A Novel Treatment for Keratoconus and Keratectasias using Nitro Technology

使用硝基技术治疗圆锥角膜和角膜扩张症的新方法

基本信息

批准号：
7754376
负责人：
DAVID C PAIK
金额：
$ 19.8万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-01-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7754376
关键词：
Animals Apoptosis Biological Biological Assay Biomechanics Blindness Cell Count Cells Clinical Trials Collagen Communities Confocal Microscopy Cornea Corneal Diseases Corneal Endothelium Corneal Topography Disease Dose Endothelial Cells Epidemiology Epithelial Epithelial Cells European Eye Eye Banks Eyedrops Female German population Glutaral Goals Grant Heart Valves Histology Human In Vitro Incubated Intellectual Property International Keratoconus Laboratories Laser In Situ Keratomileusis Legal patent Life Mechanics Methods Microscope Monitor Necrosis New Zealand Nitrites Nitro Compounds Operative Surgical Procedures Oryctolagus cuniculus Patients Penetration Peripheral Phase I Clinical Trials Physiologic Intraocular Pressure Procedures Property Rights Prosthesis Publishing Reaction Research Research Personnel Riboflavin Risk Safety Self Administration Simulate Skin Transplantation Skin graft Solutions Techniques Technology Temperature Testing Thick Tissues Toxic effect Translations Transplanted tissue Ultraviolet Rays Universities Work base catalyst cell growth cost crosslink cytotoxicity design implantation in vivo instrument member new technology novel public health relevance research study response success tonometry ultraviolet irradiation vision science young adult

项目摘要

DESCRIPTION (provided by applicant): A major breakthrough in the treatment of keratoconus and post-LASIK keratectasias has been realized. Recent work by the German group of Wollensak, Spoerl, and Seiler has shown that cross-linking corneal collagen through application of riboflavin and ultraviolet light (UVR) can limit progressive vision loss in keratoconus patients. Despite these successes, the UVR therapy poses attendant risks, particularly related to ultraviolet irradiation. As such, FDA approval for clinical trials in the US has only recently been granted. The thrust of this application is to develop an alternative method of corneal collagen cross-linking using novel technology. Our recent studies indicate that collagen cross-linking through reactions with nitrite and related agents can induce corneal changes commensurate with UVR therapy. In addition, published toxicity/mutagenicity studies suggest a good safety profile for these agents. Thus, nitro technology could be used as an "eye drop alternative" to UVR treatment. Potential advantages over current UVR therapy include the omission of ultraviolet light exposure, the ability to treat thin corneas, less patient discomfort, ease of self- administration, a dose-response effect, and the ability to treat the peripheral cornea. This project is designed to lay groundwork for rapid translation of this technology into a treatment. Specifically, we will: 1. Determine an effective way to cross-link the cornea using nitro technology under conditions simulating the human cornea (i.e. pH 7.4 and 34oC). Fresh rabbit and human eye bank corneas will be incubated under a variety of conditions that include 1. nitrite with catalysts and 2. alternative nitrite related agents. The assay of thermal shrinkage temperature will be used to determine cross-linking efficacy. Stiffness changes induced will be studied using a biaxial material tester. Specific aim 1 is designed to identify one or more "candidate eye drops" that will then be used for in vivo rabbit experiments. 2. Test the hypothesis that corneal cross-linking through nitro technology is tolerated by corneal cells. Primary cultures of rabbit endothelial cells, keratocytes, and epithelial cells will be grown in vitro. The cells will then be exposed to nitro compounds found to be efficacious for cross-linking. Cytotoxicity will be evaluated using standard apoptosis and necrosis assays. The results from this aim will establish the in vitro toxicity level of nitro compounds to corneal cells. Specific aim 3. Test the hypothesis that corneal cross-linking through nitro technology has efficacy and is safe for the living eye. 33 young adult female New Zealand white rabbits will undergo daily application (for 1-6wks) of the topical nitro solution to the right eye with the fellow eye serving as a control. The animals will undergo serial examinations using 3 non-invasive instruments, a corneal topographer, in vivo confocal microscope, and applanation tonometer. Post-mortem, biomechanical strength testing, histology, and SEM will be performed. The results of this aim will establish the in vivo efficacy and safety of this technology and will dictate the feasibility of a human phase I trial. PUBLIC HEALTH RELEVANCE This research aims to develop a simple, new treatment for diseases of corneal destabilization which include keratoconus and post-surgical keratectasias. The latter conditions are devastating, long-term (5-10 years) complications following LASIK procedures. They were previously unpredicted and are currently of unknown epidemiologic proportion.

描述（由申请人提供）：圆锥角膜和 LASIK 术后角膜扩张的治疗已实现重大突破。德国 Wollensak、Spoerl 和 Seiler 小组最近的研究表明，通过应用核黄素和紫外线 (UVR) 交联角膜胶原蛋白可以限制圆锥角膜患者进行性视力丧失。尽管取得了这些成功，但紫外线疗法也带来了随之而来的风险，特别是与紫外线照射有关的风险。因此，FDA 最近才批准在美国进行临床试验。该应用的主旨是开发一种利用新技术进行角膜胶原交联的替代方法。我们最近的研究表明，通过与亚硝酸盐和相关试剂反应发生的胶原交联可以诱导与紫外线治疗相称的角膜变化。此外，已发表的毒性/致突变性研究表明这些药物具有良好的安全性。因此，硝基技术可以用作紫外线治疗的“滴眼液替代品”。与当前 UVR 疗法相比，潜在优势包括省略紫外线照射、能够治疗薄角膜、减少患者不适、易于自我给药、剂量反应效应以及能够治疗周边角膜。该项目旨在为将该技术快速转化为治疗方法奠定基础。具体来说，我们将： 1. 在模拟人类角膜的条件下（即pH 7.4和34oC），确定使用硝基技术交联角膜的有效方法。新鲜的兔子和人眼库角膜将在多种条件下孵育，包括 1. 亚硝酸盐与催化剂和 2. 替代亚硝酸盐相关试剂。热收缩温度的测定将用于确定交联功效。将使用双轴材料测试仪研究引起的刚度变化。具体目标 1 旨在确定一种或多种“候选滴眼剂”，然后将其用于兔子体内实验。 2. 检验角膜细胞能够耐受通过硝基技术进行的角膜交联的假设。兔内皮细胞、角膜细胞和上皮细胞的原代培养物将在体外生长。然后将细胞暴露于对交联有效的硝基化合物。将使用标准细胞凋亡和坏死测定来评估细胞毒性。该目标的结果将确定硝基化合物对角膜细胞的体外毒性水平。具体目标 3. 检验通过硝基技术进行角膜交联是否有效且对活体眼睛安全的假设。 33 只年轻成年雌性新西兰白兔每天（1-6 周）将局部硝基溶液涂抹在右眼上，并用另一只眼作为对照。这些动物将使用 3 种非侵入性仪器、角膜地形图仪、活体共焦显微镜和压平眼压计进行一系列检查。将进行尸检、生物力学强度测试、组织学和扫描电镜。这一目标的结果将确定该技术的体内功效和安全性，并将决定人体 I 期试验的可行性。公众健康相关性本研究旨在开发一种简单的新疗法来治疗角膜不稳定疾病，包括圆锥角膜和术后角膜扩张。后一种情况是 LASIK 手术后破坏性的长期（5-10 年）并发症。它们以前是无法预测的，目前流行病学比例未知。