Diagnostic leukapheresis reveals distinct phenotypes of NSCLC circulating tumor cells

Sep 22, 2026

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Circulating tumor cells (CTCs) support studies of tumor heterogeneity and treatment resistance, but conventional approaches in non-small cell lung cancer (NSCLC) often yield too few CTCs for comprehensive analysis. This study applies diagnostic leukapheresis (DLA) to enable screening of larger blood volumes, and introduces a two-step negative depletion strategy for CTC enrichment followed by single-cell whole-transcriptome sequencing.

LanguageEnglish

CategoryResearch & Report

FormatPDF

Pages9

Reading Time23 min

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Abstract

Background

Methods

Results

Conclusions

Background

Rieckmann et al. Molecular Cancer (2024) 23:93 [https://doi.org/10.1186/s12943-024-01984-2](https://doi.org/10.1186/s12943-024-01984-2) Molecular Cancer CORRESPONDENCE Open Access Diagnostic leukapheresis reveals distinct phenotypes of NSCLC circulating tumor cells Lisa‑Marie Rieckmann 1,2,3,4†, Michael Spohn5,6,7,8†, Lisa Ruff1,2,3,4†, David Agorku9, Lisa Becker 10,11, Alina Borchers12,13, Jenny Krause14, Roisin O’Reilly1,2,3,4, Jurek Hille8,15, Janna‑Lisa Velthaus‑Rusik8, Niklas Beumer 1,2,3,16,17, Armin Günther1,2,3,4,17, Lena Willnow1,2,3,4, Charles D. Imbusch 16, Peter Iglauer 18, Ronald Simon18, Sören Franzenburg19, Hauke Winter20, Michael Thomas20, Carsten Bokemeyer8, Nicola Gagliani13,14,21, Christian F. Krebs 12,13, Martin Sprick 10,11, Olaf Hardt9, Sabine Riethdorf15, Andreas Trumpp10,11, Nikolas H. Stoecklein22, Sven Peine23, Philipp Rosenstiel 19, Klaus Pantel 15, Sonja Loges1,2,3,4,8,15† and Melanie Janning1,2,3,4,8,15*† Abstract Background Circulating tumor cells (CTCs) hold immense promise for unraveling tumor heterogeneity and under‑ standing treatment resistance. However, conventional methods, especially in cancers like non‑small cell lung can‑ cer (NSCLC), often yield low CTC numbers, hindering comprehensive analyses. This study addresses this limitation by employing diagnostic leukapheresis (DLA) to cancer patients, enabling the screening of larger blood volumes. To leverage DLA’s full potential, this study introduces a novel approach for CTC enrichment from DLAs. Methods DLA was applied to six advanced stage NSCLC patients. For an unbiased CTC enrichment, a two‑step approach based on negative depletion of hematopoietic cells was used. Single‑cell (sc) whole‑transcriptome sequencing was performed, and CTCs were identified based on gene signatures and inferred copy number variations. Results Remarkably, this innovative approach led to the identification of unprecedented 3,363 CTC transcriptomes. The extensive heterogeneity among CTCs was unveiled, highlighting distinct phenotypes related to the epithelial‑ mesenchymal transition (EMT) axis, stemness, immune responsiveness, and metabolism. Comparison with sc tran‑ scriptomes from primary NSCLC cells revealed that CTCs encapsulate the heterogeneity of their primary counterparts while maintaining unique CTC‑specific phenotypes. Conclusions In conclusion, this study pioneers a transformative method for enriching CTCs from DLA, resulting in a sub‑ stantial increase in CTC numbers. This allowed the creation of the first‑ever single‑cell whole transcriptome in‑depth char‑ acterization of the heterogeneity of over 3,300 NSCLC‑CTCs. The findings not only confirm the diagnostic value of CTCsin monitoring tumor heterogeneity but also propose a CTC‑specific signature that can be exploited for targeted CTC‑ directed therapies in the future. This comprehensive approach signifies a major leap forward, positioning CTCs as a key player in advancing our understanding of cancer dynamics and paving the way for tailored therapeutic interventions. Keywords Circulating tumor cells, Non‑small cell lung cancer, Single cell RNA sequencing, Intratumor heterogeneity †Lisa‑Marie Rieckmann, Michael Spohn, Lisa Ruff, Sonja Loges and Melanie Janning contributed equally to this work. *Correspondence: Melanie Janning melanie.janning@dkfz‑heidelberg.de Full list of author information is available at the end of the article © The Author(s) 2024. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the materia

FAQs

What problem does the study address about circulating tumor cells in NSCLC?

Conventional CTC methods often recover too few CTCs in NSCLC, limiting comprehensive analyses of tumor heterogeneity and treatment resistance.

How was CTC enrichment performed from diagnostic leukapheresis samples?

The study used a two-step approach based on negative depletion of hematopoietic cells to enrich CTCs for unbiased analysis.

What were the main findings from single-cell whole-transcriptome sequencing?

The approach identified 3,363 CTC transcriptomes and revealed distinct CTC phenotypes linked to EMT, stemness, immune responsiveness, and metabolism, including CTC-specific characteristics compared with primary NSCLC cells.

Sep 22, 2026

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