Regulators of Epidermal Growth and Differentiation

表皮生长和分化的调节剂

基本信息

批准号：
9197267
负责人：
GEORGE L SEN
金额：
$ 34.1万
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：
2015
资助国家：
美国
起止时间：
2015-02-17 至 2019-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9197267
关键词：
3-Dimensional 5&apos -exoribonuclease Adaptor Signaling Protein Address Affinity Chromatography Basal Cell Basal cell carcinoma Basement membrane Binding Cell Compartmentation Cell physiology Cells Dermal Differentiation and Growth Disease Enzymes Epidermis Equilibrium Foundations Gene Expression Genetic Transcription Homeostasis Human Immune Immunoprecipitation Lead Mass Spectrum Analysis Mediating Messenger RNA Methods Modeling Molecular Mus Natural regeneration Population Process Proteins Psoriasis Publishing RNA RNA Binding RNA Degradation Recruitment Activity Regulator Genes Research Design Role Skin Squamous cell carcinoma Stem cells Stratum Basale Tissues Transcript Water Work Yeasts chronic wound clinically relevant design exosome experimental study genetic element insight knock-down loss of function mRNA Transcript Degradation next generation sequencing novel premature prevent progenitor public health relevance self-renewal skin disorder therapy development transcription factor

项目摘要

DESCRIPTION (provided by applicant): Epidermal homeostasis requires a proper balance between proliferation and differentiation which when altered can lead to hyperproliferative disorders such as squamous cell carcinomas. Regulators of this process may include RNA degradation enzymes such as the exosome which is composed of 11 subunits. We have recently shown that exosome subunits such as EXOSC9, EXOSC7, and EXOSC10 are essential for maintaining epidermal self-renewal by promoting the mRNA degradation of differentiation specific transcription factors in progenitor cells. It is currently unknown the function of the other 8 subunits of the exosome as well as their associated adaptor proteins. Objective/hypothesis: This proposal seeks to understand the gene regulatory mechanisms involved in maintaining normal epidermal homeostasis. We hypothesize that exosome subunits exist in epidermal cells in subcomplexes with distinct functions. Subcomplexes such as EXOSC9, EXOSC7, and EXOSC10 are necessary to maintain progenitor function while other subcomplexes are necessary for differentiation. We also hypothesize that adaptor proteins recruit distinct exosome subcomplexes to target RNAs to either maintain self-renewal or to promote epidermal differentiation. Specific Aims: (1) To determine the role of the exosome subunits in epidermal homeostasis and (2) to determine the role of exosome associated adaptor proteins in epidermal homeostasis. Study Design: To study epidermal homeostasis in a more clinically relevant setting, we established new methods to introduce specific combinations of genetic elements into 3- dimensionally intact human skin, containing human epidermal cells in the context of human dermal stroma and basement membrane, regenerated on immune deficient mice. By using this model, we can perform loss of function experiments on exosome subunits and adaptor proteins 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 exosome subunits as well as adaptor proteins.

描述（由申请人提供）：表皮稳态需要增殖和分化之间的适当平衡，当这种平衡发生改变时，可能导致过度增殖性疾病，例如鳞状细胞癌。这一过程的调节者可能包括 RNA 降解酶，例如由 11 个亚基组成的外泌体。我们最近发现，EXOSC9、EXOSC7 和 EXOSC10 等外泌体亚基通过促进祖细胞中分化特异性转录因子的 mRNA 降解，对于维持表皮自我更新至关重要。目前尚不清楚外泌体的其他 8 个亚基及其相关接头蛋白的功能。目的/假设：本提案旨在了解维持正常表皮稳态所涉及的基因调控机制。我们假设外泌体亚基存在于表皮细胞中具有不同功能的亚复合体中。 EXOSC9、EXOSC7 和 EXOSC10 等子复合物是维持祖细胞功能所必需的，而其他子复合物是分化所必需的。我们还假设衔接蛋白招募不同的外泌体亚复合物来靶向RNA，以维持自我更新或促进表皮分化。具体目标：(1) 确定外泌体亚基在表皮稳态中的作用；(2) 确定外泌体相关衔接蛋白在表皮稳态中的作用。研究设计：为了在更临床相关的环境中研究表皮稳态，我们建立了新方法，将遗传元件的特定组合引入三维完整的人类皮肤中，其中包含在人类真皮基质和基底膜背景下的人类表皮细胞，在免疫缺陷小鼠。通过使用该模型，我们可以对再生人类皮肤中的外泌体亚基和衔接蛋白进行功能丧失实验，以表征它们在表皮生长和分化中的作用。我们还将使用 RNA 免疫沉淀，然后进行新一代测序来确定与外泌体亚基以及接头蛋白相关的 RNA。