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

事实证明，生物传感器在生物医学研究、医疗保健和制药学中具有极其重要的价值，并有望成为现代医学诊断的重要组成部分。最近，循环外切体(在细胞内形成的微小囊状结构，包含细胞中的一些蛋白质、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。尽管已报道在所有主要体液中都存在外切体，但由于外切体亚群的高度变异性、少量的货物分子以及外切体纯化的低回收率和重复性，外切体的有效检测和分子分析仍然具有特别大的挑战性。康奈尔大学(Ithaca)和布拉格光子学和电子研究所(IPE)之间的这一国际合作项目将通过开发一种新的生物传感方法来应对这些挑战，该方法能够同时检测和量化外显体和外显体货物分子，以充分利用外显体所包含的丰富信息。从生物医学诊断到基于外切体的新型药物载体的表征，外切体和外切体货物生物标记物的同时识别和定量有着广泛的应用。该项目为美国和捷克STEM领域的下一代劳动力提供培训机会。来自代表性不足群体的研究生和本科生将接受这一高度跨学科的研究项目的培训。研究生每年夏天将访问合作实验室，以促进合作并获得在不同环境中进行研究的经验。该项目的目标是将位于美国和捷克共和国的两个研究机构聚集在一起，开发一种新的生物传感方法，能够同时检测和量化外体和外体货物分子，以充分利用外体所包含的丰富信息。生物传感平台将基于光学激发的表面等离子体，能够快速而灵敏地检测生物分子，并实时研究生物分子之间的相互作用，而不需要标记。该项目的具体目标是：(1)开发一种新的基于等离子生物传感器的方法来捕获、定量和分析选定的外切体；(2)开发在等离子生物传感器中捕获的外切体的受控裂解以及对从裂解的外切体中释放的miRNA(MicroRNAs)和蛋白质货物分子进行超灵敏检测的方法；以及(3)将这些方法整合到一个单一的生物传感平台中，用于选择性地捕获和剖析外切体以及检测多个外切体货物miRNAs和蛋白质。该项目将促进利用外来体作为新兴生物标志物以及生物传感中的纳米科学和纳米技术的基础知识。这项工作的结果不仅将在生物传感领域产生重大影响，而且将在医疗诊断、细胞生物学、生物材料、表面化学和等离子体领域产生重大影响。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.