Single-cell profiling quantifies the dose gap between population-average and complete pY-STAT1/3 suppression by JAK inhibitors
- 1Institute of Fundamental Technological Research, Polish Academy of Sciences, Warsaw, Poland
Abstract
Inhibitor potency is commonly assessed from population-average responses, but how much single-cell signaling remains under strong average suppression is less clear. We measured nuclear STAT1 and STAT3 tyrosine phosphorylation in two human monocytic cell lines stimulated with IFN-{gamma}; and IL-10 and treated with six JAK-pathway inhibitors. Across 24 profiles, we compared arithmetic-mean inhibition and inhibition of the mean of log10-transformed single-cell responses (log-mean) with the fractions of cells leaving the stimulated range (partial inhibition) or entering the non-stimulated range (complete inhibition). At the lowest tested doses where arithmetic-mean or log-mean inhibition reached at least 80%, median fractions remaining above the non-stimulated range were 76% and 37%, respectively. Thus, strong average suppression frequently left substantial fractions of cells with tangibly elevated pY-STAT responses. Log-mean inhibition generally approximated partial-inhibition occupancy, whereas the apparent IC50 for complete-inhibition occupancy was a median 2.4-fold higher than the log-mean IC50 and 7.4-fold higher than the arithmetic-mean IC50. These relationships provide an empirical guide for estimating cell fractions from conventional inhibition curves.
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BibTeX
@misc{zakrzewska2026singlecell,
title = {{Single-cell profiling quantifies the dose gap between population-average and complete pY-STAT1/3 suppression by JAK inhibitors}},
author = {K. E. Zakrzewska and J. Walczak and K. Nienałtowski and P. Topolewski and C. M. Wagner and A. Braniewska and T. Rygiel and R. Moriggl and M. Komorowski},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.26.759076},
url = {https://www.biorxiv.org/content/10.64898/2026.10.26.759076v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Single-cell profiling quantifies the dose gap between population-average and complete pY-STAT1/3 suppression by JAK inhibitors
AU - K. E. Zakrzewska
AU - J. Walczak
AU - K. Nienałtowski
AU - P. Topolewski
AU - C. M. Wagner
AU - A. Braniewska
AU - T. Rygiel
AU - R. Moriggl
AU - M. Komorowski
PY - 2026
DA - 2026/10/26
PB - bioRxiv
DO - 10.64898/2026.10.26.759076
UR - https://www.biorxiv.org/content/10.64898/2026.10.26.759076v1
AB - Inhibitor potency is commonly assessed from population-average responses, but how much single-cell signaling remains under strong average suppression is less clear. We measured nuclear STAT1 and STAT3 tyrosine phosphorylation in two human monocytic cell lines stimulated with IFN-{gamma}; and IL-10 and treated with six JAK-pathway inhibitors. Across 24 profiles, we compared arithmetic-mean inhibition and inhibition of the mean of log10-transformed single-cell responses (log-mean) with the fractions of cells leaving the stimulated range (partial inhibition) or entering the non-stimulated range (complete inhibition). At the lowest tested doses where arithmetic-mean or log-mean inhibition reached at least 80%, median fractions remaining above the non-stimulated range were 76% and 37%, respectively. Thus, strong average suppression frequently left substantial fractions of cells with tangibly elevated pY-STAT responses. Log-mean inhibition generally approximated partial-inhibition occupancy, whereas the apparent IC50 for complete-inhibition occupancy was a median 2.4-fold higher than the log-mean IC50 and 7.4-fold higher than the arithmetic-mean IC50. These relationships provide an empirical guide for estimating cell fractions from conventional inhibition curves.
ER -