Investigating the role of TRPC3 in the biogenesis of extracellular vesicles
type: CreativeWork
description: Cells within multicellular organisms transcribe, package and release nano-sized spheres called extracellular vesicles (EVs) that carry an array of biomolecules that can be transferred between cells. EV-mediated communication contributes to a wide range of physiological and pathological processes including cancer progression, yet the molecular mechanisms underlying their formation and release remain incompletely characterised. Elevated intracellular calcium (Ca2+) induces EV release in a variety of cell types - with preliminary data to suggest the Ca2+ permeable transient receptor potential (TRP) channel TRPC3 may facilitate Ca2+-mediated EV biogenesis. TRPC3 contributes to the progression of ovarian cancer (OC) in vitro and in vivo, but its role in EV biogenesis and how this influences the pathogenesis of OC is unknown. This thesis therefore aimed to investigate the role of TRPC3 and TRPC3-mediated EV communication in ovarian cells, and how TRPC3 activation, Ca2+ influx and upstream GPCR stimulation influence EV biogenesis and cargo. Initial experiments detected TRPC3 expression across a panel of ovarian cell lines, with SKOV3 chosen as the principal line of study. Activating TRPC3 amplified Ca2+ signalling in SKOV3 cells, and significantly increased cell growth and migration, whilst cell invasion was unchanged. In contrast, inhibiting TRPC3 attenuated histamine-induced Ca2+ signalling, indicating that TRPC3 is activated downstream of histamine GPCRs. To enable specific detection and analysis of SKOV3-EVs, EVs were extracted in serum-free media which yielded a purer population of EVs that present typical markers and morphology. TRPC3 activation induced the release of EVs that promoted recipient SKOV3 cell growth, but bore no effect on SKOV3 motility or invasiveness. Preliminary results also indicated that TRPC3 and its upstream regulator mGluR1 are incorporated into SKOV3-EVs. Using live single-cell imaging, histamine, Ca2+, and TRPC3 activation were shown to stimul…
abstract: Cells within multicellular organisms transcribe, package and release nano-sized spheres called extracellular vesicles (EVs) that carry an array of biomolecules that can be transferred between cells. EV-mediated communication contributes to a wide range of physiological and pathological processes including cancer progression, yet the molecular mechanisms underlying their formation and release remain incompletely characterised. Elevated intracellular calcium (Ca2+) induces EV release in a variety of cell types - with preliminary data to suggest the Ca2+ permeable transient receptor potential (TRP) channel TRPC3 may facilitate Ca2+-mediated EV biogenesis. TRPC3 contributes to the progression of ovarian cancer (OC) in vitro and in vivo, but its role in EV biogenesis and how this influences the pathogenesis of OC is unknown. This thesis therefore aimed to investigate the role of TRPC3 and TRPC3-mediated EV communication in ovarian cells, and how TRPC3 activation, Ca2+ influx and upstream GPCR stimulation influence EV biogenesis and cargo. Initial experiments detected TRPC3 expression across a panel of ovarian cell lines, with SKOV3 chosen as the principal line of study. Activating TRPC3 amplified Ca2+ signalling in SKOV3 cells, and significantly increased cell growth and migration, whilst cell invasion was unchanged. In contrast, inhibiting TRPC3 attenuated histamine-induced Ca2+ signalling, indicating that TRPC3 is activated downstream of histamine GPCRs. To enable specific detection and analysis of SKOV3-EVs, EVs were extracted in serum-free media which yielded a purer population of EVs that present typical markers and morphology. TRPC3 activation induced the release of EVs that promoted recipient SKOV3 cell growth, but bore no effect on SKOV3 motility or invasiveness. Preliminary results also indicated that TRPC3 and its upstream regulator mGluR1 are incorporated into SKOV3-EVs. Using live single-cell imaging, histamine, Ca2+, and TRPC3 activation were shown to stimul…
author: Padbury, Elise
language: en
date published: 2022
date modified: 2026-07-24T03:43:03Z
provider: Oxford Brookes University
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: Oxford Brookes University

PropertyValue
property ID: DOI
value: 10.24384/pnzn-sa59