Endogenous PROM1 activation redistributes liver cancer cells along a preexisting differentiation axis

Schneider, T.; Kaneko, K.; Vogl, M.; Utpatel, K.; Immel, A.; Hirsch, D.; Raggi, C.; Spinola Lasso, E.; Evert, M.; Feng, G.-S.; Calvisi, D. F.; Scheiter, A.

doi:10.64898/2026.11.01.763386

bioRxiv · 10.64898/2026.11.01.763386

Endogenous PROM1 activation redistributes liver cancer cells along a preexisting differentiation axis

Schneider, T.·Kaneko, K.·Vogl, M.·Utpatel, K.·Immel, A.·Hirsch, D.·Raggi, C.·Spinola Lasso, E.·Evert, M.·Feng, G.-S.·Calvisi, D. F.·Scheiter, A.

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Abstract

CD133 is widely used as a marker of stem-like cells in hepatocellular carcinoma, yet its functional contribution remains incompletely defined. In PLC/PRF/5 cells, acute CRISPR activation of endogenous PROM1 suppresses hepatic programs, whereas PROM1 knockdown elicits an opposing hepatocyte-identity response, together defining a PROM1-responsive transcriptional axis. Single-cell profiling shows that this axis is preexisting and tracks endogenous PROM1 heterogeneity, separating a hepatocyte-like, PROM1-low compartment from a PROM1-enriched, less hepatocyte-like compartment. Sustained PROM1 activation reduces the hepatocyte-like compartment and shifts the population toward intermediate positions along this axis. Related transcriptional patterns are detectable in an independent HuH-7 dataset and a human HCC case. Endogenous ALFA tagging identifies candidate CD133-associated proteins. Together, these findings support CD133 as an active contributor to tumor-cell state redistribution, extending its role beyond a mere marker of stem-like populations.

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Schneider, T., Kaneko, K., Vogl, M., Utpatel, K., Immel, A., Hirsch, D., Raggi, C., Spinola Lasso, E., Evert, M., Feng, G.-S., Calvisi, D. F., Scheiter, A.. 2026-11-03. Endogenous PROM1 activation redistributes liver cancer cells along a preexisting differentiation axis. https://doi.org/10.64898/2026.11.01.763386

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