Multiplexed analysis of exosomes in cancer nano therapy

癌症纳米疗法中外泌体的多重分析

基本信息

批准号：
9487955
负责人：
RALPH WEISSLEDER, MD, PHD
金额：
$ 38.9万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-15 至 2021-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9487955
关键词：
Affinity Biological Assay Biological Testing Blood Circulation Blood Platelets Cancer Patient Cells Clinic Clinical Core Biopsy Detection Diagnostic Procedure Drug resistance Elements Endothelial Cells Erythrocytes Evaluation Event Film Generations Goals Gold Image Immunocompetent Leukocytes Ligands Malignant Neoplasms Malignant neoplasm of ovary Measures Mesothelial Cell Messenger RNA Methods Microfluidics Modeling Molecular Molecular Analysis Molecular Profiling Monitor Mus Nanotechnology Noise Outcome Parents Patients Periodicity Pharmaceutical Preparations Printing Process Protein Analysis Proteins RNA Retrieval Sampling Series Serum Signal Transduction Specimen Surface System Systems Biology Techniques Technology Therapeutic Therapeutic Trials Time Tumor-Derived Vesicle anticancer research base cancer cell cancer therapy chemotherapy clinical practice cost cost effective design drug efficacy exosome experimental study human model improved innovation insight lithography macrophage method development mouse model nano nanoencapsulated nanoparticle nanoplasmonic nanotherapeutic nanotherapy new technology peripheral blood point of care point-of-care diagnostics protein biomarkers protein expression protein profiling public health relevance rapid technique response scale up sensor therapeutic nanoparticles transmission process tumor tumor initiation tumor progression validation studies

项目摘要

DESCRIPTION (provided by applicant): Detection of reliable markers of therapy response and drug resistance remains a major clinical challenge in ovarian cancer. This is particularly true for a series of nanotherapeutics entering the clinical arena. One little explored opportunity lies in the analysis of exosomes, 50-200 nm sized vesicles continuously shed into the circulation. Importantly such tumor derived exosomes are abundant (1011 vesicles/ml of peripheral blood in cancer patients), are fairly stable over time and contain proteins reflective of those found in parent tumors. A current major challenge and opportunity is the development of methods for rapidly determining the abundance and composition of cMVs from clinical samples. We have recently developed a highly sensitive, nanotechnology-based, point-of-care diagnostic method termed nPLEX (nano Plasmonic Exosome) which comprises periodic nanohole arrays fabricated in an opaque gold film for transmission plasmon imaging (Nat Biotechnol 2014;32, 490-5). We have shown that i) such analyses are exquisitely sensitive, ii) allow profiling of dozens of proteins on and inside exosomes thus allowing cell of origin studies and iii) that nPLEX analysis allows sophisticated therapy assessment. The goal of this application is to advance the nPLEX technology by i) expanding it to allow parallel profiling of protein and RNA, ii) validating it as a therapeutic read-out of nanoparticle therapeutics in mouse models and in the clinic. We hypothesize that the approach will be more sensitive and comprehensive in exosomal analysis than is currently possible and allow treatment evaluation. The proposed integrated profiling method has the potential to transform nano therapeutic trials, cancer research and clinical practice. It will enable objective therapeutic read-outs, events that occur before conventional clinical metrics. It will facilitate extensive profiling of exosome in paucicellular clinical specimens, significantly reduce costs, and can be easily combined with other downstream analyses.

描述（由申请人提供）：检测治疗反应和耐药性的可靠标志物仍然是卵巢癌的主要临床挑战。一系列纳米治疗药物进入临床领域，其中一个鲜为人知的机会在于外泌体的分析，50-200 nm 大小的囊泡不断流入循环系统，重要的是此类肿瘤来源的外泌体非常丰富（癌症患者的外周血中有 1011 个囊泡）。），随着时间的推移相当稳定，并且含有反映亲本肿瘤中发现的蛋白质的蛋白质，当前的主要挑战和机遇是开发快速确定临床 cMV 丰度和组成的方法。我们最近开发了一种高度灵敏、基于纳米技术的即时诊断方法，称为 nPLEX（纳米等离子体外泌体），它包含在不透明金膜中制造的周期性纳米孔阵列，用于传输等离子体成像（Nat Biotechnol 2014；32， 490-5) 我们已经证明，i) 此类分析非常灵敏，ii) 可以对外泌体上和内部的数十种蛋白质进行分析，从而允许细胞进行分析。 iii) nPLEX 分析可实现复杂的治疗评估该应用的目标是通过 i) 扩展 nPLEX 技术以允许对蛋白质和 RNA 进行并行分析，ii) 验证其作为治疗读数。纳米颗粒疗法在小鼠模型和我们追求的是，该方法在外泌体分析方面将比目前的方法更加灵敏和全面，并且可以进行治疗评估，并且有望改变纳米治疗试验、癌症研究和临床实践。它将有助于对少细胞临床样本中的外泌体进行广泛的分析，显着降低成本，并且可以轻松地与其他下游分析相结合。