权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Role of 14-3-3o in development and repair of corneal epithelium

14-3-3o在角膜上皮发育和修复中的作用

基本信息

批准号：
8517123
负责人：
Qiutang Li
金额：
$ 34.2万
依托单位：
UNIVERSITY OF LOUISVILLE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-01 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8517123
关键词：
Affect Age Aging Alleles Anterior Binding Calcium Cell Culture Techniques Cell Cycle Arrest Cell Differentiation process Cell Proliferation Cell surface Cells Complex Cornea Corneal Diseases Corneal Injury Corneal Opacity Defect Development Differentiation Antigens EGF gene Embryo Embryonic Development Environment Epidermal Growth Factor Receptor Epithelial Epithelial Cells Epithelium Eye Eye Part Failure Family Family member Gene Activation Gene Dosage Gene Targeting Genes Genetic Crosses Genetic Models Heterozygote Homeostasis Individual Inflammation Injury Invaded Knock-out Link Lipids MEKs Mediating Molecular Molecular Genetics Mus Mutant Strains Mice Mutation Pathway interactions Pattern Phenotype Phosphoproteins Phosphoserine Motif Phosphotransferases Play Prevention strategy Process Proliferating Property Protein Binding Protein Family Proteins Receptor Signaling Role Series Signal Pathway Signal Transduction Site Stem cells Testing Tissues Transgenic Mice Tumor Suppressor Proteins Undifferentiated Vision Vitamin A Wound Healing base blind corneal epithelium design eye dryness in vivo limbal meibomian gland mutant neovascularization notch protein null mutation prevent progenitor programs receptor receptor expression repaired research study response to injury self-renewal

项目摘要

Exposure of the corneal epithelium to the environment leaves this important barrier susceptible to injury. Accordingly, the corneal epithelium has a remarkable self-renewal capacity. This self renewal is mediated by a reservoir of limbal stem cells which migrate to sites of injury where they proliferate and differentiate to replace the damaged epithelium. This proposal is directed toward understanding the role of 14-3-3¿ in corneal epithelial differentiation. 14-3-3 is a family of proteins that bind phosphoproteins and regulate their subcellular localization. 14-3-3¿ is the epithelial-specific family member, and using a mouse genetic model we demonstrate its crucial role in corneal epithelial differentiation Mutation of both copies of the gene leads to defective corneal epithelial development in the embryo. Heterozygous mutation allows for normal embryonic development; however, the corneal epithelial differentiation in response to injury remains impaired. Thus, 14-3-3¿ gene dosage distinguishes corneal epithelial development embryologically from corneal epithelial differentiation following injury. This defect in differentiation leads to outgrowth of undifferentiated cells which eventually form an opaque corneal plaque with associated inflammation and neovascularization. This plaque forms spontaneously as the heterozygous mice age, and we provide evidence that this is linked to a dry eye-like effect resulting from a defective meibomian gland which fails to secrete lipid. Notch1 is essential for corneal epithelial differentiation via its activation of genes important for both differentiation (including genes in the vitamin A pathway) as well as cell cycle arrest, and we have found that 14-3-3¿ mutant cells fail to activate Notch1. Expression of activated Notch1 restores differentiation in mutant cells, and accordingly the defective differentiation phenotype and the resulting corneal plaque formation closely resemble that seem with tissue-specific knockout of the Notch1 gene. We link 14-3-3¿ to cell surface expression of the EGF receptor (EGFR) and thus to the Erk signaling pathway which is activated by binding of EGF to this receptor. This is important because EGF signaling represses Notch1, and this signaling must be extinguished for Notch1 expression and subsequent corneal epithelial differentiation. We propose studies to further characterize the linkage between 14-3-3¿, EGFR, and Notch1 (and its target genes) in corneal epithelia differentiation. These studies involve both cell culture and genetic cross using 14-3-3¿ mutant mice.

