{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T02:19:03Z","timestamp":1787019543939,"version":"build-2736575974"},"reference-count":40,"publisher":"American Society of Hematology","issue":"16","content-domain":{"domain":["ashpublications.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2025,10,16]]},"abstract":"<jats:sec>\n                    <jats:title>Abstract<\/jats:title>\n                    <jats:p>Chronic lymphocytic leukemia (CLL) is the most common chronic blood cancer in adults. Active NOTCH signaling in CLL is associated with poorer prognosis. Importantly, patients with CLL with NOTCH1 noncoding mutations in the 3\u2032 untranslated region (3\u2032UTR) manifested with a more aggressive disease course even compared with those with mutations in the NOTCH1 coding region. Here, we comprehensively characterize a cryptic splice acceptor site in the 3\u2032UTR of the NOTCH1 gene being converted into a stronger site. The functional consequences of the resulting NOTCH1 protein variants depend on the exact localization of the splice site, the used open reading frame, and the appearance of the next stop codon. The most frequent 3\u2032UTR mutation (g.139390152, A&amp;gt;G) generates a novel NOTCH1 protein, lacking the PEST domain but expressing an altered C terminus consisting of 68 amino acids. Mechanistically, we demonstrate that this splice variant (NOTCH1 152) is transcriptionally less active and dysregulates the regular ubiquitination-dependent degradation of the wild-type NICD (NOTCH1 intracellular domain) in trans. Thus, the NOTCH1 152 variant acts as a \u201csponge\u201d protein in a novel mechanism of oncogenic NOTCH signaling activation, explaining the detrimental disease outcome of patients with CLL with noncoding NOTCH1 mutations. We propose that the detection of NOTCH1 152 protein by specific antibodies is a useful prognostic marker for patients with CLL.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1182\/blood.2025028529","type":"journal-article","created":{"date-parts":[[2025,7,10]],"date-time":"2025-07-10T17:55:17Z","timestamp":1752170117000},"page":"1936-1949","update-policy":"https:\/\/doi.org\/10.1182\/blood.2019cm0000","source":"Crossref","is-referenced-by-count":4,"title":["A noncoding mutation in the\n                    <i>NOTCH1<\/i>\n                    gene initiates oncogenic NOTCH signaling via wild-type NICD stabilization in CLL"],"prefix":"10.1182","volume":"146","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-4434-4341","authenticated-orcid":false,"given":"Min","family":"Guo","sequence":"first","affiliation":[{"name":"1Department of Internal Medicine I, University Hospital Ulm, Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tugba","family":"Memis","sequence":"additional","affiliation":[{"name":"1Department of Internal Medicine I, University Hospital Ulm, Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alena Sophie","family":"Ehrmann","sequence":"additional","affiliation":[{"name":"2Department of Internal Medicine III, University Hospital Ulm, Ulm, Germany"},{"name":"3German Cancer Research Center, Bridging Group Mechanisms of Leukemogenesis, Heidelberg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anselm","family":"Pittrof","sequence":"additional","affiliation":[{"name":"4Department of Hematology, Oncology and Cancer Immunology, Charit\u00e9\u2013Universit\u00e4tsmedizin, Campus Benjamin Franklin, Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4327-811X","authenticated-orcid":false,"given":"Bernd","family":"Baumann","sequence":"additional","affiliation":[{"name":"5Institute of Physiological Chemistry, University of Ulm, Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesca","family":"Ferrante","sequence":"additional","affiliation":[{"name":"6Institute of Biochemistry, Justus Liebig University Giessen, Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eugen","family":"Tausch","sequence":"additional","affiliation":[{"name":"2Department