protease Inhibitor; Protein Isoforms; Proteins; RNA Interference; Regulation; Regulatory Pathway; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Skin; Stratification; Stratified Epithelium; Structural Protein; Tandem Repeat Sequences; Tissues; Translating; Work; antimicrobial; antimicrobial peptide; epidermis cell; experimental study; extracellular; gene function; intercellular communication; keratinocyte; keratinocyte differentiation; novel; progenitor; protein function; public health relevance; sensor; skin disorder; skin organogenesis; transcriptome

 DESCRIPTION (provided by applicant): The epidermis consists of a distinct cell layers that work in concert to execute a wide variety of essential functions. Keratinocytes comprise the majority of the cells of the epidermis, and form distinct strata from the basal proliferative layerto the outermost cornified layer. Keratinocytes of all strata secrete a variety of protein factors tha have critical roles in the proper function of the epidermis, including structural proteins, antimicrobial peptides, proteases and protease inhibitors, and cell-cell signaling molecules. Despite the recognized role of extracellular proteins in epidermal homeostasis, a comprehensive evaluation of these secreted factors-the keratinocyte "secretome"- has not been defined. The proposal describes the first comprehensive catalog of secreted epidermal proteins as identified by mass spectrometry. Analysis of the secretome of progenitor and differentiated keratinocytes reveals a number of novel factors whose function in the epidermis is unknown. Preliminary experiments using genetic depletion of candidate factors revealed a novel essential roles for the proteins calmodulin-like 5 (CALML5) and suprabasin (SBSN) in epidermal differentiation. The first aim of this proposal describes the use of genomic and genetic techniques, including analysis of whole transcriptome data from CALML5-depleted tissue and gene set enrichment analysis, to predict and analyze the role of CALML5 among known epidermal regulators. Next, the functional regions of CALML5 are determined using mutagenesis in a genetic rescue assay. [The second aim focuses on the function of SBSN, a protein expressed in differentiated epidermal tissue that is produced in three distinct isoforms. These isoforms are differentially enriched in the intracellular and extracellular compartments, raising the possibility that they may have discrete functions inside and outside the cell. Genetic experiments are proposed to define which isoform(s) are required for differentiation, and to determine the subcellular compartments that these isoforms are biologically active.] Together, the aims of this project are to discover and characterize new essential secreted regulators of skin homeostasis, which can provide a deeper understanding to the mechanisms of skin development and reveal new potential targets to treat skin disease.

 描述（由申请人提供）：表皮由不同的细胞层组成，这些细胞层协同工作以执行各种基本功能。角质形成细胞构成表皮的大部分细胞，并形成从基底增殖层到最外层皮质层的不同层。所有层的角质形成细胞分泌多种蛋白质因子，其在表皮的正常功能中具有关键作用，包括结构蛋白、抗微生物肽、蛋白酶和蛋白酶抑制剂以及细胞-细胞信号传导分子。尽管公认的作用，细胞外蛋白在表皮内稳态，这些分泌因子的角质形成细胞的“分泌组”的综合评价尚未确定。 该提案描述了第一个通过质谱鉴定的分泌性表皮蛋白的综合目录。对祖细胞和分化的角质形成细胞的分泌组的分析揭示了许多新的因子，其在表皮中的功能是未知的。利用候选因子的遗传耗竭的初步实验揭示了蛋白质钙调素样5（CALML 5）和超盆（SBSN）在表皮分化中的新的重要作用。 该提案的第一个目的描述了使用基因组和遗传技术，包括分析来自CALML 5缺失组织的全转录组数据和基因集富集分析，以预测和分析CALML 5在已知表皮调节因子中的作用。接下来，在遗传拯救测定中使用突变来确定CALML 5的功能区域。 [The第二个目标集中于SBSN的功能，SBSN是一种在分化的表皮组织中表达的蛋白质，以三种不同的同种型产生。这些异构体在细胞内和细胞外区室中差异富集，提高了它们可能在细胞内外具有离散功能的可能性。建议进行遗传实验来定义分化需要哪些亚型，并确定这些亚型具有生物活性的亚细胞区室。] 总之，该项目的目的是发现和表征皮肤稳态的新的必需分泌调节剂，这可以提供对皮肤发育机制的更深入理解，并揭示治疗皮肤疾病的新的潜在靶点。