{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,28]],"date-time":"2026-01-28T07:20:33Z","timestamp":1769584833355,"version":"3.49.0"},"reference-count":29,"publisher":"Springer Science and Business Media LLC","issue":"9","license":[{"start":{"date-parts":[[2022,7,28]],"date-time":"2022-07-28T00:00:00Z","timestamp":1658966400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2022,7,28]],"date-time":"2022-07-28T00:00:00Z","timestamp":1658966400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100007903","name":"Bloodwise","doi-asserted-by":"publisher","award":["15012"],"award-info":[{"award-number":["15012"]}],"id":[{"id":"10.13039\/501100007903","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007903","name":"Bloodwise","doi-asserted-by":"publisher","award":["15012"],"award-info":[{"award-number":["15012"]}],"id":[{"id":"10.13039\/501100007903","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007903","name":"Bloodwise","doi-asserted-by":"publisher","award":["15012"],"award-info":[{"award-number":["15012"]}],"id":[{"id":"10.13039\/501100007903","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007903","name":"Bloodwise","doi-asserted-by":"publisher","award":["15012"],"award-info":[{"award-number":["15012"]}],"id":[{"id":"10.13039\/501100007903","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007903","name":"Bloodwise","doi-asserted-by":"publisher","award":["15012"],"award-info":[{"award-number":["15012"]}],"id":[{"id":"10.13039\/501100007903","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007903","name":"Bloodwise","doi-asserted-by":"publisher","award":["15012"],"award-info":[{"award-number":["15012"]}],"id":[{"id":"10.13039\/501100007903","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007903","name":"Bloodwise","doi-asserted-by":"publisher","award":["15012"],"award-info":[{"award-number":["15012"]}],"id":[{"id":"10.13039\/501100007903","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007903","name":"Bloodwise","doi-asserted-by":"publisher","award":["15012"],"award-info":[{"award-number":["15012"]}],"id":[{"id":"10.13039\/501100007903","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Leukemia"],"published-print":{"date-parts":[[2022,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Pathogenesis in chronic lymphocytic leukemia (CLL) is strongly linked to the potential for leukemic cells to migrate to and proliferate within lymph-nodes. Previous in vivo studies suggest that all leukemic cells participate in cycles of migration and proliferation. In vitro studies, however, have shown heterogeneous migration patterns.<\/jats:p><jats:p>To investigate tumor subpopulation kinetics, we performed in vivo isotope-labeling studies in ten patients with <jats:italic>IgVH<\/jats:italic>-mutated CLL (M-CLL). Using deuterium-labeled glucose, we investigated proliferation in sub-populations defined by CXCR4\/CD5 and surface (sIgM) expression. Mathematical modeling was performed to test the likelihood that leukemic cells exist as distinct sub-populations or as a single population with the same proliferative capacity. Further labeling studies in two patients with M-CLL commencing idelalisib investigated the effect of B-cell receptor (BCR) antagonists on sub-population kinetics.<\/jats:p><jats:p>Modeling revealed that data were more consistent with a model comprising distinct sub-populations (<jats:italic>p<\/jats:italic>\u2009=\u20090.008) with contrasting, characteristic kinetics. Following idelalisib therapy, similar labeling suppression across all sub-populations suggested that the most proliferative subset is the most sensitive to treatment. As the quiescent sub-population precedes treatment, selection likely explains the persistence of such residual non-proliferating populations during BCR-antagonist therapy. These findings have clinical implications for discontinuation of long-term BCR-antagonist treatment in selected patients.