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改变了远处器官中的微环境 细胞这些预先确定的微环境，称为“转移前小生境”， 并使癌细胞成功转移。最近的研究表明， 具有不同整联蛋白的外泌体，以在远处器官中准备转移前小生境， 与肺转移有关，与肝转移有关，与脑转移有关（Hoshino等， Nature 2015，527，329）。这些发现指出，来自于人的血浆外泌体的整合素谱可能与来自于人的血浆外泌体的整合素谱有关。 癌症患者可用于预测转移和向器官扩散。为了确定这是否 知识可以付诸实践，需要进一步评估。评价外泌体生物标志物在临床中的应用 样本并在临床环境中使用，需要简单但高灵敏度和高特异性的技术 但目前缺乏。这是因为外泌体很小（低于200 nm），并且肿瘤来源的外泌体 与来自各种组织和造血细胞的大量非肿瘤外泌体背景混合， 使得临床样品中外来体生物标志物的定量检测具有挑战性。 本项目的目标是验证和扩展外泌体整合素的转移预测深度 通过提供以前被集中到批量测定中的单个外泌体景观。我们提出了一个简单的双重 成像单囊泡技术（DISVT），其能够检测靶向表面蛋白标记物， 个体外泌体和定量血浆样品中的靶特异性囊泡亚型。DISVT 直接从稀释的血浆中捕获外泌体到多孔金室载玻片上， 外泌体与膜染料和激光激发荧光成像，并检测感兴趣的表面蛋白 使用光散射金纳米颗粒和单颗粒水平的暗场成像。使用人类表皮 生长因子受体2（HER 2）阳性乳腺癌作为疾病模型，我们的初步研究 结果表明，该方法能较好地早期检测BC， 打开了在单个囊泡水平上探测和定量外泌体生物标志物以供临床使用的可能性。 使用DISVT，我们建议定量表征来自以下患者的血浆样品中携带整合素的外泌体： 在诊断后3 - 5年内发生转移性复发的局部晚期BC患者， 与一组患者对照组和健康对照组进行比较。我们将基于八种整合素标志物进行表征 对器官转移预测的潜力。如果成功，这个项目将1）导致一个简单的， 强大的SVT，可用于基础囊泡研究和临床应用，2）在很大程度上扩展了我们的理解 外泌体整合素在预测乳腺癌转移和向器官性方面的临床价值， 局部晚期、晚期和早期患者的单个外泌体整合素谱图。成果 可以加速外泌体生物标志物的翻译，用于癌症诊断、诊断和监测。