NSF-GACR: An Optical Biosensing Platform for Simultaneous Detection and Quantification of Exosomes and Exosomal Cargo Biomarkers

生物传感器已被证明在生物医学研究、医疗保健和制药方面极具价值，并有望成为现代医学诊断的一个组成部分。最近，循环外泌体（细胞内形成的微小囊状结构，含有细胞的一些蛋白质，DNA和RNA）已成为医学诊断的有希望的生物标志物，因为它们包含有关其起源细胞的信息。尽管外显体已被报道存在于所有主要的体液中，但由于外显体亚群的高度可变性、少量的货物分子以及外显体纯化的低回收率和可重复性，外显体的有效检测和分子分析仍然特别具有挑战性。康奈尔大学（Ithaca）和布拉格光子学和电子学研究所（IPE）之间的国际合作项目将通过开发一种新的生物传感方法来解决这些挑战，该方法可以同时检测和定量外泌体和外泌体货物分子，以充分利用外泌体中包含的丰富信息。外泌体和外泌体货物生物标志物的同时鉴定和定量具有广泛的应用，从生物医学诊断到基于外泌体的新药物递送载体的表征。该项目为美国和捷克共和国STEM领域的下一代劳动力提供培训机会。来自弱势群体的研究生和本科生将在这个高度跨学科的研究项目中接受培训。研究生将于每年夏天参观合作实验室，以促进合作并获得在多样化环境中进行研究的经验。该项目的目标是将两个研究机构（一个位于美国，另一个位于捷克共和国）联合起来，开发一种新的生物传感方法，可以同时检测和定量外泌体和外泌体货物分子，以便充分利用外泌体中包含的丰富信息。该生物传感平台将基于光激发表面等离子体，能够快速灵敏地检测生物分子，并实时研究生物分子相互作用，而无需标记。该项目的具体目标是：(1)开发一种基于等离子体生物传感器的捕获，定量和分析选定外泌体的新方法；(2)开发等离子体生物传感器捕获的外泌体的控制裂解方法，以及从裂解的外泌体中释放的miRNA （microrna）和蛋白质货物分子的超灵敏检测方法；(3)将这些方法整合到一个单一的生物传感平台中，用于选择性捕获和分析外泌体以及检测多种外泌体货物mirna和蛋白质。该项目将推进利用外泌体作为新兴生物标志物的基础知识，以及生物传感领域的纳米科学和纳米技术。这项工作的结果将不仅在生物传感领域，而且在医学诊断、细胞生物学、生物材料、表面化学和等离子体学等领域产生重大影响。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Biosensors have proven extremely valuable in biomedical research, healthcare and pharmaceutics and are envisioned to become an integral part of modern medical diagnostics. Recently, circulating exosomes (tiny sac-like structures that were formed inside cells and contain some of the cell's proteins, DNA, and RNA) have emerged as promising biomarkers for medical diagnosis because they contain information about their cells of origin. Despite exosomes having been reported to be present in all major body fluids, effective detection and molecular analysis of exosomes remains particularly challenging due to the high variability in exosome subpopulations, small amounts of cargo molecules, and low recovery and reproducibility of exosome purification. This international collaborative project between Cornell University (Ithaca) and the Institute of Photonics and Electronics, Prague (IPE) will address these challenges by developing a new biosensing methodology that enables simultaneous detection and quantification of exosomes and exosomal cargo molecules to take full advantage of the richness of information contained in exosomes. Simultaneous identification and quantification of exosomes and exosomal cargo biomarkers have broader applications from biomedical diagnosis to the characterization of new drug delivery carriers based on exosomes. This project provides training opportunities for the next generation workforce in STEM fields in the US and the Czech Republic. Graduate and undergraduate students from underrepresented groups will receive training in this highly interdisciplinary research project. Graduate students will visit the collaborative labs each summer to foster the collaboration and gain experience on conducting research in a diverse environment.The goal of this project is to bring together two research institutions, one located in the US and the other in the Czech Republic, to develop a new biosensing methodology that enables simultaneous detection and quantification of exosomes and exosomal cargo molecules in order to take full advantage of the richness of information contained in exosomes. The biosensing platform will be based on optically excited surface plasmons that enable rapid and sensitive detection of biomolecules and investigation of biomolecular interactions in real time without the need for labeling. The specific objectives of the project are: (1) to develop a novel method for the capture, quantification, and profiling of selected exosomes based on a plasmonic biosensor; (2) to develop methods for controlled lysis of exosomes captured in the plasmonic biosensor and for the ultrasensitive detection of miRNA (microRNAs) and protein cargo molecules released from the lysed exosomes; and (3) to integrate these methods into a single biosensing platform for selective capture and profiling of exosomes and detection of multiple exosomal cargo miRNAs and proteins. The project will advance fundamental knowledge of utilizing exosomes as emerging biomarkers and nanoscience and nanotechnology in biosensing. The results of this work will make significant impacts not only in the field of biosensing but also in the fields of medical diagnosis, cell biology, biomaterials, surface chemistry and plasmonics.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.