AUTHOR=Kong Say Li , Liu Xingliang , Tan Swee Jin , Tai Joyce A. , Phua Ler Yee , Poh Huay Mei , Yeo Trifanny , Chua Yong Wei , Haw Yu Xuan , Ling Wen Huan , Ng Raymond Chee Hui , Tan Tira J. , Loh Kiley Wei Jen , Tan Daniel Shao-Weng , Ng Quan Sing , Ang Mei Kim , Toh Chee Keong , Lee Yi Fang , Lim Chwee Teck , Lim Tony Kiat Hon , Hillmer Axel M. , Yap Yoon Sim , Lim Wan-Teck TITLE=Complementary Sequential Circulating Tumor Cell (CTC) and Cell-Free Tumor DNA (ctDNA) Profiling Reveals Metastatic Heterogeneity and Genomic Changes in Lung Cancer and Breast Cancer JOURNAL=Frontiers in Oncology VOLUME=11 YEAR=2021 URL=https://www.frontiersin.org/journals/oncology/articles/10.3389/fonc.2021.698551 DOI=10.3389/fonc.2021.698551 ISSN=2234-943X ABSTRACT=Introduction

Circulating tumor cells (CTCs) and cell-free tumor DNA (ctDNA) are tumor components present in circulation. Due to the limited access to both CTC enrichment platforms and ctDNA sequencing in most laboratories, they are rarely analyzed together.

Methods

Concurrent isolation of ctDNA and single CTCs were isolated from lung cancer and breast cancer patients using the combination of size-based and CD45-negative selection method via DropCell platform. We performed targeted amplicon sequencing to evaluate the genomic heterogeneity of CTCs and ctDNA in lung cancer and breast cancer patients.

Results

Higher degrees of genomic heterogeneity were observed in CTCs as compared to ctDNA. Several shared alterations present in CTCs and ctDNA were undetected in the primary tumor, highlighting the intra-tumoral heterogeneity of tumor components that were shed into systemic circulation. Accordingly, CTCs and ctDNA displayed higher degree of concordance with the metastatic tumor than the primary tumor. The alterations detected in circulation correlated with worse survival outcome for both lung and breast cancer patients emphasizing the impact of the metastatic phenotype. Notably, evolving genetic signatures were detected in the CTCs and ctDNA samples during the course of treatment and disease progression.

Conclusions

A standardized sample processing and data analysis workflow for concurrent analysis of CTCs and ctDNA successfully dissected the heterogeneity of metastatic tumor in circulation as well as the progressive genomic changes that may potentially guide the selection of appropriate therapy against evolving tumor clonality.