权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Schlemm’s canal on a chip: A platform for screening a novel class of glaucoma medications

施莱姆氏管芯片：用于筛选新型青光眼药物的平台

基本信息

批准号：
10475283
负责人：
C ROSS ETHIER
金额：
$ 21.19万
依托单位：
GEORGIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10475283
关键词：
3-Dimensional Abbreviations Animal Model Anti-glaucoma Agent Apical Aqueous Humor Biological Biological Assay Blindness Blood-Aqueous Barrier Cell Separation Cell physiology Cells Cellular Assay Clinical Cultured Cells Data Detection Development Drainage procedure Elements Endothelial Cells Endothelium Ensure Eye FDA approved Functional disorder Future Generations Glaucoma Glossary Goals Harvest Height Human Hyperemia Impairment Knowledge Liquid substance Magnetism Measures Mechanics Membrane Microspheres Molecular Morphology Ocular Hypertension Outcome Outcome Measure Pathway interactions Patients Permeability Pharmaceutical Preparations Physiologic Intraocular Pressure Physiological Physiology Play Process Protocols documentation Published Comment Research Research Peer Review Research Personnel Resistance Rho-associated kinase Risk Risk Factors Screening procedure Stimulus Stretching Structure of sinus venosus of sclera Surface System Techniques Technology Therapeutic Tissues Trabecular meshwork structure Tracer Vacuole Vision aqueous automated image analysis base chromatin immunoprecipitation design drug development drug discovery experience fluid flow functional outcomes image processing improved in vivo insight kinase inhibitor light microscopy monolayer novel optic nerve disorder pre-clinical pressure rapid detection rapid testing response scale up screening side effect superparamagnetism therapeutic target tool

项目摘要

Glaucoma is a major cause of blindness. A major risk factor for glaucoma, and the only treatable risk factor, is elevated intraocular pressure (IOP). IOP is largely determined by the function of the tissues of the conventional outflow pathway, specifically the trabecular meshwork and the inner wall of Schlemm's canal (SC), and increased fluid flow resistance in these tissues causes elevated IOP. The inner wall of SC has been largely ignored as a target for IOP-lowering therapies, in part because our understanding of this tissue's function has been poor. However, recent research findings have greatly improved our understanding and motivate this proposal. The goal of this R21 is to optimize, characterize and validate a rapid and accurate assay system that interrogates the fluid conductivity of cultured SC cells, motivated by the knowledge that increasing SC cellular conductivity will lower IOP. The long-term vision is to use this assay system to serve as a pre-clinical screening tool for drug discovery and thereby accelerate discovery of a novel class of “SC-active” compounds for treating ocular hypertension in glaucoma patients. We expect such SC-active compounds to have fewer side effects compared to other IOP-lowering compounds, including avoidance of conjunctival hyperemia. Because the hydraulic conductivity of SC cells is normally controlled by the formation of membrane-delimited fluid pathways (“pores”), the assay system specifically focuses on detection of such pores in a high-content manner. More particularly, the focus is on intracellular pores (I pores), thought to form an under-exploited secondary aqueous outflow pathway. The key elements of this assay system include: (1) primary SC cell isolation from non-glaucomatous and glaucomatous human eyes; (2) substrate micropatterning to control cultured cell spread/height and enhance assay rigor; (3) delivery of focal mechanical stretch to the apical surface of cultured SC cells using superparamagnetic microspheres and external magnets; (4) a novel fluorescent assay that, together with automated light microscopy and image processing tools, allows the rapid detection of pores that form in SC cells in a manner that minimizes introduction of inadvertent bias; and (5) a high-content format in which cells are cultured in 96 well plates, allowing relatively rapid testing of the effects of agents on SC cells. There is a high degree of rigor in the proposal, which is underpinned by a significant body of peer-reviewed research and strong preliminary data. The applicants have extensive experience in culturing and characterizing human primary SC cells, and the proposal includes a variety of positive and negative controls to validate the assay system. Further, the multi-well assay format has the advantage of naturally allowing multiple technical and biological replicates on each 96 well plate, enabling characterization of repeatability and good statistical rigor. It is expected, as suggested by the proposal's strong preliminary data, that the proposed assay system will be a powerful pre-clinical tool which can in future be used to screen for SC-active compounds and which will also enhance our understanding of SC endothelial cell physiology and pathophysiology.

青光眼是导致失明的主要原因。青光眼的一个主要危险因素，也是唯一可治疗的危险因素，是眼内压（IOP）升高。IOP在很大程度上由常规眼组织的功能决定。流出通道，特别是小梁网和Schlemm管（SC）的内壁，并增加这些组织中的流体流动阻力导致IOP升高。SC的内壁在很大程度上被忽视，降眼压治疗的靶点，部分原因是我们对这一组织的功能了解不多。然而，最近的研究结果大大提高了我们的理解和激励这一建议。该R21的目标是优化、表征和验证快速准确的检测系统，培养的SC细胞的流体电导率，由增加SC细胞电导率的知识激发，会降低眼压长期的愿景是使用该分析系统作为药物的临床前筛选工具发现并由此加速发现用于治疗眼部疾病的新型“SC活性”化合物，青光眼患者的高血压我们预计这种SC活性化合物的副作用更少，其他降低IOP的化合物，包括避免结膜充血。由于SC细胞的水力传导率通常由膜界定的形成控制，流体通道（“孔”）中的孔，该测定系统特别关注于检测高含量流体通道中的这种孔。方式更具体地，重点是细胞内孔（I孔），其被认为形成未被充分利用的细胞内孔。二次房水流出途径。该检测系统的关键要素包括：（1）原代SC细胞分离来自非青光眼和青光眼的人眼;（2）基底微图案化以对照培养的细胞（3）将焦点机械拉伸递送到培养的细胞的顶端表面; 使用超顺磁性微球和外部磁体的SC细胞;（4）一种新的荧光测定法，与自动光学显微镜和图像处理工具一起，可以快速检测以最小化无意偏差的引入的方式在SC单元中形成;以及（5）高内容格式，所述细胞在96孔板中培养，允许相对快速地测试试剂对SC细胞的作用。该提案高度严谨，并得到了大量同行审查的支持。研究和强大的初步数据。申请人在培养和表征方面具有丰富的经验人原代SC细胞，该提案包括各种阳性和阴性对照，以验证分析系统此外，多孔测定形式具有自然地允许多种技术和方法的优点。在每个96孔板上进行生物学重复，使得能够表征重复性和良好的统计学严谨性。正如该提案的强有力的初步数据所表明的那样，预计所提出的测定系统将是一个强大的临床前工具，可在未来用于筛选SC活性化合物，增强我们对SC内皮细胞生理学和病理生理学的理解。