Extracellular vesicles from lung adenocarcinoma cells induce activation of different CAF subtypes
department or school: Medicine, Faculty of
data provider: DSpace
degree campus: UBCV
creator: Trejo Vazquez, Jessica Angelina
issued: 2024-05-16T20:36:21Z
issued: 2024
related degree: Master of Science - MSc
degree grantor: University of British Columbia
description: Lung cancer is the leading cause of cancer deaths worldwide, largely due to metastasis. Its main subtype is lung adenocarcinoma (LUAD). Communication between cancer cells and cells within the tumor microenvironment, such as fibroblasts, can promote metastasis. This interaction can be mediated by extracellular vesicles (EVs) transferring bioactive cargo. Thus, this study aimed to identify the impact of EVs derived from LAC cells on lung fibroblasts. For EV isolation, conditioned media derived from H2073 and H1437 LAC cell lines were ultracentrifuged multiple times after debris removal. Nanoparticle Tracking Analysis was used for quantification and size characterization and EV-associated markers were assessed by western blot (WB). For lung fibroblast activation, H6013 primary adult cells were treated with 1xPBS, TGFβ (10 ng/μL) or EVs (7.13x10¹¹) every 24 hours over 3 days. Protein (for WB) and RNA (for gene expression profiling) were harvested 48 hours after treatment. RNA was extracted using the Qiagen miRNeasy kit and sequenced by Illumina NextSeq2000. Most vesicles produced by both cell lines were between 50 and 250 nm in diameter. Fibroblasts treated with TGFβ or EVs had an increase in CAF markers (mainly α-SMA), especially in the EV group compared to the PBS treatment group. The activated fibroblasts shared differentially expressed genes in the TGFβ and EV treatments; however, there was an increase in some of the genes associated with epithelial-mesenchymal transition and cell cycle only in the EV-treated group, indicating unique features of CAF activation and the same was seen for the pathway analysis. Therefore, LUAD EVs have a role in CAF activation by using both similar and unique pathways to TGFβ activation. Further understanding of the genes distinct to the EV-treated fibroblasts could lead to the identification of novel genes involved in CAF activation, and thus aid in the development of novel therapeutics that could improve patient outcomes.
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/88329
note: EXTRACELLULAR VESICLES FROM LUNG ADENOCARCINOMA CELLS INDUCE ACTIVATION OF DIFFERENT CAF SUBTYPES by Jessica Angelina Trejo Vazquez B.Sc., National Polytechnic Institute, 2018 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in THE FACULTY OF GRADUATE AND POSTDOCTORAL STUDIES (Interdisciplinary Oncology) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) May 2024 © Jessica Angelina Trejo Vazquez, 2024 ii The following individuals certify that they have read and recommend to the Faculty of Graduate and Postdoctoral Studies for acceptance the thesis entitled: Extracellular vesicles from lung adenocarcinoma cells induce activation of different CAF subtypes submitted by Jessica Angelina Trejo Vazquez in partial fulfilment of the requirements for the degree of Master of Science in Interdisciplinary Oncology Examining Committee: Dr. Cathie Garnis, Associate Professor, Department of Surgery, UBC Supervisor Dr. Karla Williams, Associate Professor, Faculty of Pharmaceutical Sciences, UBC Supervisory Committee Member Dr. Tony Ng, Clinical Associate Professor, Pathology and Laboratory Medicine, UBC Supervisory Committee Member Dr. William Lockwood, Associate Professor, Pathology and Laboratory Medicine, UBC Additional Examiner Additional Supervisory Committee Members: Dr. Kevin Bennewith, Associate Professor, Pathology and Laboratory Medicine, UBC Supervisory Committee Member iii Abstract Lung cancer is the leading cause of cancer deaths worldwide, largely due to metastasis. Its main subtype is lung adenocarcinoma (LUAD). Communication between cancer cells and cells within the tumor microenvironment, such as fibroblasts, can promote metastasis. This interaction can be mediated by extracellular vesicles (EVs) transferring bioactive cargo. Thus, this study aimed to identify the impact of EVs derived from LAC cells on lung fibroblasts. For EV isolation, conditioned media derived from H2073 and H1437 LAC cell lines were ultracentrifuged…
has type: Thesis/Dissertation
date issued: 2024-11
is shown at: 10.14288/1.0443548
language: eng
degree discipline: Interdisciplinary Oncology
provider: Vancouver : University of British Columbia Library
publisher: University of British Columbia
rights: Attribution-NonCommercial-NoDerivatives 4.0 International
rights URI: http://creativecommons.org/licenses/by-nc-nd/4.0/
scholar level: Graduate
contributor: Garnis, Cathie
type: Text
identifier URI: http://hdl.handle.net/2429/88329