Assay development to discover therapeutics against human cytomegalovirus

开发检测方法以发现针对人类巨细胞病毒的治疗方法

基本信息

批准号：
8667395
负责人：
Domenico Tortorella
金额：
$ 33.71万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-01 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8667395
关键词：
Acquired Immunodeficiency Syndrome Affect Attenuated Autoimmune Process Back Biological Assay Biology Cell Cycle Cell Nucleus Cell physiology Cells Chemicals Chimera organism Cidofovir Clinical Congenital Abnormality Cytomegalovirus Cytomegalovirus Infections DNA Development Disease Drug Interactions Drug resistance Drug toxicity Elderly Foscarnet Ganciclovir Gene Expression Generations Genetic Transcription Goals HIV Health Immediate-Early Genes Immunocompromised Host Individual Infection Lead Licensing Life Cycle Stages Luciferases Measures Medical Molecular Morbidity - disease rate Newborn Infant Organ Transplantation Patients Pharmaceutical Preparations Phase Poison Polymerase Population Prevention Production Proliferating Proteins Protocols documentation Reagent Reporting Retinitis Severities Signal Transduction Specificity Staging System Therapeutic Transplant Recipients United States United States National Academy of Sciences Valganciclovir Variant Viral Viral Load result Viral Proteins Virion Virus Virus Assembly Virus Diseases assay development base congenital cytomegalovirus cost effective therapy env Gene Products fomivirsen high throughput screening inhibitor/antagonist latent persistent infection monocyte mortality mutant nervous system disorder novel older patient particle prevent protein expression public health relevance resistant strain response

项目摘要

DESCRIPTION (provided by applicant): Human cytomegalovirus (HCMV) causes latent and persistent infections in 60-90% of the population. Virus proliferation significantly increases the morbidity and/or mortality in immunocompromised individuals such as newborns, organ transplant recipients, AIDS patients, as well as the elderly. HCMV is a significant health challenge requiring the development of a multi-facet approach for the generation of effective and safe treatments to limit HCMV proliferation. The current treatments against HCMV are anti-viral drugs such as ganciclovir, valganciclovir, foscarnet, and cidofovir that target the viral polymerase. A major draw-back to these drugs is their toxicity, drug-drug interactions, and the development of drug resistance strains of the virus upon prolonged treatment. The current application proposes to develop and utilize high-throughput screening assays to identify lead compounds that limit HCMV proliferation and dissemination in response to PA-10-213, 'Development of Assays for High-Throughput screening for use in Probe and Pre-therapeutic Discovery.' Our central hypothesis is that targeting multiple stages of HCMV proliferation would significantly reduce HCMV disease through the effective inhibition of the HCMV life cycle. Thus, we propose to discover compounds that block different steps of HCMV proliferation through the development of high-throughput screening assays. These assays will utilize HCMV variants that express chimeric viral proteins fused to yellow fluorescent proteins or luciferase. In addition, secondary assays to determine the viral and cellular specificity for patient-derived clinical strais of hit compounds and the compound's general mode of action are routine protocols in the lab. The development of robust high-throughput screens for the identification of anti-HCMV drugs is an essential first step to generating effective therapies against HCMV and the associated diseases. Potential lead compounds may target various steps of the HCMV life cycle such as viral entry, viral protein expression, viral egress, and virus particle release from the infected cll. These compounds in combination with current therapy as a multi-drug cocktail would likely be very effective at treating patients with a high HCMV viral load and limit HCMV associated diseases.

描述（由申请人提供）：人类巨细胞病毒（HCMV）在60-90％的人群中引起潜在感染和持续感染。病毒增殖可显着增加免疫功能低下的个体（例如新生儿，器官移植受者，艾滋病患者以及老年人）的发病率和/或死亡率。 HCMV是一项重大的健康挑战，需要开发多面方法，以生成有效和安全的治疗，以限制HCMV的增殖。目前针对HCMV的治疗方法是针对病毒聚合酶的抗病毒药物，例如Ganciclovir，valganciclovir，falganciclovir，foscarnet和cidofovir。这些药物的主要吸引力是它们的毒性，药物相互作用以及长期治疗后病毒耐药性菌株的发展。当前的应用程序建议开发和利用高通量筛选测定法，以识别限制HCMV增殖和响应PA-10-213的铅化合物，“开发用于用于探针和治疗前发现的高通量筛选的测定法。我们的核心假设是，针对HCMV增殖的多个阶段将通过有效抑制HCMV生命周期大大减少HCMV疾病。因此，我们建议发现通过开发高通量筛选测定法阻断HCMV增殖不同步骤的化合物。这些测定将利用与黄色荧光蛋白或荧光素酶一起表达嵌合病毒蛋白的HCMV变体。此外，确定HIT化合物的患者衍生临床strais的病毒和细胞特异性以及该化合物的一般作用方式是实验室中的常规方案。用于鉴定抗HCMV药物的强大高通量筛查的开发是生成针对HCMV和相关疾病有效疗法的重要第一步。潜在的铅化合物可能针对HCMV生命周期的各个步骤，例如病毒蛋白表达，病毒表达，病毒出口和病毒颗粒从感染的CLL释放。这些化合物与当前疗法作为多药鸡尾酒结合使用，可能非常有效地治疗高HCMV病毒载量并限制HCMV相关疾病的患者。