Decoding Individual Exosomes in Cancer

解码癌症中的个体外泌体

• 批准号: 10440265
• 负责人: GREGORY W FARIS
• 金额: $ 22.63万
• 依托单位: NUMENTUS TECHNOLOGIES INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10440265
• 关键词: Address; Biological; Biology; Blood; Blood specimen; Breast Cancer Cell; Breast Cancer cell line; Caliber; Cancer Biology; Cancer Prognosis; Cardiovascular Diseases; Cell Line; Cells; Clinical; Clinical Management; Communication; Detection; Development; Diagnosis; Diagnostic; Disease; Dyes; Gel; Goals; Heat-Shock Proteins 70; Heterogeneity; Homeostasis; Human; Image; Immune response; In Situ; Individual; Label; Lead; Learning; Life; Lipid Bilayers; Lipids; Liposomes; Liquid substance; Malignant Neoplasms; Mammary Neoplasms; Masks; Medicine; Membrane Proteins; Methods; MicroRNAs; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Normal Cell; Nucleic Acids; Oncogenic; Outcome; Pathology; Patients; Physiological Processes; Physiology; Plasma; Polymerase Chain Reaction; Process; Protein Analysis; Proteins; Reading; Recurrence; Research; Reverse Transcription; Sampling; Screening for cancer; Serum; Signal Transduction; Speed; Technology; Testing; Therapeutic; Tissues; Tumor Burden; Tumor-Derived; Work; amplification detection; base; cancer cell; cell type; clinically relevant; design; exosome; extracellular vesicles; fluorescence imaging; imaging platform; imaging system; improved; intercellular communication; liquid biopsy; malignant breast neoplasm; malignant phenotype; nanomedicine; nanoparticle; nanoscale; new technology; novel; novel diagnostics; particle; polyacrylamide gels; prognostic; prognostic tool; prognostic value; stem cell biology; therapeutic nanoparticles; tumor

PROJECT SUMMARY There is increasing recognition that extracellular vesicles (EVs)¾micrometer- or nanometer-sized lipid particles containing protein and nucleic acid cargoes (information)¾are highly promising for new diagnostics/prognostics and even have therapeutic value. There is, however, an unmet need for methods able to solve a fundamental problem confounding the exploitation of EV information in biology and medicine¾EVs are naturally highly heterogeneous particles. In this pilot (R21) project, our experimental goal is to address the need for analyzing EV heterogeneity (subpopulations) by focusing on particles called exosomes. Specifically, our new technology platform is designed to address the problem of resolving bulk exosomal subpopulations by directly correlating surface protein and nucleic acid (microRNA or miRNA) cargoes of single exosomes by performing highly multiplexed fluorescence imaging analysis. Our ultimate goal is to develop a unique imaging platform for the high-throughput, high-content analysis of the protein and nucleic acid cargoes of single exosomes (and other EVs) obtained from any biological sample. This new platform could enable novel diagnostic/prognostic “liquid biopsy” tests for managing cancers as well as other pathologies, such as neurodegenerative and cardiovascular diseases. To achieve our experimental goal, in the work for Aims 1 and 2 we will develop and optimize our imaging platform for experimental multiplexed analysis of human breast cancer cell exosomes and their miRNA cargoes. These Aims will validate our platform for in situ miRNA analysis of single exosomes from cancer cells with documented exosomal miRNA signatures. During Aim 3, we will use the validated platform to correlate the surface protein display and miRNA cargo of individual exosomes released by human breast cancer cell lines representing early- and late-stage cancers and compare these with a normal cell line control. Aim 3 will test the capability of our platform to resolve exosomal subpopulations in an original bulk sample, based on correlated signals for surface proteins and miRNA cargoes of single exosomes. We propose that our novel imaging platform has the potential to become a new diagnostic/prognostic tool for the clinical management of breast cancer, other tumor types, and other pathologies in which EV/exosomal analysis could provide clinically useful information. Recent research demonstrates that breast-cancer-cell- derived exosomes can transport cargoes that promote oncogenic or malignant phenotypes. Thus, exosomes released from breast tumors could carry information with critical diagnostic/prognostic value, which could be sampled from blood or other patient fluids (a “liquid biopsy”). This capability may also assist in developing exosome-based therapies for human tumor types.

项目概要 人们越来越认识到细胞外囊泡 (EV) 是微米或纳米大小的脂质 含有蛋白质和核酸货物的颗粒（信息）在新产品中非常有前景 然而，对能够进行诊断/预测甚至具有治疗价值的方法的需求尚未得到满足。 解决生物学和医学中电动汽车信息利用的基本问题：电动汽车 是天然高度异质的粒子，在这个试点 (R21) 项目中，我们的实验目标是解决 需要通过关注称为外泌体的颗粒来分析 EV 异质性（亚群）。 我们的新技术平台旨在通过以下方式解决大量外泌体亚群的解析问题 通过直接关联单个外泌体的表面蛋白和核酸（microRNA 或 miRNA）货物 我们的最终目标是开发独特的成像。 用于对单一蛋白质和核酸货物进行高通量、高内涵分析的平台 从任何生物样本中获得的外泌体（和其他 EV）可以实现新颖的功能。 用于管理癌症以及其他病理的诊断/预后“液体活检”测试，例如 神经退行性疾病和心血管疾病。 为了实现我们的实验目标，在目标 1 和 2 的工作中，我们将开发和优化我们的成像 人乳腺癌细胞外泌体及其 miRNA 实验多重分析平台 这些目标将验证我们对癌细胞单个外泌体进行原位 miRNA 分析的平台。 在目标 3 中，我们将使用经过验证的平台来关联记录的外泌体 miRNA 特征。 人乳腺癌细胞释放的单个外泌体的表面蛋白展示和 miRNA 货物 Aim 3 将代表早期和晚期癌症的细胞系，并将其与正常细胞系对照进行比较。 测试我们的平台解析原始批量样品中外泌体亚群的能力，基于 单个外泌体表面蛋白和 miRNA 货物的相关信号。 我们认为我们的新型成像平台有潜力成为一种新的诊断/预后工具 用于乳腺癌、其他肿瘤类型以及 EV/外泌体的其他病理的临床管理 分析可以提供临床上有用的信息。最近的研究表明乳腺癌细胞。 衍生的外泌体可以运输促进致癌或恶性表型的货物。 从乳腺肿瘤释放的信息可能携带具有关键诊断/预后价值的信息，这可能是 从血液或其他患者体液中取样（“液体活检”）此功能也可能有助于发展。 针对人类肿瘤类型的外泌体疗法。