{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T13:35:57Z","timestamp":1790516157590,"version":"4.1.0"},"reference-count":33,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,9,24]],"date-time":"2021-09-24T00:00:00Z","timestamp":1632441600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,9,24]],"date-time":"2021-09-24T00:00:00Z","timestamp":1632441600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100012504","name":"Royal Melbourne Hospital Foundation","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100012504","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Hairy cell leukaemia (HCL) is a rare CD20+ B cell malignancy characterised by rare \u201chairy\u201d B cells and extensive bone marrow (BM) infiltration. Frontline treatment with the purine analogue cladribine (CDA) results in a highly variable response duration. We hypothesised that analysis of the BM tumour microenvironment would identify prognostic biomarkers of response to CDA. HCL BM immunology pre and post CDA treatment and healthy controls were analysed using Digital Spatial Profiling to assess the expression of 57 proteins using an immunology panel. A bioinformatics pipeline was developed to accommodate the more complex experimental design of a spatially resolved study. Treatment with CDA was associated with the reduction in expression of HCL tumour markers (CD20, CD11c) and increased expression of myeloid markers (CD14, CD68, CD66b, ARG1). Expression of HLA-DR, STING, CTLA4, VISTA, OX40L were dysregulated pre- and post-CDA. Duration of response to treatment was associated with greater reduction in tumour burden and infiltration by CD8 T cells into the BM post-CDA. This is the first study to provide a high multiplex analysis of HCL BM microenvironment demonstrating significant immune dysregulation and identify biomarkers of response to CDA. With validation in future studies, prospective application of these biomarkers could allow early identification and increased monitoring in patients at increased relapse risk post CDA.<\/jats:p>","DOI":"10.1038\/s41598-021-98536-1","type":"journal-article","created":{"date-parts":[[2021,9,29]],"date-time":"2021-09-29T20:06:05Z","timestamp":1632945965000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Dissection of the bone marrow microenvironment in hairy cell leukaemia identifies prognostic tumour and immune related biomarkers"],"prefix":"10.1038","volume":"11","author":[{"given":"Rachel M.","family":"Koldej","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashvind","family":"Prabahran","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chin Wee","family":"Tan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashley P.","family":"Ng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melissa J.","family":"Davis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David S.","family":"Ritchie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,9,24]]},"reference":[{"key":"98536_CR1","doi-asserted-by":"publisher","first-page":"62","DOI":"10.1038\/s41408-020-0328-z","volume":"10","author":"J Paillassa","year":"2020","unstructured":"Paillassa, J. et al. Analysis of a cohort of 279 patients with hairy-cell leukemia (HCL): 10 years of follow-up. Blood Cancer J. 10, 62. https:\/\/doi.org\/10.1038\/s41408-020-0328-z (2020).","journal-title":"Blood Cancer J."},{"key":"98536_CR2","doi-asserted-by":"publisher","first-page":"2869","DOI":"10.1080\/10428194.2019.1608536","volume":"60","author":"RJ Kreitman","year":"2019","unstructured":"Kreitman, R. J. Hairy cell leukemia: present and future directions. Leuk. Lymphoma 60, 2869\u20132879. https:\/\/doi.org\/10.1080\/10428194.2019.1608536 (2019).","journal-title":"Leuk. Lymphoma"},{"key":"98536_CR3","doi-asserted-by":"publisher","first-page":"172","DOI":"10.1016\/0046-8177(92)90239-y","volume":"23","author":"R Stroup","year":"1992","unstructured":"Stroup, R. & Sheibani, K. Antigenic phenotypes of hairy cell leukemia and monocytoid B-cell lymphoma: an immunohistochemical evaluation of 66 cases. Hum. Pathol. 23, 172\u2013177. https:\/\/doi.org\/10.1016\/0046-8177(92)90239-y (1992).","journal-title":"Hum. Pathol."