Abstract 555: A workflow for enrichment and whole genome amplification (WGA) of circulating tumor cells (CTCs) for next generation sequencing

Abstract

Abstract Metastasis is the leading cause of cancer deaths. Circulating tumor cells (CTCs) are thought to be responsible for the seeding of metastases. These tumor cells are heterogeneous with variation occurring at the single-cell level. Using a flexible micro spring array (FMSA) device for label-free mechanical enrichment of CTCs from whole blood samples, we established a workflow from isolation followed by whole genome amplification (WGA) and finally sequencing of circulating tumor cells. Briefly, circulating tumor cells were collected from the FMSA using the Zeiss Laser Capture Microdissection System. Isolated cells underwent whole genome amplification using a commercial WGA kit. The quality of the amplified genomic DNA was analyzed using the TaqMan® Real-Time PCR Assays. Specific loci were then amplified with the Ion AmpliSeq™ Cancer Hotspot Panel v2 and sequenced using the Ion PGM™ System. These studies were performed to understand the genomic heterogeneity observed in circulating tumor cells. Citation Format: Merisa Nisic, Sijie Hao, Ramdane Harouaka, Si-Yang Zheng. A workflow for enrichment and whole genome amplification (WGA) of circulating tumor cells (CTCs) for next generation sequencing. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 555. doi:10.1158/1538-7445.AM2015-555