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.

项目摘要 人们越来越认识到细胞外蔬菜（EV）¾microter-或纳米级脂质 含有蛋白质和核酸货物的颗粒（信息）¾非常有希望 诊断/预后，甚至具有治疗价值。但是，有未满足的方法 解决一个基本问题，以混淆生物学和Medicine¾EV中电动汽车信息的开发 是天然高度异质的颗粒。在这个飞行员（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）。具体来说， 我们的新技术平台旨在解决通过通过 直接将单个外泌体的表面蛋白和核酸（microRNA或miRNA）货物相关联 进行高度多重的荧光成像分析。我们的最终目标是开发独特的成像 对单个蛋白质和核酸货物的高通量，高含量分析的平台 从任何生物样品获得的外泌体（和其他EV）。这个新平台可以使小说 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，我们将发展和优化我们的想象力 实验性多路复用人类乳腺癌细胞外泌体及其miRNA的平台 货物。这些目标将验证我们的平台，以对癌细胞的单个外泌体进行原位miRNA分析 带有记录的外泌体miRNA签名。在AIM 3期间，我们将使用经过验证的平台进行关联 人类乳腺癌细胞释放的单个外泌体的表面蛋白显示和miRNA货物 代表早期和晚期癌症的线，并将其与正常的细胞线对照进行比较。目标3意志 基于 单个外泌体的表面蛋白和miRNA货物的相关信号。 我们建议我们的新型成像平台有可能成为一种新的诊断/预后工具 用于乳腺癌的临床管理，其他肿瘤类型和其他病理，其中EV/外泌体 分析可以提供临床上有用的信息。最近的研究表明，乳腺癌 - 衍生的外泌体可以运输促进致癌或恶性表型的货物。那，外泌体 从乳腺肿瘤中释放的可能具有关键诊断/预后价值的信息，这可能是 从血液或其他患者液体中取样（“液体活检”）。此功能也可能有助于发展 基于外部的人类肿瘤类型的疗法。