Development of a FISH-Based HPV-Associated Cancer Detection Test

开发基于 FISH 的 HPV 相关癌症检测测试

• 批准号: 7669927
• 负责人: Jane Houldsworth
• 金额: $ 9.96万
• 依托单位: CANCER GENETICS, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7669927
• 关键词: 20q; Abnormal Cell; Anogenital region; Anus; Appearance; Biological Assay; Biological Markers; Cancer Detection; Caring; Cell Nucleus; Cervical; Cervical Cancer Screening; Collaborations; Color; Colposcopy; Country; Cytology; Data Set; Detection; Development; Event; Fluorescent in Situ Hybridization; Genetic; Genomics; Goals; Gold; HPV-High Risk; Human; Human Papillomavirus; Human papilloma virus infection; Incidence; Infection; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Manuals; Materials Testing; Medical; Molecular; National Cancer Institute; Nature; Oncogenic; Oncogenic Viruses; Output; Pap smear; Patients; Performance; Phase; Population; Populations at Risk; Premalignant; Procedures; Recommendation; Residual state; Resources; Role; Scanning; Screening for cancer; Screening procedure; Sensitivity and Specificity; Severities; Specimen; System; Testing; Triage; Unnecessary Procedures; Validation; Woman; base; cancer genetics; cancer prevention; cervical and anal cancer; cervical cancer prevention; cost effectiveness; follow-up; improved; programs; public health relevance; success; tumor progression

DESCRIPTION (provided by applicant): A causative role of high-risk human papillomavirus (HPV) types in human cancer has been established, particularly for those of the anogenital region including cervical and anal cancers. The etiologic course of such cancers is thought to involve HPV infection, persistence of infection, progression to an immediate precursor of cancer, and finally invasion (the latter two steps requiring additional host oncogenic events). Despite the success of cervical cancer screening programs based on the appearance of abnormal cells in cytology specimens and HPV-type, there is a great need to identify additional biomarkers to increase the sensitivity with which precancer/cancer are detected, so providing additional triage of the approximately 2,000,000 women each year in the US who undergo colposcopy for follow-up of abnormal cytology. Gain of 3q and to a lesser extent gain of 5p and 20q, are genomic abnormalities commonly detected in HPV-associated cancers, and as such represent potential biomarkers of HPV-associated cancer progression. In the current application, Cancer Genetics, Inc. proposes to develop and validate a fluorescence in situ hybridization (FISH)-based HPV- associated cancer detection test (FHACT) to detect the three genomic abnormalities in cervical and anal cytology specimens. Phase I; Specific Aim 1: Confirmation of gain of 3q, 5p, and 20q in cervical precancerous and cancer cytology specimens. 200 cervical cytology specimens will be submitted to four- color FISH using probes to detect gain of 3q (red), 5p (green), and 20q (gold), to confirm association with cytologic severity and to provide an initial estimate of the sensitivity for precancer/cancer detection. If the sensitivity is comparable to that for cytology alone, then Phase II will be initiated. Phase II; Specific Aim 1: Validation of automated scanning for the FHACT in cervical cytology specimens. The scanning/scoring procedure for the FHACT will be automated in order to adapt the assay for increased output. Phase II; Specific Aim 2: Optimization of the FHACT in 1,000 cervical cytology specimens. The sensitivity and specificity of the FHACT will be determined for the entire dataset of 1,000 cervical cytology specimens and compared with that by cytology alone, after optimization of cut-offs, probe combinations, and numbers of nuclei to score. Phase II; Specific Aim 3: Optimization of the FHACT in 1,000 anal cytology specimens. Comparable studies to those above will be performed for anal cytology specimens, where standard-of-care screening programs have as yet to be developed for at-risk populations. Specimens will be obtained from Drs. Castle and Wentzensen, at the National Cancer Institute who are leading studies aimed at improving cancer prevention screening programs and identifying biomarkers of HPV-associated cancers. The overall goal of the project then is to develop and validate a robust, sensitive, and specific FISH-based test that together with standard cytology and HPV-typing will significantly contribute to more accurate detection of precancer/cancer, impacting standard-of-care recommendations in HPV-associated cancer screening programs. PUBLIC HEALTH RELEVANCE: Despite the success of cervical cancer prevention screening programs, there are many women in the US, who annually undergo medical procedures based on an abnormal Pap test and oncogenic virus test, which are later found to be unnecessary and costly, due to the lack of the sensitivity of these tests. Thus, in the current proposal a genetic-based precancer and cancer detection test is to be developed and validated, that uses residual Pap or anal test material. Implementation of this test into cervical and anal cancer prevention screening programs should provide an additional triage to identify those patients who require medical follow-up versus who do not, thereby reducing the performance of unnecessary and costly medical procedures, ultimately impacting population coverage of such programs in low resource countries.

