Digital Nanoplasmonic Quantification of Tumor-derived Extracellular Vesicles in Plasma Microsamples

血浆微样品中肿瘤源性细胞外囊泡的数字纳米等离子体定量

• 批准号: 10684737
• 负责人: Tony Y. Hu
• 金额: $ 56.62万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-25 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10684737
• 关键词: Address; Affect; Benign; Binding; Biological Assay; Biological Markers; Blood specimen; CA-19-9 Antigen; CD44 gene; CLIA certified; Cancer Detection; Cells; Circulation; Clinical; Complex; Detection; Development; Diagnosis; Diagnostic; Disease; Early Diagnosis; Early treatment; Excision; Exhibits; Family history of; Guidelines; Image; Individual; Inherited; Laboratories; Lesion; Longitudinal Studies; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of pancreas; Manuals; Methods; Microfabrication; Modeling; Monitor; Mus; Neoadjuvant Therapy; Neoplasm Metastasis; Non-Malignant; Operative Surgical Procedures; Pancreas; Pancreatitis; Patients; Performance; Pilot Projects; Plasma; Predictive Cancer Model; Procedures; Proteins; Protocols documentation; Reader; Reagent; Regimen; Reporting; Reproducibility; Research; Resectable; Resistance; Resolution; Sampling; Sensitivity and Specificity; Serum; Source; Specificity; Surface; Syndrome; TACSTD1 gene; Therapeutic; Time; Treatment outcome; Tumor stage; Tumor-Derived; Urine; Validation; Variant; Vesicle; Work; advanced disease; antibody detection; cancer diagnosis; cancer initiation; cancer stem cell; cancer therapy; candidate validation; clinical translation; cohort; design; detection assay; digital; disorder control; early screening; extracellular vesicles; fabrication; human disease; human model; improved; improved outcome; longitudinal analysis; minimally invasive; mouse model; nanoparticle; nanoplasmonic; novel; novel marker; pancreatic cancer model; pancreatic cancer patients; pre-clinical; premalignant; response; sample collection; success; translational impact; treatment response; tumor; tumor initiation

ABSTRACT Extracellular vesicles (EVs) hold great potential as novel biomarkers for minimally invasive detection of early stage tumors, since tumors abundantly secrete EVs that accumulate in the circulation and these EVs can transport factors that regulate tumor initiation, progression, and metastasis. However, most current EV analysis methods require pre-isolation of EVs prior to analysis and are low-throughput and impractical for clinical use. We recently developed a rapid, robust, isolation-free and inexpensive assay that directly quantifies tumor-derived EVs in small volumes (1~5 µL) of serum or plasma. In this assay, EVs that bind probes specific for two different EV target proteins produce a distance-regulated nanoplasmon-enhanced scattering (nPES) effect that allows sensitive detection of specific EVs. In a pilot study we used a nPES assay for a pancreatic cancer (PC)- associated EV marker to distinguish PC cases from non-malignant controls (patients with pancreatitis and healthy individuals) with high reproducibility, specificity, and sensitivity. This assay also differentiated PC tumor stages and tumor responses to neoadjuvant, outperforming CA19-9, a biomarker widely used for PC therapy assessment. Our nPES assay platform has multiple features required for research and clinical translation: 1) It is rapid, high-throughput, and inexpensive; 2) it does not employ EV isolation, avoiding a major source of EV assay variation; 3) it robustly and reproducibly quantifies EV biomarkers from small volumes of serum, plasma or urine, allowing its use in longitudinal analysis of mouse models of human disease; and 4) it can be readily adapted to diagnose and monitor cancers that express other EV biomarkers. Based on the success of our pilot study, we propose to develop and validate an automated and highly reproducible nPES EV assay to allow rapid and accurate PC diagnosis in clinical settings. We hypothesize that a nPES-based digital EV reader will equal or outperform the analytical performance of our current manual assay. We will build a diagnostic EV assay model for early PC detection by examining the ability of proteins reported to be enriched on the surface of EVs derived from PC stem cells or PC-initiating cells (e.g., CD44, CD133 and EpCAM) to diagnose patients with early stage PC and to differentiate them from patients with pre-malignant pancreatic lesions, hereditary syndromes or family history of PC, and individuals with normal pancreases. Specifically, we propose to: 1) Development and fabrication of Chip-nPES platform to achieve single EV resolution; 2) automate and refine our nPES-based digital EV reader to enhance assay reliability and reproducibility. We will also select and validate candidate EV capture and detection antibodies for PC diagnosis; 3) establish and evaluate a diagnostic model that integrates EV biomarkers with known cancer-associations; and 4) perform a pre-clinical validation of this assay in a third-party laboratory. The successful results of this work would have a significant translational impact in cancer management, through reliable and accessible screens for early detection of pancreatic cancer.

