Novel Modular Vascular Patterning Assay for HTS

HTS 的新型模块化血管模式分析

• 批准号: 7527007
• 负责人: ROYCE MOHAN
• 金额: $ 28.96万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7527007
• 关键词: 3-Dimensional; Address; Adoption; Age related macular degeneration; Arthritis; Biological; Biological Assay; Biology; Blood Vessels; Cardiovascular Diseases; Cell Line; Characteristics; Chemicals; Class; Complex; Computer software; Condition; Controlled Study; Development; Diabetes Mellitus; Differentiation and Growth; Disease; Diversity Library; Endothelial Cells; Funding; Goals; Grant; Growth; Growth Factor; Hand; Health; Heart Diseases; Homeostasis; Human; Hyperglycemia; Hypoxia; Image; Image Analysis; Impaired wound healing; Individual; Inflammatory; Lead; Life; Malignant Neoplasms; Manuals; Mind; Modeling; Multicellular Process; Multiple Organ Failure; Numbers; Optics; Pathway interactions; Pattern; Pharmaceutical Preparations; Preclinical Drug Evaluation; Principal Investigator; Process; Production; Quality Control; Research; Robotics; Running; Screening procedure; Signal Transduction; Simulate; Site; Stimulus; Stroke; System; Technology; Therapeutic; United States Food and Drug Administration; United States National Institutes of Health; Validation; Vascular Endothelial Cell; Wound Healing; angiogenesis; drug discovery; high throughput screening; human disease; improved; innovation; novel; programs; protein function; prototype; response; small molecule libraries; tool

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）：人类衬里血管的血管内皮细胞是对各种人类疾病的炎症和增生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。它响应炎症和血管生成刺激的不当激活可能会成为一种致病成分，从而加剧了癌症的生长和传播，并有助于衰弱的关节炎，与年龄相关的黄斑变性以及与糖尿病（如糖尿病）的潜在疾病相关的多个器官衰竭。另一方面，血管生成受损也同样具有致病性，并通过减慢伤口愈合并导致心脏病和中风来折磨受害者。总体而言，鉴于血管生成信号系统的复杂性，由于血管生长的失调而引起的这些主要人类健康负担需要通过在药物发现方面进行的更加协调的努力来解决。生物学测定法对血管生成疾病的过程进行建模可以帮助药物发现和疾病靶标识别过程。但是，目前，很少有测定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理/途径。考虑到这一点，我们建议继续开发高素质高吞吐量筛查（HC-HTS）血管模式测定法，我们最近通过先前资助的NIH Roadmap Initiative R21赠款将其确认为药物筛查工具。在此R01提案中，我们计划扩大三维内皮细胞发芽测定法（3D-ECSA）的规模和范围，以促进其对HTS范式的采用。我们的创新方法带来了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产生，2）高内容图像分析软件在HTS条件下与3D-ECSA一起使用，以及3）开发了针对免疫蛋白酶体作为一种化学启动的工具，以验证3D-ECSA的化学剂。这些目标的成功成就不仅将为我们提供大规模生物学的宝贵工具，而且还将有助于提出技术进步，以确定蛋白质功能的新型化学探针和药物铅的新型化学探针，以挽救生命的治疗疗法。