Dataset Information

Powering single-cell genomics to unravel circulating tumour cell subpopulations in non-small cell lung cancer patients.

ABSTRACT:

Background

Circulating tumour cells (CTCs) are attractive "liquid biopsy" candidates that could provide insights into the different phenotypes of tumours present within a patient. The epithelial-to-mesenchymal transition (EMT) of CTCs is considered a critical step in tumour metastasis; however, it may confound traditional epithelial feature-based CTC isolation and detection. We applied single-cell copy number alteration (CNA) analysis for the identification of genomic alterations to confirm the neoplastic nature of circulating cells with only mesenchymal phenotypes.

Methods

We isolated CTCs from blood samples collected from 46 NSCLC patients using the Parsortix system. Enriched cells were subjected to immunofluorescent staining for CTC identification using a multi-marker panel comprising both epithelial and mesenchymal markers. A subset of isolated CTCs was subjected to whole genome amplification (WGA) and low-pass whole-genome sequencing (LP-WGS) for the analysis of copy number alterations (CNAs).

Results

CTCs were detected in 16/46 (34.8%) patients, inclusive of CK⁺/EpCAM⁺ CTCs (3/46, 6.5%) and Vim⁺ CTCs (13/46, 28.3%). Clusters of Vim⁺ cells were detected in 8 samples, which constitutes 50% of the total number of NSCLC patients with CTCs. No patients had detectable hybrid CK⁺/EpCAM⁺/Vim⁺ cells. All of the tested CK⁺/EpCAM⁺ CTCs and 7/8 Vim⁺ CTCs or CTC clusters carried CNAs confirming their neoplastic nature. Notably, the Vim⁺ cluster with no CNAs was characterised by spindle morphology and, therefore, defined as normal mesenchymal circulating cells.

Conclusion

Our results revealed that CK-negative, vimentin-expressing cells represent a large proportion of CTCs detected in NSCLC patients, which are likely missed by standard epithelial-marker-dependent CTC categorisation.

SUBMITTER: Acheampong E

PROVIDER: S-EPMC10097753 | biostudies-literature | 2023 May

REPOSITORIES: biostudies-literature

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Publications

Powering single-cell genomics to unravel circulating tumour cell subpopulations in non-small cell lung cancer patients.

Acheampong Emmanuel E Morici Michael M Abed Afaf A Bowyer Samantha S Asante Du-Bois DB Lin Weitao W Millward Michael M Gray Elin S ES Beasley Aaron B AB

Journal of cancer research and clinical oncology 20220728 5

<h4>Background</h4>Circulating tumour cells (CTCs) are attractive "liquid biopsy" candidates that could provide insights into the different phenotypes of tumours present within a patient. The epithelial-to-mesenchymal transition (EMT) of CTCs is considered a critical step in tumour metastasis; however, it may confound traditional epithelial feature-based CTC isolation and detection. We applied single-cell copy number alteration (CNA) analysis for the identification of genomic alterations to conf ...[more]

PMID: 35896898

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Project description:BackgroundOver the past decade, numerous reports describe the generation and increasing utility of non-small-cell lung cancer (NSCLC) patient-derived xenografts (PDX) from tissue biopsies. While PDX have proven useful for genetic profiling and preclinical drug testing, the requirement of a tissue biopsy limits the available patient population, particularly those with advanced oligometastatic disease. Conversely, 'liquid biopsies' such as circulating tumour cells (CTCs) are minimally invasive and easier to obtain. Here, we present a clinical case study of a NSCLC patient with advanced metastatic disease, a never smoker whose primary tumour was EGFR and ALK wild-type. We demonstrate for the first time, tumorigenicity of their CTCs to generate a patient CTC-derived eXplant (CDX).Patients and methodsCTCs were enriched at diagnosis and again 2 months later during disease progression from 10 ml blood from a 48-year-old NSCLC patient and implanted into immunocompromised mice. Resultant tumours were morphologically, immunohistochemically, and genetically compared with the donor patient's diagnostic specimen. Mice were treated with cisplatin and pemetrexed to assess preclinical efficacy of the chemotherapy regimen given to the donor patient.ResultsThe NSCLC CDX expressed lung lineage markers TTF1 and CK7 and was unresponsive to cisplatin and pemetrexed. Examination of blood samples matched to that used for CDX generation revealed absence of CTCs using the CellSearch EpCAM-dependent platform, whereas size-based CTC enrichment revealed abundant heterogeneous CTCs of which ∼80% were mesenchymal marker vimentin positive. Molecular analysis of the CDX, mesenchymal and epithelial CTCs revealed a common somatic mutation confirming tumour origin and showed CDX RNA and protein profiles consistent with the predominantly mesenchymal phenotype.ConclusionsThis study shows that the absence of NSCLC CTCs detected by CellSearch (EpCAM(+)) does not preclude CDX generation, highlighting epithelial to mesenchymal transition and the functional importance of mesenchymal CTCs in dissemination of this disease.

Dataset Information

Powering single-cell genomics to unravel circulating tumour cell subpopulations in non-small cell lung cancer patients.

Background

Methods

Results

Conclusion

Publications

Powering single-cell genomics to unravel circulating tumour cell subpopulations in non-small cell lung cancer patients.

Similar Datasets

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