Novel Approaches to Detect Virus-Cancer Associations

描述（由申请方提供）：确定病毒，因果因素 在15%-20%的人类癌症中，人们普遍怀疑它与更多的癌症有关。 该项目的长期目标是开发方法， 筛选肿瘤和其他组织样品的基因组或部分基因组， 许多病毒，使用这些方法来测试新的病毒-癌症 关联，并确定其统计学和临床意义。 特别是，这一高度合作的提案描述了一个新的， 独特灵活的无掩模光刻技术将用于 生产和精制高密度寡核苷酸微阵列， 灵敏地检测所有已知人类肿瘤病毒的所有基因或转录物。 为了比较，将PCR扩增的病毒探针阵列点在载玻片上， 也将受到考验。 额外的实验将系统地优化 用于特异性、灵敏性和稳健性的提取和扩增方法 临床组织中低拷贝数病毒RNA和DNA序列的检测 样品 由此产生的工具和方法将应用于相关的 临床样本，以测试和解决两个井的新兴连接- 特征性的人肿瘤病毒、人乳头瘤病毒（HPV）和Epstein- 巴尔病毒（EBV）分别与口腔癌和乳腺癌有关，并与 将微阵列结果与其他最先进的病毒检测方法进行比较 方法应用于相同的组织样本。 这些研究将提供证据- 微阵列用于检测和表征病毒的基本测试 癌症的贡献，并解决重大的，目前的问题，在病毒 特定肿瘤的病因学。 同时微阵列筛选许多 病毒或病毒基因型增强了病原学和诊断能力 对珍贵组织样本的研究，提供了发现 未被怀疑的病毒-癌症联系，并允许分析病毒的表达， 选择细胞基因以提供肿瘤的分子特征， 评估病毒对肿瘤的潜在影响。 的方法 开发的也将适用于探测病毒参与其他 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.