Targeted proteomics of MUC16 to enable early detection of ovarian cancer recurrence

MUC16 的靶向蛋白质组学可实现卵巢癌复发的早期检测

基本信息

批准号：
10543477
负责人：
Rebecca Jean Whelan
金额：
$ 16.73万
依托单位：
UNIVERSITY OF KANSAS LAWRENCE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-01-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10543477
关键词：
Anxiety Area Biological Assay Biological Markers CA-125 Antigen Cancer Patient Caregivers Charge Clinical Collaborations Community Clinical Oncology Program Core Protein Data Detection Diagnostic Diagnostic tests Disease Early Diagnosis Epitopes FDA approved Funding Future Glycopeptides Glycoproteins Goals Gynecologic Oncology Immunoassay Individual Institution Intervention Lesion Link Malignant Neoplasms Malignant neoplasm of ovary Mass Spectrum Analysis Messenger RNA Methods Modernization Molecular Monitor Mucin 1 protein Mucins Parents Patients Peptide Library Peptide Mapping Peptides Periodicals Peritoneal Fluid Polysaccharides Population Post-Translational Protein Processing Process Proteins Proteomics Protocols documentation Publications RNA Splicing Reaction Recovery Recurrence Recurrent Malignant Neoplasm Recurrent disease Reporting Research Research Personnel Resources Risk Sampling Schedule Screening for Ovarian Cancer Serous Serum Site Tandem Repeat Sequences Testing Thinking Tumor Debulking United States National Institutes of Health Variant Visit Woman cancer biomarkers cancer cell cancer recurrence cancer survival chemotherapy computerized tools diagnostic biomarker diagnostic tool early detection biomarkers effective therapy experimental study fighting follow-up glycosylation high reward high risk improved in silico molecular dynamics novel novel diagnostics ovarian neoplasm overexpression treatment strategy tumor

项目摘要

High-grade serous ovarian cancer (HGSOC) is a deadly disease, in large part because most cases recur, and little can be done after that point. New diagnostic tools are urgently needed to improve HGSOC management and detect recurrence before clinical presentation. MUC16 is overexpressed on ovarian cancer cells and bears the CA125 epitope that is currently detected in clinical assays. The value of CA125 titers for surveillance is an active area of debate within the gynecologic oncology community. Some studies report no survival benefit, while others point to higher quality secondary cytoreductive surgery if action is taken quickly. Successfully fighting HGSOC requires that new assays are developed for more reliable and sensitive detection of recurrent disease. Our goal is to develop a new biomarker for HGSOC recurrence by fundamentally rethinking MUC16. Instead of considering MUC16 as the carrier of the CA125 epitope, we recognize that MUC16 itself is present as multiple diverse proteoforms. Because of variation in mRNA splicing, post-translational cleavage, and post- translational modifications, the population of MUC16 molecules present in an individual is heterogeneous. MUC16 is abundantly glycosylated, and glycan profiles of MUC16 derived from cancer cells is an untapped wealth of diagnostic information that is now lost using the existing peptide-based CA125 immunoassay. The goal of this project is to develop a method to enrich MUC16 from the serum of women undergoing treatment for HGSOC and analyze MUC16 using targeted (glyco)proteomics. We hypothesize that the molecular diversity of MUC16 revealed using modern bioanalytical and computational tools will enable novel diagnostics to detect HGSOC resurgence earlier than is currently possible. This hypothesis will be investigated by pursuing three research aims. In Aim 1 (Develop methods to quantitate MUC16 (glyco)peptide libraries using mass spectrometry), MUC16 from banked peritoneal fluid of HGSOC patients will be used to develop an optimized novel targeted mass spectrometry method for identification of MUC16 glycopeptides and peptides before and after deglycosylation. A parallel reaction monitoring (PRM) inclusion list will be generated by examination of in silico digest data and experimental MS/MS data. In Aim 2 (Develop a microscale separation method to enrich MUC16 from serum), we will adapt an immunoaffinity-free protocol that we developed to enrich MUC16 from peritoneal fluid to isolate the mucin from serum samples. A cartridge-based format will be used to capture MUC16 based on its negative charge and specific glycosylation. Finally, in Aim 3: (Develop pilot data on longitudinal variations in MUC16 (glyco)peptide libraries in patients with HGSOC before and during treatment), longitudinal serum samples from HGSOC patients will be enriched for MUC16 using the microscale protocol. Samples will be proteolyzed and analyzed by nLC-PRM. Quantitative (glyco)peptide maps for MUC16 from each patient will be compared longitudinally. The (glyco)peptides that deviate most between samples will be determined as potential diagnostic markers for early detection of recurring HGSOC in a future study.

高级别浆液性卵巢癌 (HGSOC) 是一种致命的疾病，很大程度上是因为大多数病例会复发，并且在那之后就无能为力了。迫切需要新的诊断工具来改进 HGSOC 管理并在临床表现前检测复发。 MUC16 在卵巢癌细胞和熊上过度表达目前在临床检测中检测到的 CA125 表位。 CA125 滴度的监测价值是妇科肿瘤学界争论的活跃领域。一些研究报告没有生存获益，而其他人则指出，如果迅速采取行动，二次细胞减灭术的质量会更高。成功地对抗 HGSOC 需要开发新的检测方法，以更可靠、更灵敏地检测复发性疾病。我们的目标是通过从根本上重新思考 MUC16，开发一种新的 HGSOC 复发生物标志物。我们不认为MUC16是CA125表位的载体，而是认识到MUC16本身存在作为多种不同的蛋白质形式。由于 mRNA 剪接、翻译后切割和翻译后的变异由于翻译修饰，个体中存在的 MUC16 分子群是异质的。 MUC16 具有丰富的糖基化，来自癌细胞的 MUC16 聚糖谱尚未开发使用现有的基于肽的 CA125 免疫测定法，现在会丢失大量的诊断信息。这该项目的目标是开发一种方法，从接受治疗的女性血清中富集 MUC16 HGSOC 并使用靶向（糖）蛋白质组学分析 MUC16。我们假设分子多样性 MUC16 揭示使用现代生物分析和计算工具将使新的诊断方法能够检测 HGSOC 的复苏比目前可能的更早。该假设将通过追求三个研究目的。目标 1（开发使用质量数定量 MUC16（糖）肽库的方法光谱法），来自 HGSOC 患者腹膜液库的 MUC16 将用于开发优化的用于在之前和之后鉴定MUC16糖肽和肽的新型靶向质谱方法去糖基化后。平行反应监测 (PRM) 包含列表将通过检查生成硅片摘要数据和实验 MS/MS 数据。目标 2（开发一种微尺度分离方法来富集 MUC16（来自血清），我们将采用我们开发的无免疫亲和方案来富集来自血清的 MUC16 腹膜液以从血清样本中分离出粘蛋白。基于盒式磁带的格式将用于捕获 MUC16 基于其负电荷和特异性糖基化。最后，在目标 3 中：（开发试点数据 HGSOC 患者治疗前和治疗期间 MUC16（糖）肽库的纵向变化），将使用微量方案对 HGSOC 患者的纵向血清样本进行 MUC16 富集。样品将通过 nLC-PRM 进行蛋白水解和分析。 MUC16 的定量（糖）肽图谱对每位患者进行纵向比较。样品之间差异最大的（糖）肽是在未来的研究中被确定为早期检测复发性 HGSOC 的潜在诊断标志物。