{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:05:55Z","timestamp":1760058355869,"version":"build-2065373602"},"reference-count":42,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2025,3,27]],"date-time":"2025-03-27T00:00:00Z","timestamp":1743033600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Science Center in Poland","award":["NCN 2018\/31\/B\/NZ6\/03361","DS 462"],"award-info":[{"award-number":["NCN 2018\/31\/B\/NZ6\/03361","DS 462"]}]},{"DOI":"10.13039\/501100010621","name":"Medical University of Lublin","doi-asserted-by":"publisher","award":["NCN 2018\/31\/B\/NZ6\/03361","DS 462"],"award-info":[{"award-number":["NCN 2018\/31\/B\/NZ6\/03361","DS 462"]}],"id":[{"id":"10.13039\/501100010621","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JCM"],"abstract":"<jats:p>Background\/Objectives: Chronic myeloid leukemia (CML) is characterized by the presence of the BCR::ABL1 fusion gene, most commonly in the e14a2 or e13a2 variants. Studies show that the transcript type in CML may be important for achieving treatment-free remission (TFR). This study aimed to immunologically characterize CML patients with e13a2 and e14a2 transcripts to search for differences that may contribute to achieving remission in patients after therapy withdrawal. Methods: Using multicolor flow cytometry, we analyzed the differences in the immune system at the time of imatinib discontinuation and the early stage of TFR in fifty-one CML patients with different transcripts. RQ-PCR and ddPCR were used to monitor the dynamics of BCR::ABL1 transcript changes. The patients were grouped using principal component analysis (PCA) based on the percentage of detected immune cells that were classified as populations consistently selected by the MCFS-ID algorithm from randomly selected data. Results: PCA separated CML patients into two groups defined by k-means clustering, indicating significant heterogeneity within the studied population. We found a significant association between Cluster metrics (Cluster 1 and 2) and BCR::ABL1 transcript types (e13a2 or e14a2) (p = 0.003, 95% CI: 0.026\u20130.595, OR = 0.14, Fisher test). The e13a2 transcript was less frequent in Cluster 2 than in Cluster 1, while e14a2 was more common in Cluster 2. Additionally, patients grouped into Cluster 1 had significantly higher percentages of the PD1 expressing populations cDC PD1+, CD56dimCD16+PD1+, CD8+PD1+, CD4+PD1+, and CD19+PD1+, as identified by the MCFS-ID algorithm, compared to patients in Cluster 2. Conclusions: Our results suggest that immunological differences may be related to the BCR::ABL1 transcript type, which could affect the number of active CML cells represented by the BCR::ABL1 transcript amount and thus may determine molecular recurrence.<\/jats:p>","DOI":"10.3390\/jcm14072304","type":"journal-article","created":{"date-parts":[[2025,3,28]],"date-time":"2025-03-28T03:35:28Z","timestamp":1743132928000},"page":"2304","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The PD1 Molecule May Contribute to Lower Treatment-Free Remission Rates in Patients with Chronic Myeloid Leukemia with the e13a2 Transcript"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4244-1213","authenticated-orcid":false,"given":"Paulina","family":"Kwa\u015bnik","sequence":"first","affiliation":[{"name":"Department of Experimental Hematooncology, Medical University of Lublin, 20-093 Lublin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6156-6217","authenticated-orcid":false,"given":"Micha\u0142","family":"Kie\u0142bus","sequence":"additional","affiliation":[{"name":"Department of Experimental Hematooncology, Medical University of Lublin, 20-093 Lublin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4417-7831","authenticated-orcid":false,"given":"Joanna","family":"Zaleska","sequence":"additional","affiliation":[{"name":"Department of Experimental Hematooncology, Medical University of Lublin, 20-093 Lublin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dorota","family":"Link-Lenczowska","sequence":"additional","affiliation":[{"name":"Diagnostic Department of Hematology and Genetics, The University Hospital in Krak\u00f3w, 30-688 Krak\u00f3w, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9105-1767","authenticated-orcid":false,"given":"Magdalena","family":"Zawada","sequence":"additional","affiliation":[{"name":"Diagnostic Department of Hematology and Genetics, The University Hospital in Krak\u00f3w, 30-688 Krak\u00f3w, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hubert","family":"Wysogl\u0105d","sequence":"additional","affiliation":[{"name":"Department of Hematology, University Hospital in Krak\u00f3w, 30-688 Krak\u00f3w, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tomasz","family":"Sacha","sequence":"additional","affiliation":[{"name":"Department of Hematology, University Hospital in Krak\u00f3w, 30-688 Krak\u00f3w, Poland"},{"name":"Department