Single Cell Copy Number Variation Analysis (CNV) of Circulating Tumor Cells (CTCs) in Neuroendocrine Tumor (NET) Patients

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Introduction: The identification and characterization of CTCs as part of a minimally invasive “liquid biopsy” provides an opportunity to explore NET biology, identify therapeutic targets and investigate tumour heterogeneity.

Aim(s): To perform the first genome-wide analysis of CNV in NET CTCs.

Materials and methods: Peripheral blood samples from metastatic NET patients were enriched for CTCs using the EpCAM-dependent CellSearch and EpCAM-independent, size-based Parsortix platforms. Enriched samples were fluorescently labelled with DAPI, CK, +/- EpCAM and CD45 antibodies to enable identification and recovery of single CTCs and white blood cells (WBC) using DEPArray technology. Whole genome amplification was performed on individual cells, followed by quality-control PCR to assess amplified DNA quality. Low pass whole genome sequencing was undertaken and the CNV profile of CTCs analysed to identify chromosomal gains and losses. WBC were used as controls.

Conference: 15th Annual ENETSConcerence (2018)

Presenting Author: Childs A

Authors: Childs A, Vesely C, Rizzo F, Ensell L, Lowe H,

Keywords: CTCs, tumor heterogeneity, CNV,