{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T00:00:21Z","timestamp":1768521621070,"version":"3.49.0"},"posted":{"date-parts":[[2025,4,2]]},"group-title":"Pharmacology and Toxicology","reference-count":49,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,4,2]],"date-time":"2025-04-02T00:00:00Z","timestamp":1743552000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,4,2]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Single-cell studies of signal transduction have revealed complex temporal dynamics that determine downstream biological function. For example, the stimulus-specific dynamics of the transcription factor NF\u03baB specify stimulus-specific gene expression programs, and loss of specificity leads to disease. Thus, it is intriguing to consider drugs that may restore signaling specificity in disease contexts, or reduce activity but maintain signaling specificity to avoid unwanted side effects. However, while steady-state dose-response relationships have been the focus of pharmacological studies, there are no established methods for quantifying drug impact on stimulus-response signaling dynamics. Here we evaluated how drug treatments affect the stimulus-specificity of NF\u03baB activation dynamics and its ability to accurately code ligand identity and dose. Specifically, we simulated the dynamic NF\u03baB trajectories in response to 15 stimuli representing various immune threats under treatment of 10 representative drugs across 20 dosage levels. To quantify the effects on coding capacity, we introduced a Stimulus Response Specificity (SRS) score and a stimulus confusion score. We constructed stimulus confusion maps by employing epsilon network clustering in the trajectory space and in various dimensionally reduced spaces: canonical polyadic decomposition (CPD), functional principal component analysis (fPCA), and NF\u03baB signaling codons (i.e. established, informative dynamic features). Our results indicated that the SRS score and the stimulus confusion map based on signaling codons are best-suited to quantify stimulus-specific NF\u03baB dynamics confusion under pharmacological perturbations. Using these tools we found that temporal coding capacity of the NF\u03baB signaling network is generally robust to a variety of pharmacological perturbations, thereby enabling the targeting of stimulus-specific dynamics without causing broad side-effects.<\/jats:p>","DOI":"10.1101\/2025.03.31.645599","type":"posted-content","created":{"date-parts":[[2025,4,2]],"date-time":"2025-04-02T05:50:38Z","timestamp":1743573038000},"source":"Crossref","is-referenced-by-count":0,"title":["A computational workflow for assessing drug effects on temporal signaling dynamics reveals robustness in stimulus-specific NF\u03baB signaling"],"prefix":"10.64898","author":[{"given":"Emily R.","family":"Bozich","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5740-2428","authenticated-orcid":false,"given":"Xiaolu","family":"Guo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jennifer L.","family":"Wilson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5607-3845","authenticated-orcid":false,"given":"Alexander","family":"Hoffmann","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025040404000727000_2025.03.31.645599v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2013.02.005"},{"key":"2025040404000727000_2025.03.31.645599v1.2","doi-asserted-by":"publisher","DOI":"10.1126\/scisignal.281pe44"},{"key":"2025040404000727000_2025.03.31.645599v1.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.gde.2010.09.007"},{"key":"2025040404000727000_2025.03.31.645599v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2021.04.011"},{"key":"2025040404000727000_2025.03.31.645599v1.5","doi-asserted-by":"publisher","DOI":"10.1038\/nature09145"},{"issue":"6","key":"2025040404000727000_2025.03.31.645599v1.6","first-page":"638","article-title":"NF-\u03baB Dynamics Discriminate between TNF Doses in Single Cells","volume":"5","year":"2017","journal-title":"cels"},{"key":"2025040404000727000_2025.03.31.645599v1.7","doi-asserted-by":"crossref","unstructured":"Rahman SMT , Singh A , Lowe S , Aqdas M , Jiang K , Narayanan HV , et al. Co-imaging of RelA and c-Rel reveals features of NF-\u03baB signaling for ligand discrimination. Cell Reports [Internet]. 2024 Mar 26 [cited 2025 Feb 23];43(3). 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