<\/jats:p>","DOI":"10.1038\/s41375-022-01656-4","type":"journal-article","created":{"date-parts":[[2022,7,28]],"date-time":"2022-07-28T11:04:18Z","timestamp":1659006258000},"page":"2233-2241","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Identification of proliferative and non-proliferative subpopulations of leukemic cells in CLL"],"prefix":"10.1038","volume":"36","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4377-8413","authenticated-orcid":false,"given":"Kirsty M.","family":"Cuthill","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yan","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9795-1280","authenticated-orcid":false,"given":"Andrea","family":"Pepper","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3786-5545","authenticated-orcid":false,"given":"Lies","family":"Boelen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8443-9595","authenticated-orcid":false,"given":"Eve","family":"Coulter","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5911-3160","authenticated-orcid":false,"given":"Becca","family":"Asquith","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8046-0386","authenticated-orcid":false,"given":"Stephen","family":"Devereux","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3014-7148","authenticated-orcid":false,"given":"Derek C.","family":"Macallan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2022,7,28]]},"reference":[{"key":"1656_CR1","doi-asserted-by":"publisher","first-page":"4191","DOI":"10.1182\/blood-2012-05-433540","volume":"120","author":"A Schuh","year":"2012","unstructured":"Schuh A, Becq J, Humphray S, Adrian A, Burns A, Clifford R, et al. Monitoring chronic lymphocytic leukemia progression by whole genome sequencing reveals heterogeneous clonal evolution patterns. Blood. 2012;120:4191\u20136.","journal-title":"Blood"},{"key":"1656_CR2","doi-asserted-by":"publisher","first-page":"4117","DOI":"10.1182\/blood-2012-09-452805","volume":"120","author":"CJ Wu","year":"2012","unstructured":"Wu CJ. CLL clonal heterogeneity: an ecology of competing subpopulations. Blood. 2012;120:4117\u20138. d.","journal-title":"Blood"},{"key":"1656_CR3","doi-asserted-by":"publisher","first-page":"714","DOI":"10.1016\/j.cell.2013.01.019","volume":"152","author":"DA Landau","year":"2013","unstructured":"Landau DA, Carter SL, Stojanov P, McKenna A, Stevenson K, Lawrence M, et al. Evolution and impact of subclonal mutations in chronic lymphocytic leukemia. Cell. 2013;152:714\u201326.","journal-title":"Cell"},{"key":"1656_CR4","doi-asserted-by":"publisher","first-page":"5173","DOI":"10.1182\/blood-2007-08-108605","volume":"111","author":"PEM Patten","year":"2008","unstructured":"Patten PEM, Buggins AGS, Richards J, Wotherspoon A, Salisbury J, Mufti G, et al. CD38 expression in chronic lymphocytic leukemia is regulated by the tumor microenvironment. Blood. 2008;111:5173\u201381.","journal-title":"Blood"},{"key":"1656_CR5","doi-asserted-by":"publisher","first-page":"6387","DOI":"10.1158\/0008-5472.CAN-08-4750","volume":"69","author":"A Vlad","year":"2009","unstructured":"Vlad A, Deglesne PA, Letestu R, Saint-Georges S, Chevalier N, Baran-Marszak F, et al. Down-regulation of CXCR4 and CD62L in chronic lymphocytic leukemia cells is triggered by B-cell receptor ligation and associated with progressive disease. Cancer Res. 2009;69:6387\u201395.","journal-title":"Cancer Res"},{"key":"1656_CR6","doi-asserted-by":"publisher","first-page":"4832","DOI":"10.1182\/blood-2009-05-219634","volume":"114","author":"C Calissano","year":"2009","unstructured":"Calissano C, Damle RN, Hayes G, Murphy E, Hellerstein M, Moreno C, et al. In vivo intraclonal and interclonal kinetic heterogeneity in B-cell chronic lymphocytic leukemia. Blood. 2009;114:4832\u201342.","journal-title":"Blood"},{"key":"1656_CR7","doi-asserted-by":"publisher","first-page":"1374","DOI":"10.2119\/molmed.2011.00360","volume":"17","author":"C Calissano","year":"2011","unstructured":"Calissano C, Damle RN, Marsilio S, Yan X, Yanocopoulos S, Hayes G, et al. Intraclonal complexity in chronic lymphocytic leukemia: fractions enriched in recently born\/divided and older\/quiescent cells. Mol Med. 2011;17:1374\u201382.","journal-title":"Mol Med"},{"key":"1656_CR8","doi-asserted-by":"publisher","first-page":"1340","DOI":"10.1038\/leu.2017.11","volume":"31","author":"TM Herndon","year":"2017","unstructured":"Herndon TM, Chen S-S, Saba NS, Valdez J, Emson C, Gatmaitan M, et al. Direct in vivo evidence for increased proliferation of CLL cells in lymph nodes compared to bone marrow and peripheral blood. Leukemia. 2017;31:1340\u20137.","journal-title":"Leukemia"},{"key":"1656_CR9","doi-asserted-by":"publisher","first-page":"1810","DOI":"10.1182\/blood-2013-09-527853","volume":"123","author":"JA Woyach","year":"2014","unstructured":"Woyach JA, Smucker K, Smith LL, Lozanski A, Zhong Y, Ruppert A, et al. Prolonged lymphocytosis during ibrutinib therapy is associated with distinct molecular characteristics and does not indicate a suboptimal response to therapy. Blood. 