{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,4]],"date-time":"2026-11-04T05:59:47Z","timestamp":1793771987750,"version":"4.6.1"},"reference-count":19,"publisher":"Springer Science and Business Media LLC","issue":"9","license":[{"start":{"date-parts":[[2021,9,21]],"date-time":"2021-09-21T00:00:00Z","timestamp":1632182400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,9,21]],"date-time":"2021-09-21T00:00:00Z","timestamp":1632182400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Blood Cancer J."],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The BCL2-inhibitor, Venetoclax (VEN), has shown significant anti-leukemic efficacy in combination with the DNMT-inhibitor, Azacytidine (AZA). To explore the mechanisms underlying the selective sensitivity of mutant leukemia cells to VEN and AZA, we used cell-based isogenic models containing a common leukemia-associated mutation in the epigenetic regulator <jats:italic>ASXL1<\/jats:italic>. KBM5 cells with CRISPR\/Cas9-mediated correction of the <jats:italic>ASXL1<\/jats:italic><jats:sup><jats:italic>G710X<\/jats:italic><\/jats:sup> mutation showed reduced leukemic growth, increased myeloid differentiation, and decreased <jats:italic>HOXA<\/jats:italic> and <jats:italic>BCL2<\/jats:italic> gene expression in vitro compared to uncorrected KBM5 cells. Increased expression of the anti-apoptotic gene, <jats:italic>BCL2<\/jats:italic>, was also observed in bone marrow CD34+ cells from <jats:italic>ASXL1<\/jats:italic> mutant MDS patients compared to CD34+ cells from wild-type MDS cases. ATAC-sequencing demonstrated open chromatin at the <jats:italic>BCL2<\/jats:italic> promoter in the ASXL1 mutant KBM5 cells. BH3 profiling demonstrated increased dependence of mutant cells on <jats:italic>BCL2<\/jats:italic>. Upon treatment with VEN, mutant cells demonstrated increased growth inhibition. In addition, genome-wide methylome analysis of primary MDS samples and isogenic cell lines demonstrated increased gene-body methylation in <jats:italic>ASXL1<\/jats:italic> mutant cells, with consequently increased sensitivity to AZA. These data mechanistically link the common leukemia-associated mutation <jats:italic>ASXL1<\/jats:italic> to enhanced sensitivity to VEN and AZA via epigenetic upregulation of <jats:italic>BCL2<\/jats:italic> expression and widespread alterations in DNA methylation.<\/jats:p>","DOI":"10.1038\/s41408-021-00541-0","type":"journal-article","created":{"date-parts":[[2021,9,21]],"date-time":"2021-09-21T19:21:09Z","timestamp":1632252069000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":62,"title":["ASXL1 mutations are associated with distinct epigenomic alterations that lead to sensitivity to venetoclax and azacytidine"],"prefix":"10.1038","volume":"11","author":[{"given":"Nora E.","family":"Rahmani","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nandini","family":"Ramachandra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Srabani","family":"Sahu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nadege","family":"Gitego","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"Lopez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kith","family":"Pradhan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tushar D.","family":"Bhagat","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shanisha","family":"Gordon-Mitchell","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8821-0210","authenticated-orcid":false,"given":"Bianca Rivera","family":"Pena","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammad","family":"Kazemi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keshav","family":"Rao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1328-3682","authenticated-orcid":false,"given":"Orsi","family":"Giricz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shahina Bano","family":"Maqbool","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raul","family":"Olea","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongmei","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1436-6460","authenticated-orcid":false,"given":"Jinghang","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4724-1720","authenticated-orcid":false,"given":"Hamid","family":"Dolatshad","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vickram","family":"Tittrea","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dharamveer","family":"Tatwavedi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shalini","family":"Singh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juseong","family":"Lee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8245-0771","authenticated-orcid":false,"given":"Tianyu","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ulrich","family":"Steidl","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5366-8288","authenticated-orcid":false,"given":"Aditi","family":"Shastri