Specific Detection of Cervical Cancers Using Cytometry-Based Molecular Diagnostic

使用基于细胞计数的分子诊断对宫颈癌进行特异性检测

• 批准号: 8327306
• 负责人: Vincent Jo Davisson
• 金额: $ 40.49万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-08 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8327306
• 关键词: Abnormal Cell; Automation; Biological Assay; Biological Markers; Biopsy; Blinded; CDKN2A gene; Cancer Etiology; Cancerous; Capillary Electrophoresis; Cell Separation; Cells; Cervical; Cervical dysplasia; Classification; Clinical; Colposcopy; Cytology; Cytometry; DNA analysis; Detection; Developed Countries; Diagnosis; Diagnostic; Diagnostic Procedure; Diagnostic Specificity; Disease; Dysplasia; Emerging Technologies; Evaluation; Flow Cytometry; Frequencies; Future; Genotype; Goals; Grant; Gynecologic; HPV-High Risk; Human Papillomavirus; Immune; Immunofluorescence Immunologic; Incidence; Infection; Label; Lesion; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Methodology; Methods; Molecular; Monitor; Morbidity - disease rate; Mucous Membrane; Pap smear; Pathology; Patients; Phase; Plague; Play; Population; Predictive Value; Premalignant; Preparation; Protein Analysis; Proteins; Protocols documentation; Relative (related person); Research Personnel; Resolution; Risk; Role; Sampling; Screening procedure; Sensitivity and Specificity; Slide; Solutions; Sorting - Cell Movement; Specificity; Specimen; Staging; Staining method; Stains; Techniques; Technology; Test Result; Testing; Translating; Translations; Triage; United States; Woman; Work; base; blind; cancer cell; clinical efficacy; clinically significant; efficacy research; genotyping technology; improved; malignant breast neoplasm; meetings; mortality; new technology; novel strategies; protein expression; public health relevance; research study; standard of care; success

DESCRIPTION (provided by applicant): Cervical cancer is second to breast cancer as the most common form of malignancy in both incidence and mortality for women worldwide. The population-wide utilization of screening cervical cytology (Pap tests) has been associated with a dramatic decrease in morbidity and mortality from cervical cancer in the United States and in other industrialized nations. Despite this success, the cytologic diagnosis of cervical lesions is plagued by a persistent problem of low specificity for clinically significant high-grade lesions in patients with low-grade cytologic abnormalities. As a result, over four million women each year receive a cytologic diagnosis that requires further evaluation to rule out the possibility of high-grade dysplasia or cancer. In most cases, further evaluation does not identify underlying high-grade lesions in patients with low-grade cytologic abnormalities. Although HPV testing can play an important role for the triage of some patients, it is not useful for several cytologic diagnoses. Complicating the situation is that simple detection of high risk HPVs does not predict an underlying high grade lesion, since infections do not indicate clinically significant cervical lesions. The long-term goal of this project is to apply emerging technology to develop a high-throughput cell-based analysis with suitable specificity to identify high grade premalignant and malignant lesions of the cervical mucosa. The methods to be used in this project will employ, test and validate the approach of cytometry-based molecular diagnostics to detect false negative cervical specimens. Under the guise of the previous grant phase, an application of protein expression of p16INK4a and Mcm5 (cervical cancer biomarkers) with high- throughput flow cytometry and cell sorting has been used to identify and capture the rare cancerous cells in cervical specimens. Furthermore, a multiplexed HPV genotyping assay has been implemented to analyze the rare cells isolated in this approach. Importantly, the work flow has been implemented using common sample preparation with current pathology testing protocols. The technology and methodology being applied in this application will be implemented using an integrated workflow, with substantial automation, to assess feasibility of further translation to accommodate clinical need and to improve the standard of care worldwide. The ultimate goal is to establish a primary assay with potential to supplant slide based cervical cytology with greater sensitivity, less subjectivity, and less labor intensiveness. PUBLIC HEALTH RELEVANCE: This proposed project will apply new technology, for detection of abnormal cervical cells, to clinical samples from previous Pap tests. We will use automated analysis of protein content of cells to isolate abnormal cells, after which automated DNA analysis will determine whether cancer-causing human papillomavirus types are present. These results will be compared to the Pap smear and biopsy results of the same samples in order to determine how well our test can detect early stages of cervical cancer.

描述（由申请人提供）：对于全世界女性来说，无论是发病率还是死亡率，宫颈癌都是仅次于乳腺癌的最常见的恶性肿瘤。在美国和其他工业化国家，宫颈细胞学筛查（巴氏试验）在全民范围内的使用与宫颈癌发病率和死亡率的急剧下降有关。尽管取得了这一成功，但宫颈病变的细胞学诊断仍然受到低度细胞学异常患者临床上显着的高级病变特异性低的持续问题的困扰。因此，每年有超过 400 万女性接受细胞学诊断，需要进一步评估以排除高度不典型增生或癌症的可能性。在大多数情况下，进一步评估并不能识别出具有低度细胞学异常的患者潜在的高级病变。尽管 HPV 检测对于某些患者的分诊可以发挥重要作用，但对于某些细胞学诊断并无用处。使情况变得复杂的是，简单检测高危 HPV 并不能预测潜在的高级病变，因为感染并不表明有临床意义的宫颈病变。该项目的长期目标是应用新兴技术开发具有适当特异性的高通量细胞分析，以识别宫颈粘膜的高级别癌前病变和恶性病变。该项目使用的方法将采用、测试和验证基于细胞计数的分子诊断方法来检测假阴性宫颈标本。在先前资助阶段的幌子下，应用 p16INK4a 和 Mcm5（宫颈癌生物标志物）的蛋白质表达以及高通量流式细胞术和细胞分选来识别和捕获宫颈标本中的罕见癌细胞。此外，还实施了多重 HPV 基因分型测定来分析这种方法中分离出的稀有细胞。重要的是，工作流程是使用常见的样品制备和当前的病理学测试协议来实现的。该应用程序中应用的技术和方法将使用集成的工作流程来实施，并具有高度自动化，以评估进一步翻译的可行性，以满足临床需求并提高全球护理标准。最终目标是建立一种初级检测方法，有可能取代基于玻片的宫颈细胞学检测，具有更高的灵敏度、更少的主观性和更少的劳动强度。 公共健康相关性：该拟议项目将应用新技术，检测异常宫颈细胞，以及以前巴氏试验的临床样本。我们将使用细胞蛋白质含量的自动分析来分离异常细胞，然后自动 DNA 分析将确定是否存在致癌的人乳头瘤病毒类型。这些结果将与相同样本的巴氏涂片和活检结果进行比较，以确定我们的测试对早期宫颈癌的检测效果。

项目成果

High-throughput secondary screening at the single-cell level.

单细胞水平的高通量二次筛选。

• 发表时间: 2013-02
• 作者: Robinson, J Paul;Patsekin, Valery;Holdman, Cheryl;Ragheb, Kathy;Sturgis, Jennifer;Fatig, Ray;Avramova, Larisa V;Rajwa, Bartek;Davisson, V Jo;Lewis, Nicole;Narayanan, Padma;Li, Nianyu;Qualls Jr, C W
• 通讯作者: Qualls Jr, C W