ABSTRACT

Extracellular vesicles (EVs) are lipid bi-layered nanoparticles shed from the cells that carry RNA, DNA, transmembrane and cytosolic proteins. The variety in EV size, cargo and origin attracted researchers to decipher the mechanisms that have been involved in packaging, secretion, uptake and roles of EVs on cells in vivo and in vitro, lightening the path for biomarker studies for diagnosis, prognosis, therapy and therapy monitoring. They are one of the many means that cells use to communicate with neighboring and distant cells and tissues. With improvements in next-generation sequencing technologies and increased resolution of mass spectrometry for proteomic analysis, EVs have been shown to take role in angiogenesis, epithelial-to-mesenchymal transition, stemness in cancer, malignancy, metastasis and drug resistance.

Although exosomes show unprecedented promising advantages over other biotargets in the circulation for clinical use, a major challenge rapidly emerging in the field of EV utilization for clinical and non-clinical applications is the absence of reproducible, inexpensive and robust tools for efficient sorting and isolation of EV populations at a high yield. The field lacks a clear consensus over an optimum approach or a tool for isolation of EVs avoiding contamination with many other proteins and such other biostructures and reproducible procedures for downstream analysis of EV cargo and content. Existing approaches for EV isolation include a variety of methods. Additionally, methods for the exosome-derived analyte isolation, library preparation for sequencing, and downstream analysis including genomic, proteomic and metabolic analysis are highly varied. Hence, there is a need for well-developed experimental tools, interlaboratory evaluations and in-depth descriptions of experimental steps and designs to ensure reliable, robust and reproducible experiments and tools. In this talk, we will describe a new technique, i.e., Exosome Total Isolation Chip (ExoTIC), that is developed in our lab to isolate EVs and EV subpopulations from a variety of sample types including plasma and culture media. We will present further downstream genomic and proteomic analysis of these EVs focusing on applications in cancer and cardiovascular disorders.

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