{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T12:00:45Z","timestamp":1774958445228,"version":"3.50.1"},"reference-count":50,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2023,3,15]],"date-time":"2023-03-15T00:00:00Z","timestamp":1678838400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003196","name":"Italian Ministry of Health","doi-asserted-by":"publisher","award":["RC-2022-2773350"],"award-info":[{"award-number":["RC-2022-2773350"]}],"id":[{"id":"10.13039\/501100003196","id-type":"DOI","asserted-by":"publisher"}]},{"name":"AIL Pesaro ONLUS (Visani)","award":["RC-2022-2773350"],"award-info":[{"award-number":["RC-2022-2773350"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Primary myelofibrosis (PMF) is a chronic myeloproliferative neoplasm (MPN) characterized by progressive bone marrow sclerosis, extra-medullary hematopoiesis, and possible transformation to acute leukemia. In the last decade, the molecular pathogenesis of the disease has been largely uncovered. Particularly, genetic and genomic studies have provided evidence of deregulated oncogenes in PMF as well as in other MPNs. However, the mechanisms through which transformation to either the myeloid or lymphoid blastic phase remain obscure. Particularly, it is still debated whether the disease has origins in a multi-potent hematopoietic stem cells or instead in a commissioned myeloid progenitor. In this study, we aimed to shed light upon this issue by using next generation sequencing (NGS) to study both myeloid and lymphoid cells as well as matched non-neoplastic DNA of PMF patients. Whole exome sequencing revealed that most somatic mutations were the same between myeloid and lymphoid cells, such findings being confirmed by Sanger sequencing. Particularly, we found 126\/146 SNVs to be the e same (including JAK2V617F), indicating that most genetic events likely to contribute to disease pathogenesis occurred in a non-commissioned precursor. In contrast, only 9\/27 InDels were similar, suggesting that this type of lesion contributed instead to disease progression, occurring at more differentiated stages, or maybe just represented \u201cpassenger\u201d lesions, not contributing at all to disease pathogenesis. In conclusion, we showed for the first time that genetic lesions characteristic of PMF occur at an early stage of hematopoietic stem cell differentiation, this being in line with the possible transformation of the disease in either myeloid or lymphoid acute leukemia.<\/jats:p>","DOI":"10.3390\/cancers15061785","type":"journal-article","created":{"date-parts":[[2023,3,16]],"date-time":"2023-03-16T02:40:11Z","timestamp":1678934411000},"page":"1785","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Use of Next Generation Sequencing to Define the Origin of Primary Myelofibrosis"],"prefix":"10.3390","volume":"15","author":[{"given":"Giuseppe","family":"Visani","sequence":"first","affiliation":[{"name":"Hematology and Stem Cell Transplantation, AORMIN, 61121 Pesaro, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maryam","family":"Etebari","sequence":"additional","affiliation":[{"name":"Department of Medical Biotechnology, School of Paramedical Sciences, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh 33787-95196, Iran"},{"name":"Research Center of Advanced Technologies in Medicine, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh 33787-95196, Iran"},{"name":"Department of Medical Science and Surgery (DIMEC), University of Bologna, 40126 Bologna, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fabio","family":"Fuligni","sequence":"additional","affiliation":[{"name":"The Hospital for Sick Children, Toronto, ON M5G 1X8, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antonio","family":"Di Guardo","sequence":"additional","affiliation":[{"name":"Department of Medical Science and Surgery (DIMEC), University of Bologna, 40126 Bologna, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alessandro","family":"Isidori","sequence":"additional","affiliation":[{"name":"Hematology and Stem Cell Transplantation, AORMIN, 61121 Pesaro, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Federica","family":"Loscocco","sequence":"additional","affiliation":[{"name":"Hematology and Stem Cell Transplantation, AORMIN, 61121 Pesaro, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stefania","family":"Paolini","sequence":"additional","affiliation":[{"name":"Biobank of Research, IRCCS Azienda Ospedaliero-Universitaria di Bologna, 40138 Bologna, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mohsen","family":"Navari","sequence":"additional","affiliation":[{"name":"Department of Medical Biotechnology, School of Paramedical Sciences, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh 33787-95196, Iran"},{"name":"Research Center of Advanced Technologies in Medicine, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh 33787-95196, Iran"},{"name":"Bioinformatics Research Center, Mashhad University of Medical Sciences, Mashhad 91778-99191, Iran"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9432-0595","authenticated-orcid":false,"given":"Pier