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Interspecies interaction and uptake mechanism of extracellular vesicles with tropism to B cells

Grant number: 22/04146-9
Support type:Scholarships abroad - Research Internship - Doctorate (Direct)
Effective date (Start): September 19, 2022
Effective date (End): September 18, 2023
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal researcher:Heloisa Sobreiro Selistre de Araújo
Grantee:Bianca Cruz Pachane
Supervisor abroad: Kenneth W. Witwer
Home Institution: Centro de Ciências Biológicas e da Saúde (CCBS). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil
Research place: Johns Hopkins University (JHU), United States  
Associated to the scholarship:21/01983-4 - Influence of hypoxic extracellular vesicles in cell invasion and tumour microenvironment on Breast Cancer, BP.DD

Abstract

Extracellular vesicles (EVs) are nano-scaled particles secreted by cells that offer stable transport and storage of biomolecules. Their structure allows them to be taken up by other cells, thus turning EVs as potential candidates for new therapeutical approaches, particularly in drug delivery. To fully comprehend how EVs perform in a therapeutic setting, knowledge on cargo loading mechanisms, interaction processes, dosage, administration and vesicular origin need to be built further. Earlier results from Witwer Lab identified a great involvement of white blood cells in the clearance of human EVs from the blood of nonhuman primates, with particular emphasis in B cells, who are critical to immune response and antibody expression. B cells also promote homeostasis maintenance, activation of secondary lymphoid organs, development of CD4+ T cells and inflammatory immune responses. EVs are great modulators of the immune system, aiding in cytokine and growth factor transport, antigen presentation and pleiotropic effects on complement and coagulation, mostly via miRNA signaling. In this project, we aim to understand the molecular mechanisms of EV tropism to B cells in vitro. To test our hypothesis, we intend to separate EVs from cells in culture using differential ultracentrifugation coupled with filtration and SEC, then proceed to characterize them as suggested by the MISEV2018 guidelines. Interaction assays will be performed with primary cell cultures from primates (both human and nonhuman) via imaging cell cytometry and confocal microscopy analysis. We also suggest RNAseq and stem-loop RT-qPCR to identify miRNA profile that might explain B cell tropism. (AU)

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