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daichi","family":"Inoue","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Omar","family":"Abdel-Wahab","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6122-0221","authenticated-orcid":false,"given":"Andrea","family":"Pellagatti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6319-8331","authenticated-orcid":false,"given":"Evripidis","family":"Gavathiotis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4330-2928","authenticated-orcid":false,"given":"Jacqueline","family":"Boultwood","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7592-7693","authenticated-orcid":false,"given":"Amit","family":"Verma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,9,21]]},"reference":[{"key":"541_CR1","doi-asserted-by":"publisher","first-page":"2795","DOI":"10.1038\/s41375-019-0612-8","volume":"33","author":"BA Jonas","year":"2019","unstructured":"Jonas BA, Pollyea DA. How we use venetoclax with hypomethylating agents for the treatment of newly diagnosed patients with acute myeloid leukemia. Leukemia 2019;33:2795\u2013804.","journal-title":"Leukemia"},{"key":"541_CR2","doi-asserted-by":"publisher","first-page":"362","DOI":"10.1158\/2159-8290.CD-13-0609","volume":"4","author":"R Pan","year":"2013","unstructured":"Pan R, Hogdal LJ, Benito JM, Bucci D, Han L, Borthakur G, et al. Selective BCL-2 inhibition by ABT-199 causes on-target cell death in acute myeloid leukemia. Cancer Discov. 2013;4:362\u201375.","journal-title":"Cancer Discov"},{"key":"541_CR3","doi-asserted-by":"publisher","first-page":"329","DOI":"10.1016\/j.stem.2012.12.013","volume":"12","author":"ED Lagadinou","year":"2013","unstructured":"Lagadinou ED, Sach A, Callahan K, Rossi RM, Neering SJ, Minhajuddin M, et al. BCL-2 inhibition targets oxidative phosphorylation and selectively eradicates quiescent human leukemia stem cells. Cell Stem Cell. 2013;12:329\u201341.","journal-title":"Cell Stem Cell"},{"key":"541_CR4","doi-asserted-by":"publisher","first-page":"4326","DOI":"10.1182\/bloodadvances.2019000937","volume":"3","author":"DA Pollyea","year":"2019","unstructured":"Pollyea DA, Amaya M, Strati P, Konopleva MY. Venetoclax for AML: changing the treatment paradigm. Blood Adv. 2019;3:4326\u201335.","journal-title":"Blood Adv"},{"key":"541_CR5","doi-asserted-by":"publisher","first-page":"355ra117","DOI":"10.1126\/scitranslmed.aag1180","volume":"8","author":"BZ Carter","year":"2016","unstructured":"Carter BZ, Mak PY, Mu H, Zhou H, Mak DH, Schober W, et al. Combined targeting of BCL-2 and BCR-ABL tyrosine kinase eradicates chronic myeloid leukemia stem cells. Sci Transl Med. 2016;8:355ra117.","journal-title":"Sci Transl Med"},{"issue":"Mar","key":"541_CR6","doi-asserted-by":"publisher","first-page":"316","DOI":"10.1016\/j.stem.2012.12.011","volume":"12","author":"DJ Goff","year":"2013","unstructured":"Goff DJ, Court Recart A, Sadarangani A, Chun HJ, Barrett CL, Krajewska M, et al. A Pan-BCL2 inhibitor renders bone-marrow-resident human leukemia stem cells sensitive to tyrosine kinase inhibition. Cell Stem Cell. 2013;12(Mar):316\u201328. 7","journal-title":"Cell Stem Cell"},{"key":"541_CR7","doi-asserted-by":"publisher","first-page":"1062","DOI":"10.1038\/leu.2010.20","volume":"24","author":"J Boultwood","year":"2010","unstructured":"Boultwood J, Perry J, Pellagatti A, Fernandez-Mercado M, Fernandez-Santamaria C, Calasanz MJ, et al. Frequent mutation of the polycomb-associated gene ASXL1 in the myelodysplastic syndromes and in acute myeloid leukemia. Leukemia 2010;24:1062\u20135.","journal-title":"Leukemia"},{"key":"541_CR8","doi-asserted-by":"publisher","first-page":"788","DOI":"10.1111\/j.1365-2141.2009.07697.x","volume":"145","author":"V Gelsi-Boyer","year":"2009","unstructured":"Gelsi-Boyer V, Trouplin V, Ad\u00e9la\u00efde J, Bonansea J, Cervera N, Carbuccia N, et al. Mutations of polycomb-associated gene ASXL1 in myelodysplastic syndromes and chronic myelomonocytic leukaemia. Br J Haematol. 2009;145:788\u2013800.","journal-title":"Br J Haematol"},{"key":"541_CR9","doi-asserted-by":"publisher","first-page":"2477","DOI":"10.1056\/NEJMoa1409405","volume":"371","author":"G Genovese","year":"2014","unstructured":"Genovese G, K\u00e4hler AK, Handsaker RE, Lindberg J, Rose SA, Bakhoum SF, et al. clonal hematopoiesis and blood-cancer risk inferred from blood DNA sequence. N Engl J Med. 2014;371:2477\u201387.","journal-title":"N Engl J Med"},{"key":"541_CR10","doi-asserted-by":"crossref","unstructured":"Yannakou CK, Kate Jones, Michelle McBean, Ella R Thompson, Georgina L Ryland, Ken Doig. ASXL1 c.1934dup;p.Gly646Trpfs*12\u2014a true somatic alteration requiring a new approach. Blood Cancer J. 2017;7:1-4.","DOI":"10.1038\/s41408-017-0025-8"},{"key":"541_CR11","doi-asserted-by":"publisher","first-page":"180","DOI":"10.1016\/j.ccr.2012.06.032","volume":"22","author":"O