TY - JOUR
T1 - Exploring sialyl-Tn expression in microfluidic-isolated circulating tumour cells
T2 - A novel biomarker and an analytical tool for precision oncology applications
AU - Neves, Manuel
AU - Azevedo, Rita
AU - Lima, Luís
AU - Oliveira, Marta I.
AU - Peixoto, Andreia
AU - Ferreira, Dylan
AU - Soares, Janine
AU - Fernandes, Elisabete
AU - Gaiteiro, Cristiana
AU - Palmeira, Carlos
AU - Cotton, Sofia
AU - Mereiter, Stefan
AU - Campos, Diana
AU - Afonso, Luís Pedro
AU - Ribeiro, Ricardo
AU - Fraga, Avelino
AU - Tavares, Ana
AU - Mansinho, Hélder
AU - Monteiro, Eurico
AU - Videira, Paula A.
AU - Freitas, Paulo P.
AU - Reis, Celso A.
AU - Santos, Lúcio Lara
AU - Dieguez, Lorena
AU - Ferreira, José Alexandre
N1 - Funding:
info:eu-repo/grantAgreement/FCT/FARH/SFRH%2FBD%2F105355%2F2014/PT#
info:eu-repo/grantAgreement/FCT/POR_NORTE/SFRH%2FBD%2F111242%2F2015/PT#
info:eu-repo/grantAgreement/FCT/POR_NORTE/SFRH%2FBD%2F103571%2F2014/PT#
info:eu-repo/grantAgreement/FCT/POR_NORTE/SFRH%2FBD%2F127327%2F2016/PT#
info:eu-repo/grantAgreement/FCT/FARH/SFRH%2FBPD%2F101827%2F2014/PT#
info:eu-repo/grantAgreement/FCT/OE/SFRH%2FBPD%2F111048%2F2015/PT#
info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/PEst-OE%2FSAU%2FUI0776%2F2011/PT#
info:eu-repo/grantAgreement/FCT/9471 - RIDTI/PTDC%2FBBB-EBI%2F0567%2F2014/PT#
info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UID%2FBIM%2F04293%2F2013/PT#
FCT is co-financed by European Social Fund (ESF) under Human Potential Operation Programme (POPH) from National Strategic Reference Framework (NSRF) .
Portuguese Oncology Institute of Porto Research Centre (CI-IPOP-29-2014; CI-IPOP-58-2015) and PhD Programs in Biomedicine and Pathology and Molecular Pathology of ICBAS-University of Porto . This work was also supported by FEDER funds through the Operational Programme for Competitiveness Factors-COMPETE (POCI-01-0145-FEDER-016585; POCI-01-0145-FEDER-007274);and the project NORTE-01-0145-FEDER-000029, co-funded through the NORTE-45-2015-02 (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). The authors are also gratefully thanked the Portuguese Digestive Cancer Investigation Group (GICD) for its support.
PY - 2019/3/25
Y1 - 2019/3/25
N2 - Circulating tumour cells (CTCs) originating from a primary tumour, lymph nodes and distant metastases hold great potential for liquid biopsies by providing a molecular fingerprint for disease dissemination and its temporal evolution through the course of disease management. CTC enumeration, classically defined on the basis of surface expression of Epithelial Cell Adhesion Molecule (EpCAM) and absence of the pan-leukocyte marker CD45, has been shown to correlate with clinical outcome. However, existing approaches introduce bias into the subsets of captured CTCs, which may exclude biologically and clinically relevant subpopulations. Here we explore the overexpression of the membrane protein O-glycan sialyl-Tn (STn) antigen in advanced bladder and colorectal tumours, but not in blood cells, to propose a novel CTC isolation technology. Using a size-based microfluidic device, we show that the majority (>90%) of CTCs isolated from the blood of patients with metastatic bladder and colorectal cancers express the STn antigen, supporting a link with metastasis. STn+ CTC counts were significantly higher than EpCAM-based detection in colorectal cancer, providing a more efficient cell-surface biomarker for CTC isolation. Exploring this concept, we constructed a glycan affinity-based microfluidic device for selective isolation of STn+ CTCs and propose an enzyme-based strategy for the recovery of viable cancer cells for downstream investigations. Finally, clinically relevant cancer biomarkers (transcripts and mutations) in bladder and colorectal tumours, were identified in cells isolated by microfluidics, confirming their malignant origin and highlighting the potential of this technology in the context of precision oncology.
AB - Circulating tumour cells (CTCs) originating from a primary tumour, lymph nodes and distant metastases hold great potential for liquid biopsies by providing a molecular fingerprint for disease dissemination and its temporal evolution through the course of disease management. CTC enumeration, classically defined on the basis of surface expression of Epithelial Cell Adhesion Molecule (EpCAM) and absence of the pan-leukocyte marker CD45, has been shown to correlate with clinical outcome. However, existing approaches introduce bias into the subsets of captured CTCs, which may exclude biologically and clinically relevant subpopulations. Here we explore the overexpression of the membrane protein O-glycan sialyl-Tn (STn) antigen in advanced bladder and colorectal tumours, but not in blood cells, to propose a novel CTC isolation technology. Using a size-based microfluidic device, we show that the majority (>90%) of CTCs isolated from the blood of patients with metastatic bladder and colorectal cancers express the STn antigen, supporting a link with metastasis. STn+ CTC counts were significantly higher than EpCAM-based detection in colorectal cancer, providing a more efficient cell-surface biomarker for CTC isolation. Exploring this concept, we constructed a glycan affinity-based microfluidic device for selective isolation of STn+ CTCs and propose an enzyme-based strategy for the recovery of viable cancer cells for downstream investigations. Finally, clinically relevant cancer biomarkers (transcripts and mutations) in bladder and colorectal tumours, were identified in cells isolated by microfluidics, confirming their malignant origin and highlighting the potential of this technology in the context of precision oncology.
KW - Cancer
KW - Circulating tumour cells
KW - Glycosylation
KW - Lab-on-a-chip
KW - Liquid biopsy
KW - Microfluidics
UR - http://www.scopus.com/inward/record.url?scp=85054441529&partnerID=8YFLogxK
U2 - 10.1016/j.nbt.2018.09.004
DO - 10.1016/j.nbt.2018.09.004
M3 - Article
AN - SCOPUS:85054441529
SN - 1871-6784
VL - 49
SP - 77
EP - 87
JO - New Biotechnology
JF - New Biotechnology
ER -