Novel nano-plasmonic technology for quantitative analysis of cancer exosomes

 描述(由申请人提供)：这项建议的主要目标是开发和应用一种新型的传感平台来检测和分子图谱外切体，用于监测多形性胶质母细胞瘤(GBM)的肿瘤进展和治疗反应。外切体是癌细胞主动脱落的膜结合磷脂纳米囊泡，该平台将被用来确定血清中外切体的丰度和组成。候选人Im HyungSoon在开发新的传感平台方面拥有丰富的经验，具有工程背景，现在的目标是过渡到生物医学癌症研究。通过培训，他将进一步发展进行自主翻译研究的技能集，并为癌症外显体研究领域做出重大贡献。这项工作将在马萨诸塞州总医院和哈佛医学院的系统生物学中心进行，由拉尔夫·韦斯莱德博士博士指导。为了扩大专业知识，他还聘请了Xandra Breakefield和Antonio Chiocca教授，Xandra Breakefield是国际公认的外体生物学专家，Antonio Chiocca教授是一位对翻译工作感兴趣的神经外科肿瘤学家，需要额外的指导和专业知识。顾问委员会的这些成员将促进贤顺的工作向独特和独立的调查方向过渡，并使他能够作为一名独立的生物医学癌症研究人员发展成功的职业生涯。该研究计划旨在首先开发一种微阵列纳米等离子体外切体传感平台(NPLEx)，用于血清中外切体蛋白质和核酸的多重图谱分析。外切体作为一种循环生物标记物具有独特的优势，但其分子组成的量化一直是具有挑战性的，特别是在临床环境中。这项工作源于使用等离子体纳米孔以无标记的方式灵敏地检测外切体并量化其标记表达水平的初步研究。在具体目标1中，将结合最先进的纳米制造技术和高通量读出系统，开发具有400个传感阵列的先进NPLEx平台。在具体目标2中，将应用NPLEx平台对GBM小鼠模型中的外切体进行系列分析。治疗前后的外切体纵向评估将用于确定与肿瘤进展和治疗反应相关的特定外切体标记物，以预测治疗效果。拟议的平台将允许对肿瘤来源的外切体进行全面分析，以揭示原发和转移肿瘤的分子特征，并提供信息以指导有效的治疗方案。新的NPLEx系统可能是一个促进癌症研究和临床实践的变革性工具。

 DESCRIPTION (provided by applicant): The broad goal of this proposal is to develop and apply a novel sensing platform to detect and molecularly profile exosomes for monitoring tumor progression and treatment response in glioblastoma multiforme (GBM). Exosomes are membrane-bound phospholipid nanovesicles actively shed by cancer cells and the platform will be used to determine the abundance and composition of exosomes in serum. The candidate, Hyungsoon Im, has extensive experience in developing new sensing platforms with a background in engineering and now aims to transition to biomedical cancer research. Through the training, he will further develop skill sets to conduct independent translational research and make significant contribution to the field of cancer exosome research. This work will be conducted in the Center for Systems Biology at Massachusetts General Hospital and Harvard Medical School under the mentorship of Dr. Ralph Weissleder MD, PhD. To expand expertise, he has also brought in Xandra Breakefield, an internationally recognized expert in exosome biology and Prof. Antonio Chiocca, a neurosurgical oncologist with interest in translational work for additional mentoring and expertise. These members of the advisory committee will facilitate the transition of Hyungsoon's work toward a unique and independent line of investigation and allow him to develop a successful career as an independent biomedical cancer researcher. The research plan aims to first develop a microarray nano-plasmonic exosome sensing platform (nPLEX) for multiplexed profiling of exosome proteins and nucleic acids in serum. Exosomes have unique advantages as a circulating biomarker, but the quantification of their molecular composition has been challenging especially in clinical settings. This work stems from preliminary research using plasmonic nanoholes for sensitive detection of exosomes in a label-free manner and quantification of their marker expression levels. In specific aim 1, an advanced nPLEX platform with 400 sensing arrays will be developed by integrating with state-of-the- art nanofabrication techniques and a high-throughput readout system. In specific aim 2, the nPLEX platform will be applied for serial analyses of exosomes in GBM mouse models. Longitudinal assessment of exosomes pre- and post-treatment will be used to identify specific exosomes markers that correlate with tumor progression and treatment response to predict therapeutic efficacy. The proposed platform would allow comprehensive analyses of tumor-derived exosomes to shed light on molecular profiles of primary and metastatic tumors and provide information to guide efficient treatment schemes. The new nPLEX system could be a transformative tool facilitating cancer research and clinical practice.