},{"key":"98536_CR4","doi-asserted-by":"publisher","first-page":"275","DOI":"10.1002\/cyto.b.21785","volume":"96","author":"DA Salem","year":"2019","unstructured":"Salem, D. A. et al. Differential expression of CD43, CD81, and CD200 in classic versus variant hairy cell leukemia. Cytometry Part B Clin. Cytom. 96, 275\u2013282. https:\/\/doi.org\/10.1002\/cyto.b.21785 (2019).","journal-title":"Cytometry Part B Clin. Cytom."},{"key":"98536_CR5","doi-asserted-by":"publisher","first-page":"385","DOI":"10.1046\/j.1365-2257.2001.00417.x","volume":"23","author":"L Tytherleigh","year":"2001","unstructured":"Tytherleigh, L., Taparia, M. & Leahy, M. F. Detection of hairy cell leukaemia in blood and bone marrow using multidimensional flow cytometry with CD45-PECy5 and SS gating. Clin. Lab. Haematol. 23, 385\u2013390. https:\/\/doi.org\/10.1046\/j.1365-2257.2001.00417.x (2001).","journal-title":"Clin. Lab. Haematol."},{"key":"98536_CR6","doi-asserted-by":"publisher","first-page":"115","DOI":"10.1038\/s41408-020-00384-1","volume":"10","author":"P Kumar","year":"2020","unstructured":"Kumar, P. et al. Hairy cell leukemia expresses programmed death-1. Blood Cancer J. 10, 115. https:\/\/doi.org\/10.1038\/s41408-020-00384-1 (2020).","journal-title":"Blood Cancer J."},{"key":"98536_CR7","first-page":"600","volume":"53","author":"RA Knight","year":"1983","unstructured":"Knight, R. A., Worman, C. P. & Cawley, J. C. Defective autologous and allogeneic mixed lymphocyte reactions in hairy cell leukaemia. Clin. Exp. Immunol. 53, 600\u2013604 (1983).","journal-title":"Clin. Exp. Immunol."},{"key":"98536_CR8","doi-asserted-by":"publisher","first-page":"1525","DOI":"10.1182\/blood.V75.7.1525.1525","volume":"75","author":"L Trentin","year":"1990","unstructured":"Trentin, L. et al. Mechanisms accounting for the defective natural killer activity in patients with hairy cell leukemia. Blood 75, 1525\u20131530 (1990).","journal-title":"Blood"},{"key":"98536_CR9","doi-asserted-by":"publisher","first-page":"256","DOI":"10.1006\/clim.1999.4794","volume":"93","author":"L van de Corput","year":"1999","unstructured":"van de Corput, L., Falkenburg, J. H., Kester, M. G., Willemze, R. & Kluin-Nelemans, J. C. Impaired expression of CD28 on T cells in hairy cell leukemia. Clin. Immunol. 93, 256\u2013262. https:\/\/doi.org\/10.1006\/clim.1999.4794 (1999).","journal-title":"Clin. Immunol."},{"key":"98536_CR10","first-page":"46","volume":"7","author":"FA van der Horst","year":"1993","unstructured":"van der Horst, F. A., van der Marel, A., den Ottolander, G. J. & Kluin-Nelemans, H. C. Decrease of memory T helper cells (CD4+ CD45R0+) in hairy cell leukemia. Leukemia 7, 46\u201350 (1993).","journal-title":"Leukemia"},{"key":"98536_CR11","doi-asserted-by":"publisher","first-page":"1254","DOI":"10.1038\/sj.leu.2401476","volume":"13","author":"D Raspadori","year":"1999","unstructured":"Raspadori, D. et al. Long-lasting decrease of CD4+\/CD45RA+ T cells in HCL patients after 2-chlorodeoxyadenosine (2-CdA) treatment. Leukemia 13, 1254\u20131257. https:\/\/doi.org\/10.1038\/sj.leu.2401476 (1999).","journal-title":"Leukemia"},{"key":"98536_CR12","doi-asserted-by":"publisher","first-page":"1051","DOI":"10.1016\/j.hoc.2006.06.012","volume":"20","author":"E Matutes","year":"2006","unstructured":"Matutes, E. Immunophenotyping and differential diagnosis of hairy cell leukemia. Hematol. Oncol. Clin. North Am. 20, 1051\u20131063. https:\/\/doi.org\/10.1016\/j.hoc.2006.06.012 (2006).","journal-title":"Hematol. Oncol. Clin. North Am."},{"key":"98536_CR13","doi-asserted-by":"publisher","first-page":"543","DOI":"10.1111\/bjh.14524","volume":"177","author":"PA Thompson","year":"2017","unstructured":"Thompson, P. A. & Ravandi, F. How I manage patients with hairy cell leukaemia. Br. J. Haematol. 177, 543\u2013556. https:\/\/doi.org\/10.1111\/bjh.14524 (2017).","journal-title":"Br. J. Haematol."},{"key":"98536_CR14","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.iotech.2020.02.001","volume":"5","author":"R Koldej","year":"2020","unstructured":"Koldej, R. & Ritchie, D. High multiplex analysis of the immune microenvironment in bone marrow trephine samples using GeoMXTM digital spatial profiling. Immuno-Oncol. Technol. 5, 1\u20139. https:\/\/doi.org\/10.1016\/j.iotech.2020.02.001 (2020).","journal-title":"Immuno-Oncol. Technol."