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However, conventional methods, especially in cancers like non-small cell lung cancer (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\u2019s full potential, this study introduces a\u00a0novel approach for CTC enrichment from DLAs.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>DLA was applied to six advanced\u00a0stage 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.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>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 transcriptomes from primary NSCLC cells revealed that CTCs encapsulate the heterogeneity of their primary counterparts while maintaining unique CTC-specific phenotypes.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>In conclusion, this study pioneers a transformative method for enriching CTCs from DLA, resulting in a substantial increase in CTC numbers. This allowed the creation of the first-ever single-cell whole transcriptome in-depth characterization of the heterogeneity of over 3,300 NSCLC-CTCs. The findings not only confirm the diagnostic value of CTCs in 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.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12943-024-01984-2","type":"journal-article","created":{"date-parts":[[2024,5,8]],"date-time":"2024-05-08T00:01:39Z","timestamp":1715126499000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":30,"title":["Diagnostic leukapheresis reveals distinct phenotypes of NSCLC circulating tumor 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KG. CB: honoraria and\/or advisory board from AstraZeneca, Bayer, Berlin Chemie, BMS, Merck, Roche, Novartis, Sanofi, Lilly\/ImClone and travel expenses from BMS, Merck, Pfizer and Sanofi. SL received funding from BerGenBio, Bristol Myers Squibb, Eli Lilly, Roche Pharma and ADC Therapeutics; consulting fees from BerGenBio, BMS, Boehringer Ingelheim, Eli Lilly, Roche Pharma, Medac, Sanofi Aventis, Novartis, AstraZeneca, Pfizer, Takeda, Amgen, Bayer, Janssen and Merck; money for presentations\/speakers bureaus from BerGenBio, BMS, Boehringer Ingelheim, Eli Lilly, Roche Pharma, Medac, Sanofi Aventis, Novartis, AstraZeneca, Pfizer, Takeda, Amgen, Bayer, Jannssen and Merck; travel support from BerGenBio, BMS, Boehringer Ingelheim, Eli Lilly, Roche Pharma, Medac, Sanofi Aventis, Novartis, AstraZeneca, Pfizer, Takeda, Amgen, Bayer, Janssen and Merck. MJ: received research support (material) from Miltenyi and funding from Bristol Myers Squibb; speakers honoraria and \/or advisory board from Roche, Amgen, AstraZeneca, Novartis, Takeda and travel support from AstraZeneca.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"93"}}