{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2024,9,10]],"date-time":"2024-09-10T13:52:30Z","timestamp":1725976350136},"reference-count":48,"publisher":"American Society of Hematology","issue":"16","content-domain":{"domain":["ashpublications.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2021,8,24]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Although ibrutinib improves the overall survival of patients with chronic lymphocytic leukemia (CLL), some patients still develop resistance, most commonly through point mutations affecting cysteine residue 481 (C481) in Bruton\u2019s tyrosine kinase (BTKC481S and BTKC481R). To enhance our understanding of the biological impact of these mutations, we established cell lines that overexpress wild-type or mutant BTK in in\u00a0vitro and in\u00a0vivo models that mimic ibrutinib-sensitive and -resistant CLL. MEC-1 cell lines stably overexpressing wild-type or mutant BTK were generated. All cell lines coexpressed GFP, were CD19+ and CD23+, and overexpressed BTK. Overexpression of wild-type or mutant BTK resulted in increased signaling, as evidenced by the induction of p-BTK, p-PLC\u03b32, and p-extracellular signal\u2013related kinase (ERK) levels, the latter further augmented upon IgM stimulation. In all cell lines, cell cycle profiles and levels of BTK expression were similar, but the RNA sequencing and reverse-phase protein array results revealed that the molecular transcript and protein profiles were distinct. To mimic aggressive CLL, we created xenograft mouse models by transplanting the generated cell lines into Rag2\u2212\/\u2212\u03b3c\u2212\/\u2212 mice. Spleens, livers, bone marrow, and peripheral blood were collected. All mice developed CLL-like disease with systemic involvement (engraftment efficiency, 100%). We observed splenomegaly, accumulation of leukemic cells in the spleen and liver, and macroscopically evident necrosis. CD19+ cells accumulated in the spleen, bone marrow, and peripheral blood. The overall survival duration was slightly lower in mice expressing mutant BTK. Our cell lines and murine models mimicking ibrutinib-resistant CLL will serve as powerful tools to test reversible BTK inhibitors and novel, non\u2013BTK-targeted therapeutics.<\/jats:p>","DOI":"10.1182\/bloodadvances.2020003821","type":"journal-article","created":{"date-parts":[[2021,8,20]],"date-time":"2021-08-20T15:38:43Z","timestamp":1629473923000},"page":"3134-3146","update-policy":"http:\/\/dx.doi.org\/10.1182\/blood.2019cm0000","source":"Crossref","is-referenced-by-count":5,"title":["Development and characterization of prototypes for in\u00a0vitro and in\u00a0vivo mouse models of ibrutinib-resistant CLL"],"prefix":"10.1182","volume":"5","author":[{"given":"Burcu","family":"Aslan","sequence":"first","affiliation":[{"name":"Department of Experimental Therapeutics,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gorkem","family":"Kismali","sequence":"additional","affiliation":[{"name":"Department of Experimental Therapeutics,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lisa S.","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Experimental Therapeutics,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"LaKesla R.","family":"Iles","sequence":"additional","affiliation":[{"name":"Department of Experimental Therapeutics,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mikhila","family":"Mahendra","sequence":"additional","affiliation":[{"name":"TRACTION Platform, Therapeutics Discovery Division,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Peoples","sequence":"additional","affiliation":[{"name":"TRACTION Platform, Therapeutics Discovery Division,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mihai","family":"Gagea","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine and Surgery,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Natalie W.","family":"Fowlkes","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine and Surgery,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaofeng","family":"Zheng","sequence":"additional","affiliation":[{"name":"Department of Bioinformatics and Computational Biology, and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jing","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Bioinformatics and Computational Biology, and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christopher P.","family":"Vellano","sequence":"additional","affiliation":[{"name":"TRACTION Platform, Therapeutics Discovery Division,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joseph R.","family":"Marszalek","sequence":"additional","affiliation":[{"name":"TRACTION Platform, Therapeutics Discovery Division,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Teresa Sabrina","family":"Bertilaccio","sequence":"additional","affiliation":[{"name":"Department of Experimental Therapeutics,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Varsha","family":"Gandhi","sequence":"additional","affiliation":[{"name":"Department of Experimental Therapeutics,"},{"name":"Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, TX"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"234","published-online":{"date-parts":[[2021,8,20]]},"reference":[{"issue":"16","key":"2021082015375810100_B1","doi-asserted-by":"crossref","first-page":"4313","DOI":"10.1182\/blood-2011-06-338855","article-title":"B-cell receptor signaling in chronic lymphocytic 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, GaidanoG, TrentinL, . Half of chronic lymphocytic leukemia patients relapsing under ibrutinib carry btk and plcg2 mutations: a European Research Initiative on CLL (Eric) real-world study [abstract]. Presented at the Annual Meeting of the European Hematology Association. 17 June 2018. 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