抽象的 细胞外囊泡（EV）作为新型生物标志物具有巨大的潜力，可用于早期微创检测 阶段肿瘤，因为肿瘤大量分泌 EV，并在循环中积聚，这些 EV 可以 调节肿瘤发生、进展和转移的转运因子然而，大多数当前的 EV 分析。 该方法需要在分析之前预先分离 EV，并且通量低且不适合临床使用。 最近开发了一种快速、稳健、免分离且廉价的检测方法，可直接量化肿瘤来源的 小体积 (1~5 µL) 血清或血浆中的 EV 在此测定中，EV 与两种不同的特异性探针结合。 EV 靶蛋白产生距离调节的纳米等离子体增强散射 (nPES) 效应，允许 在一项试点研究中，我们使用 nPES 检测胰腺癌 (PC)。 相关的 EV 标记物可将 PC 病例与非恶性对照（患有胰腺炎和 健康个体）具有高重现性、特异性和敏感性。该测定还可以区分 PC 肿瘤。 分期和肿瘤对新辅助治疗的反应，优于广泛用于 PC 治疗的生物标志物 CA19-9 我们的 nPES 检测平台具有研究和临床转化所需的多项功能：1) 它 快速、高通量且廉价；2) 不采用 EV 隔离，避免了 EV 的主要来源 测定变异；3) 能够稳健且可重复地定量少量血清、血浆中的 EV 生物标志物 或尿液，使其可用于人类疾病小鼠模型的纵向分析；4）它可以很容易地进行； 基于我们试点的成功，适用于诊断和监测表达其他 EV 生物标志物的癌症。 研究中，我们建议开发并验证一种自动化且高度可重复的 nPES EV 测定，以实现快速 我们发现基于 nPES 的数字 EV 阅读器将等同于临床环境中的准确 PC 诊断。 或超越我们当前手动检测的分析性能，我们将建立一个诊断 EV 检测模型。 通过检查据报道在 EV 衍生表面上富集的蛋白质的能力，进行早期 PC 检测 来自 PC 干细胞或 PC 起始细胞（例如 CD44、CD133 和 EpCAM）来诊断早期患者 PC 并将其与患有癌前胰腺病变、遗传综合征或家族的患者区分开来 具体来说，我们建议： 1) 发展和胰腺病史。 制造 Chip-nPES 平台以实现单一 EV 分辨率 2) 自动化和完善我们基于 nPES 的技术； 数字 EV 读取器，以提高检测的可靠性和重现性。我们还将选择和验证候选者。 用于 PC 诊断的 EV 捕获和检测抗体；3) 建立和评估诊断模型 将 EV 生物标志物与已知的癌症关联相结合；4) 对该测定进行临床前验证 这项工作的成功结果将对第三方实验室产生重大的转化影响。 癌症管理，通过可靠且易于使用的筛查来早期发现胰腺癌。

项目成果

A gradient of force in the endocytic coat is revealed by new coiled-coil force sensors

新型卷绕线圈力传感器揭示了内吞层中的力梯度

• 发表时间: 2024-09-14
• 作者: Yuan Rena;Jie Yangc;Y. Zhanga;J. Berroa
• 通讯作者: J. Berroa

Identifying the Phenotypes of Tumor-Derived Extracellular Vesicles Using Size-Coded Affinity Microbeads.

使用尺寸编码亲和微珠鉴定肿瘤来源的细胞外囊泡的表型。

• DOI: 10.1021/jacs.2c10042
• 发表时间: 2022-12-17
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Jiacheng Wu;Zhun Lin;Zhengyu Zou;Siping Liang;Minhao Wu;Tony Y. Hu;Yuanqing Zhang
• 通讯作者: Yuanqing Zhang