Ex vivo culture of circulating tumour cells derived from non-small cell lung cancer.

Kapeleris, J; Kulasinghe, A; Warkiani, ME; Oleary, C; Vela, I; Leo, P; Sternes, P; O'Byrne, K; Punyadeera, C

Ex vivo culture of circulating tumour cells derived from non-small cell lung cancer.

Kapeleris, J Kulasinghe, A Warkiani, ME Oleary, C Vela, I Leo, P Sternes, P O'Byrne, K Punyadeera, C

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Publisher:: AME Publishing
Publication Type:: Journal Article
Citation:: Translational Lung Cancer Research, 2020, 9, (5), pp. 1795-1809
Issue Date:: 2020-10

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Field	Value	Language
dc.contributor.author	Kapeleris, J
dc.contributor.author	Kulasinghe, A
dc.contributor.author	Warkiani, ME
dc.contributor.author	Oleary, C
dc.contributor.author	Vela, I
dc.contributor.author	Leo, P
dc.contributor.author	Sternes, P
dc.contributor.author	O'Byrne, K
dc.contributor.author	Punyadeera, C
dc.date.accessioned	2021-03-19T08:26:20Z
dc.date.available	2021-03-19T08:26:20Z
dc.date.issued	2020-10
dc.identifier.citation	Translational Lung Cancer Research, 2020, 9, (5), pp. 1795-1809
dc.identifier.issn	2218-6751
dc.identifier.issn	2226-4477
dc.identifier.uri	http://hdl.handle.net/10453/147388
dc.description.abstract	<h4>Background</h4>Tumour tissue-based information is limited. Liquid biopsy can provide valuable real-time information through circulating tumour cells (CTCs). Profiling and expanding CTCs may provide avenues to study transient metastatic disease.<h4>Methods</h4>Seventy non-small cell lung cancer (NSCLC) patients were recruited. CTCs were enriched using the spiral microfluidic chip and a RosetteSep™ using bloods from NSCLC patients. CTC cultures were carried out using the Clevers media under hypoxic conditions. CTCs were characterized using immunofluorescence and mutation-specific antibodies for samples with known mutation profiles. Exome sequencing was used to characterized CTC cultures.<h4>Results</h4>CTCs (>2 cells) were detected in 38/70 (54.3%) of patients ranging from 0 to 385 CTCs per 7.5 mL blood. In 4/5 patients where primary tumours harboured an EGFR exon 19 deletion, this EGFR mutation was also captured in CTCs. ALK translocation was confirmed on CTCs from a patient harbouring an ALK-rearrangement in the primary tumour. Short term CTC cultures were successfully generated in 9/70 NSCLC patients. Whole exome sequencing (WES) confirmed the presence of somatic mutations in the CTC cultures with mutational signatures consistent with NSCLC.<h4>Conclusions</h4>We were able to detect CTCs in >50% of NSCLC patients. NSCLC patients with >2 CTCs had a poor prognosis. The short-term CTC culture success rate was 12.9%. Further optimization of this culture methodology may provide a means by which to expand CTCs derived from NSCLC patient's bloods. CTC cultures allow for expansion of cells to a critical mass, allowing for functional characterization of CTCs with the goal of drug sensitivity testing and the creation of CTC cell lines.
dc.format	Print
dc.language	eng
dc.publisher	AME Publishing
dc.relation.ispartof	Translational Lung Cancer Research
dc.relation.isbasedon	10.21037/tlcr-20-521
dc.rights	info:eu-repo/semantics/openAccess
dc.subject	1103 Clinical Sciences, 1112 Oncology and Carcinogenesis
dc.title	Ex vivo culture of circulating tumour cells derived from non-small cell lung cancer.
dc.type	Journal Article
utslib.citation.volume	9
utslib.location.activity	China
utslib.for	1103 Clinical Sciences
utslib.for	1112 Oncology and Carcinogenesis
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology
pubs.organisational-group	/University of Technology Sydney/Strength - CHT - Health Technologies
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology/School of Biomedical Engineering
pubs.organisational-group	/University of Technology Sydney/Strength - IBMD - Initiative for Biomedical Devices
utslib.copyright.status	open_access	*
pubs.consider-herdc	true
dc.date.updated	2021-03-19T08:26:18Z
pubs.issue	5
pubs.publication-status	Published
pubs.volume	9
utslib.citation.issue	5

Abstract:

Background

Tumour tissue-based information is limited. Liquid biopsy can provide valuable real-time information through circulating tumour cells (CTCs). Profiling and expanding CTCs may provide avenues to study transient metastatic disease.

Methods

Seventy non-small cell lung cancer (NSCLC) patients were recruited. CTCs were enriched using the spiral microfluidic chip and a RosetteSep™ using bloods from NSCLC patients. CTC cultures were carried out using the Clevers media under hypoxic conditions. CTCs were characterized using immunofluorescence and mutation-specific antibodies for samples with known mutation profiles. Exome sequencing was used to characterized CTC cultures.

Results

CTCs (>2 cells) were detected in 38/70 (54.3%) of patients ranging from 0 to 385 CTCs per 7.5 mL blood. In 4/5 patients where primary tumours harboured an EGFR exon 19 deletion, this EGFR mutation was also captured in CTCs. ALK translocation was confirmed on CTCs from a patient harbouring an ALK-rearrangement in the primary tumour. Short term CTC cultures were successfully generated in 9/70 NSCLC patients. Whole exome sequencing (WES) confirmed the presence of somatic mutations in the CTC cultures with mutational signatures consistent with NSCLC.

Conclusions

We were able to detect CTCs in >50% of NSCLC patients. NSCLC patients with >2 CTCs had a poor prognosis. The short-term CTC culture success rate was 12.9%. Further optimization of this culture methodology may provide a means by which to expand CTCs derived from NSCLC patient's bloods. CTC cultures allow for expansion of cells to a critical mass, allowing for functional characterization of CTCs with the goal of drug sensitivity testing and the creation of CTC cell lines.

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http://hdl.handle.net/10453/147388