Novel Approaches to Detect Virus-Cancer Associations

说明(申请人提供)：病毒已确定，致病因素 在15%-20%的人类癌症中，被广泛怀疑参与的更多。 这个项目的长期目标是开发方法来同时 筛选肿瘤和其他组织样本中的基因组或部分基因组 许多病毒，使用这些方法来检测新的病毒-癌症 并确定它们的统计和临床意义。 特别是，这份高度协作的提案描述了一个新的、 独特的柔性、无掩膜光刻技术将用于 生产和提炼高密度寡核苷酸芯片，能够 灵敏地检测所有已知人类肿瘤病毒的所有基因或转录本。 作为对比，在玻璃片上发现的聚合酶链式反应扩增的病毒探针阵列 也将接受测试。其他实验将系统地优化 特异、灵敏、稳健的提取和扩增方法 临床组织中低拷贝数病毒RNA和DNA序列的检测 样本。所产生的工具和方法将应用于相关的 临床样本测试和解决两口井的新出现的联系- 表征人类肿瘤病毒、人乳头瘤病毒(HPV)和爱泼斯坦- Barr病毒(EBV)，分别与口腔癌和乳腺癌有关，并 将微阵列结果与其他最先进的病毒检测结果进行比较 方法应用于相同的组织样本。这些研究将提供证据- 基因芯片用于病毒检测和鉴定的原理测试 对癌症的贡献，并解决病毒中的重大当前问题 特定肿瘤的病因学。同时进行多个微阵列筛查 病毒或病毒基因型别增强了病因学和诊断学的力量 对珍贵组织样本的研究，提供了发现 意外的病毒-癌症链接，并允许病毒和癌症的简档表达 选定的细胞基因，以提供肿瘤的分子签名并 评估病毒对该肿瘤的潜在贡献。这些方法 开发的也将适用于其他涉及病毒的探测器 慢性或急性疾病。

DESCRIPTION (provided by applicant): Viruses are established, causal factors in 15%-20% of human cancers and are widely suspected to be involved in more. The long-range goals of this project are to develop methods to simultaneously screen tumor and other tissue samples for the genomes or partial genomes of many viruses, to use these methods to test for novel virus-cancer associations, and to determine their statistical and clinical significance. In particular, this highly collaborative proposal describes how a new, uniquely flexible, maskless photolithography technology will be used to produce and refine high density oligonucleotide microarrays able to sensitively detect all genes or transcripts of all known human tumor viruses. For comparison, arrays of PCR-amplified viral probes spotted on glass slides will also be tested. Additional experiments will systematically optimize extraction and amplification methods for specific, sensitive, and robust detection of viral RNA and DNA sequences at low copy number in clinical tissue samples. The resulting tools and methodology will be applied to relevant clinical samples to test and resolve the emerging connections of two well- characterized human tumor viruses, human papilloamavirus (HPV) and Epstein- Barr virus (EBV), with oral cancers and breast cancers, respectively, and to compare the microarray results with other state-of-the-art virus detection methods applied to the same tissue samples. These studies will provide proof- of-principle tests of microarray use in detecting and characterizing viral contributions to cancer and address significant, current issues in the viral etiology of specific tumors. Simultaneous microarray screening for many viruses o viral genotypes amplifies the power of etiologic and diagnostic studies on precious tissue samples, offers the potential to discover unsuspected virus-cancer links, and allows profiling expression of viral and selected cell genes to provide the molecular signatures of a tumor and to evaluate the potential contribution of the virus to that tumor. The methods developed also will be applicable to probe for virus involvement in other chronic or acute disorders.