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

• 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
• 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

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.

高级浆液卵巢癌（HGSOC）是一种致命疾病，在很大程度上是因为大多数情况复发，并且 在那之后几乎无法完成。迫切需要新的诊断工具来改善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表达 目前在临床测定中检测到的CA125表位。 CA125滴度对监视的值是 妇科肿瘤学社区中辩论的活跃领域。一些研究报告没有生存益处， 而其他人则指出，如果采取行动迅速采取行动，则指出了更高的二级细胞减少手术。成功地 战斗HGSOC要求开发新测定 疾病。我们的目标是通过从根本上重新思考MUC16来开发HGSOC复发的新生物标志物。 我们意识到MUC16本身存在，而不是将MUC16视为CA125表位的载体 作为多种蛋白质类型。由于mRNA剪接的变化，翻译后的裂解和后 翻译修饰，个体中存在的MUC16分子的种群是异质的。 MUC16大量糖基化，源自癌细胞的MUC16的聚糖谱是未开发的 现在使用现有的基于肽的CA125免疫测定法丢失的大量诊断信息。这 该项目的目标是开发一种方法，以富集MUC16的妇女血清接受治疗 HGSOC使用靶向（GlyCO）蛋白质组学分析MUC16。我们假设分子多样性 使用现代生物分析和计算工具揭示的MUC16将使新型诊断能够检测 HGSOC的复苏比目前可能更早。该假设将通过追求三个 研究目的。在AIM 1中（开发使用质量量化MUC16（Glyco）肽库的方法 光谱法），来自HGSOC患者的腹膜腹膜液的MUC16将用于开发优化 新颖的靶向质谱法，用于鉴定MUC16糖肽和肽之前和 脱糖基化后。通过检查In将生成平行反应监测（PRM）包含列表 硅摘要数据和实验性MS/MS数据。在AIM 2中（开发一种微观分离方法以丰富 MUC16来自血清），我们将适应一种无免疫疗法的方案，我们开发了以富集Muc16 腹膜液可将粘蛋白与血清样品分离。基于墨盒的格式将用于捕获 MUC16基于其负电荷和特异性糖基化。最后，在AIM 3中：（开发有关 在治疗前和治疗期间，HGSOC患者的MUC16（Glyco）肽库的纵向变化）， 来自HGSOC患者的纵向血清样品将使用微观方案富含MUC16。 样品将通过NLC-PRM进行蛋白水解和分析。 MUC16的定量（Glyco）肽图 每个患者将进行纵向比较。在样品之间偏离最大的（Glyco）肽将是 确定为潜在的诊断标记物，用于在未来的研究中早期检测重复的HGSOC。