Development and clinical validation of a microfluidic-based platform for CTC enrichment and downstream molecular analysis

Songhua Cai; Youjun Deng; Zhe Wang; Junyu Zhu; Chujian Huang; Longde Du; Chunguang Wang; Xiangyang Yu; Wenyi Liu; Chenglin Yang; Zhe Wang; Lixu Wang; Kai Ma; Rui Huang; Xiaoyu Zhou; Heng Zou; Wenchong Zhang; Yan Huang; Zhi Li; Tiaoping Qin; Tao Xu; Xiaotong Guo; Zhentao Yu

doi:10.3389/fonc.2023.1238332

Development and clinical validation of a microfluidic-based platform for CTC enrichment and downstream molecular analysis

Front Oncol. 2023 Oct 2:13:1238332. doi: 10.3389/fonc.2023.1238332. eCollection 2023.

Authors

Songhua Cai¹, Youjun Deng¹, Zhe Wang², Junyu Zhu³, Chujian Huang¹, Longde Du¹, Chunguang Wang¹, Xiangyang Yu¹, Wenyi Liu¹, Chenglin Yang¹, Zhe Wang¹, Lixu Wang¹, Kai Ma¹, Rui Huang⁴, Xiaoyu Zhou⁵, Heng Zou⁵, Wenchong Zhang⁵, Yan Huang⁵, Zhi Li⁵, Tiaoping Qin⁵, Tao Xu⁵, Xiaotong Guo¹, Zhentao Yu¹

Affiliations

¹ Department of Thoracic Surgery, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital and Shenzhen Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Shenzhen, China.
² Department of Oncology, The First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou, China.
³ Institute of Cancer Control, Cancer Hospital of Xinjiang Medical University, Urumqi, China.
⁴ Shenzhen Futian Research Institute, City University of Hong Kong, Shenzhen, China.
⁵ Department of Medical Affairs, Cellomics (ShenZhen) Limited, Shenzhen, China.

Abstract

Background: Although many CTC isolation and detection methods can provide information on cancer cell counts, downstream gene and protein analysis remain incomplete. Therefore, it is crucial to develop a technology that can provide comprehensive information on both the number and profile of CTC.

Methods: In this study, we developed a novel microfluidics-based CTC separation and enrichment platform that provided detailed information about CTC.

Results: This platform exhibits exceptional functionality, achieving high rates of CTC recovery (87.1%) and purification (∼4 log depletion of WBCs), as well as accurate detection (95.10%), providing intact and viable CTCs for downstream analysis. This platform enables successful separation and enrichment of CTCs from a 4 mL whole-blood sample within 15 minutes. Additionally, CTC subtypes, selected protein expression levels on the CTC surface, and target mutations in selected genes can be directly analyzed for clinical utility using immunofluorescence and real-time polymerase chain reaction, and the detected PD-L1 expression in CTCs is consistent with immunohistochemical assay results.

Conclusion: The microfluidic-based CTC enrichment platform and downstream molecular analysis together provide a possible alternative to tissue biopsy for precision cancer management, especially for patients whose tissue biopsies are unavailable.

Keywords: CTC counts; CTC separation platforms; circulating tumor cells; downstream molecular analysis; microfluidic chip.

Grants and funding

This study was supported by funding as follows, Shenzhen High-level Hospital Construction Fund, Shenzhen Key Medical Discipline Construction Fund (grant no. SZXK075), Sanming Project of Medicine in Shenzhen (grant no. SZSM201612097), Cancer Hospital, Chinese Academy of Medical Sciences, Shenzhen Center/Shenzhen Cancer Hospital Research Project (grant no. SZ2020QN009), Shenzhen Innovation and Entrepreneurship Program (grant no. JSGG20210802153402006), Shenzhen Innovation and Entrepreneurship Program (grant no. JCYJ20220530153615034) and Guangzhou Science and Technology Key Point Project (grant no. 202103000041).