Surface exosome integrin profiling to predict organotropic metastasis of breast cancer

表面外泌体整合素分析预测乳腺癌的器官转移

• 批准号: 10654221
• 负责人: Thang Ba Hoang
• 金额: $ 41.55万
• 依托单位: UNIVERSITY OF MEMPHIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10654221
• 关键词: Acceleration; Biological; Biological Assay; Biological Markers; Body Fluids; Breast Cancer Detection; Breast Cancer Model; Breast Cancer Patient; Breast cancer metastasis; CD81 gene; Cancer Diagnostics; Cancer Etiology; Cancer Patient; Cells; Cessation of life; Clinical; Code; Consumption; Country; Darkness; Detection; Diagnosis; Disease model; Distant; Distant Metastasis; Dyes; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor Receptor; Evaluation; Fluorescence; Goals; Gold; Hematopoietic; Human; Image; Image Analysis; Individual; Integrin beta4; Integrins; Knowledge; Label; Lasers; Malignant Neoplasms; Maps; Membrane; Membrane Proteins; Metastatic Neoplasm to the Liver; Metastatic Neoplasm to the Lung; Metastatic malignant neoplasm to brain; Metastatic/Recurrent; Methods; Molecular; Monitor; Nature; Neoplasm Metastasis; Organ; Outcome; Patients; Plasma; Process; Provider; Pythons; Research; Risk; Sampling; Slide; Specificity; Surface; Technology; Tissues; Translations; Tumor-Derived; United States; Validation; Vesicle; Woman; advanced breast cancer; antibody conjugate; cancer biomarkers; cancer cell; cyanine dye 5; design; effective therapy; exosome; fluorescence imaging; human subject; image processing; imaging approach; imaging system; interest; light scattering; lipophilicity; malignant breast neoplasm; microscopic imaging; molecular subtypes; mortality; nanoGold; nanomedicine; neoplastic cell; optical imaging; particle; prognostic; protein biomarkers; protein profiling; technology development; tumor; undergraduate student

PROJECT SUMMARY It is well established that tumors modify microenvironments in distant organs before the arrival of cancer cells. These predetermined microenvironments, termed ‘pre-metastatic niches’, encourage the outgrowth of the incoming cancer cells and make metastasis sucessful. Recent studies have shown that tumors send exosomes with distinct integrins to prepare pre-metastatic niches in distant organs, with 64 and 61 associated with lung metastasis, v5 associated with liver metastasis, and IIb3 with brain metastasis (Hoshino et al., Nature 2015, 527, 329). These findings point to the possibility that the integrin profiles of plasma exosomes from cancer patients could be used to predict metastasis and organotropic dissemination. To establish whether this knowledge can be put to practice use, further evaluation is required. To evaluate exosomal biomarkers in clinical samples and use it in the clinical settings, facile but high sensitivity and high specificity technologies are needed but currently lacking. This is because exosomes are small (lower than 200 nm) and the tumor-derived exosomes are mixed with a vast background of non-tumor exosomes from various tissues and hematopoietic cells, which makes quantitative detection of exosomal biomarkers in clinical samples challenging. The goal of this project is to validate and extend the depth of exosomal integrins for metastasis prediction by providing single exosome landscapes that were previously lumped into bulk assays. We propose a facile dual imaging single vesicle technology (DISVT) that is capable of detecting targeted surface protein markers on individual exosomes and quantifying the target-specific vesicle subtypes in plasma samples. The DISVT captures exosomes directly from diluted plasma onto a multi-well gold chamber slide, localize the individual exosomes with membrane dye and laser excited fluorescence imaging, and detect surface proteins of interests using light scattering gold nanoparticles and dark field imaging at single particle level. Using human epidermal growth factor receptor 2 (HER2)-positive breast cancer as the disease model, our preliminary studies have shown that this DISVT, but not traditional enzyme-linked immunosorbent assay, can detect BC at early-stage, opening the possibilities to probe and quantify exosomal biomarkers at single vesicle level for clinical use. Using DISVT, we propose to quantitatively characterize integrin-carrying exosomes in plasma samples from locally-advanced BC patients who developed metastatic recurrence within three to five years of diagnosis and compare with a group of patient controls and healthy control. We will characterize eight integrin markers based on their potential for organotropic metastasis prediction. If successful, this project will 1) lead to a facile and robust SVT that can be used for basic vesicle research and clinical use, and 2) largely expand our understanding of the clinical value of exosomal integrins for the prediction of BC metastasis and organotropism by providing a map of single exosome integrin profiles for locally-advanced, advanced, and early-stage patients. The outcomes may accelerate the translation of exosomal biomarkers for cancer diagnostics, prognostics, and monitoring.

项目摘要 众所周知，肿瘤在癌症到来之前会改变远处器官的微环境 细胞。这些预定的微环境被称为“前迁移壁ches”，鼓励了生长 传入的癌细胞并使转移成功。最近的研究表明肿瘤发送 具有独特整合素的外泌体以制备远处器官中的te-64和61相关的遥不可及的壁ni 与肺转移，与肝转移相关的V5和脑转移的IIB3（Hoshino等， 自然2015，527，329）。这些发现指出了血浆外泌体的整联蛋白轮廓的可能性 癌症患者可用于预测转移和器官传播。确定是否 知识可以练习使用，需要进一步评估。评估临床中的外泌体生物标志物 样品并在临床环境中使用，需要敏感但高灵敏度和高特异性技术 但目前缺乏。这是因为外泌体小（低于200 nm）和肿瘤衍生的外泌体 与来自各种组织和造血细胞的非肿瘤外泌体的广泛背景混合在一起， 在临床样品挑战中对外泌体生物标志物进行定量检测。 该项目的目的是验证和扩展外泌体整合素的深度进行转移预测 通过提供以前被批量测定的单个外部景观。我们提出了一个轻松的双重 成像单囊泡技术（DISVT），能够在 单个外泌体并量化等离子体样品中靶标特异性囊泡亚型。 disvt 将外泌体直接从稀释的血浆中捕获到多孔金室滑梯上 具有膜染料和激光激发荧光成像的外泌体，并检测感兴趣的表面蛋白 使用光散射金纳米颗粒和单个颗粒水平的暗场成像。使用人表皮 生长因子受体2（HER2）阳性乳腺癌作为疾病模型，我们的初步研究具有 表明这种DESVT但没有传统的酶相关的免疫吸附测定可以在早期检测BC， 打开在单个囊泡水平上探测和量化外泌体生物标志物的可能性，以供临床使用。 使用DESVT，我们建议定量地表征来自等离子体样品中的整联蛋白携带外泌体 在诊断的三到五年内发生转移性复发的本地进取的卑诗省患者 与一组患者对照和健康对照进行比较。我们将表征八个基于整联蛋白标记的 关于它们进行器官转移预测的潜力。如果成功的话，该项目将1）导致一个便利和 可用于基本囊泡研究和临床用途的强大SVT，2）在很大程度上扩展了我们的理解 外泌体整合素在预测BC转移和有机体的临床价值中，通过提供 用于本地进化，高级和早期患者的单个外泌体整合素剖面图。结果 可能会加速用于癌症诊断，预后和监测的外泌体生物标志物的翻译。