Paolo","family":"Piccaluga","sequence":"additional","affiliation":[{"name":"Department of Medical Science and Surgery (DIMEC), University of Bologna, 40126 Bologna, Italy"},{"name":"Biobank of Research, IRCCS Azienda Ospedaliero-Universitaria di Bologna, 40138 Bologna, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,3,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1703","DOI":"10.1038\/s41375-022-01613-1","article-title":"The 5th edition of the World Health Organization Classification of Haematolymphoid Tumours: Myeloid and Histiocytic\/Dendritic Neoplasms","volume":"36","author":"Khoury","year":"2022","journal-title":"Leukemia"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2891","DOI":"10.1182\/blood-2009-08-236596","article-title":"Two routes to leukemic transformation after a JAK2 mutation-positive myeloproliferative neoplasm","volume":"115","author":"Beer","year":"2010","journal-title":"Blood"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1118","DOI":"10.1016\/j.mayocp.2017.05.010","article-title":"Leukemic Transformation in Myeloproliferative Neoplasms: A Literature Review on Risk, Characteristics, and Outcome","volume":"92","author":"Yogarajah","year":"2017","journal-title":"Mayo Clin. Proc."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.cancergen.2017.10.001","article-title":"B-cell acute lymphoblastic leukemia with +der(1)t(1;19) (p13;p13.1) arising in the setting of CALR exon 9 mutated essential thrombocythemia","volume":"218\u2013219","author":"Hilal","year":"2017","journal-title":"Cancer Genet."},{"key":"ref_5","first-page":"6545861","article-title":"Acute Lymphoblastic Leukemia Arising in CALR Mutated Essential Thrombocythemia","volume":"2016","author":"Langabeer","year":"2016","journal-title":"Case Rep. Hematol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2068","DOI":"10.1158\/1055-9965.EPI-09-0353","article-title":"Increased risk of lymphoid neoplasms in patients with Philadelphia chromosome-negative myeloproliferative neoplasms","volume":"18","author":"Vannucchi","year":"2009","journal-title":"Cancer Epidemiol. Biomark. Prev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1002\/ccr3.1264","article-title":"Acute lymphoblastic leukemia secondary to myeloproliferative neoplasms or after lenalidomide exposure","volume":"6","author":"Alhuraiji","year":"2018","journal-title":"Clin. Case Rep."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"96","DOI":"10.5858\/2005-129-96-ALLTIA","article-title":"Acute lymphoblastic leukemic transformation in a patient with chronic idiopathic myelofibrosis and paroxysmal nocturnal hemoglobinuria: A case report and review of the literature","volume":"129","author":"Shaheen","year":"2005","journal-title":"Arch. Pathol. Lab. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e186","DOI":"10.1016\/j.leukres.2009.05.005","article-title":"Primary myelofibrosis with concurrent precursor T-cell lymphoblastic lymphoma of the spleen in a 26-year-old patient","volume":"33","author":"Sever","year":"2009","journal-title":"Leuk. Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1016\/j.prp.2012.04.007","article-title":"Transformation of primary myelofibrosis with 20q- in Philadelphia-positive acute lymphoblastic leukemia: Case report and review of literature","volume":"208","author":"Jurisic","year":"2012","journal-title":"Pathol. Res. Pract."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1016\/j.leukres.2015.08.004","article-title":"Three-fold higher frequency of circulating chronic lymphocytic leukemia-like B-cell clones in patients with Ph-Myeloproliferative neoplasms","volume":"39","author":"Miltiades","year":"2015","journal-title":"Leuk. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"8520","DOI":"10.1200\/JCO.2004.00.9316","article-title":"Pathogenesis of myelofibrosis with myeloid metaplasia","volume":"23","author":"Tefferi","year":"2005","journal-title":"J. Clin. Oncol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1007\/s11899-022-00685-1","article-title":"Molecular Pathogenesis of Myeloproliferative Neoplasms","volume":"17","author":"Rolles","year":"2022","journal-title":"Curr. Hematol. Malig. Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1779","DOI":"10.1056\/NEJMoa051113","article-title":"A Gain-of-Function Mutation of JAK2 in Myeloproliferative Disorders","volume":"352","author":"Kralovics","year":"2005","journal-title":"N. Engl. J. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1182\/blood-2006-03-007146","article-title":"Evidence that the JAK2 G1849T (V617F) mutation occurs in a lymphomyeloid progenitor in polycythemia vera and idiopathic myelofibrosis","volume":"109","author":"Delhommeau","year":"2007","journal-title":"Blood"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1007\/s00277-016-2785-9","article-title":"Primary myelofibrosis and its targeted therapy","volume":"96","author":"Shantzer","year":"2017","journal-title":"Ann. Hematol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1182\/blood-2016-10-695940","article-title":"Genetic basis and molecular pathophysiology of classical myeloproliferative neoplasms","volume":"129","author":"Vainchenker","year":"2017","journal-title":"Blood"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1056\/NEJMoa065202","article-title":"JAK2 Exon 12 Mutations in Polycythemia Vera and Idiopathic Erythrocytosis","volume":"356","author":"Scott","year":"2007","journal-title":"N. Engl. J. