The role of extracellular vesicles in keratoconus pathogenesis

项目摘要/摘要 圆锥角膜是一种慢性终身角膜营养不良，全世界约有1：400人受到影响， 包括雄性和雌性。治疗晚期圆锥角膜患者一直是一个挑战。 角膜移植仍然是治疗的金标准，然而，术后有很大的风险 并发症是存在的。目前，圆锥角膜的诊断是通过专门的设备和测试，如裂隙灯 和Pentacam®。早期诊断，不规则散光会使事情复杂化，对于 圆锥角膜的治疗。到目前为止，还没有生物标记物测试因此推动了对 协助诊断、管理和/或治疗圆锥角膜的新方法。 细胞外小泡(EVS)是由细胞释放的脂双层限定的颗粒，它可以提供一种 以非侵入性方式实时拍摄整个细胞/组织/器官的快照。电动汽车可以调节生理 包括蛋白质、miRNAs、DNA片段、非编码RNA和类脂。 因此，电动汽车有望发现未来的生物标记物。在角膜的背景下 疾病/营养不良、泪液和泪液EV(TEV)是进行生物标志物研究的最佳候选对象。 令人惊讶的是，很少有研究报道(只有干眼症和干燥综合征)研究电视电视。 我们的初步数据为圆锥角膜患者中tEV的存在提供了强有力的证据 表型与健康的同龄人不同。这项拟议的临床研究的目的是要揭示 圆锥角膜病理生物学中的TEV，并发现与年龄、性别和疾病相关的生物标记物 严肃性。在我们的初步研究中，我们招募了一小批健康的圆锥角膜患者， 并组建了一支强大的临床医生/采集点小组，帮助我们在 建议的期限。我们的数据提示圆锥角膜患者的tEV有显著的表型差异。 患者与健康对照组比较。因此，我们提出了一种集成的多方面的“指纹”方法 这将包括TeV物理特征、蛋白质组学和miRNA图谱。我们相信，我们的战略可以 为圆锥角膜的诊断、治疗甚至治疗指明了未来的方向。 与公共健康相关-KC是导致全球范围内视力障碍的主要临床问题。长的- 我们研究的术语含义对KC患者很重要，因为它可以建立新的治疗方式。 归根结底，有效和安全地管理/治疗所有KC患者的能力至关重要。建议的工作是新颖的， 翻译的，临床相关的，与NEI的目标和研究重点一致的，以了解KC并开发 新的治疗选择，以减轻全球范围内的疾病负担。

PROJECT SUMMARY/ABSTRACT Keratoconus is a chronic, lifelong corneal dystrophy and affects approximately 1:400 people worldwide, including both males and females. Treating patients with advanced keratoconus has always been a challenge. Corneal transplantation remains the gold standard for treatment, however, significant risk of postoperative complications exist. Presently, keratoconus is diagnosed with specialized equipment and tests such as slit-lamp and Pentacam®. Early diagnosis, where irregular astigmatism can complicate things, is critical for the management of keratoconus. To-date, there are no biomarker tests which has consequently driven demand for novel approaches to assist in the diagnosis, management and/or treatment of keratoconus. Extracellular vesicles (EVs) are lipid bilayer-delimited particles released by cells, which may provide a real-time snapshot of the entire cell/tissue/organ in a non-invasive way. EVs can regulate physiological processes and contain components including proteins, miRNAs, DNA fragments, non-coding RNAs and lipids. Therefore, EVs hold promise for the discovery of future biomarkers. In the context of corneal diseases/dystrophies, tear fluid and tear EVs (tEVs) are the best candidate for biomarker investigations. Surprisingly, very few studies have been reported (only Dry eye and Sjogren’s syndrome) investigating tEVs. Our preliminary data provides strong evidence for the existence of tEVs in keratoconus patients that are phenotypically different from their healthy counterparts. The proposed clinical studies aim to unravel the role of tEVs in keratoconus pathobiology and discover relevant biomarkers associated with age, sex, and disease severity. During our preliminary studies, we have recruited a small cohort of healthy and keratoconus patients, and have put together a strong group of clinicians/collection sites that will assist us with sample collection during the proposed period. Our data suggests significant phenotypic differences in the tEVs derived from keratoconus patients, compared to healthy controls. We, therefore, propose an integrated multi-faceted “fingerprint” approach that will include tEV physical characterization, proteomics, and miRNA profiling. We believe that our strategy can shed light on future directions in this field for keratoconus diagnosis, management, and even treatment. Relevance to Public Health – KC is a major clinical problem resulting in visual impairment worldwide. The long- term implications of our study are important for KC patients since it could establish novel treatment modalities. Ultimately, the ability to effectively, and safely, manage/treat all KC patients, is vital. The proposed work is novel, translational, clinically relevant, and in line with NEI’s goals and research priorities to understand KC and develop novel treatment options to reduce the burden of the disorder worldwide.