角膜上皮暴露于环境使这一重要屏障变得脆弱以致受伤。因此，角膜上皮具有显着的自我更新能力。这个自己更新是由角膜缘干细胞库介导的，这些干细胞迁移到受伤部位它们增殖并分化以取代受损的上皮。这个提案是针对以了解 14-3-3¿ 在角膜上皮分化中的作用。 14-3-3 是一个家庭结合磷蛋白并调节其亚细胞定位的蛋白质。 14-3-3¿ 是上皮特异性家族成员，并使用小鼠遗传模型，我们证明了其至关重要在角膜上皮分化中的作用该基因的两个拷贝的突变导致缺陷胚胎中角膜上皮的发育。杂合突变允许正常胚胎发育；然而，角膜上皮因损伤而分化仍然受损。因此，14-3-3¿基因剂量区分角膜上皮发育胚胎学上来自损伤后角膜上皮分化。这个缺陷在分化导致未分化细胞的生长，最终形成不透明的细胞角膜斑块与相关的炎症和新生血管形成。此牌匾形成随着杂合子小鼠年龄的增长，这一现象会自发发生，我们提供的证据表明，这与由于睑板腺有缺陷，无法分泌脂质，导致类似干眼症的效果。 Notch1 通过激活重要的基因而对角膜上皮分化至关重要分化（包括维生素 A 途径中的基因）以及细胞周期停滞，以及我们发现14-3-3¿突变细胞无法激活Notch1。激活的Notch1的表达恢复突变细胞的分化，从而恢复有缺陷的分化表型由此产生的角膜斑块形成与组织特异性的情况非常相似 Notch1 基因的敲除。我们将 14-3-3¿ 与 EGF 受体的细胞表面表达联系起来 (EGFR) 以及 Erk 信号通路，该通路通过 EGF 与其结合而被激活受体。这很重要，因为 EGF 信号传导会抑制 Notch1，并且该信号传导必须是 Notch1 表达和随后的角膜上皮分化消失。我们提出研究进一步表征 14-3-3¿、EGFR 和 Notch1（以及其靶基因）在角膜上皮分化中的作用。这些研究涉及细胞培养和使用 14-3-3¿ 突变小鼠进行遗传杂交。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Study of corneal epithelial progenitor origin and the Yap1 requirement using keratin 12 lineage tracing transgenic mice.

DOI：
10.1038/srep35202
发表时间：
2016-10-13
期刊：
Scientific reports
影响因子：
4.6
作者：
Kasetti RB;Gaddipati S;Tian S;Xue L;Kao WW;Lu Q;Li Q
通讯作者：
Li Q

IKK1 control of epidermal differentiation is modulated by notch signaling.

IKK1 对表皮分化的控制由 Notch 信号传导调节。

DOI：
10.1016/j.ajpath.2010.12.021
发表时间：
2011
期刊：
The American journal of pathology
影响因子：
0
作者：
Xin,Ying;Lu,Qingxian;Li,Qiutang
通讯作者：
Li,Qiutang

IKK2 inhibition using TPCA-1-loaded PLGA microparticles attenuates laser-induced choroidal neovascularization and macrophage recruitment.

DOI：
10.1371/journal.pone.0121185
发表时间：
2015
期刊：
PloS one
影响因子：
3.7
作者：
Gaddipati S;Lu Q;Kasetti RB;Miller MC;Lu Q;Trent JO;Kaplan HJ;Li Q
通讯作者：
Li Q

14-3-3σ regulates keratinocyte proliferation and differentiation by modulating Yap1 cellular localization.

14-3-3σ通过调节YAP1细胞定位来调节角质形成细胞的增殖和分化。

DOI：
10.1038/jid.2015.42
发表时间：
2015-06
期刊：
JOURNAL OF INVESTIGATIVE DERMATOLOGY
影响因子：
6.5
作者：
Sambandam, Sumitha A. T.;Kasetti, Ramesh B.;Xue, Lei;Dean, Douglas C.;Lu, Qingxian;Li, Qiutang
通讯作者：
Li, Qiutang

Notch signaling promotes the corneal epithelium wound healing