描述（由申请人提供）：已经确定高危人类乳头瘤病毒（HPV）类型在人类癌症中的致病作用，特别是对于包括子宫颈癌和肛门癌在内的肛门生殖器区域的癌症。这类癌症的病因学过程被认为包括HPV感染、持续感染、发展为癌症的直接前兆，以及最后的侵袭（后两个步骤需要额外的宿主致癌事件）。尽管基于细胞学标本中异常细胞的外观和hpv类型的宫颈癌筛查项目取得了成功，但仍然非常需要识别额外的生物标志物来提高检测癌前病变/癌症的敏感性，因此为美国每年约200万接受阴道镜检查以进行异常细胞学随访的妇女提供额外的分类。3q的增益和较小程度的5p和20q的增益是hpv相关癌症中常见的基因组异常，因此代表了hpv相关癌症进展的潜在生物标志物。在当前的应用中，Cancer Genetics, Inc.提议开发并验证基于荧光原位杂交（FISH）的HPV相关癌症检测测试（FHACT），以检测宫颈和肛门细胞学标本中的三种基因组异常。第一阶段;特异性目的1：确认宫颈癌前和癌细胞学标本中3q、5p和20q的增加。200例宫颈细胞学标本将提交给四色FISH，使用探针检测3q（红色），5p（绿色）和20q（金色）的增益，以确认与细胞学严重程度的关联，并提供癌前/癌症检测敏感性的初步估计。如果灵敏度与单独细胞学的灵敏度相当，则将启动II期。第二阶段;具体目标1：验证FHACT在宫颈细胞学标本中的自动扫描。FHACT的扫描/评分程序将实现自动化，以适应增加产量的分析。第二阶段;具体目标2：优化FHACT在1000例宫颈细胞学标本中的应用。FHACT的敏感性和特异性将在1000个宫颈细胞学标本的整个数据集上确定，并与细胞学单独进行比较，优化截止点、探针组合和细胞核数量进行评分。第二阶段;具体目标3：优化FHACT在1000个肛门细胞学标本中的应用。将对肛门细胞学标本进行与上述类似的研究，其中标准护理筛查计划尚未为高危人群制定。标本将从博士。Castle和Wentzensen是美国国家癌症研究所的研究员，他们领导的研究旨在改善癌症预防筛查项目，并识别hpv相关癌症的生物标志物。该项目的总体目标是开发和验证一种强大、敏感和特异性的基于fish的测试，该测试与标准细胞学和hpv分型一起，将大大有助于更准确地检测癌前/癌，影响hpv相关癌症筛查项目的标准护理建议。公共卫生相关性：尽管宫颈癌预防筛查项目取得了成功，但在美国仍有许多妇女每年接受基于异常巴氏涂片检查和致癌病毒检查的医疗程序，后来发现由于这些检查缺乏敏感性，这些检查是不必要的，而且费用高昂。因此，在目前的建议中，将开发和验证一种基于基因的癌前和癌症检测测试，该测试使用残留的Pap或肛门测试材料。在宫颈癌和肛门癌预防筛查项目中实施这一测试将提供额外的分类，以确定哪些患者需要进行医疗随访，哪些不需要，从而减少不必要和昂贵的医疗程序的执行，最终影响资源匮乏国家此类项目的人口覆盖率。