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Role of keratin intermediate filaments in skin epithelial differentiation.

角蛋白中间丝在皮肤上皮分化中的作用。

基本信息

批准号：
10462643
负责人：
Pierre Coulombe
金额：
$ 42.25万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-09 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10462643
关键词：
Accounting Adaptor Signaling Protein Affect Alleles Architecture Automobile Driving Binding Binding Sites Biochemistry Bullous Congenital Ichthyosiform Erythroderma Cell physiology Cells Cessation of life Code Coupled Crystallization Cysteine Cytokeratin filaments Cytoplasm Cytoskeletal Filaments Cytoskeletal Proteins Defect Disease Disulfides Ectoderm Epidermis Epidermolysis Bullosa Simplex Epithelial Epithelial Cells Equilibrium Exhibits Family Fostering Functional disorder Gene Expression Gene Family Genes Growth Hair follicle structure Homeostasis Human Impairment Individual Intermediate Filament Gene Intermediate Filament Proteins Intermediate Filaments Keratin Knock-in Knockout Mice Link Maintenance Maps Mechanics Mediating Medicine Metabolism Modification Mutation Phenotype Phosphotransferases Plant Roots Play Pliability Population Process Property Protein Isoforms Proteins Regulation Research Role Signal Transduction Skin Societies Source Specific qualifier value Specificity Stress Structure Stuttering Surface Sweat Glands Testing Therapeutic Tissues Tooth structure Transcriptional Regulation Transgenic Mice Translations appendage base disease-causing mutation disulfide bond high throughput screening in vivo insight keratin 5 keratinocyte keratinocyte differentiation live cell imaging mechanical signal migration mouse model notch protein pachyonychia congenita progenitor protein function response scaffold skin disorder transcriptomics

项目摘要

ABSTRACT Genetically-determined mutations that affect the sequence of keratin proteins account for a large number of skin epithelial disorders, many of which are rooted in defective cellular mechanics. Keratins are the most abundant proteins in surface epithelia like epidermis, where they primarily occur as intricate cytoplasmic networks of 10 nm wide intermediate filaments (IFs). Keratins are encoded by an evolutionarily conserved family of 54 genes subdivided into two major types (I and II). Pairwise regulation of type I and II keratin genes in epithelial cells reflects a strict heteropolymerization requirement during IF assembly. Specific pairs of keratin genes are regulated in an epithelial tissue-type and differentiation-specific fashion. One of the many roles so far elucidated for keratin IFs is to act as resilient yet pliable scaffolds that endow epithelial cells and tissues with the ability to sustain various stresses. A newly emerging, exciting role for keratin proteins is to regulate epithelial differentiation through the modulation of Hippo and Notch signaling. We recently discovered that keratin-dependent disulfide bonding plays a crucial role towards the intracellular organization and steady-state dynamics of keratin filaments in progenitor keratinocytes of epidermis, with an associated impact on the balance between proliferation and differentiation. This role entails a delicate interplay between disulfide bonding mediated by a specific residue known as the “stutter cysteine” in keratin 14 (K14), the adaptor protein 14-3-3sigma, and YAP1, a terminal effector of Hippo signaling. Since the stutter cysteine in K14 is conserved in several additional type I keratins expressed in skin epithelia we propose, as an overarching hypothesis, that keratins act as general regulators of Hippo signaling with an associated impact on differentiation and homeostasis in skin epithelia. In Aim 1, we will test the hypothesis that the stutter cysteine in K10 is responsible for proper regulation of YAP1 and Hippo signaling in the differentiating layers of epidermis, with direct relevance for the keratin disorder epidermolytic hyperkeratosis. In Aim 2 we will map the binding interface between 14-3-3sigma and each of K14 and K10, define the basis for the regulation the keratin/14-3-3 interactions, and identify the source of the signal that drives K14- and K10-dependent disulfide bonding in keratinocytes. We will also conduct a high-throughput screen comparing gene expression, at the single cell level, in control vs. Krt14 C373A transgenic mice, which exhibit an epidermal differentiation defect coupled to misregulation of YAP1. In Aim 3, finally, we will test the hypothesis that the stutter cysteine in K17 regulates YAP1 and Hippo signaling and the balance between proliferation and differentiation in hair follicles, sweat glands, tooth, and possibly in other appendages, with relevance for the disorder pachyonychia congenita. The proposed body of work lies on a robust premise and is poised to significantly advance our understanding of the significance of keratin protein function and regulation in vivo and the pathophysiology of keratin mutation-based skin diseases.

摘要