Modeling cancer cell-derived extracellular vesicles using synthetic nanoparticles

Abstract

Metastasis represents a major challenge in cancer care, with the liver being a common site for cancer to spread. Extracellular vesicles (EVs), small membrane-bound vesicles released by cells, play a crucial role in this process. Cancer derived EVs deliver important cargo, such as proteins, to recipient cells that then prepare metastatic sites for colonization by circulating tumor cells (a process referred to as premetastatic niche formation). However, studying EVs is difficult because they are highly heterogenous and difficult to isolate in large amounts. To circumvent these challenges, the aim of this thesis was to create synthetic vesicles (SVs) using liposomes to replicate the properties and functions of cancer cell-derived EVs, with a focus on encapsulation of EV proteins. Two methods were used: 1) specific EV candidate proteins (e.g. heat shock protein 70 (HSP70) and CD63) were commercially sourced and encapsulated into liposomes, and 2) whole proteins were extracted directly from EVs isolated from cancer cells and encapsulated into liposomes. SVs were produced using a microfluidic approach, designed to match the size and zeta potential of natural EVs. The SVs were characterized for their size and surface charge using nanoparticle tracking analysis (NTA) and zeta potential measurements. Protein loading efficiency was evaluated through biochemical assays, including stain-free SDS-PAGE and the micro-BCA protein quantification assay. Proteomic profiling via mass spectrometry identified the encapsulated proteins. Cellular uptake studies were performed using live cell imaging to quantify the internalization of SVs by recipient cells. The findings demonstrate that engineered SVs can successfully incorporate EV-associated proteins and exhibit properties comparable to natural EVs, offering a scalable system to investigate the role of EV proteins in cancer metastasis. This approach bridges nanotechnology and cancer biology, laying the groundwork for designing targeted, EV-inspired delivery systems for future therapeutic applications.

Date	7 Jul 2025
Original language	American English
Awarding Institution	École de technologie supérieure
Supervisor	Vahé Nerguizian (Supervisor) & Julia Valdemarin Burnier (Co-supervisor)