Lucia Languino's Biography

CANCER CELL TARGETING VIA Small EXTRACELLULAR VESICLES

Tuesday, 14 December 2021 at 09:00

Add to Calendar ▼2021-12-13 14:20:002021-12-13 15:20:00Europe/LondonWelcome and Introduction by Conference Co-Chair -- Extracellular Vesicles and Cellular CommunicationExtracellular Vesicles (EVs): Technologies and Biological Investigations in Coronado Island, CaliforniaCoronado Island, CaliforniaSELECTBIOenquiries@selectbiosciences.com

Cancer cells crosstalk with the tumor microenvironment by releasing small extracellular vesicles (sEVs). sEVs, isolated from cancer cell culture media, express the epithelial-specific alphaVbeta6 integrin.  After confirming the fidelity of the sEV preparations by electron microscopy, density gradient, and immunoblotting, we determined that the alphaVbeta6 integrin is actively packaged into sEVs isolated from cancer cells.  sEVs mediate protein transfer of alphaVbeta6 integrin to microvascular endothelial cells and increase the number of their junctions and tubules.  We also show that alphaVbeta6 is transferred from cancer cells to monocytes by sEVs and that sEVs, purified via density gradients, promote M2 polarization.  In addition, as evaluated by our proteomic analysis, alphaVbeta6 down-regulation causes a significant increase in donor cancer cells, and their sEVs, of two molecules that have a tumor suppressive role, STAT1 and MX1/2.  De novo alphaVbeta6 expression in an alphaVbeta6-negative recipient cell is not a result of a change in mRNA levels but is a consequence of sEV-mediated transfer of this integrin.

We have now selected alphaVbeta6, which is expressed on the cell surface in many cancers but absent in normal tissues, as candidate for therapeutic targeting via sEVs.  The benefits of sEV-mediated cancer therapy are: low immunogenicity, ability to infiltrate biological barriers and ‘targetability’.  We demonstrate an efficient strategy to therapeutically target alphaVbeta6 by using short interfering RNA (siRNA) loaded into sEVs.  We first demonstrate that fluorescently labeled siRNAs can be efficiently loaded into sEVs by electroporation.  By confocal microscopy, we show efficient internalization of these siRNA-loaded sEVs into recipient cells.  We then show that sEV-mediated delivery of alphaVbeta6 -targeting siRNA into cancer cells specifically downregulates expression and function of alphaVbeta6.  Overall, this study shows that sEVs from cancer cells may contribute to a horizontal propagation of integrin-associated phenotypes from cancer cells to the tumor microenvironment.

Welcome and Introduction by Conference Co-Chair -- Extracellular Vesicles and Cellular Communication

Monday, 13 December 2021 at 14:20

Add to Calendar ▼2021-12-13 14:20:002021-12-13 15:20:00Europe/LondonWelcome and Introduction by Conference Co-Chair -- Extracellular Vesicles and Cellular CommunicationExtracellular Vesicles (EVs): Technologies and Biological Investigations in Coronado Island, CaliforniaCoronado Island, CaliforniaSELECTBIOenquiries@selectbiosciences.com

This conference is focused on current Technologies and Biological Investigations on Extracellular Vesicles. Extracellular vesicles are nano-sized membranous structures released by cells into the extracellular space, and easily detectable in body fluids.  Extracellular vesicles are highly heterogenous; large extracellular vesicles are plasma membrane-derived extracellular vesicles, while small extracellular vesicles are of endosomal or non-endosomal origin and are secreted upon fusion with the plasma membrane. Extracellular vesicle biological functions contribute to cell-cell communications in physiological and pathological conditions by carrying unique cargo (proteins, lipids, mRNAs and miRNAs) that modifies the functional state of the recipient cells. Recent investigations have clearly demonstrated the therapeutic and diagnostic potential of extracellular vesicles.  The therapeutic potential of extracellular vesicles is of great interest especially because these vesicles can be engineered to achieve specific targeting.  Currently, more than 200 trials that utilize extracellular vesicles are already registered in clinicaltrials.gov in the US. In this conference, emerging biological topics and novel technologies will be presented to stimulate discussion between areas of research of microfluidics, organoids and extracellular vesicles.