2014;123:1810\u20137.","journal-title":"Blood"},{"key":"1656_CR10","doi-asserted-by":"publisher","first-page":"563","DOI":"10.1182\/blood-2016-01-683128","volume":"128","author":"M Pasikowska","year":"2016","unstructured":"Pasikowska M, Walsby E, Apollonio B, Cuthill K, Phillips E, Coulter E, et al. Phenotype and immune function of lymph node and peripheral blood CLL cells are linked to transendothelial migration. Blood. 2016;128:563\u201373.","journal-title":"Blood"},{"key":"1656_CR11","unstructured":"Gruber M, Bozic I, Leshchiner I, Livitz D, Stevenson K, Rassenti L, et al. Growth dynamics in naturally progressing chronic lymphocytic leukaemia. Nature. June 2019:1\u201320."},{"key":"1656_CR12","doi-asserted-by":"publisher","first-page":"240","DOI":"10.1111\/j.1365-2141.2008.07348.x","volume":"143","author":"J Defoiche","year":"2008","unstructured":"Defoiche J, Debacq C, Asquith B, Zhang Y, Burny A, Bron, D et al. Reduction of B cell turnover in chronic lymphocytic leukaemia. Br J Haematol. 2008;143:240\u20137.","journal-title":"Br J Haematol"},{"key":"1656_CR13","doi-asserted-by":"publisher","first-page":"4424","DOI":"10.1182\/blood-2006-11-056648","volume":"109","author":"C Ian Mockridge","year":"2007","unstructured":"Ian Mockridge C, Potter KN, Wheatley I, Neville LA, Packham G, Stevenson FK. Reversible anergy of sIgM-mediated signaling in the two subsets of CLL defined by VH-gene mutational status. Blood. 2007;109:4424\u201331.","journal-title":"Blood"},{"key":"1656_CR14","doi-asserted-by":"publisher","first-page":"497","DOI":"10.3324\/haematol.2017.176164","volume":"103","author":"EM Coulter","year":"2018","unstructured":"Coulter EM, Pepper A, Mele S, Folarin N, Townsend W, Cuthill K, et al. In vitroand in vivoevidence for uncoupling of B-cell receptor internalization and signaling in chronic lymphocytic leukemia. Haematologica. 2018;103:497\u2013505.","journal-title":"Haematologica"},{"key":"1656_CR15","doi-asserted-by":"publisher","first-page":"5446","DOI":"10.1182\/blood-2007-06-093906","volume":"111","author":"M Hallek","year":"2008","unstructured":"Hallek M, Cheson BD, Catovsky D, Caligaris-Cappio F, Dighiero G, Dohner H, et al. Guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: a report from the International Workshop on Chronic Lymphocytic Leukemia updating the National Cancer Institute\u2013Working Group 1996 guidelines. Blood. 2008;111:5446\u201356.","journal-title":"Blood"},{"key":"1656_CR16","doi-asserted-by":"publisher","first-page":"1313","DOI":"10.1038\/nprot.2009.117","volume":"4","author":"DC Macallan","year":"2009","unstructured":"Macallan DC, Asquith B, Zhang Y, de Lara C, Ghattas H, Defoiche J, et al. Measurement of proliferation and disappearance of rapid turnover cell populations in human studies using deuterium-labeled glucose. Nat Protoc. 2009;4:1313\u201327.","journal-title":"Nat Protoc"},{"key":"1656_CR17","doi-asserted-by":"publisher","first-page":"3045","DOI":"10.1038\/nprot.2007.420","volume":"2","author":"R Busch","year":"2007","unstructured":"Busch R, Neese RA, Awada M, Hayes GM, Hellerstein MK. Measurement of cell proliferation by heavy water labeling. Nat Protoc. 2007;2:3045\u201357.","journal-title":"Nat Protoc"},{"key":"1656_CR18","doi-asserted-by":"publisher","first-page":"e89904","DOI":"10.1172\/jci.insight.89904","volume":"2","author":"JA Burger","year":"2017","unstructured":"Burger JA, Li KW, Keating MJ, Sivina M, Amer A, Garg N, et al. Leukemia cell proliferation and death in chronic lymphocytic leukemia patients on therapy with the BTK inhibitor ibrutinib. JCI Insight. 2017;2:e89904.","journal-title":"JCI Insight"},{"key":"1656_CR19","doi-asserted-by":"publisher","first-page":"755","DOI":"10.1172\/JCI23409","volume":"115","author":"BT Messmer","year":"2005","unstructured":"Messmer BT, Messmer D, Allen SL, Kolitz J, Kudalkar P, Cesar D, et al. In vivo measurements document the dynamic cellular kinetics of chronic lymphocytic leukemia B cells. J Clin Invest. 2005;115:755\u201364.","journal-title":"J Clin Invest"},{"key":"1656_CR20","doi-asserted-by":"publisher","first-page":"2664","DOI":"10.1182\/blood-2013-02-485425","volume":"122","author":"V Coelho","year":"2013","unstructured":"Coelho V, Krysov S, Steele A, Hidalgo M, Johnson P, Chana P, et al. Identification in CLL of circulating intraclonal subgroups with varying B-cell receptor expression and function. Blood. 2013;122:2664\u201372.","journal-title":"Blood"},{"key":"1656_CR21","doi-asserted-by":"publisher","first-page":"563","DOI":"10.1182\/blood-2010-05-284984","volume":"117","author":"Y Herishanu","year":"2011","unstructured":"Herishanu Y, P\u00e9rez-Gal\u00e1n P, Liu D, Biancotto A, Pittaluga S, Vire B, et al. The lymph node microenvironment promotes B-cell receptor signaling, NF-kappaB activation, and tumor proliferation in chronic lymphocytic leukemia. Blood. 