of Hematology, Jagiellonian University Medical College in Krak\u00f3w, 31-008 Krak\u00f3w, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0135-4030","authenticated-orcid":false,"given":"Krzysztof","family":"Giannopoulos","sequence":"additional","affiliation":[{"name":"Department of Experimental Hematooncology, Medical University of Lublin, 20-093 Lublin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2191","DOI":"10.1002\/ajh.27443","article-title":"Chronic Myeloid Leukemia: 2025 Update on Diagnosis, Therapy, and Monitoring","volume":"99","author":"Jabbour","year":"2024","journal-title":"Am. J. Hematol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"966","DOI":"10.1038\/s41375-020-0776-2","article-title":"European LeukemiaNet 2020 Recommendations for Treating Chronic Myeloid Leukemia","volume":"34","author":"Hochhaus","year":"2020","journal-title":"Leukemia"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"747","DOI":"10.1016\/S1470-2045(18)30192-X","article-title":"Discontinuation of Tyrosine Kinase Inhibitor Therapy in Chronic Myeloid Leukaemia (EURO-SKI): A Prespecified Interim Analysis of a Prospective, Multicentre, Non-Randomised, Trial","volume":"19","author":"Saussele","year":"2018","journal-title":"Lancet Oncol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1016\/S1470-2045(10)70233-3","article-title":"Discontinuation of Imatinib in Patients with Chronic Myeloid Leukaemia Who Have Maintained Complete Molecular Remission for at Least 2 Years: The Prospective, Multicentre Stop Imatinib (STIM) Trial","volume":"11","author":"Mahon","year":"2010","journal-title":"Lancet Oncol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1182\/blood-2013-02-483750","article-title":"Safety and Efficacy of Imatinib Cessation for CML Patients with Stable Undetectable Minimal Residual Disease: Results from the TWISTER Study","volume":"122","author":"Ross","year":"2013","journal-title":"Blood"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1200\/JCO.2012.48.5797","article-title":"Loss of Major Molecular Response as a Trigger for Restarting Tyrosine Kinase Inhibitor Therapy in Patients with Chronic-Phase Chronic Myelogenous Leukemia Who Have Stopped Imatinib after Durable Undetectable Disease","volume":"32","author":"Rousselot","year":"2014","journal-title":"J. Clin. Oncol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"856","DOI":"10.1002\/ajh.26550","article-title":"Treatment-Free Remission in Patients with Chronic Myeloid Leukemia Following the Discontinuation of Tyrosine Kinase Inhibitors","volume":"97","author":"Haddad","year":"2022","journal-title":"Am. J. Hematol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1229","DOI":"10.1038\/s41375-021-01238-w","article-title":"Improving Outcomes in Chronic Myeloid Leukemia through Harnessing the Immunological Landscape","volume":"35","author":"Hsieh","year":"2021","journal-title":"Leukemia"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"642","DOI":"10.1007\/s12185-021-03117-7","article-title":"Treatment-Free Remission and Immunity in Chronic Myeloid Leukemia","volume":"113","author":"Ureshino","year":"2021","journal-title":"Int. J. Hematol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1108","DOI":"10.1038\/leu.2016.360","article-title":"Increased Proportion of Mature NK Cells Is Associated with Successful Imatinib Discontinuation in Chronic Myeloid Leukemia","volume":"31","author":"Ilander","year":"2017","journal-title":"Leukemia"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1038\/leu.2017.9","article-title":"Expression of the CTLA-4 Ligand CD86 on Plasmacytoid Dendritic Cells (PDC) Predicts Risk of Disease Recurrence after Treatment Discontinuation in CML","volume":"31","author":"Inselmann","year":"2017","journal-title":"Leukemia"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1368","DOI":"10.3324\/haematol.2017.165001","article-title":"Natural Killer-Cell Counts Are Associated with Molecular Relapse-Free Survival after Imatinib Discontinuation in Chronic Myeloid Leukemia: The IMMUNOSTIM Study","volume":"102","author":"Rea","year":"2017","journal-title":"Haematologica"},{"doi-asserted-by":"crossref","unstructured":"Kwa\u015bnik, P., Zaleska, J., Link-Lenczowska, D., Zawada, M., Wysogl\u0105d, H., Ochrem, B., Bober, G., Wasilewska, E., Hus, I., and Szarejko, M. (2024). High Level of CD8+PD-1+ Cells in Patients with Chronic Myeloid Leukemia Who Experienced Loss of MMR after Imatinib Discontinuation. Cells, 13.","key":"ref_13","DOI":"10.3390\/cells13080723"},{"doi-asserted-by":"crossref","unstructured":"Chen, Y., Zou, J., Cheng, F., and Li, W. (2021). Treatment-Free Remission in Chronic Myeloid Leukemia and New Approaches by Targeting Leukemia Stem Cells. Front. Oncol., 11.","key":"ref_14","DOI":"10.3389\/fonc.2021.769730"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1038\/s41375-023-02074-w","article-title":"Single-Cell