A paradigm shift for ovarian cancer biomarkers: Utilizing routine Pap tests as liquid biopsies for the development of targeted mass spectrometry-based proteomic assays for early detection

卵巢癌生物标志物的范式转变：利用常规巴氏试验作为液体活检，开发基于质谱的靶向蛋白质组学检测，以进行早期检测

• 批准号: 10462771
• 负责人: AMY PATRICE SKUBITZ
• 金额: $ 52.73万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-05 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10462771
• 关键词: Abnormal Cell; Alcohols; Benign; Bioinformatics; Biological Assay; Biological Markers; Cancer Etiology; Cells; Cervical; Cervical Cancer Screening; Cessation of life; Clinic; Clinical; Collection; Coupled; Data; Data Analyses; Detection; Development; Diagnosis; Diagnostic; Diagnostic Reagent Kits; Disease; Early Diagnosis; Fixatives; Future; General Population; Goals; Gynecologic; Gynecologic Surgical Procedures; Hand; Home; Human Papillomavirus; Institutional Review Boards; Label; Lesion; Liquid substance; Malignant Female Reproductive System Neoplasm; Malignant neoplasm of cervix uteri; Malignant neoplasm of ovary; Mammalian Oviducts; Mass Spectrum Analysis; Measurement; Methods; Monitor; Pap smear; Patients; Peptides; Performance; Physicians; Positioning Attribute; Proteins; Proteomics; Reaction; Residual state; Resources; Sampling; Savings; Screening for Ovarian Cancer; Screening procedure; Self Administration; Sensitivity and Specificity; Serous; Site; Source; Survival Rate; Swab; Techniques; Testing; Translating; Tumor Tissue; United States; Uterus; Vagina; Woman; base; biomarker discovery; cancer biomarkers; cancer cell; cancer diagnosis; candidate marker; cohort; cost effective; detection assay; differential expression; experimental study; fimbria; high risk population; home test; improved; liquid biopsy; protein biomarkers; reproductive tract; sample collection; screening; tumor

PROJECT SUMMARY/ABSTRACT Ovarian cancer is the 5th leading cause of cancer deaths in women in the U.S. Earlier detection is the key to increased survival for women with ovarian cancer, yet a screening tool that is both sensitive and specific enough for use in the general population has yet to be developed. In contrast, screening for cervical cancer by Pap tests has been routinely performed for over 50 years. In the liquid-based Pap test, cells are collected from the cervical opening and placed into an alcohol-based fixative and then examined for abnormal cells. Since ovarian cancer cells have been observed in Pap tests, ovarian cancer peptide biomarkers may also be present; yet Pap samples have not been rigorously examined for diagnostic peptides. Our central hypothesis is that proteins shed by ovarian cancer cells can be detected in the Pap test fixative and on cervical-vaginal swabs by Mass Spectrometry (MS)-based proteomics. The Pap test fixative and swabs are ideal for biomarker discovery since they are derived from a site proximal to the ovarian cancer (i.e. proteins may be secreted or shed from the tumor and flow through the fallopian tube into the uterus and out the cervical opening). Recently, ovarian cancer precursor lesions have been identified in the fimbria of the fallopian tube, strengthening the hypothesis that ovarian cancer proteins will be found in the lower genital tract, perhaps even at early stages. In preliminary studies, candidate biomarkers were successfully identified by MS-based proteomics in Pap test fixatives from women with ovarian cancer, and the levels of biomarker peptides were quantified in cases vs. benign and healthy controls, demonstrating the feasibility of this approach. The long- term goal of this project is to develop a noninvasive screening test that can be incorporated into a routine Pap test or a self-administered home test, so that women can be screened simultaneously for cervical and ovarian cancer. The objective of this study is to verify ovarian cancer biomarker peptides identified in Pap test samples and extend the results to cervical swabs, using our collection of biospecimens from over 600 patients and controls. Aim #1 will build on our preliminary studies to prioritize ovarian cancer biomarker candidates found in liquid-based Pap test samples by performing Tandem Mass Tag™ 11-plex isobaric labeling, 2D LC-MS/MS, and bioinformatics integration. In Aim #2, the most robust candidate biomarkers will be quantified by developing a targeted Selected Reaction Monitoring (SRM)-MS assay coupled with a multi-protein classifier, which will then be used to test hundreds of Pap test samples from women with ovarian cancer, other gynecologic cancers, and benign gynecologic conditions, as well as healthy women. In Aim #3, proteins eluted from cervical swabs will be tested to determine whether they are also detected by our SRM-MS assay. Results from this study may be extended to the early detection of ovarian cancer in women in high risk groups and eventually for screening the general population, thereby shifting the paradigm for how women are screened for ovarian cancer and improving survival rates for the >22,000 women diagnosed with ovarian cancer each year.

项目概要/摘要 卵巢癌是美国女性癌症死亡的第五大原因，早期发现是预防癌症的关键 提高卵巢癌女性的生存率，同时是一种既敏感又特异的筛查工具 尚待开发出足以供一般人群使用的产品。相比之下，宫颈癌筛查 巴氏试验已作为常规检查进行了 50 多年。在基于液体的巴氏试验中，细胞是从 宫颈开口并放入酒精固定剂中，然后检查是否有异常细胞。自从 巴氏试验中已观察到卵巢癌细胞，卵巢癌肽生物标志物也可能是 展示;然而巴氏涂片样本尚未经过严格的诊断肽检查。我们的中心假设 是可以在巴氏试验固定剂和宫颈阴道上检测到卵巢癌细胞脱落的蛋白质 通过基于质谱 (MS) 的蛋白质组学检测拭子。巴氏试验固定剂和拭子是生物标志物的理想选择 发现，因为它们源自靠近卵巢癌的部位（即蛋白质可能被分泌或 从肿瘤中脱落并通过输卵管流入子宫并从宫颈口流出）。 最近，在输卵管伞部发现了卵巢癌前兆病变， 加强了卵巢癌蛋白将在下生殖道中发现的假设，甚至可能 在早期阶段。在初步研究中，基于 MS 的方法成功鉴定了候选生物标志物 卵巢癌女性巴氏试验固定剂中的蛋白质组学，以及生物标志物肽的水平 对病例与良性健康对照进行量化，证明了这种方法的可行性。长- 该项目的长期目标是开发一种可纳入常规巴氏涂片检查的无创筛查测试 测试或自行进行的家庭测试，以便女性可以同时筛查宫颈和卵巢 癌症。本研究的目的是验证巴氏试验样本中鉴定出的卵巢癌生物标志物肽 并将结果扩展到宫颈拭子，使用我们收集的 600 多名患者的生物样本， 控制。目标#1将建立在我们的初步研究的基础上，优先考虑在以下领域发现的卵巢癌候选生物标志物： 通过执行 Tandem Mass Tag™ 11 重同量异位标记、2D LC-MS/MS、 和生物信息学整合。在目标#2中，最稳健的候选生物标志物将通过以下方式进行量化 开发与多蛋白分类器相结合的靶向选择反应监测 (SRM)-MS 检测， 然后将用于测试来自患有卵巢癌和其他癌症的女性的数百个巴氏试验样本 妇科癌症、良性妇科疾病以及健康女性。在目标 #3 中，蛋白质被洗脱 我们将对宫颈拭子中的病毒进行测试，以确定我们的 SRM-MS 检测是否也能检测到它们。结果 这项研究可能会扩展到高危女性卵巢癌的早期检测和 最终用于对一般人群进行筛查，从而改变女性筛查方式的范式 卵巢癌，并提高每年超过 22,000 名被诊断患有卵巢癌的女性的生存率。