A High Throughput Protein Complementation Assay for Inhibitors of NEMO-K13 Intera

NEMO-K13 Intera 抑制剂的高通量蛋白质互补测定

• 批准号: 8100494
• 负责人: Preet M. Chaudhary
• 金额: $ 26.09万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8100494
• 关键词: Acquired Immunodeficiency Syndrome; Apoptosis; Biological; Biological Assay; Biological Process; CASP8 and FADD-like apoptosis regulating protein; CASP8 gene; Cell Line; Cell Proliferation; Cells; Clinical; Complex; Development; Disease; Firefly Luciferases; Genome; Goals; Growth Factor; HIV; Human; Human Herpesvirus 8; Hydrocarbons; Immune response; Immunocompromised Host; Immunosuppression; Immunotherapy; Infection; Inflammatory Response; Jurkat Cells; Kaposi Sarcoma; Large-Cell Immunoblastic Lymphoma; Lead; Link; Luciferases; Lymphoma; Lymphoproliferative Disorders; Malignant Neoplasms; Molecular; Morphologic artifacts; Multicentric Angiofollicular Lymphoid Hyperplasia; NF-kappa B; Names; Non-Hodgkin' s Lymphoma; Open Reading Frames; Pathogenesis; Pathway interactions; Patients; Peptides; Phosphotransferases; Physiological; Play; Protein Fragment; Proteins; Regimen; Reporter; Reproducibility; Screening procedure; Solid; Subgroup; Testing; Toxic effect; Viral; Viral Proteins; Withdrawal; base; caspase-8; cytokine; domain mapping; drug development; effusion; genetic regulatory protein; high throughput screening; inhibitor/antagonist; metaplastic cell transformation; novel; outcome forecast; public health relevance; reconstitution; small molecule; therapeutic target; young adult

DESCRIPTION (provided by applicant): Human Herpesvirus 8 (HHV8, also known as KSHV) is one of the commonest causes of malignancies among young adults in parts of the world and has been associated with Kaposi's sarcoma, primary effusion lymphoma (PEL) and multicentric Castleman's disease (MCD). The prognosis of patients with HHV8-associated lymphoproliferative disorders is extremely poor due to their immunocompromised status and there is an urgent need for less toxic therapies for the treatment of these malignancies. We have discovered that K13, a small protein encoded by HHV8, directly interacts with the NEMO/IKK3 subunit of the IkB kinase (IKK) complex to activate the NF-kB pathway and utilizes this pathway to promote cellular survival, proliferation, transformation and cytokine secretion. The above studies have established NF-kB pathway as an important therapeutic target for the treatment of HHV8-associated malignancies. However, since NF-kB pathway plays a key role in normal immune and inflammatory response, global inhibitors of this pathway are likely to lead to severe immunosuppression, thus limiting their potential clinical utility in HHV8-infected patients. To circumvent this problem, we propose to develop a high throughput screening (HTS) assay for isolating small molecule inhibitors of K13-NEMO interaction. It is hoped that such inhibitors will specifically block K13-induced NF-kB without interfering with the physiological activation of this pathway during normal immune and inflammatory response. Furthermore, specific inhibitors of K13-NEMO interaction will serve as useful pharmacological probes to understand the various biological activities of K13. PUBLIC HEALTH RELEVANCE: Infection with the Human Herpesvirus 8 (HHV8) has been linked to a number of human cancers. In this project, we propose to develop a high throughput screening assay for compounds that can block the interaction of K13, a small protein encoded by HHV8, with the cellular regulatory protein NEMO. It is hoped that such compounds will not only lead to a better understanding of the biological functions of K13 but also serve as lead compounds for the development of targeted therapies for HHV8-associated malignancies.

描述（由申请人提供）：人类疱疹病毒 8 型（HHV8，也称为 KSHV）是世界部分地区年轻人中最常见的恶性肿瘤之一，与卡波西肉瘤、原发性渗出性淋巴瘤 (PEL) 和多中心性淋巴瘤有关卡斯尔曼病（MCD）。由于 HHV8 相关淋巴增殖性疾病患者的免疫功能低下，其预后极差，迫切需要毒性较小的疗法来治疗这些恶性肿瘤。我们发现K13是HHV8编码的小蛋白，直接与IkB激酶（IKK）复合物的NEMO/IKK3亚基相互作用，激活NF-kB通路，并利用该通路促进细胞存活、增殖、转化和细胞因子分泌。上述研究已确立NF-kB通路作为治疗HHV8相关恶性肿瘤的重要治疗靶点。然而，由于NF-kB通路在正常免疫和炎症反应中发挥着关键作用，该通路的整体抑制剂可能会导致严重的免疫抑制，从而限制了它们在HHV8感染患者中的潜在临床应用。为了解决这个问题，我们建议开发一种高通量筛选（HTS）测定法来分离 K13-NEMO 相互作用的小分子抑制剂。希望此类抑制剂能够特异性阻断 K13 诱导的 NF-kB，而不干扰正常免疫和炎症反应过程中该通路的生理激活。此外，K13-NEMO 相互作用的特异性抑制剂将作为有用的药理学探针来了解 K13 的各种生物活性。 公共卫生相关性：人类疱疹病毒 8 型 (HHV8) 感染与多种人类癌症有关。在这个项目中，我们建议开发一种高通量筛选方法，寻找能够阻断 K13（一种由 HHV8 编码的小蛋白）与细胞调节蛋白 NEMO 相互作用的化合物。希望此类化合物不仅能够更好地了解 K13 的生物学功能，而且能够作为开发 HHV8 相关恶性肿瘤靶向治疗的先导化合物。