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）：宫颈癌第二位是乳腺癌是全球女性发病率和死亡率最常见的恶性肿瘤形式。筛查宫颈细胞学筛查（PAP检验）的全部利用与美国和其他工业化国家的宫颈癌的发病率和死亡率显着降低有关。尽管取得了成功，但颈椎病变的细胞学诊断仍受到低度细胞学异常患者临床显着高级病变的持续特异性持续问题的困扰。结果，每年有超过四百万女性接受细胞学诊断，需要进一步评估以排除高级发育不良或癌症的可能性。在大多数情况下，进一步的评估不能识别出低度细胞学异常患者的潜在高级病变。尽管HPV测试对于某些患者的分类可以发挥重要作用，但对于多种细胞学诊断没有用。使情况复杂化的是，高风险HPV的简单检测不会预测潜在的高级病变，因为感染并未表明临床上显着的宫颈病变。该项目的长期目标是应用新兴技术以适当的特异性开发基于高通量的细胞分析，以识别颈椎粘膜的高级前态和恶性病变。该项目中要使用的方法将采用，测试和验证基于细胞术的分子诊断方法检测假阴性宫颈样品的方法。在上一个赠款阶段的幌子下，使用高吞吐量流式细胞仪和细胞分选的p16INK4A和MCM5（宫颈癌生物标志物）的蛋白质表达应用来识别和捕获颈椎标本中的罕见癌细胞。此外，已经实施了多重的HPV基因分型测定，以分析在这种方法中分离的稀有细胞。重要的是，使用当前病理测试方案的常见样品制备实施了工作流程。本应用程序中应用的技术和方法将使用具有实质性自动化的集成工作流程实施，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水平。最终目标是建立一个主要的测定，其潜力具有更高的敏感性，较少的主观性和较少的劳动力强度，以取代基于滑动的宫颈细胞学。 公共卫生相关性：该拟议的项目将应用新技术，以检测异常的宫颈细胞，用于以前的PAP测试中的临床样本。我们将使用对细胞蛋白质含量的自动分析来分离异常细胞，之后自动DNA分析将确定是否存在引起癌症的人乳头瘤病毒类型。这些结果将与同一样品的子宫颈抹片检查和活检结果进行比较，以确定我们的测试能够检测到宫颈癌的早期阶段。

项目成果

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

• DOI: 10.1177/2211068212456978
• 发表时间: 2013-02
• 期刊: Journal of laboratory automation
• 作者: Robinson JP;Patsekin V;Holdman C;Ragheb K;Sturgis J;Fatig R;Avramova LV;Rajwa B;Davisson VJ;Lewis N;Narayanan P;Li N;Qualls CW Jr
• 通讯作者: Qualls CW Jr