{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:24:17Z","timestamp":1787027057775,"version":"build-2736575974"},"reference-count":49,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,4,6]],"date-time":"2023-04-06T00:00:00Z","timestamp":1680739200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100011019","name":"Nemzeti Kutat\u00e1si Fejleszt\u00e9si \u00e9s Innov\u00e1ci\u00f3s Hivatal","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100011019","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002332","name":"Semmelweis Egyetem","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002332","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Pharmacol."],"abstract":"<jats:p>\n                    Tyrosine kinases are crucial signaling components of diverse biological processes and are major therapeutic targets in various malignancies and immune-mediated disorders. A critical step of development of novel tyrosine kinase inhibitors is the transition from the confirmation of the\n                    <jats:italic>in vitro<\/jats:italic>\n                    effects of drug candidates to the analysis of their\n                    <jats:italic>in vivo<\/jats:italic>\n                    efficacy. To facilitate this transition, we have developed a rapid\n                    <jats:italic>in vivo<\/jats:italic>\n                    assay for the analysis of the effect of oral tyrosine kinase inhibitors on basal tyrosine phosphorylation of circulating mouse neutrophils. The assay uses a single drop of peripheral blood without sacrificing the mice. Flow cytometry using intracellular staining by fluorescently labeled anti-phosphotyrosine antibodies revealed robust basal tyrosine phosphorylation in resting circulating neutrophils. This signal was abrogated by the use of isotype control antibodies or by pre-saturation of the anti-phosphotyrosine antibodies with soluble phosphotyrosine amino acids or tyrosine-phosphorylated peptides. Basal tyrosine phosphorylation was dramatically reduced in neutrophils of triple knockout mice lacking the Src-family tyrosine kinases Hck, Fgr, and Lyn. Neutrophil tyrosine phosphorylation was also abrogated by oral administration of the Abl\/Src-family inhibitor dasatinib, a clinically used anti-leukemic agent. Detailed dose-response and kinetic studies revealed half-maximal reduction of neutrophil tyrosine phosphorylation by 2.9\u00a0mg\/kg dasatinib, with maximal reduction observed 2\u00a0h after inhibitor administration. Taken together, our assay allows highly efficient analysis of the\n                    <jats:italic>in vivo<\/jats:italic>\n                    effect of orally administered tyrosine kinase inhibitors, and may be used as a suitable alternative to other existing approaches.\n                  <\/jats:p>","DOI":"10.3389\/fphar.2023.1056154","type":"journal-article","created":{"date-parts":[[2023,4,6]],"date-time":"2023-04-06T00:30:01Z","timestamp":1680741001000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Analysis of intracellular tyrosine phosphorylation in circulating neutrophils as a rapid assay for the in vivo effect of oral tyrosine kinase inhibitors"],"prefix":"10.3389","volume":"14","author":[{"given":"Krisztina","family":"Futosi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bogl\u00e1rka","family":"Bajza","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dorottya","family":"Deli","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e1s","family":"Erd\u00e9lyi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simon","family":"Tusn\u00e1dy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Attila","family":"M\u00f3csai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,4,6]]},"reference":[{"key":"B1","first-page":"1","article-title":"Tyrosine kinases","volume-title":"xPharm: The comprehensive Pharmacology reference","author":"Blaukat","year":"2007"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.4172\/2153-2435.1000560","article-title":"In vivo studies for drug development via oral delivery: Challenges, animal models and techniques","volume":"8","author":"Brake","year":"2017","journal-title":"Pharm. 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