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1016\/j.ccr.2005.03.023","article-title":"Activating mutation in the tyrosine kinase JAK2 in polycythemia vera, essential thrombocythemia, and myeloid metaplasia with myelofibrosis","volume":"7","author":"Levine","year":"2005","journal-title":"Cancer Cell"},{"key":"ref_20","first-page":"300","article-title":"Circulating cell-free DNA improves the molecular characterisation of Ph-negative myeloproliferative neoplasms","volume":"192","author":"Gibert","year":"2020","journal-title":"Br. J. Haematol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"745","DOI":"10.1111\/j.1365-2141.2007.06497.x","article-title":"The JAK2 V617F mutation involves B- and T-lymphocyte lineages in a subgroup of patients with Philadelphia-chromosome negative chronic myeloproliferative disorders","volume":"136","author":"Larsen","year":"2007","journal-title":"Br. J. Haematol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4508","DOI":"10.1182\/blood-2004-08-3238","article-title":"Constitutive mobilization of CD34+ cells into the peripheral blood in idiopathic myelofibrosis may be due to the action of a number of proteases","volume":"105","author":"Xu","year":"2005","journal-title":"Blood"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1699","DOI":"10.1182\/blood-2004-06-2485","article-title":"The constitutive mobilization of bone marrow-repopulating cells into the peripheral blood in idiopathic myelofibrosis","volume":"105","author":"Xu","year":"2005","journal-title":"Blood"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"768","DOI":"10.3324\/haematol.2014.118463","article-title":"CD133 marks a stem cell population that drives human primary myelofibrosis","volume":"100","author":"Triviai","year":"2015","journal-title":"Haematologica"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1038\/s41586-020-2864-x","article-title":"Single-cell mutation analysis of clonal evolution in myeloid malignancies","volume":"587","author":"Miles","year":"2020","journal-title":"Nature"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Visani, G., Sapienza, M.R., Isidori, A., Tripodo, C., Laginestra, M.A., Righi, S., Sacchetti, C.A.S., Gazzola, A., Mannu, C., and Rossi, M. (2011). SNPs array karyotyping reveals a novel recurrent 20p13 amplification in primary myelofibrosis. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0027560"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1754","DOI":"10.1093\/bioinformatics\/btp324","article-title":"Fast and accurate short read alignment with Burrows-Wheeler transform","volume":"25","author":"Li","year":"2009","journal-title":"Bioinformatics"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1297","DOI":"10.1101\/gr.107524.110","article-title":"The genome analysis toolkit: A MapReduce framework for analyzing next-generation DNA sequencing data","volume":"20","author":"McKenna","year":"2010","journal-title":"Genome Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1038\/nbt.1754","article-title":"Integrative genomics viewer","volume":"29","author":"Robinson","year":"2011","journal-title":"Nat. Biotechnol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1011","DOI":"10.1038\/srep01011","article-title":"Managing biological complexity across orthologs with a visual knowledgebase of documented biomolecular interactions","volume":"2","author":"Vanburen","year":"2012","journal-title":"Sci. Rep."},{"key":"ref_31","first-page":"167","article-title":"Use of IGK gene rearrangement analysis for clonality assessment of lymphoid malignancies: A single center experience","volume":"1","author":"Mannu","year":"2011","journal-title":"Am. J. Blood Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1186\/s12929-016-0269-9","article-title":"Zinc finger proteins in cancer progression","volume":"23","author":"Jen","year":"2016","journal-title":"J. Biomed. Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1111\/apm.12692","article-title":"IFI16 reduced expression is correlated with unfavorable outcome in chronic lymphocytic leukemia","volume":"125","author":"Piccaluga","year":"2017","journal-title":"Apmis"},{"key":"ref_34","first-page":"1151","article-title":"The role of neuregulin4 and HER4 in gastrointestinal malignant lymphoma","volume":"4","author":"Kataoka","year":"2011","journal-title":"Mol. Med. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"44735","DOI":"10.18632\/oncotarget.8203","article-title":"Molecular and clinical significance of fibroblast growth factor 2 (FGF2\/bFGF) in malignancies of solid and hematological cancers for personalized therapies","volume":"7","author":"Akl","year":"2016","journal-title":"Oncotarget"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"627","DOI":"10.3892\/ijmm.2015.2267","article-title":"Autocrine motility factor receptor promotes the proliferation of human acute monocytic leukemia THP-1 cells","volume":"36","author":"Wang","year":"2015","journal-title":"Int. J. Mol. Med."