Diagnostic leukapheresis reveals distinct phenotypes of NSCLC circulating tumor cells.

BACKGROUND: Circulating tumor cells (CTCs) hold immense promise for unraveling tumor heterogeneity and understanding treatment resistance. 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'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 transcriptomes 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 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.

Suspect graveyard burial (South Armagh, N. Ireland): Combined search, forensic anthropology and radiocarbon dating.

Geophysical investigation of a former convent graveyard for conversion to a community centre identified an unrecorded, unmarked burial below a later burial. Archaeological excavation confirmed the presence of skeletonized human remains, considered by police as a possible clandestine burial. Mortuary examination indicated the remains belonged to a mature adult female. To determine whether the deceased could be a recorded missing person, radiocarbon dating was undertaken on a femur and a rib bone. This is not always straightforward, and results showed two possible ages due to intercepts on either side of the nuclear weapons testing spike in atmospheric 14C; however, the later dated burial allowed us to constrain the date of a rib to CE 1959. This study demonstrates that dating a second tissue with a longer turnaround time, such as a femur, can help to constrain which side of the bomb spike is most probable. This paper documents in one work the search, scene and sample and then advances this to resolution by anthropological analysis and radiocarbon dating of human remains.