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Clinical Studies

Leukapheresis increases circulating tumour cell yield in non-small cell lung cancer, counts related to tumour response and survival

British Journal of Cancer volume 126, pages 409–418 (2022)Cite this article

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Abstract

Background

Circulating tumour cells (CTCs) can be used to monitor cancer longitudinally, but their use in non-small cell lung cancer (NSCLC) is limited due to low numbers in the peripheral blood. Through diagnostic leukapheresis (DLA) CTCs can be obtained from larger blood volumes.

Methods

Patients with all stages of NSCLC were selected. One total body blood volume was screened by DLA before and after treatment. Peripheral blood was drawn pre- and post DLA for CTC enumeration by CellSearch. CTCs were detected in the DLA product (volume equalling 2 × 108 leucocytes) and after leucocyte depletion (RosetteSep, 9 mL DLA product). Single-cell, whole-genome sequencing was performed on isolated CTCs.

Results

Fifty-six patients were included. Before treatment, CTCs were more often detected in DLA (32/55, 58%) than in the peripheral blood (pre-DLA: 18/55, 33%; post DLA: 13/55, 23%, both at p < 0.01). CTCs per 7.5 mL DLA product were median 9.2 times (interquartile range = 5.6–24.0) higher than CTCs in 7.5 mL blood. RosetteSEP did not significantly improve CTC detection (pretreatment: 34/55, 62%, post treatment: 16/34, 47%) and CTCs per mL even decreased compared to DLA (p = 0.04)..

Patients with advanced-stage disease with DLA-CTC after treatment showed fewer tumour responses and shorter progression-free survival (PFS) than those without DLA-CTC (median PFS, 2.0 vs 12.0 months, p < 0.01). DLA-CTC persistence after treatment was independent of clinical factors associated with shorter PFS (hazard ratio (HR) = 5.8, 95% confidence interval (CI), 1.4–35.5, p = 0.02). All evaluable CTCs showed aneuploidy.

Conclusions

DLA detected nine times more CTCs than in the peripheral blood. The sustained presence of CTCs in DLA after treatment was associated with therapy failure and shortened PFS.

Trial registration

The study was approved by the Medical Ethical Committee (NL55754.042.15) and was registered in the Dutch trial register (NL5423).

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Fig. 1: Patients and apheresis procedures.
Fig. 2: Proportion of non-small cell lung cancer patients with CTCs detected in either blood or apheresis product before and after treatment.
Fig. 3: Circulating tumour cells in the blood and diagnostic leukapheresis product of non-small cell lung cancer patients per volume or corrected for the number of lymphocytes.
Fig. 4: Single-cell whole-genome sequencing of isolated circulating tumour cells from non-small cell lung cancer patients derived from leukapheresis product.
Fig. 5: Persistent CTCs after treatment in advanced-stage non-small cell lung cancer and their relation with tumour response and survival.
Fig. 6: Two non-small cell lung cancer cases showing an association between measured CTC counts and response by imaging.

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Data availability

Data obtained in this study are available upon reasonable request from the corresponding author.

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Acknowledgements

During the study, R. Smith, J. Wheeler and J. Ladtkow (Terumo BCT, Lakewood Co, USA) provided advice and key insights into the apheresis procedure and technology. Procedures were run with the assistance of the personnel from Sanquin (Sanquin, Bloedvoorziening, Groningen, The Netherlands). We are very grateful for all of their expertise and efforts.

Funding

The authors are part of the CANCER-ID consortium, which has received support from the Innovative Medicines Initiative (IMI) Joint Undertaking under grant agreement no. 115749. Additional funding from the Dutch cancer funds (KWF subsidy no. 10491 from 2016) was received. Funding sources had no influence on data gathering, analysing, interpreting or the writing and publishing of the manuscript.

Author information

Authors and Affiliations

  1. Department of Pulmonary Diseases, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands

    Menno Tamminga, T. Jeroen N. Hiltermann & Harry J. M. Groen

  2. Department of Medical Cell BioPhysics, Faculty of Sciences and Technology, University of Twente, Enschede, The Netherlands

    Kiki C. Andree, Anouk Mentink & Leon W. M. M. Terstappen

  3. European Research Institute for the Biology of Ageing, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands

    Hilda van den Bos, Diana C. J. Spierings & Peter Lansdorp

  4. Terry Fox Laboratory, BC Cancer Agency, Vancouver, BC, V5Z 1L3, Canada

    Peter Lansdorp

  5. Department of Medical Genetics, University of British Columbia, Vancouver, BC, V6T 1Z4, Canada

    Peter Lansdorp

  6. Department of Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands

    Wim Timens & Ed Schuuring

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Contributions

Study concepts: LWMMT and HJMG. Study design: HJMG, MT, TJNH, KCA and ES. Data acquisition: MT and HJMG. Quality control of data and algorithms: MT, ES, HJMG and TJNH. Data analysis and interpretation: MT, TJNH and JMG. Statistical analysis: MT and HJMG. Manuscript preparation: MT. Manuscript editing: KCA, HvdB, TJNH, AM, DCJS, PL, WT, ES, LWMMT and HJMG. Manuscript review: PL, WT, ES, LWMMT and HJMG.

Corresponding author

Correspondence to Harry J. M. Groen.

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The authors declare no competing interests.

Ethics approval and consent to participate

The study was approved by the Medical Ethical Committee (NL55754.042.15) and was registered in the Dutch trial register (NL5423). Informed consent was obtained from all patients.

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Tamminga, M., Andree, K.C., van den Bos, H. et al. Leukapheresis increases circulating tumour cell yield in non-small cell lung cancer, counts related to tumour response and survival. Br J Cancer 126, 409–418 (2022). https://doi.org/10.1038/s41416-021-01634-0

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