},{"key":"98536_CR15","doi-asserted-by":"publisher","first-page":"497","DOI":"10.3109\/10428199609093449","volume":"21","author":"S Rutella","year":"1996","unstructured":"Rutella, S. et al. Flow cytometric detection of CD44 (H-CAM) in hairy cell leukemia. Leuk. Lymphoma 21, 497\u2013500. https:\/\/doi.org\/10.3109\/10428199609093449 (1996).","journal-title":"Leuk. Lymphoma"},{"key":"98536_CR16","doi-asserted-by":"publisher","first-page":"274","DOI":"10.1182\/bloodadvances.2019000792","volume":"4","author":"O Bruck","year":"2020","unstructured":"Bruck, O. et al. Immune profiles in acute myeloid leukemia bone marrow associate with patient age, T-cell receptor clonality, and survival. Blood Adv. 4, 274\u2013286. https:\/\/doi.org\/10.1182\/bloodadvances.2019000792 (2020).","journal-title":"Blood Adv."},{"key":"98536_CR17","doi-asserted-by":"publisher","DOI":"10.3390\/cancers13020186","author":"M Bauer","year":"2021","unstructured":"Bauer, M. et al. Altered spatial composition of the immune cell repertoire in association to CD34(+) blasts in myelodysplastic syndromes and secondary acute myeloid leukemia. Cancers (Basel) https:\/\/doi.org\/10.3390\/cancers13020186 (2021).","journal-title":"Cancers (Basel)"},{"key":"98536_CR18","doi-asserted-by":"publisher","first-page":"180","DOI":"10.1159\/000203677","volume":"97","author":"M Schirmer","year":"1997","unstructured":"Schirmer, M., Hilbe, W., Geisen, F., Thaler, J. & Konwalinka, G. T cells and natural killer cells after treatment of hairy cell leukaemia with 2-chlorodeoxyadenosine. Acta Haematol. 97, 180\u2013183. https:\/\/doi.org\/10.1159\/000203677 (1997).","journal-title":"Acta Haematol."},{"key":"98536_CR19","doi-asserted-by":"publisher","first-page":"3672","DOI":"10.1182\/blood.V83.12.3672.3672","volume":"83","author":"G Juliusson","year":"1994","unstructured":"Juliusson, G., Lenkei, R. & Liliemark, J. Flow cytometry of blood and bone marrow cells from patients with hairy cell leukemia: phenotype of hairy cells and lymphocyte subsets after treatment with 2-chlorodeoxyadenosine. Blood 83, 3672\u20133681 (1994).","journal-title":"Blood"},{"key":"98536_CR20","doi-asserted-by":"publisher","first-page":"2906","DOI":"10.1182\/blood.V83.10.2906.2906","volume":"83","author":"JF Seymour","year":"1994","unstructured":"Seymour, J. F., Kurzrock, R., Freireich, E. J. & Estey, E. H. 2-chlorodeoxyadenosine induces durable remissions and prolonged suppression of CD4+ lymphocyte counts in patients with hairy cell leukemia. Blood 83, 2906\u20132911 (1994).","journal-title":"Blood"},{"key":"98536_CR21","doi-asserted-by":"publisher","first-page":"2199","DOI":"10.1002\/acn3.51206","volume":"7","author":"T Moser","year":"2020","unstructured":"Moser, T. et al. Long-term peripheral immune cell profiling reveals further targets of oral cladribine in MS. Ann. Clin. Transl. Neurol. 7, 2199\u20132212. https:\/\/doi.org\/10.1002\/acn3.51206 (2020).","journal-title":"Ann. Clin. Transl. Neurol."},{"key":"98536_CR22","doi-asserted-by":"publisher","first-page":"175628641985498","DOI":"10.1177\/1756286419854986","volume":"12","author":"O Stuve","year":"2019","unstructured":"Stuve, O. et al. Effects of cladribine tablets on lymphocyte subsets in patients with multiple sclerosis: an extended analysis of surface markers. Ther. Adv. Neurol. Disord. 12, 1756286419854986. https:\/\/doi.org\/10.1177\/1756286419854986 (2019).","journal-title":"Ther. Adv. Neurol. Disord."},{"key":"98536_CR23","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1016\/j.jns.2013.06.003","volume":"332","author":"K Mitosek-Szewczyk","year":"2013","unstructured":"Mitosek-Szewczyk, K. et al. Impact of cladribine therapy on changes in circulating dendritic cell subsets, T cells and B cells in patients with multiple sclerosis. J. Neurol. Sci. 332, 35\u201340. https:\/\/doi.org\/10.1016\/j.jns.2013.06.003 (2013).","journal-title":"J. Neurol. Sci."},{"key":"98536_CR24","doi-asserted-by":"publisher","first-page":"347","DOI":"10.1016\/j.intimp.2013.11.027","volume":"18","author":"SH Kraus","year":"2014","unstructured":"Kraus, S. H. et al. Cladribine exerts an immunomodulatory effect on human and murine dendritic cells. Int. Immunopharmacol. 18, 347\u2013357. https:\/\/doi.org\/10.1016\/j.intimp.2013.11.027 (2014).","journal-title":"Int. Immunopharmacol."