2011;117:563\u201374.","journal-title":"Blood"},{"key":"1656_CR22","doi-asserted-by":"publisher","first-page":"553","DOI":"10.1182\/blood-2015-11-682997","volume":"128","author":"B Guo","year":"2016","unstructured":"Guo B, Zhang L, Chiorazzi N, Rothstein TL. IL-4 rescues surface IgM expression in chronic lymphocytic leukemia. Blood. 2016;128:553\u201362.","journal-title":"Blood"},{"key":"1656_CR23","doi-asserted-by":"publisher","first-page":"3015","DOI":"10.1182\/blood-2015-11-682906","volume":"127","author":"MM Aguilar-Hernandez","year":"2016","unstructured":"Aguilar-Hernandez MM, Blunt MD, Dobson R, Yeomans A, Thirdborough S, Larrayoz M, et al. IL-4 enhances expression and function of surface IgM in CLL cells. Blood. 2016;127:3015\u201325.","journal-title":"Blood"},{"key":"1656_CR24","doi-asserted-by":"publisher","first-page":"3607","DOI":"10.1182\/blood-2013-12-544569","volume":"123","author":"E Walsby","year":"2014","unstructured":"Walsby E, Buggins A, Devereux S, Jones C, Pratt G, Brennan P, et al. Development and characterization of a physiologically relevant model of lymphocyte migration in chronic lymphocytic leukemia. Blood. 2014;123:3607\u201317.","journal-title":"Blood"},{"key":"1656_CR25","doi-asserted-by":"publisher","first-page":"2188","DOI":"10.1038\/leu.2014.122","volume":"28","author":"SEM Herman","year":"2014","unstructured":"Herman SEM, Niemann CU, Farooqui M, Jones J, Mustafa R, Lipsky A, et al. Ibrutinib-induced lymphocytosis in patients with chronic lymphocytic leukemia: correlative analyses from a phase II study. Leukemia. 2014;28:2188\u201396.","journal-title":"Leukemia"},{"key":"1656_CR26","doi-asserted-by":"publisher","first-page":"2503","DOI":"10.1158\/1078-0432.CCR-18-1286","volume":"25","author":"S Drennan","year":"2019","unstructured":"Drennan S, Chiodin G, D\u2019Avola A, Tracy I, Johnson P, Trenton L, et al. Ibrutinib therapy releases leukemic surface IgM from antigen drive in chronic lymphocytic leukemia patients. Clin Cancer Res. 2019;25:2503\u201312.","journal-title":"Clin Cancer Res"},{"key":"1656_CR27","doi-asserted-by":"publisher","first-page":"389","DOI":"10.1038\/371389a0","volume":"371","author":"JG Cyster","year":"1994","unstructured":"Cyster JG, Hartley SB, Goodnow CC. Competition for follicular niches excludes self-reactive cells from the recirculating B-cell repertoire. Nature. 1994;371:389\u201395.","journal-title":"Nature"},{"key":"1656_CR28","doi-asserted-by":"publisher","DOI":"10.1186\/gm451","volume":"5","author":"DA Landau","year":"2013","unstructured":"Landau DA, Wu CJ. Chronic lymphocytic leukemia: molecular heterogeneity revealed by high-throughput genomics. Genome Med. 2013;5:47.","journal-title":"Genome Med"},{"key":"1656_CR29","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1182\/blood-2019-126824","volume":"134","author":"TD Shanafelt","year":"2019","unstructured":"Shanafelt TD, Wang V, Kay NE, Hanson C, O\u2019Brien S, Barrientos J, et al. Ibrutinib and Rituximab provides superior clinical outcome compared to FCR in younger patients with Chronic Lymphocytic Leukemia (CLL): extended follow-up from the E1912 trial. Blood. 2019;134:33\u201333.","journal-title":"Blood"}],"container-title":["Leukemia"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41375-022-01656-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41375-022-01656-4","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41375-022-01656-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,8,26]],"date-time":"2022-08-26T10:04:44Z","timestamp":1661508284000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41375-022-01656-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,28]]},"references-count":29,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2022,9]]}},"alternative-id":["1656"],"URL":"https:\/\/doi.org\/10.1038\/s41375-022-01656-4","relation":{},"ISSN":["0887-6924","1476-5551"],"issn-type":[{"value":"0887-6924","type":"print"},{"value":"1476-5551","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,7,28]]},"assertion":[{"value":"15 November 2021","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"4 July 2022","order":2,"name":"revised","label":"Revised","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"12 July 2022","order":3,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"28 July 2022","order":4,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}}]}}