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 被诊断为诊断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，为检测和 在恶性肿瘤的早期阶段去除病变。这也产生了改善的迫切需求 诊断，因为目前有80％的患者患有不必要的甲状腺切除术 结节是良性的。当前的护理标准包括通过超声来收集组织活检 指导性细针吸入：这些样品通过细胞学测试，该细胞学寻找异常细胞， 以及遗传风险标记。这些诊断测试的表现不佳，因此三分之一 结节仍被归类为不确定的，并被去除以进行病理诊断。我们 假设蛋白水解失调，这是一种侵入性的酶活性 癌症可能会产生来自良性和恶性结节的FNA组织中区分活性水平。 Alaunus Biosciences，Inc。诊断管道利用底物分析技术 在UCSF的Craik实验室中开发，该平台允许创新的蛋白质组学 生物标志物发现。使用此技术，我们以前已经确定并表征了 在恶性前胰腺囊肿中显着增加的专有蛋白酶活性； 然后，我们开发了荧光底物测定法，以使用少量 液体活检（囊肿液）。在这个阶段，我的建议我们旨在将我们的发现管道应用于 病理确认的甲状腺癌并匹配正常组织的回顾性队列，以鉴定 特定于恶性组织的蛋白酶活性。在AIM 1中，我们将介绍甲状腺肿瘤特异性 蛋白水解信号，并识别负责观察到的候选酶标记 蛋白水解活性。在AIM 2中，我们将设计并合理优化特定的荧光基材 这些蛋白酶可用于快速检测FNA样品中的恶性结节。带领 候选底物将通过 根据当前的临床标准指南对原型测定进行基准测试。 我们的目标是开发快速测定，以改善患者分层，而不是当前标准诊断 标记和指导临床决策以避免不必要的手术干预。 如果成功，该建议将为可操作的诊断测试奠定基础 改善了不确定甲状腺结节的风险分层，并改变了 这些具有挑战性的病变。