of Internal Medicine III, University Hospital Ulm, Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kirsten","family":"Fischer","sequence":"additional","affiliation":[{"name":"7German CLL Study Group, Department I of Internal Medicine, University Hospital of Cologne, Cologne, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2116-5536","authenticated-orcid":false,"given":"Hartmut","family":"D\u00f6hner","sequence":"additional","affiliation":[{"name":"2Department of Internal Medicine III, University Hospital Ulm, Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4325-5452","authenticated-orcid":false,"given":"Tilman","family":"Borggrefe","sequence":"additional","affiliation":[{"name":"6Institute of Biochemistry, Justus Liebig University Giessen, Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephan","family":"Stilgenbauer","sequence":"additional","affiliation":[{"name":"2Department of Internal Medicine III, University Hospital Ulm, Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0929-1794","authenticated-orcid":false,"given":"Ulrich","family":"Pannicke","sequence":"additional","affiliation":[{"name":"8Institute for Transfusion Medicine, University Hospital Ulm, Ulm, Germany"},{"name":"9Department of Pediatrics and Adolescent Medicine, University Hospital Ulm, Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Klaus","family":"Schwarz","sequence":"additional","affiliation":[{"name":"8Institute for Transfusion Medicine, University Hospital Ulm, Ulm, Germany"},{"name":"10Institute for Clinical Transfusion Medicine and Immunogenetics Ulm, German Red Cross Blood Service Baden-W\u00fcrttemberg\u2013Hessen, Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0227-7188","authenticated-orcid":false,"given":"Daniel","family":"Mertens","sequence":"additional","affiliation":[{"name":"2Department of Internal Medicine III, University Hospital Ulm, Ulm, Germany"},{"name":"3German Cancer Research Center, Bridging Group Mechanisms of Leukemogenesis, Heidelberg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2923-5765","authenticated-orcid":false,"given":"Franz","family":"Oswald","sequence":"additional","affiliation":[{"name":"1Department of Internal Medicine I, University Hospital Ulm, Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"234","reference":[{"issue":"1","key":"2025101611221520000_bib1","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1186\/s12938-021-00973-6","article-title":"The global burden and attributable risk factors of chronic lymphocytic leukemia in 204 countries and territories from 1990 to 2019: analysis based on the global burden of disease study 2019","volume":"21","author":"Yao","year":"2022","journal-title":"Biomed Eng Online"},{"issue":"11","key":"2025101611221520000_bib2","doi-asserted-by":"crossref","first-page":"918","DOI":"10.1001\/jama.2023.1946","article-title":"Diagnosis and treatment of chronic lymphocytic leukemia: a review","volume":"329","author":"Shadman","year":"2023","journal-title":"JAMA"},{"issue":"12","key":"2025101611221520000_bib3","doi-asserted-by":"crossref","first-page":"1679","DOI":"10.1002\/ajh.26367","article-title":"Chronic lymphocytic leukemia: 2022 update on diagnostic and therapeutic procedures","volume":"96","author":"Hallek","year":"2021","journal-title":"Am J Hematol"},{"key":"2025101611221520000_bib4","unstructured":"National Comprehensive Cancer Network\n          . Chronic Lymphocytic Leukemia\/Small Lymphocytic Lymphoma (Version 2.2024). 2024. Accessed 16 July 2025. https:\/\/www.nccn.org\/guidelines\/guidelines-detail?category=1&id=1478."