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"4773","DOI":"10.3748\/wjg.v24.i42.4773","article-title":"Ubiquitin-like modifier activating enzyme 2 promotes cell migration and invasion through Wnt\/\u03b2-catenin signaling in gastric cancer","volume":"24","author":"Li","year":"2018","journal-title":"World J. Gastroenterol."},{"key":"ref_38","first-page":"186","article-title":"Predicting pathogenic genes for primary myelofibrosis based on a system-network approach","volume":"17","author":"Xu","year":"2018","journal-title":"Mol. Med. Rep."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"S105","DOI":"10.1016\/j.clml.2016.02.031","article-title":"Prognosis of Primary Myelofibrosis in the Genomic Era","volume":"16","author":"Bose","year":"2016","journal-title":"Clin. Lymphoma Myeloma Leuk."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"258","DOI":"10.3324\/haematol.10527","article-title":"B-, T-, and NK-cell lineage involvement in JAK2V617F-positive patients with idiopathic myelofibrosis","volume":"92","author":"Bogani","year":"2007","journal-title":"Haematologica"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Farina, M., Bernardi, S., Polverelli, N., D\u2019Adda, M., Malagola, M., Bosio, K., Re, F., Almici, C., Dunbar, A., and Levine, R.L. (2021). Comparative mutational profiling of hematopoietic progenitor cells and circulating endothelial cells (Cecs) in patients with primary myelofibrosis. Cells, 10.","DOI":"10.3390\/cells10102764"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"G\u0103man, M.-A., Cozma, M.-A., Dobric\u0103, E.-C., Cre\u021boiu, S.M., G\u0103man, A.M., and Diaconu, C.C. (2021). Liquid biopsy and potential liquid biopsy-based biomarkers in philadelphia-negative classical myeloproliferative neoplasms: A systematic review. Life, 11.","DOI":"10.3390\/life11070677"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"454","DOI":"10.3324\/haematol.2010.033779","article-title":"Increased risk of lymphoid neoplasm in patients with myeloproliferative neoplasm: A study of 1,915 patients","volume":"96","author":"Rumi","year":"2011","journal-title":"Haematologica"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"822","DOI":"10.1016\/j.leukres.2015.05.002","article-title":"Association of lymphoid malignancies and Philadelphia-chromosome negative myeloproliferative neoplasms: Clinical characteristics, therapy and outcome","volume":"39","author":"Masarova","year":"2015","journal-title":"Leuk. Res."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2203","DOI":"10.1038\/s41375-018-0027-y","article-title":"Second malignancies in patients with myeloproliferative neoplasms: A population-based cohort study of 9379 patients","volume":"32","author":"Landtblom","year":"2018","journal-title":"Leukemia"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1469","DOI":"10.3892\/ol.2015.4040","article-title":"Coexistence of follicular lymphoma and an unclassifiable myeloproliferative neoplasm in a treatment-na\u00efve patient: A case report","volume":"11","author":"Jeong","year":"2016","journal-title":"Oncol. Lett."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1182\/blood-2017-10-810739","article-title":"Aggressive B-cell lymphomas in patients with myelofibrosis receiving JAK1\/2 inhibitor therapy","volume":"132","author":"Porpaczy","year":"2018","journal-title":"Blood"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.3109\/10428194.2015.1092527","article-title":"Chronic lymphocytic leukemia and myeloproliferative neoplasms concurrently diagnosed: Clinical and biological characteristics","volume":"57","author":"Todisco","year":"2016","journal-title":"Leuk Lymphoma."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1159\/000342484","article-title":"Simultaneous and sequential concurrent myeloproliferative and lymphoproliferative neoplasms","volume":"129","author":"Hauck","year":"2013","journal-title":"Acta Haematol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"2647","DOI":"10.1038\/s41375-022-01700-3","article-title":"Compartment-specific mutational landscape of clonal hematopoiesis","volume":"36","author":"Hartmann","year":"2022","journal-title":"Leukemia"}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/15\/6\/1785\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:56:03Z","timestamp":1760122563000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/15\/6\/1785"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,15]]},"references-count":50,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2023,3]]}},"alternative-id":["cancers15061785"],"URL":"https:\/\/doi.org\/10.3390\/cancers15061785","relation":{},"ISSN":["2072-6694"],"issn-type":[{"value":"2072-6694","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3,15]]}}}