The Role of MicroRNAs in Corneal Epithelial Homeostasis

MicroRNA 在角膜上皮稳态中的作用

基本信息

批准号：
8759983
负责人：
ROBERT M LAVKER
金额：
$ 43.98万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
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：
2010-06-01 至 2019-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8759983
关键词：
3-Dimensional Affect Age related macular degeneration Anterior Apoptosis Area Attenuated Autophagocytosis Behavior Biochemical Biological Biological Assay Biology Blood Vessels Bromodeoxyuridine Cell Communication Cell Cycle Cell Proliferation Cell Survival Cell physiology Cell-Cell Adhesion Cells Child Clinical Connexin 43 Cornea Cultured Cells Data Development Diabetic Retinopathy Disease Endothelial Cells Environment Epithelial Epithelial Cells Epithelium Extravasation Eye Family Funding Gene Targeting Goals Homeostasis Human Immigration Immunoblotting Injury Investments Kinetics Knowledge Maintenance Measures Methods MicroRNAs Molecular Morphogenesis Natural regeneration Pathogenesis Patients Pattern Pericytes Physiological Platelet-Derived Growth Factor Play Process Production Property Proteins RNA Interference RNA Splicing Regenerative Medicine Regulation Reporting Role Site Source Stem cell transplant Stem cells Structure Surface System Techniques Testing Tissues Transmission Electron Microscopy Transplantation Treatment Protocols Tube Vacuole Vascular Endothelial Growth Factor Receptor-1 Vascular Endothelial Growth Factors Vision Visual Acuity angiogenesis base cell motility corneal epithelial stem cells corneal epithelium gene therapy glycogen metabolism improved in vivo inhibitor/antagonist innovation keratinocyte light transmission limbal loss of function matrigel migration neovascularization novel novel strategies ocular surface posters prevent progenitor public health relevance retinal angiogenesis self-renewal stem cell biology tissue regeneration

项目摘要

DESCRIPTION (provided by applicant): The anterior surface of the eye functions as a barrier to the external environment and protects the delicate underlying structures from injury, in part, through the elaboration of the limbal and corneal epithelia. As self-renewing tissues, these epithelia are governed by stem cells, which play a crucial role in tissue homeostasis, regeneration, transplantation, gene therapy and in the pathogenesis of several anterior ocular surface diseases. Equally important for proper vision is the need for corneal transparency, which is achieved through avascularity. It is well-accepted that the limbal epithelium is the site f the corneal epithelial stem cells; however, major questions remain unresolved concerning how the limbal epithelium is regulated. Likewise, our understanding of factors that control angiogenesis is incomplete. microRNAS (miRNAs) are a major class of regulatory molecules that are part of the RNAi silencing machinery. While some studies have been directed towards deciphering the roles of miRNAs in the corneal epithelium, little is known about the miRNA signature in the stem cell-enriched limbal epithelium. We have recently discovered that miRs-103/107 are limbal-preferred. Furthermore, we have evidence that miRs-103/107 function to insure proper limbal epithelial cell-cell contact, autophagy and impact on cell cycle quiescence. Until recently, it was believed that miR-184, the most abundant corneal epithelial miRNA, functioned to attenuate miR-205, which insured proper cell migration and cell survival. We now have evidence that miR-184 may directly prevent corneal epithelial angiogenesis. Proper vision requires both a stabile limbal epithelial and corneal clarity; therefore we propose to focus on: (1 The roles of miRs-103/107 in assuring the integrity of the limbal epithelium and (2) how miR-184 functions to maintain corneal avascularity. To accomplish these goals, we will capitalize on our ability to elucidate miRNA target proteins and modulate miRNA and target protein levels in submerged cultures of human limbal and corneal epithelial keratinocytes and human microvascular endothelial cells. We will manipulate these cultured cells to form either 3-D organotypic rafts, or endothelial tubes, which mimic the in vivo tissues. We will assess the functional consequences of such miRNA and protein modulations with a combination of biochemical, molecular biological, cell biological and physiological approaches. By focusing on the biology of these miRNAs and their target proteins, our proposal represents a novel approach to understand: (i) how the limbal epithelium is regulated; and (ii) what contributes to the maintenance of corneal avascularity. Such knowledge has relevance to stem cell biology and clinically to ex vivo corneal epithelial transplantation. This proposal also has clinical implicatins beyond just corneal avascularity and may impact on pathological retinal angiogenesis. Ultimately our studies will provide rationales for the development of innovative treatment regimens focused on the use of either: (i) inhibitors of specific miRNAs or their targets; or (ii) delivery of miRNAs in patients with diseases that affect the ocular anterior segmental epithelia.

描述（由申请人提供）：眼睛的前表面充当外部环境的障碍，并通过详细阐述缘层和角膜上皮来保护精致的基础结构免受损伤。作为自我更新组织，这些上皮受到干细胞的控制，干细胞在组织稳态，再生，移植，基因治疗以及几种前眼表面疾病的发病机理中起着至关重要的作用。对于正确的视力，同样重要的是需要角膜透明度，这是通过血管性实现的。众所周知的是，缘缘上皮是角膜上皮干细胞的部位。但是，关于如何调节缘缘上皮的方式仍未解决。同样，我们对控制血管生成因素的理解是不完整的。 MicroRNA（miRNA）是RNAi沉默机制的一部分的主要调节分子。虽然一些研究是针对破译miRNA在角膜上皮中的作用的，但对富含干细胞富含干细胞的边缘上皮的miRNA特征知之甚少。我们最近发现，miRS-103/107是偏爱的。此外，我们有证据表明，miRS-103/107的功能可确保适当的缘缘上皮细胞 - 细胞接触，自噬和对细胞周期静止的影响。直到最近，人们认为Mir-184是最丰富的角膜上皮miRNA，可衰减miR-205，这为适当的细胞迁移和细胞存活提供了保证。现在，我们有证据表明miR-184可以直接预防角膜上皮血管生成。适当的视力需要稳定的边缘上皮和角膜清晰度。因此，我们建议专注于：（1 miRS-103/107在确保边缘上皮完整性方面的作用，以及（2）miR-184如何发挥保持角膜血管性的作用。为实现这些目标，我们将利用我们的能力来阐明我们阐明miRNA目标蛋白质，并在人类和人类的角膜含量和人类的含量蛋白质中，并在人含量的蛋白质中含有人类的含量和靶标蛋白质含量，并具有较高的人类含量。微血管内皮细胞。提案代表了一种新的理解方法：（i）层缘上皮如何受到调节；（ii）有助于维持角膜血管性。该建议还具有临床上的含义，而不仅仅是角膜血管性，并且可能影响病理视网膜血管生成。最终，我们的研究将为开发专注于使用的创新治疗方案提供理由：（i）特定miRNA或其靶标的抑制剂；（ii）在影响眼前节段性上皮的疾病患者中递送miRNA。