An improved CTC isolation scheme for pairing with downstream genomics: Demonstrating clinical utility in metastatic prostate, lung and pancreatic cancer. Issue 1 (28th September 2016)

• Record Type: Journal Article
• Authors: Premasekharan, Gayatri; Gilbert, Elizabeth; Okimoto, Ross A.; Hamirani, Ashiya; Lindquist, Karla J.; Ngo, Vy T.; Roy, Ritu; Hough, Jeffrey; Edwards, Matthew; Paz, Rosa; Foye, Adam; Sood, Riddhi; Copren, Kirsten A.; Gubens, Matthew; Small, Eric J.; Bivona, Trever G.; Collisson, Eric A.; Friedlander, Terence W.; Paris, Pamela L.
• Is Part Of: Cancer letters. Volume 380:Issue 1(2016)
• Page Start: 144
• Page End: 152
• Publication Date: 2016-09-28
• Subjects: Circulating tumor cells -- High-purity -- Cell adhesion matrix -- Fluorescence-activated cell sorting -- Copy number -- Sequencing; aCGH array comparative genomic hybridization -- CAM cell adhesion matrix -- CK cytokeratin -- CTCs circulating tumor cells -- EGFR epidermal growth factor receptor -- EMT epithelial to mesenchymal transition -- EpCAM epithelial cell adhesion molecule -- FACS fluorescence-activated cell sorting -- iCTCs invasive circulating tumor cells -- mCRPC metastatic castration resistant prostate cancer -- mNSCLC metastatic non-small cell lung cancer -- mPDAC metastatic pancreatic cancer -- NGS next-generation sequencing -- PSA prostate-specific antigen -- PSMA prostate-specific membrane antigen -- WGA whole genome amplification; Cancer -- Periodicals; Neoplasms -- Periodicals; Cancer -- Périodiques; Electronic journals; 616.994
• DOI: 10.1016/j.canlet.2016.06.017
• Languages: English
• ISSNs: 0304-3835

Abstract

Highlights: A two-step CTC enrichment process to improve the purity of CTCs is reported. Three different tumor types – prostate, lung and pancreas cancers – were evaluated. Average iCTC purity from spiked-in cancer cell lines increased from 0.2 to 79.1%. Average iCTC purity from metastatic cancer patients increased from 0.3 to 80.1%. High purity iCTCs were compatible with various downstream genomic analyses. Abstract: Improvements in technologies to yield purer circulating tumor cells (CTCs) will enable a broader range of clinical applications. We have previously demonstrated the use of a commercially available cell-adhesion matrix (CAM) assay to capture invasive CTCs (iCTCs). To improve the purity of the isolated iCTCs, here we used fluorescence-activated cell sorting (FACS) in combination with the CAM assay (CAM + FACS). Our results showed an increase of median purity from the CAM assay to CAM + FACS for the spiked-in cell lines and patient samples analyzed from three different metastatic cancer types: castration resistant prostate cancer (mCRPC), non-small cell lung cancer (mNSCLC) and pancreatic ductal adenocarcinoma cancer (mPDAC). Copy number profiles for spiked-in mCRPC cell line and mCRPC patient iCTCs were similar to expected mCRPC profiles and a matched biopsy. A somatic epidermal growth factor receptor (EGFR) mutation specific to mNSCLC was observed in the iCTCs recovered from EGFR + mNSCLC cell lines and patient samples. Next-generation sequencing (NGS) of spiked-in pancreatic cancer cell line and mPDAC patient iCTCs showed mPDAC common mutations. CAM + FACS iCTC enrichment enables multiple downstream genomic characterizations across different tumor types.