Development of activity-based biomarkers for risk stratification of indeterminate thyroid nodules

开发基于活动的生物标志物，用于不确定甲状腺结节的风险分层

• 批准号: 10080280
• 负责人: GISELLE M KNUDSEN
• 金额: $ 26.47万
• 依托单位: ALAUNUS BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10080280
• 关键词: Benchmarking; Benign; Biochemical; Biological Assay; Biological Markers; Biological Sciences; Biopsy; Biopsy Specimen; Cancer Patient; Cells; Classification; Clinic; Clinical; Clinical Management; Coupled; Cyst Fluid; Cytology; Detection; Development; Diagnosis; Diagnostic; Diagnostic Imaging; Diagnostic tests; Early Diagnosis; Engineering; Enzymes; Excision; Fine needle aspiration biopsy; Fluorescence; Fluorescence Resonance Energy Transfer; Fluorogenic Substrate; Foundations; Future; Generations; Genetic Risk; Goals; Guidelines; Laboratories; Lead; Lesion; Libraries; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of thyroid; Mass Spectrum Analysis; Molecular Analysis; Monitor; Nodule; Normal tissue morphology; Operative Surgical Procedures; Pancreatic Cyst; Pathologic; Patient risk; Patients; Peptide Hydrolases; Peptide Library; Performance; Phase; Population; Proteolysis; Proteomics; Resected; Retrospective cohort; Risk Marker; Risk stratification; Sampling; Scanning; Sensitivity and Specificity; Small Business Innovation Research Grant; Stratification; Technology; Testing; Thyroid Gland; Thyroid Nodule; Thyroidectomy; Tissue Sample; Tissues; Ultrasonography; United States; Validation; base; biomarker discovery; cancer invasiveness; clinical decision-making; clinical practice; cohort; cost; design; diagnostic assay; diagnostic biomarker; enzyme activity; experience; follow-up; imaging modality; improved; innovation; lead candidate; lead optimization; liquid biopsy; mortality; patient stratification; premalignant; prognostic; prototype; screening; standard of care; thyroid neoplasm; tool; tumor

Abstract A dramatic increase in the number of patients being diagnosed with incidental thyroid nodules through different diagnostic imaging modalities has created a unique opportunity to detect and remove lesions at early stages of malignancy. This also creates a critical need for improved diagnostics, as 80% of patients currently undergo unnecessary thyroidectomies because their nodules were benign. Current standard of care includes collecting a tissue biopsy by ultrasound- guided fine needle aspiration: these samples are tested by cytology, which looks for aberrant cells, and for genetic risk markers. These diagnostic tests perform poorly, consequently a third of all nodules are still classified as indeterminate and are removed for a pathological diagnosis. We hypothesized that dysregulated proteolysis, a type of enzyme activity that is a hallmark of invasive cancer, might yield discriminating levels of activity in FNA tissue from benign and malignant nodules. The Alaunus Biosciences, Inc. diagnostic pipeline takes advantage of a substrate profiling technology developed in the Craik Laboratory at UCSF, a platform that allows innovative proteomics-based biomarker discovery. Using this technology, we have previously identified and characterized a proprietary set of protease activities that are significantly increased in pre-malignant pancreatic cysts; we then developed fluorogenic substrate assays to monitor these activities using small volumes of liquid biopsy (cyst fluid). In this Phase I proposal we aim to apply our discovery pipeline to a retrospective cohort of pathologically-confirmed thyroid cancer and matched normal tissue, to identify protease activities that are specific to malignant tissue. In Aim 1, we will profile thyroid tumor-specific proteolytic signatures, and identify candidate enzyme markers responsible for the observed proteolytic activity. In Aim 2, we will design and rationally optimize specific fluorogenic substrates for these proteases that can be used to rapidly detect malignant nodules in FNA samples. Lead candidate substrates will proceed to a robust validation of their clinical utility as diagnostic tools, by benchmarking a prototype assay against the current clinical standard guidelines. Our goal is to develop a rapid assay that improves patient stratification over current standard diagnostic markers and guides clinical decision-making to avoid unnecessary surgical intervention. If successful, this proposal will lay the groundwork for an actionable diagnostic test that will enable improved risk stratification of indeterminate thyroid nodules, and transform the clinical management of these challenging lesions.

抽象的 被诊断患有偶发甲状腺结节的患者数量急剧增加 通过不同的诊断成像方式创造了一个独特的机会来检测和 在恶性肿瘤的早期阶段去除病变。这也迫切需要改进 诊断，因为目前 80% 的患者接受了不必要的甲状腺切除术，因为他们的 结节是良性的。当前的护理标准包括通过超声收集组织活检 引导细针抽吸：这些样本经过细胞学检测，寻找异常细胞， 以及遗传风险标记。这些诊断测试表现不佳，因此三分之一 结节仍被分类为不确定，并被切除以进行病理诊断。我们 假设蛋白水解失调，这是一种酶活性，是侵入性的标志 癌症，可能会在 FNA 组织中产生良性和恶性结节的区分活性水平。 Alaunus Biosciences, Inc. 诊断管道利用底物分析技术 加州大学旧金山分校 Craik 实验室开发的平台，允许基于蛋白质组学的创新 生物标志物的发现。使用这项技术，我们之前已经识别并表征了 一组专有的蛋白酶活性在癌前胰腺囊肿中显着增加； 然后，我们开发了荧光底物测定法，使用小体积的 液体活检（囊肿液）。在第一阶段提案中，我们的目标是将我们的发现管道应用于 病理证实的甲状腺癌和匹配的正常组织的回顾性队列，以确定 恶性组织特有的蛋白酶活性。在目标 1 中，我们将分析甲状腺肿瘤特异性 蛋白水解特征，并确定负责观察到的候选酶标记 蛋白水解活性。在目标2中，我们将设计并合理优化特定的荧光底物 这些蛋白酶可用于快速检测 FNA 样本中的恶性结节。带领 候选底物将继续对其作为诊断工具的临床效用进行强有力的验证，方法是 根据当前临床标准指南对原型测定进行基准测试。 我们的目标是开发一种快速检测方法，在当前标准诊断的基础上改善患者分层 标记物并指导临床决策，以避免不必要的手术干预。 如果成功，该提案将为可行的诊断测试奠定基础，从而使 改进了不确定甲状腺结节的风险分层，并改变了甲状腺结节的临床管理 这些具有挑战性的病变。