Implementation of a methodology for extracellular vesicle extraction from cell lines and human plasma

Exosomes are vesicles with a diameter ranging from 30 to 100 nm whose synthesis is mediated by the endosomal system. The invagination of the cell membrane generates the early endosome, which matures into the late endosome and forms multivessel bodies (CMV), which fuse with the cell membrane to release the vesicles into the extracellular environment. These vesicles are composed of a bi-lipid membrane rich in cholesterol, ceramides, and phosphatidylserine (PS) residues. Inside they can store DNA, RNA, MicroRNAs, proteins, and lipids. Exosomes have a variety of functions in different settings ranging from molecule transport and cell-cell communication at the local or systemic level to immune regulation.

Exosomes play a fundamental role in the tumor microenvironment by favoring communication between tumor cells or between these tumor cells and immune system cells, which is why exosome secretion is considered to be a mechanism by which the tumor favors its own development by transporting immune system modulating molecules such as HLA-G, oncoproteins and growth factors. Additionally, horizontal transfer of various types of RNA has also been described as giving greater aggressiveness to tumor cells. Considering the importance of exosomes in the development and progression of cancer, this project is proposed in order to generate in our laboratory, a methodology for the extraction and characterization of extracellular vesicles, with particular interest in exosomes, establishing the basis for the development of more robust research with these cellular structures and thus contribute to advancing the understanding of cancer biology.

- The active participation of two undergraduate students in biology with interest in becoming involved in basic research associated with cell culture, molecular markers and gene expression is expected. At the end of the work, students are expected to present their undergraduate work in written and oral communication. One of the students will work on the characterization of exosomes by microscopy, immunoassays and flow cytometry and the other student will be in charge of the characterization of RNAs transported by exosomes.

- Linking of a young researcher who will be trained to support the experimental work with cell lines this with the funds requested within the proposal.

- Attachment of a research assistant by the INC to support the work related to the treatment of plasma samples and chromatography.

- The intention is to articulate the activities of those involved in the research project in such a way as to strengthen the research and develop skills that are in line with the scientific method among the young participants