},{"key":"98536_CR25","doi-asserted-by":"publisher","first-page":"31","DOI":"10.3109\/10428199809050927","volume":"30","author":"L Van De Corput","year":"1998","unstructured":"Van De Corput, L., Falkenburg, J. H. & Kluin-Nelemans, J. C. T-cell dysfunction in hairy cell leukemia: an updated review. Leuk. Lymphoma 30, 31\u201339. https:\/\/doi.org\/10.3109\/10428199809050927 (1998).","journal-title":"Leuk. Lymphoma"},{"key":"98536_CR26","doi-asserted-by":"publisher","first-page":"674","DOI":"10.1038\/nature07317","volume":"455","author":"H Ishikawa","year":"2008","unstructured":"Ishikawa, H. & Barber, G. N. STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling. Nature 455, 674\u2013678. https:\/\/doi.org\/10.1038\/nature07317 (2008).","journal-title":"Nature"},{"key":"98536_CR27","first-page":"1056","volume":"52","author":"S Vedantham","year":"1992","unstructured":"Vedantham, S., Gamliel, H. & Golomb, H. M. Mechanism of interferon action in hairy cell leukemia: a model of effective cancer biotherapy. Can. Res. 52, 1056\u20131066 (1992).","journal-title":"Can. Res."},{"key":"98536_CR28","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0191629","volume":"13","author":"LC Gandolfo","year":"2018","unstructured":"Gandolfo, L. C. & Speed, T. P. RLE plots: Visualizing unwanted variation in high dimensional data. PLoS ONE 13, e0191629. https:\/\/doi.org\/10.1371\/journal.pone.0191629 (2018).","journal-title":"PLoS ONE"},{"key":"98536_CR29","doi-asserted-by":"publisher","first-page":"R25","DOI":"10.1186\/gb-2010-11-3-r25","volume":"11","author":"MD Robinson","year":"2010","unstructured":"Robinson, M. D. & Oshlack, A. A scaling normalization method for differential expression analysis of RNA-seq data. Genome Biol. 11, R25. https:\/\/doi.org\/10.1186\/gb-2010-11-3-r25 (2010).","journal-title":"Genome Biol."},{"key":"98536_CR30","doi-asserted-by":"publisher","first-page":"e47","DOI":"10.1093\/nar\/gkv007","volume":"43","author":"ME Ritchie","year":"2015","unstructured":"Ritchie, M. E. et al. limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res. 43, e47\u2013e47. https:\/\/doi.org\/10.1093\/nar\/gkv007 (2015).","journal-title":"Nucleic Acids Res."},{"key":"98536_CR31","doi-asserted-by":"publisher","first-page":"2067","DOI":"10.1093\/bioinformatics\/bti270","volume":"21","author":"GK Smyth","year":"2005","unstructured":"Smyth, G. K., Michaud, J. & Scott, H. S. Use of within-array replicate spots for assessing differential expression in microarray experiments. Bioinformatics 21, 2067\u20132075. https:\/\/doi.org\/10.1093\/bioinformatics\/bti270 (2005).","journal-title":"Bioinformatics"},{"key":"98536_CR32","doi-asserted-by":"publisher","first-page":"R29","DOI":"10.1186\/gb-2014-15-2-r29","volume":"15","author":"CW Law","year":"2014","unstructured":"Law, C. W., Chen, Y., Shi, W. & Smyth, G. K. voom: precision weights unlock linear model analysis tools for RNA-seq read counts. Genome Biol. 15, R29. https:\/\/doi.org\/10.1186\/gb-2014-15-2-r29 (2014).","journal-title":"Genome Biol."},{"key":"98536_CR33","doi-asserted-by":"publisher","first-page":"765","DOI":"10.1093\/bioinformatics\/btp053","volume":"25","author":"DJ McCarthy","year":"2009","unstructured":"McCarthy, D. J. & Smyth, G. K. Testing significance relative to a fold-change threshold is a TREAT. Bioinformatics 25, 765\u2013771. https:\/\/doi.org\/10.1093\/bioinformatics\/btp053 (2009).","journal-title":"Bioinformatics"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-98536-1.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-98536-1","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-98536-1.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,3]],"date-time":"2022-12-03T16:03:37Z","timestamp":1670083417000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-98536-1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,9,24]]},"references-count":33,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["98536"],"URL":"https:\/\/doi.org\/10.1038\/s41598-021-98536-1","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,9,24]]},"assertion":[{"value":"18 May 2021","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"8 September 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"24 September 2021","order":3,"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":"Competing interests"}}],"article-number":"19056"}}