Molecular biology of human polyomaviruses

人类多瘤病毒的分子生物学

基本信息

批准号：
8937953
负责人：
Christopher Buck
金额：
$ 52.88万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8937953
关键词：
Address Adult Affect Age-Years Antibodies Antibody Formation Area BRAF gene Biological Markers Biology Capsid Proteins Carcinoma Cell Nucleus Cell-Matrix Junction Cells Collaborations Cultured Cells DNA Development Disease Exanthema Genome Genomic DNA Heparitin Sulfate Human Human Papillomavirus Human Virus Incidence Individual Infection Light Local Microbicides Malignant Neoplasms Merkel Cells Merkel cell carcinoma Methods Minor Modeling Molecular Biology Neurosecretory Systems Oncogenic Viruses Papillomavirus Pathway interactions Patients Plasmids Play Polyomavirus Polyomavirus Infections Polysaccharides Population Process Production Reporter Research Role Serum Skin Skin Cancer Structure Surface Technology The Sun Vaccines Viral Virion Virus Virus-like particle Work base deep sequencing experience genetic analysis high throughput screening human disease human subject immunogenic immunosuppressed inhibitor/antagonist interest microbial novel novel virus prevent receptor success tool tumor vaccine development vector

项目摘要

Merkel cell carcinoma (MCC) is a highly aggressive form of skin cancer that typically affects older and/or immunosuppressed individuals. Although MCC is relatively rare, its incidence in the U.S. has increased dramatically in the past decade. In 2008, DNA from a previously-unidentified polyomavirus was found to be present in a substantial majority of MCC tumors. The result suggests the possibility that the virus may be an etiologic cause of MCC and perhaps other forms of cancer. In FY08, work in the Tumor Virus Molecular Biology Section was focused on the biology of the human papillomavirus (HPV) virion. In FY09 we succeeded in applying a variety of tools originally developed for HPV research to the study of the newly-discovered Merkel cell polyomavirus (MCV). As a result, our Section is well situated to rapidly answer several important questions about MCV biology. MCV-based reporter vectors are a primary technology for this project. Production of the MCV vectors relies on intracellular expression of the MCV capsid proteins, which are capable of packaging reporter plasmids of interest in the milieu of the cell nucleus. The MCV reporter vectors are capable of delivering the packaged reporter plasmid to cultured cells, likely via mechanisms that resemble the infectious entry pathway of authentic MCV. In FY10, we completed studies showing that the infectivity of MCV vectors can be inhibited by MCV-specific antibodies. This allowed us to develop a high-throughput assay for quantitative analysis of antibody responses elicited by natural infection with MCV. We found that a great majority of adults display MCV-specific antibody responses, suggesting that infection is very common. Although infection appears to be common, we found that the magnitude of normal adults' antibody responses to MCV was typically much lower than the MCV-specific antibody responses observed in MCC patients. The result suggests a model in which MCC patients have typically experienced an unusually immunogenic form of MCV infection. Extremely strong antibody responses against MCV might thus serve as a biomarker for MCV-associated diseases. A major focus of our work during FY10 was elucidation of the infectious entry pathways that MCV uses to gain entry into host cells. This work showed that MCV employs heparan sulfate as the same primary attachment factor. The result draws a surprising parallel with HPVs, which also use heparan sulfate for initial attachment to cells. The result could point the way toward development of topical microbicides that might inhibit MCV infection. Further work during FY10 revealed that apparently healthy human skin surfaces are often chronically infected with MCV, suggesting that this is the major reservoir of the virus. The work serendipitously revealed the existence of two additional skin-dwelling viruses, human polyomavirus 6 (HPyV6) and HPyV7. A major portion of the lab's work in FY11 will be aimed at discovering whether HPyV6 or HPyV7 are associated with human disease. During fiscal year 2011, our group discovered the cell-attachment receptor that MCV uses for infectious entry into host cells and shed light on an additional co-receptor moiety that plays a post-attachment role in the entry process. In collaboration with Thilo Stehle's lab, we elucidated the structure of the MCV capsid protein VP1 with a model co-receptor glycan. During FY12 and FY13 we investigated the range of cells that MCV can infect and elucidated the biology of the minor capsid protein VP2. During FY14 the lab has shifted mostly toward the study of the infectious entry and molecular biology of BKV, JCV, and HPyV7. We also maintain an interest in virus discovery.

默克尔细胞癌（MCC）是一种高度侵略性的皮肤癌，通常会影响老年和/或免疫抑制个体。尽管MCC相对罕见，但在过去十年中，其在美国的发病率显着增加。在2008年，发现来自以前未识别的多瘤病毒的DNA存在于绝大多数MCC肿瘤中。结果表明，该病毒可能是MCC以及其他形式的癌症的病因。在08财年，肿瘤病毒分子生物学部分的工作集中在人乳头瘤病毒（HPV）病毒素的生物学上。在09财年，我们成功地应用了最初为HPV研究开发的各种工具，以研究新发现的默克尔细胞多组病毒（MCV）。结果，我们的部分位置良好，可以快速回答有关MCV生物学的几个重要问题。基于MCV的记者向量是该项目的主要技术。 MCV载体的产生依赖于MCV衣壳蛋白的细胞内表达，该蛋白能够在细胞核的环境中包装感兴趣的报道质粒。 MCV报道矢量能够将包装的报告基因质粒传递到培养的细胞中，这可能是通过类似于真实MCV的感染性进入途径的机制。在FY10中，我们完成了研究表明，MCV特异性抗体可以抑制MCV载体的感染性。这使我们能够开发高通量测定法，以定量分析MCV自然感染引起的抗体反应。我们发现，大多数成年人都表现出MCV特异性抗体反应，这表明感染非常普遍。尽管感染似乎很常见，但我们发现正常成年人对MCV的抗体反应的大小通常比MCV患者观察到的MCV特异性抗体反应低得多。结果表明，MCC患者通常经历了MCV感染的异常免疫原性形式。因此，针对MCV的极强抗体反应可能是MCV相关疾病的生物标志物。我们在第10财年期间工作的主要重点是MCV用来进入宿主细胞的传染性进入途径。这项工作表明，MCV采用硫酸乙酰肝素作为相同的主要依恋因子。结果与HPV平行相似，该HPV也将硫酸乙酰肝素用于初始附着在细胞上。结果可能指出了可能抑制MCV感染的局部菌心发展的道路。在第10财年期间的进一步工作表明，显然健康的人皮表面通常会长期感染MCV，这表明这是该病毒的主要储层。这项工作偶然地揭示了存在两个其他居住的皮肤病毒，即人类多瘤病毒6（HPYV6）和HPYV7。该实验室在第11财年的大部分工作将旨在发现HPYV6或HPYV7是否与人类疾病有关。在2011财政年度，我们的小组发现了MCV用于传染性进入宿主细胞的细胞跟踪受体，并阐明了一个额外的共受体部分，该部分在进入过程中起着连接后的作用。与Thilo Stehle的实验室合作，我们用模型共受体聚糖阐明了MCV Capsid蛋白VP1的结构。在2012财年和2013财年期间，我们研究了MCV可以感染和阐明次要衣壳蛋白VP2的生物学的细胞范围。在2014财年期间，实验室主要转向BKV，JCV和HPYV7的传染性进入和分子生物学的研究。我们还对病毒发现感兴趣。