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（由适用提供）：多种物理和病理状况，例如感染，炎症和角膜疾病，例如边缘干细胞缺乏症（LSCD）和糖尿病可能会影响膜缘上皮干细胞（LESC）功能，从而导致严重的视觉问题。 LESC位于角膜角膜骨连接处的边缘基本层中的特殊壁ni内部。 LESC的健康是伤口愈合过程的关键改变，让人联想到糖尿病角膜，可能导致角膜血管和透明度的丧失，这可能导致角膜失明。当前，有必要更好地了解负责这些异常的机制，并开发特定而有效的治疗方法。正常角膜稳态和获得愈合过程中LESC增殖，迁移，粘合剂和分化的机制可能是理解角膜疾病的关键。因此，对miRNA调节机制的理解在病因和治疗各种角膜疾病（如LSCD和糖尿病性角膜病）中至关重要，这是角膜失明的主要原因。我们将采用各种分子，功能性测试和前体的有组织的角膜，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，及其在正常和脱落角膜中驱动边缘干细胞的外周血激活中的作用。我们将专注于确定其靶标，并尝试通过过度表达或对特定miRNA的沉默来尝试基因治疗，以使角膜角膜中的角膜伤口愈合和LESC标记模式正常化。我们将通过单个microRNA或它们的组合靶向糖尿病角膜上皮细胞，以改善角膜上皮的功能。这项研究可能导致生成新的细胞来源，以恢复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文化。正常和糖尿病的人类尸检将从国家疾病研究互换（NDRI）购买，这是国家供体人类研究组织的供应商。所有时间都是在我们的实验室中获得的，没有捐助者身份。