},{"issue":"7","key":"2025101611221520000_bib5","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1001\/jamaoncol.2015.0779","article-title":"Predicting prognosis in chronic lymphocytic leukemia in the contemporary era","volume":"1","author":"Nabhan","year":"2015","journal-title":"JAMA Oncol"},{"issue":"8","key":"2025101611221520000_bib6","doi-asserted-by":"crossref","first-page":"1403","DOI":"10.1182\/blood-2012-09-458265","article-title":"Integrated mutational and cytogenetic analysis identifies new prognostic subgroups in chronic lymphocytic leukemia","volume":"121","author":"Rossi","year":"2013","journal-title":"Blood"},{"issue":"14","key":"2025101611221520000_bib7","doi-asserted-by":"crossref","first-page":"E2911","DOI":"10.1073\/pnas.1702564114","article-title":"Common nonmutational NOTCH1 activation in chronic lymphocytic leukemia","volume":"114","author":"Fabbri","year":"2017","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"10","key":"2025101611221520000_bib8","doi-asserted-by":"crossref","first-page":"1631","DOI":"10.1007\/s00018-009-8668-7","article-title":"The Notch signaling pathway: transcriptional regulation at Notch target genes","volume":"66","author":"Borggrefe","year":"2009","journal-title":"Cell Mol Life Sci"},{"issue":"4","key":"2025101611221520000_bib9","doi-asserted-by":"crossref","first-page":"536","DOI":"10.1016\/j.ccell.2018.07.009","article-title":"Notch signaling in the tumor microenvironment","volume":"34","author":"Meurette","year":"2018","journal-title":"Cancer Cell"},{"key":"2025101611221520000_bib10","doi-asserted-by":"crossref","first-page":"609324","DOI":"10.3389\/fimmu.2020.609324","article-title":"Notch signaling in B cell immune responses","volume":"11","author":"Garis","year":"2020","journal-title":"Front Immunol"},{"key":"2025101611221520000_bib11","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1007\/978-3-319-89512-3_14","article-title":"CSL-associated corepressor and coactivator complexes","volume":"1066","author":"Oswald","year":"2018","journal-title":"Adv Exp Med Biol"},{"key":"2025101611221520000_bib12","doi-asserted-by":"crossref","first-page":"115624","DOI":"10.1016\/j.bcp.2023.115624","article-title":"Structure and function of the highly homologous deubiquitinases ubiquitin specific peptidase 25 and 28: Insights into their pathophysiological and therapeutic roles","volume":"213","author":"Zhou","year":"2023","journal-title":"Biochem Pharmacol"},{"issue":"10","key":"2025101611221520000_bib13","doi-asserted-by":"crossref","first-page":"RESEARCH0043","DOI":"10.1186\/gb-2001-2-10-research0043","article-title":"Characterization of alternatively spliced products and tissue-specific isoforms of USP28 and USP25","volume":"2","author":"Valero","year":"2001","journal-title":"Genome Biol"},{"key":"2025101611221520000_bib14","doi-asserted-by":"crossref","first-page":"229","DOI":"10.3389\/fonc.2018.00229","article-title":"NOTCH1 aberrations in chronic lymphocytic leukemia","volume":"8","author":"Rosati","year":"2018","journal-title":"Front Oncol"},{"issue":"11","key":"2025101611221520000_bib15","doi-asserted-by":"crossref","first-page":"684","DOI":"10.1038\/s41571-019-0239-8","article-title":"Chronic lymphocytic leukaemia: from genetics to treatment","volume":"16","author":"Bosch","year":"2019","journal-title":"Nat Rev Clin Oncol"},{"issue":"7574","key":"2025101611221520000_bib16","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1038\/nature14666","article-title":"Non-coding recurrent mutations in chronic lymphocytic leukaemia","volume":"526","author":"Puente","year":"2015","journal-title":"Nature"},{"issue":"3","key":"2025101611221520000_bib17","doi-asserted-by":"crossref","first-page":"E100","DOI":"10.1002\/ajh.26457","article-title":"Non-coding NOTCH1 mutations in chronic lymphocytic leukemia negatively impact prognosis","volume":"97","author":"Jelloul","year":"2022","journal-title":"Am J Hematol"},{"issue":"2","key":"2025101611221520000_bib18","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1038\/leu.2016.298","article-title":"Non-coding NOTCH1 mutations in chronic lymphocytic leukemia; their clinical impact in the UK CLL4 