Abdel-Wahab","year":"2012","unstructured":"Abdel-Wahab O, Adli M, LaFave LM, Gao J, Hricik T, Shih AH, et al. ASXL1 mutations promote myeloid transformation through loss of PRC2-mediated gene repression. Cancer Cell. 2012;22:180\u201393.","journal-title":"Cancer Cell"},{"key":"541_CR12","doi-asserted-by":"publisher","first-page":"44061","DOI":"10.18632\/oncotarget.6392","volume":"6","author":"S Valletta","year":"2015","unstructured":"Valletta S, Dolatshad H, Bartenstein M, Yip BH, Bello E, Gordon S, et al. ASXL1 mutation correction by CRISPR\/Cas9 restores gene function in leukemia cells and increases survival in mouse xenografts. Oncotarget 2015;6:44061\u201371.","journal-title":"Oncotarget"},{"key":"541_CR13","doi-asserted-by":"publisher","first-page":"5479","DOI":"10.1172\/JCI120156","volume":"128","author":"A Shastri","year":"2018","unstructured":"Shastri A, Choudhary G, Teixeira M, Gordon-Mitchell S, Ramachandra N, Bernard L, et al. Antisense STAT3 inhibitor decreases viability of myelodysplastic and leukemic stem cells. J Clin Investig. 2018;128:5479\u201388.","journal-title":"J Clin Investig"},{"key":"541_CR14","doi-asserted-by":"publisher","first-page":"959","DOI":"10.1038\/nmeth.4396","volume":"14","author":"MR Corces","year":"2017","unstructured":"Corces MR, Trevino AE, Hamilton EG, Greenside PG, Sinnott-Armstrong NA, Vesuna S, et al. An improved ATAC-seq protocol reduces background and enables interrogation of frozen tissues. Nat Methods. 2017;14:959\u201362.","journal-title":"Nat Methods"},{"key":"541_CR15","doi-asserted-by":"publisher","first-page":"156","DOI":"10.1016\/j.ymeth.2013.04.006","volume":"61","author":"J Ryan","year":"2013","unstructured":"Ryan J, Letai A. BH3 profiling in whole cells by fluorimeter or FACS. Methods 2013;61:156\u201364.","journal-title":"Methods"},{"key":"541_CR16","doi-asserted-by":"publisher","first-page":"1046","DOI":"10.1101\/gr.5273806","volume":"16","author":"B Khulan","year":"2006","unstructured":"Khulan B, Thompson RF, Ye K, Fazzari MJ, Suzuki M, Stasiek E, et al. Comparative isoschizomer profiling of cytosine methylation: The HELP assay. Genome Res. 2006;16:1046\u201355.","journal-title":"Genome Res"},{"issue":"Jun","key":"541_CR17","doi-asserted-by":"publisher","first-page":"1729","DOI":"10.1084\/jem.20171151","volume":"215","author":"R Nagase","year":"2018","unstructured":"Nagase R, Inoue D, Pastore A, Fujino T, Hou HA, Yamasaki N, et al. Expression of mutant Asxl1 perturbs hematopoiesis and promotes susceptibility to leukemic transformation. J Exp Med. 2018;215(Jun):1729\u201347. 4","journal-title":"J Exp Med"},{"key":"541_CR18","doi-asserted-by":"publisher","first-page":"361","DOI":"10.1038\/nbt.1533","volume":"27","author":"MP Ball","year":"2009","unstructured":"Ball MP, Li JB, Gao Y, Lee JH, LeProust EM, Park IH, et al. Targeted and genome-scale strategies reveal gene-body methylation signatures in human cells. Nat Biotechnol. 2009;27:361\u20138.","journal-title":"Nat Biotechnol"},{"key":"541_CR19","doi-asserted-by":"publisher","first-page":"216","DOI":"10.1016\/S1470-2045(18)30010-X","volume":"19","author":"CD Dinardo","year":"2018","unstructured":"Dinardo CD, Pratz KW, Letai A, Jonas BA, Wei AH, Thirman M, et al. Safety and preliminary efficacy of venetoclax with decitabine or azacitidine in elderly patients with previously untreated acute myeloid leukaemia: a non-randomised, open-label, phase 1b study. Lancet Oncol. 2018;19:216\u201328.","journal-title":"Lancet Oncol"}],"container-title":["Blood Cancer Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41408-021-00541-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41408-021-00541-0","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41408-021-00541-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,1]],"date-time":"2022-12-01T18:23:24Z","timestamp":1669919004000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41408-021-00541-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,9,21]]},"references-count":19,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2021,9]]}},"alternative-id":["541"],"URL":"https:\/\/doi.org\/10.1038\/s41408-021-00541-0","relation":{},"ISSN":["2044-5385"],"issn-type":[{"value":"2044-5385","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,9,21]]},"assertion":[{"value":"8 February 2021","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"23 February 2021","order":2,"name":"revised","label":"Revised","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"16 March 2021","order":3,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"21 September 2021","order":4,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"Amit Verma has received research funding from GSK, Incyte, Medpacto, and Eli Lilly and is a scientific advisor for Stelexis, Novartis, Acceleron, and Celgene. The remaining authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"157"}}