Protein Biomarkers for a New Human Polyomavirus in AIDS-related Malignancies

艾滋病相关恶性肿瘤中新型人类多瘤病毒的蛋白质生物标志物

• 批准号: 7691836
• 负责人: PATRICK S. MOORE
• 金额: $ 34.09万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-25 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7691836
• 关键词: AIDS related cancer; Acquired Immunodeficiency Syndrome; Antibodies; Antibody Formation; Antigens; Baculoviruses; Biological Assay; Biological Markers; Blood; Blood Tests; Carcinoma; Cell Line; Cells; Cercopithecus pygerythrus; Clinical; Common Neoplasm; Diagnostic tests; Early Diagnosis; Elderly; Ensure; Enzyme-Linked Immunosorbent Assay; Epitope Mapping; Etiology; Funding; Gene Expression Profile; Generations; Genome; Human; Human Virus; Immune; Immunocompromised Host; Immunological Diagnosis; Infection; Laboratories; Malignant Epithelial Cell; Malignant Neoplasms; Mammalian Cell; Measures; Merkel cell carcinoma; Methods; Monoclonal Antibodies; Mutate; Oncogenic Viruses; Patients; Pattern; Peptide Library; Peptide Mapping; Persons; Pharmaceutical Preparations; Polyomavirus; Polyomavirus Infections; Population; Preventive; Proteins; Reagent; Research Personnel; Role; Screening procedure; Serum; Skin Cancer; Southern Blotting; Specificity; Staining method; Stains; Techniques; Time; Tissue Microarray; Tissues; Transplantation; Vaccines; Viral; Viral Antigens; Viral Genome; Viral Proteins; Viral Tumor Antigens; Virus; Virus-like particle; Viviparous-1 protein; advanced disease; base; cross reactivity; digital; immunosuppressed; mortality; neoplastic cell; novel; patient population; sialosyl-T antigen; tumor

DESCRIPTION (provided by applicant): Merkel cell carcinoma (MCC) is a neuroectodermal, AIDS-related cancer suspected to have a viral etiology. MCC is 13 times more common than expected among AIDS patients and occurs in post-transplant and other immunosuppressed patient populations. While rare, it is the most aggressive skin cancer and only 50% of patients with advanced disease survive 9 months or longer. There are no early detection markers and it is frequently misdiagnosed with other small round cell tumors. We identified a novel polyomavirus in MCC using digital transcriptome subtraction (DTS). This agent, Merkel carcinoma polyomavirus (MCPyV), is most closely related to African green monkey lymphotropic polyomavirus (LPyV). Between 15-30% of persons have antibodies to LPyV, which has led investigators to speculate on the existence of a closely related but undiscovered human virus. MCPyV has a 5387 bp genome and expresses T antigen proteins in MCC tumors. Most MCC are positive for an integrated MCPyV that is readily detected by PCR and by Southern blotting. In contrast only 8-16% of control tissues harbor detectable viral genome. On Southern blotting, the majority of MCPyV positive tumors show a monoclonal somatic insertion pattern suggesting that the virus contributes to clonal cell expansion. We have developed reagents required to study this new virus including infected cell lines, specific antibodies and an overlapping peptide library for the VP1 protein. Preliminary studies suggest that MCPyV is a common but not ubiquitous infection that contributes to MCC when the virus mutates and integrates into the host genome. Identification of specific biomarkers for MCPyV infection is essential to determine MCPyV's role in human cancers. We propose to develop screening blood tests for MCPyV infection using established polyomavirus immunodiagnostic techniques as well as new immunodiagnostic methods. We will identify and characterize human antibody responses to MCPyV infection using cell-based IFA, MCPyV peptide mapping and virus-like particle ELISA with sera from well-defined populations. These assays will be vetted for cross-reactivity to known human polyomaviruses to ensure assay specificity. We also propose generating specific monoclonal antibodies to be used to detect viral antigens in tissue microarrays so that common tumors can be screened for infection. Expression of viral proteins can also be correlated with cellular protein biomarker expression to determine clinical course and responsiveness to treatment. This is the first proposal for funding to study a new human tumor virus found in our laboratory, MCPyV, likely to cause Merkel cell carcinoma. Merkel cell carcinoma is a cancer of the elderly, AIDS, transplant and other immunosuppressed persons. It has the highest overall mortality rate among skin cancers and there are no specific preventive measures or diagnostic tests for MCPyV infection. If funded, this proposal will develop accurate blood assays to measure antibody proteins marking MCPyV infection and to measure viral proteins in tissues. These are required to understand human infection with this virus and to develop effective vaccines and drugs against MCPyV.

描述（由申请人提供）：默克尔细胞癌（MCC）是一种神经外科，与艾滋病相关的癌症，怀疑患有病毒病因。 MCC在AIDS患者中的普遍性比预期的13倍，发生在移植后和其他免疫抑制的患者人群中。虽然很少见，但它是最具侵略性的皮肤癌，只有50％的晚期疾病患者生存了9个月或更长时间。没有早期的检测标记，并且经常被其他小圆细胞肿瘤误诊。我们使用数字转录组减法（DTS）鉴定了MCC中的新型多瘤病毒。该药物默克尔癌多瘤病毒（MCPYV）与非洲绿色猴子淋巴细胞病毒（LPYV）最密切相关。 15-30％的人对LPYV具有抗体，这导致研究人员推测存在紧密相关但未发现的人类病毒。 MCPYV具有5387 bp的基因组，并在MCC肿瘤中表达T抗原蛋白。大多数MCC对PCR和Southern印迹很容易检测到的集成MCPYV都是阳性的。相比之下，只有8-16％的对照组织具有可检测的病毒基因组。在Southern印迹上，大多数MCPYV阳性肿瘤显示出单克隆体细胞插入模式，这表明该病毒有助于克隆细胞的扩张。我们已经开发了研究这种新病毒所需的试剂，包括感染的细胞系，特定抗体和VP1蛋白的重叠肽库。初步研究表明，当病毒突变并整合到宿主基因组中时，MCPYV是一种常见但不是普遍的感染，会导致MCC。确定MCPYV感染的特定生物标志物对于确定MCPYV在人类癌症中的作用至关重要。我们建议使用已建立的多瘤病毒免疫诊断技术以及新的免疫诊断方法开发筛查MCPYV感染的筛查。我们将使用基于细胞的IFA，MCPYV肽映射和病毒样颗粒ELISA识别并表征对MCPYV感染的人类抗体反应，并具有来自定义明确的种群的血清。这些测定法将经过审查，以使其与已知的人类多余病毒的交叉反应性，以确保测定特异性。我们还建议使用特定的单克隆抗体来检测组织微阵列中的病毒抗原，以便可以筛选常见的肿瘤以感染。病毒蛋白的表达也可以与细胞蛋白生物标志物表达相关，以确定临床过程和对治疗的反应。这是研究在我们的实验室MCPYV中发现新的人类肿瘤病毒资金的第一个建议，可能会引起默克尔细胞癌。默克尔细胞癌是老年人，艾滋病，移植和其他免疫抑制人的癌症。它在皮肤癌中的总体死亡率最高，并且没有针对MCPYV感染的具体预防措施或诊断测试。如果资助，该建议将开发准确的血液测定，以测量标记MCPYV感染并测量组织中病毒蛋白的抗体蛋白。这些病毒需要了解人类感染，并开发针对MCPYV的有效疫苗和药物。