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Novel Modular Vascular Patterning Assay for HTS

HTS 的新型模块化血管模式分析

基本信息

批准号：
8243126
负责人：
ROYCE MOHAN
金额：
$ 22.32万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-01 至 2012-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8243126
关键词：
Novel Modular Vascular Patterning Assay

项目摘要

DESCRIPTION (provided by applicant): The vascular endothelial cells lining blood vessels in humans are one of the principle sites that become involved in inflammatory and proliferative responses to a diverse array of human diseases. Microvascular homeostasis is thus a vital component of human health; its inappropriate activation in response to inflammatory and angiogenic stimuli can become a pathogenic component fueling the growth and spread of cancers, and contributing to debilitating arthritis, age-related macular degeneration and multiple organ failure associated with underlying diseases such as diabetes. On the other hand, impaired angiogenesis is also equally pathogenic, and afflicts its victims by slowing down wound healing and contributing to heart diseases and stroke. Collectively, given the complexity of the angiogenesis signaling system, these major burdens of human health that arise from dysregulation of blood vessel growth need to be addressed by a more concerted effort in drug discovery. Biological assays that model the processes of angiogenic diseases can assist the process of drug discovery and disease target identification. However, currently few assays represent the complexity of the diseased microvasculature as they typically focus on one pathogenic mechanism/pathway. With this in mind, we propose to continue the development of a high content high throughput screening (HC-HTS) vascular patterning assay, which we have recently validated as drug screening tool through a previously funded NIH Roadmap Initiative R21 grant. In this R01 proposal, we plan to extend the scale and scope of the three dimensional endothelial cell sprouting assay (3D-ECSA) to promote its adoption for the HTS paradigm. Our innovative approaches bring in 1) modern automated robotic systems that allow us to improve efficiency and standardize production of spheroids, 2) high content image analysis software to use with 3D-ECSA under HTS conditions, and 3) development of a pilot scale chemical library focused on the immunoproteosome as a chemical enabling tool towards validation of the 3D-ECSA. The successful accomplishments of these goals will not only afford us a valuable tool for large scale biology, but will help bring forward a technology advancement to identify new classes of chemical probes of protein function and drug leads for life saving therapeutics.

描述（由申请人提供）：人类血管内壁的血管内皮细胞是参与对多种人类疾病的炎症和增殖反应的主要部位之一。因此，微血管稳态是人类健康的重要组成部分。它对炎症和血管生成刺激的不当激活可能成为促进癌症生长和扩散的致病成分，并导致衰弱性关节炎、年龄相关性黄斑变性以及与糖尿病等基础疾病相关的多器官衰竭。另一方面，受损的血管生成也同样具有致病性，并通过减慢伤口愈合并导致心脏病和中风来折磨受害者。总的来说，考虑到血管生成信号系统的复杂性，这些因血管生长失调而产生的人类健康的主要负担需要通过药物发现方面更加协调一致的努力来解决。模拟血管生成疾病过程的生物测定可以协助药物发现和疾病靶标识别的过程。然而，目前很少有测定能够代表患病微脉管系统的复杂性，因为它们通常专注于一种致病机制/途径。考虑到这一点，我们建议继续开发高内涵高通量筛选 (HC-HTS) 血管模式分析，我们最近通过先前资助的 NIH 路线图计划 R21 拨款将其验证为药物筛选工具。在此 R01 提案中，我们计划扩大三维内皮细胞出芽测定 (3D-ECSA) 的规模和范围，以促进其在 HTS 范式中的采用。我们的创新方法引入了 1) 现代自动化机器人系统，使我们能够提高效率并标准化球体的生产，2) 在高温超导条件下与 3D-ECSA 一起使用的高内涵图像分析软件，以及 3) 开发一个中试规模的化学库，重点关注免疫蛋白酶体作为验证 3D-ECSA 的化学支持工具。这些目标的成功实现不仅将为我们提供大规模生物学的宝贵工具，还将有助于推动技术进步，以识别新型蛋白质功能化学探针和用于挽救生命的治疗药物。