Surface Marker Identification to Capture Live Circulating Tumor Cells in Metastatic Triple-Negative Breast Cancer
Cancer Research Communications. 2026;6(1):115-129. DOI 10.1158/2767-9764.CRC-25-0536. PMID 41543394. PMCID PMC12805936.
How to cite
AMA
Lege BM, Patel KJ, Panici B, Gong P, Lewis MT, Ellis MJ, et al. Surface Marker Identification to Capture Live Circulating Tumor Cells in Metastatic Triple-Negative Breast Cancer. Cancer Res Commun. 2026;6(1):115-129. doi:10.1158/2767-9764.CRC-25-0536APA
Lege, B. M., Patel, K. J., Panici, B., Gong, P., Lewis, M. T., Ellis, M. J., et al. (2026). Surface Marker Identification to Capture Live Circulating Tumor Cells in Metastatic Triple-Negative Breast Cancer. Cancer Research Communications, 6(1), 115-129. https://doi.org/10.1158/2767-9764.CRC-25-0536BibTeX
@article{lege2026surface,
title = {Surface Marker Identification to Capture Live Circulating Tumor Cells in Metastatic Triple-Negative Breast Cancer},
author = {Lege, B. M. and Patel, K. J. and Panici, B. and Gong, P. and Lewis, M. T. and Ellis, M. J. and others},
journal = {Cancer Research Communications},
volume = {6},
number = {1},
pages = {115--129},
year = {2026},
doi = {10.1158/2767-9764.CRC-25-0536},
pmid = {41543394}
}Circulating tumor cells carry a near real-time readout of how a cancer is spreading, but they are rare, and the surface markers used to catch them were built around epithelial tumors. Triple-negative breast cancer cells lean more mesenchymal, so many slip past those markers unseen.
This team built a workflow to capture live tumor cells from blood and read their transcriptomes one cell at a time, without the fixation that degrades single-cell RNA. From those profiles they nominated four new surface markers, then showed that adding them to the standard set caught tumor cells the standard set had missed entirely — widening the net that a liquid biopsy needs to see across a diverse cell population.
Key findings
- Live CTCs picked one at a time yielded high-quality single-cell transcriptomes. From a metastatic TNBC mouse model, 37 circulating tumor cells passed quality filtering, with a median of 7,353 genes detected per cell (range 2,227 to 11,010).
- Single-cell profiling nominated four new surface markers for CTC capture. AHNAK2, CAVIN1, ODR4, and TRIML2 stained the TNBC line WHIM12 at close to 100% coverage — cells the three established markers (EpCAM, HER2, EGFR) left undetected.
- Combining new and established markers widened detection in mouse blood. In MDA-MB-231 samples, a cocktail of the new plus published markers raised flow-cytometry CTC detection to 95.2%, compared with 40% for the published cocktail alone.
CyteFinder in the methods
“Captured cells were stained on cassettes for 15 minutes (modified protocol with no fixation or permeabilization steps) using the negative selection marker CD45 and the nuclear marker Hoechst and subsequently harvested twice (with rinse) into RareCyte chamber slides (#24-1068-000) precoated with polyHEMA. GFP-tagged CTCs were then visualized and individually picked (40-μm needle #24-1117-000 and tube #22-1056-001) using a CyteFinder instrument (RareCyte) for scRNA-seq.”
— Lege et al., Cancer Research Communications (2026), Methods, “Mice”
Why it matters for CyteFinder users
If you are considering CyteFinder for rare-cell work, look at the step it carried here. The whole approach rests on reading the transcriptome of a single circulating tumor cell, and that is only possible if you can find one fluorescent cell among the blood cells around it and lift that one cell, intact, into a tube. That is the job the instrument did: GFP-tagged tumor cells were visualized on chamber slides and picked individually, then sent straight to single-cell sequencing. Two things follow for your own workflow. Because the cells are stained on the surface with no fixation or permeabilization step, the RNA is still intact when the cell reaches the tube — which is what let the authors recover a median of 7,353 genes per cell. And because each cell is picked one at a time under direct view, you decide which cells go forward, rather than pooling whatever an upstream enrichment step happened to retain. For a rare, heterogeneous population, that per-cell control is the point.
