Towards establishing Extracellular Vesicle-associated RNAs as biomarkers for HER2+ breast cancer

Abstract

Extracellular vesicles (EVs) are emerging as key players in breast cancer progression and hold immense promise as cancer biomarkers. However, difficulties in obtaining sufficient quantities of EVs for the identification of potential biomarkers hampers progress in this area. To circumvent this obstacle, we cultured BT-474 breast cancer cells in a two chambered bioreactor with CDM-HD serum replacement to significantly improve the yield of cancer cell-associated EVs and eliminate bovine EV contamination. Cancer-relevant mRNAs BIRC5 (Survivin) and YBX1 as well as long-noncoding RNAs HOTAIR, ZFAS1, and AGAP2-AS1 were detected in BT-474 EVs by quantitative RT-PCR. Bioinformatics meta-analyses showed that BIRC5 and HOTAIR RNAs were substantially upregulated compared to non-tumour breast tissue, encouraging further studies to explore their usefulness as biomarkers in patient EV samples. We contend that this effective procedure for obtaining large amounts of cancer-specific EVs will accelerate discovery of EV-associated RNA biomarkers for detection of HER2+ breast cancer.

Non-Coding RNA Signatures of B-Cell Acute Lymphoblastic Leukemia

“…In 2012, PCA3 was FDA-approved as the only urinary biomarker for prostate cancer [ 166 , 167 , 168 ]. Other lncRNAs such as HOTAIR, MALAT1, and GAS5 also show promise as cancer biomarkers [ 169 , 170 , 171 ]. In leukemia, higher GAS5 expression was shown to correlate with poor overall survival [ 172 ].…”

Investigating the consistency of extracellular vesicle production from breast cancer subtypes using CELLine adherent bioreactors

“…Many benefits are already well-reported in the literature for the use of bioreactor systems for large-scale EV production (Artuyants et al, 2021;Guerreiro et al, 2018;Hisey et al, 2020;Mitchell et al, 2008;Mittermaier, 2004;Watson et al, 2016;Watson et al, 2018;Yan & Wu, 2020;Yan et al, 2018). For many researchers, the lower serum usage, removal of concentration steps, and incredible reduction of manual labour makes these systems highly cost and resource effective.…”

Investigating the Consistency of Extracellular Vesicle Production from Breast Cancer Subtypes Using CELLine Adherent Bioreactors

“…Many benefits are already well-reported in the literature for the use of bioreactor systems for large scale EV production [16,21,22,25,26,33,43,52,53]. For many researchers, the lower serum usage, removal of concentration steps, and incredible reduction of manual labour makes these systems highly cost and resource effective.…”

“…Improvements have since been made in the culture media formulations by utilising advanced media to minimise bovine EV contamination while maintaining high levels of cell viability [26]. Several other studies have used similar culture conditions and focused on the downstream RNA and proteomics analyses which are enabled by the production of abundant EVs using these reactors [27][28][29][30][31][32][33]. One recent study demonstrated the culture of non-malignant HEK-293 cells and subsequent EV isolation, loading with siRNA, and uptake by pancreatic cancer cells [34].…”