Analysis of Extracellular Vesicles (EVs) Derived from CAR-T Cells

Maria Margarita Labastida Arzate

Affiliation: Faculty of Pharmacy and Biochemistry, Faculty of Medicine, University of Buenos Aires, Argentina; Faculty of Medicine, University of Freiburg, Germany

Keywords: Extracellular Vesicles (EVs), CAR-T Cells, Biomarkers, Surface Protein Signature, Isolation Method

Categories: Demetrios Project, News and Views, Life Sciences, Medicine

DOI: 10.17160/josha.10.4.924

Languages: English

Extracellular vesicles (EVs) conform a heterogeneous group of lipid bilayer membrane particles naturally released by cells. They vary in size, surface composition, mechanism of formation, biochemical content, and function within the human body. EVs are classified in 3 groups, depending on their size and biogenesis: exosomes (originated by the endolysosomal system, measuring 60-150nm in diameter), microvesicles (direct budding from the plasma membrane, measure 100 – 1000 nm size) and apoptotic bodies (released by dying cells, size >1μm). A very important feature of the EVs is the delivery of information by horizontal transfer between the origin cell and the recipient one, evidence suggests that the uptake of EVs by recipients’ cells can induce changes in their own characteristics and function. CAR T-cells are genetically modified T-cells that express on their surface a Chimeric Antigen T-cell Receptor (CAR) specific for a unique antigen expressed in the surface of tumor cells. Upon activation with several molecules or cytokines, normal T-cells and CART-cells can express related co-receptors and derived active T-cell biomarkers that will help them to perform a successful immunological synapsis (IS) within the human body, and to induce a successful TCR activation for a subsequent intracellular signal and immune response. For several years the study of EVs has brought to light one of their most important features, the acquisition of the characteristics originally native from the donor cell by means of the expression of specific proteins in their surface, and the subsequent phenotypical and genotypical modification on the recipient cell after the uptake of the EVs, using this property to our advantage, a successful EV protein surface analysis could lead to the identification and future use of different biomarkers expressed by them to be implemented as a diagnostic tool in the clinical field. Is in here where our work comes into play, the aim of this thesis is the analysis of extracellular vesicles (EVs) derived from CAR-T cells, by the establishment of a successful isolation method of EVs, and by the assessment of their surface protein signature, and the presence of some molecular markers derived from T-cells.

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