trial","volume":"31","author":"Larrayoz","year":"2017","journal-title":"Leukemia"},{"issue":"8","key":"2025101611221520000_bib19","doi-asserted-by":"crossref","first-page":"e305","DOI":"10.3324\/haematol.2016.162594","article-title":"Mutations in the 3' untranslated region of NOTCH1 are associated with low CD20 expression levels chronic lymphocytic leukemia","volume":"102","author":"Bittolo","year":"2017","journal-title":"Haematologica"},{"issue":"2-3","key":"2025101611221520000_bib20","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1089\/1066527041410418","article-title":"Maximum entropy modeling of short sequence motifs with applications to RNA splicing signals","volume":"11","author":"Yeo","year":"2004","journal-title":"J Comput Biol"},{"issue":"13","key":"2025101611221520000_bib21","doi-asserted-by":"crossref","first-page":"2315","DOI":"10.1093\/bioinformatics\/bty960","article-title":"A plugin for the Ensembl Variant Effect Predictor that uses MaxEntScan to predict variant spliceogenicity","volume":"35","author":"Shamsani","year":"2019","journal-title":"Bioinformatics"},{"issue":"1","key":"2025101611221520000_bib22","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1002\/ijc.28974","article-title":"Loss of cooperativity of secreted CD40L and increased dose-response to IL4 on CLL cell viability correlates with enhanced activation of NF-kB and STAT6","volume":"136","author":"Bhattacharya","year":"2015","journal-title":"Int J Cancer"},{"issue":"5","key":"2025101611221520000_bib23","doi-asserted-by":"crossref","first-page":"801","DOI":"10.1261\/rna.046342.114","article-title":"Comparison of splice sites reveals that long noncoding RNAs are evolutionarily well conserved","volume":"21","author":"Nitsche","year":"2015","journal-title":"Rna"},{"issue":"7","key":"2025101611221520000_bib24","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1038\/s41419-022-05052-9","article-title":"Hydroxylation of the NOTCH1 intracellular domain regulates Notch signaling dynamics","volume":"13","author":"Ferrante","year":"2022","journal-title":"Cell Death Dis"},{"issue":"3","key":"2025101611221520000_bib25","doi-asserted-by":"crossref","first-page":"1099","DOI":"10.1016\/j.celrep.2014.09.057","article-title":"Dual regulation of Fbw7 function and oncogenic transformation by Usp28","volume":"9","author":"Sch\u00fclein-V\u00f6lk","year":"2014","journal-title":"Cell Rep"},{"issue":"8","key":"2025101611221520000_bib26","doi-asserted-by":"crossref","first-page":"830","DOI":"10.1182\/blood-2018-09-874529","article-title":"FBXW7 mutations reduce binding of NOTCH1, leading to cleaved NOTCH1 accumulation and target gene activation in CLL","volume":"133","author":"Close","year":"2019","journal-title":"Blood"},{"issue":"suppl 1","key":"2025101611221520000_bib27","doi-asserted-by":"crossref","first-page":"4100","DOI":"10.1182\/blood-2022-158168","article-title":"NOTCH1 signaling is dysregulated in CLL by the deubiquitinase USP28 that is recurrently affected by Del(11q) and can be therapeutically targeted [abstract]","volume":"140","author":"Ehrmann","year":"2022","journal-title":"Blood"},{"issue":"8","key":"2025101611221520000_bib28","doi-asserted-by":"crossref","first-page":"1579","DOI":"10.3109\/10428194.2013.767459","article-title":"Missing the notch in NOTCH1","volume":"54","author":"Kugler","year":"2013","journal-title":"Leuk Lymphoma"},{"issue":"1","key":"2025101611221520000_bib29","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1038\/s41589-022-01113-4","article-title":"Affinity-matured DLL4 ligands as broad-spectrum modulators of Notch signaling","volume":"19","author":"Gonzalez-Perez","year":"2023","journal-title":"Nat Chem Biol"},{"issue":"3","key":"2025101611221520000_bib30","doi-asserted-by":"crossref","first-page":"784","DOI":"10.1016\/j.celrep.2017.09.066","article-title":"A B cell regulome links Notch to downstream oncogenic pathways in small B cell lymphomas","volume":"21","author":"Ryan","year":"2017","journal-title":"Cell