The Role of MicroRNAs in Normal and Diseased Corneal Epithelial Homeostasis

MicroRNA 在正常和患病角膜上皮稳态中的作用

• 批准号: 9533576
• 负责人: Mehrnoosh Saghizadeh Ghiam
• 金额: $ 43.75万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9533576
• 关键词: 3' Untranslated Regions; Adhesions; Affect; Apoptosis; Automobile Driving; Autopsy; Biological Assay; Blindness; Cell Death; Cell Maintenance; Cell Proliferation; Cell physiology; Cells; Cleaved cell; Cornea; Corneal Diseases; Corneal Injury; Corneal Opacity; Data; Debridement; Defect; Development; Diabetes Mellitus; Disease; Edema; Epidermal Growth Factor; Epithelial; Epithelial Cells; Epithelium; Etiology; Eye; Functional disorder; Funding; Future; Gene Expression; Gene Targeting; Generations; Goals; Growth; Health; Homeostasis; Human; In Vitro; Individual; Infection; Inflammation; Keratopathy; Lead; Limbus Corneae; Luciferases; Maintenance; Messenger RNA; MicroRNAs; Molecular; Natural regeneration; Organ; Organ Culture Techniques; Pathologic; Patients; Pattern; Peripheral; Physiological; Process; Property; Propidium Diiodide; Proteins; Proteomics; Recurrence; Regulation; Regulator Genes; Research; Retinal; Role; Signal Transduction; Signaling Molecule; Small Interfering RNA; Source; Stains; Stem cells; Stratum Basale; Testing; Tissues; Translations; Ulcer; Vascularization; Vision; Visual; Western Blotting; Wound Healing; annexin A5; cell motility; clinical translation; corneal epithelium; deep sequencing; diabetic; diabetic patient; differential expression; gene therapy; healing; improved; improved functioning; in vitro Assay; limbal; miRNA expression profiling; migration; neovascularization; next generation sequencing; overexpression; public health relevance; self-renewal; targeted treatment; therapeutic miRNA; transcriptome sequencing; translational study

 DESCRIPTION (provided by applicant): A wide variety of physiological and pathological conditions such as infections, inflammations and corneal diseases such as limbal stem cell deficiency (LSCD) and diabetes can affect limbal epithelial stem cells (LESC) functions resulting in serious visual problems. LESC are located within special niches in the limbal basal layer at the corneo-scleral junction of cornea. The health of LESC is a key factor in renewal of healthy and regeneration of wounded corneal epithelium. Altered wound healing process, reminiscent of diabetic cornea, can result in corneal vascularization and loss of transparency, which may lead to corneal blindness. Currently, there is a need to better understand the mechanisms responsible for these abnormalities and develop a specific and efficacious treatment. The mechanisms of LESC proliferation, migration, adhesion, and differentiation in normal cornea homeostasis and wound healing process could be key to understanding corneal diseases. Therefore, understanding of miRNA regulatory mechanisms is critically important in the etiology and treatment of various corneal diseases such as LSCD and diabetic keratopathy, which are the major causes for corneal blindness. We will employ a variety of molecular, functional tests and ex-vivo human organ-cultured corneas to understand the role of microRNAs in both the differentiated epithelial and stem cells of normal and diabetic corneas and their roles in driving peripheral activation of limbal stem cells in normal and diseased cornea. We will focus on determining their targets, and attempt gene therapy with either overexpression or silencing of specific miRNAs to normalize corneal wound healing and LESC marker patterns in diabetic cornea. We will target diabetic corneal epithelial cells by single microRNA or their combinations to improve function of corneal epithelium. This research could lead to the generation of a new cell source for restoring vision in patients with altered LESC. We will use human corneal organ culture in our studies. Normal and diabetic human autopsy eyes will be purchased from the National Disease Research Interchange (NDRI), a national supplier of donor human research tissue. All tissues are obtained in our lab without donor identity.

 描述（由申请人提供）：多种生理和病理状况，如感染、炎症和角膜疾病，如角膜缘干细胞缺乏症（LSCD）和糖尿病，可影响角膜缘上皮干细胞（LESC）功能，导致严重的视力问题。LESC位于角膜的角巩膜交界处的角膜缘基底层中的特殊龛内。LESC的健康是角膜上皮健康更新和损伤再生的关键因素。改变的伤口愈合过程，使人联想到糖尿病角膜，可导致角膜血管化和透明度丧失，这可能导致角膜失明。目前，有必要更好地了解负责这些异常的机制，并制定一个具体的和有效的治疗。LESC在正常角膜稳态和创伤愈合过程中的增殖、迁移、粘附和分化机制可能是理解角膜疾病的关键。因此，了解miRNA调控机制对于各种角膜疾病如LSCD和糖尿病性角膜病的病因学和治疗至关重要，这些疾病是角膜失明的主要原因。我们将采用各种分子，功能测试和离体人类器官培养的角膜，以了解microRNA在正常和糖尿病角膜的分化上皮细胞和干细胞中的作用，以及它们在驱动正常和患病角膜的角膜缘干细胞的外周活化中的作用。我们将专注于确定它们的靶点，并尝试通过过表达或沉默特定miRNA的基因治疗来使糖尿病角膜中的角膜伤口愈合和LESC标志物模式正常化。我们将通过单个microRNA或其组合靶向糖尿病角膜上皮细胞，以改善角膜上皮功能。这项研究可能导致产生一种新的细胞来源，用于恢复LESC改变患者的视力。我们将在我们的研究中使用人类角膜器官培养。正常和糖尿病人类尸检眼睛将从国家疾病研究交流中心（NDRI）购买，该中心是一家捐赠人类研究组织的国家供应商。所有的组织都是在我们的实验室获得的，没有供体身份。