Rep"},{"key":"2025101611221520000_bib31","doi-asserted-by":"crossref","first-page":"106827","DOI":"10.1016\/j.leukres.2022.106827","article-title":"Landscape of NOTCH1 mutations and co-occurring biomarker alterations in chronic lymphocytic leukemia","volume":"116","author":"Jelloul","year":"2022","journal-title":"Leuk Res"},{"key":"2025101611221520000_bib32","author":"Geppert","year":"2023"},{"issue":"7","key":"2025101611221520000_bib33","doi-asserted-by":"crossref","first-page":"1602","DOI":"10.1038\/leu.2015.14","article-title":"Obinutuzumab as frontline treatment of chronic lymphocytic leukemia: updated results of the CLL11 study","volume":"29","author":"Goede","year":"2015","journal-title":"Leukemia"},{"issue":"31","key":"2025101611221520000_bib34","doi-asserted-by":"crossref","first-page":"3529","DOI":"10.1200\/JCO.2017.73.3402","article-title":"Obinutuzumab or rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone in previously untreated diffuse large B-cell lymphoma","volume":"35","author":"Vitolo","year":"2017","journal-title":"J Clin Oncol"},{"issue":"1","key":"2025101611221520000_bib35","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1111\/bjh.15232","article-title":"A tale of two antibodies: obinutuzumab versus rituximab","volume":"182","author":"Freeman","year":"2018","journal-title":"Br J Haematol"},{"issue":"1","key":"2025101611221520000_bib36","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1038\/leu.2015.182","article-title":"NOTCH1 mutations associate with low CD20 level in chronic lymphocytic leukemia: evidence for a NOTCH1 mutation-driven epigenetic dysregulation","volume":"30","author":"Pozzo","year":"2016","journal-title":"Leukemia"},{"issue":"5","key":"2025101611221520000_bib37","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1101\/gad.2014311","article-title":"Reduced fidelity of branch point recognition and alternative splicing induced by the anti-tumor drug spliceostatin A","volume":"25","author":"Corrionero","year":"2011","journal-title":"Genes Dev"},{"issue":"3","key":"2025101611221520000_bib38","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1038\/s41568-022-00541-7","article-title":"RNA splicing dysregulation and the hallmarks of cancer","volume":"23","author":"Bradley","year":"2023","journal-title":"Nat Rev Cancer"},{"key":"2025101611221520000_bib39","doi-asserted-by":"crossref","first-page":"100191","DOI":"10.1074\/jbc.REV120.015203","article-title":"Frameworks for targeting RNA with small molecules","volume":"296","author":"Umuhire Juru","year":"2021","journal-title":"J Biol Chem"},{"issue":"14","key":"2025101611221520000_bib40","doi-asserted-by":"crossref","first-page":"6549","DOI":"10.1093\/nar\/gkw533","article-title":"Splice-switching antisense oligonucleotides as therapeutic drugs","volume":"44","author":"Havens","year":"2016","journal-title":"Nucleic Acids Res"}],"container-title":["Blood"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/ashpublications.org\/blood\/article-pdf\/146\/16\/1936\/2416987\/blood_bld-2025-028529-main.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/ashpublications.org\/blood\/article-pdf\/146\/16\/1936\/2416987\/blood_bld-2025-028529-main.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,16]],"date-time":"2025-10-16T15:22:28Z","timestamp":1760628148000},"score":1,"resource":{"primary":{"URL":"https:\/\/ashpublications.org\/blood\/article\/146\/16\/1936\/546139\/A-noncoding-mutation-in-the-NOTCH1-gene-initiates"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,16]]},"references-count":40,"journal-issue":{"issue":"16","published-print":{"date-parts":[[2025,10,16]]}},"URL":"https:\/\/doi.org\/10.1182\/blood.2025028529","relation":{},"ISSN":["0006-4971","1528-0020"],"issn-type":[{"value":"0006-4971","type":"print"},{"value":"1528-0020","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2025,10,16]]},"published":{"date-parts":[[2025,10,16]]}}}