Analysis of Immune Recognition in Chronic Myeloid Leukemia Patients Following Tyrosine Kinase Inhibitor Discontinuation","volume":"38","author":"Huuhtanen","year":"2024","journal-title":"Leukemia"},{"doi-asserted-by":"crossref","unstructured":"Hughes, A., and Yong, A.S.M. (2017). Immune Effector Recovery in Chronic Myeloid Leukemia and Treatment-Free Remission. Front. Immunol., 8.","key":"ref_16","DOI":"10.3389\/fimmu.2017.00469"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"434","DOI":"10.1111\/bjh.19269","article-title":"How to Improve Treatment-Free Remission Eligibility in Chronic Myeloid Leukaemia?","volume":"204","author":"Costa","year":"2024","journal-title":"Br. J. Haematol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e297","DOI":"10.3324\/haematol.2017.168740","article-title":"E14a2 BCR-ABL1 Transcript Is Associated with a Higher Rate of Treatment-Free Remission in Individuals with Chronic Myeloid Leukemia after Stopping Tyrosine Kinase Inhibitor Therapy","volume":"102","author":"Claudiani","year":"2017","journal-title":"Haematologica"},{"doi-asserted-by":"crossref","unstructured":"Chen, K., Ruan, Y., Tian, K., Xiong, P., Xia, N., Li, J., Huang, W., Cao, F., and Chen, Q. (2022). Impact of BCR-ABL1 Transcript Type on Outcome in Chronic Myeloid Leukemia Patients Treated with Tyrosine Kinase Inhibitors: A Pairwise and Bayesian Network Meta-Analysis. Front. Oncol., 12.","key":"ref_19","DOI":"10.3389\/fonc.2022.841546"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"797","DOI":"10.1002\/ajh.24774","article-title":"The BCR-ABL1 Transcript Type Influences Response and Outcome in Philadelphia Chromosome-Positive Chronic Myeloid Leukemia Patients Treated Frontline with Imatinib","volume":"92","author":"Castagnetti","year":"2017","journal-title":"Am. J. Hematol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1269","DOI":"10.1182\/blood-2015-10-674242","article-title":"Impact of BCR-ABL Transcript Type on Outcome in Patients with Chronic-Phase CML Treated with Tyrosine Kinase Inhibitors","volume":"127","author":"Jain","year":"2016","journal-title":"Blood"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1674","DOI":"10.1002\/cncr.31977","article-title":"The E13a2 BCR-ABL Transcript Negatively Affects Sustained Deep Molecular Response and the Achievement of Treatment-Free Remission in Patients with Chronic Myeloid Leukemia Who Receive Tyrosine Kinase Inhibitors","volume":"125","author":"Farina","year":"2019","journal-title":"Cancer"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"728","DOI":"10.1016\/j.clml.2017.06.009","article-title":"Influence of BCR-ABL Transcript Type on Outcome in Patients with Chronic-Phase Chronic Myeloid Leukemia Treated With Imatinib","volume":"17","author":"Pagnano","year":"2017","journal-title":"Clin. Lymphoma. Myeloma Leuk."},{"key":"ref_24","first-page":"454","article-title":"Standaryzacja Ilo\u015bciowej Oceny Ekspresji Genu BCR-ABL Metoda RQ-PCR u Chorych Na Przewlek\u0142a Bia\u0142aczke Szpikowa We Wsp\u00f3\u0142pracy z European Leukemia Net","volume":"67","author":"Sacha","year":"2010","journal-title":"Przegla\u0327d Lek."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1111\/ejh.13238","article-title":"Variant-Specific Discrepancy When Quantitating BCR-ABL1 E13a2 and E14a2 Transcripts Using the Europe Against Cancer QPCR Assay","volume":"103","author":"Kjaer","year":"2019","journal-title":"Eur. J. Haematol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1","DOI":"10.18637\/jss.v085.i12","article-title":"Rmcfs: An R Package for Monte Carlo Feature Selection and Interdependency Discovery","volume":"85","author":"Draminski","year":"2018","journal-title":"J. Stat. Softw."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"852","DOI":"10.3324\/haematol.2009.019224","article-title":"Impact of the Type of the BCR-ABL Fusion Transcript on the Molecular Response in Pediatric Patients with Chronic Myeloid Leukemia","volume":"95","author":"Suttorp","year":"2010","journal-title":"Haematologica"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1362","DOI":"10.3324\/haematol.2009.009134","article-title":"Chronic Myeloid Leukemia Patients with the E13a2 BCR-ABL Fusion Transcript Have Inferior Responses to Imatinib Compared to Patients with the E14a2 Transcript","volume":"94","author":"Lucas","year":"2009","journal-title":"Haematologica"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2887","DOI":"10.1182\/blood.V98.10.2887","article-title":"Direct Evidence That Leukemic Cells Present HLA-Associated Immunogenic Peptides Derived from the BCR-ABL B3a2 Fusion Protein","volume":"98","author":"Clark","year":"2001","journal-title":"Blood"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1166","DOI":"10.1182\/blood-2016-10-745992","article-title":"CML Patients with Deep Molecular Responses to TKI Have Restored Immune Effectors and Decreased PD-1 and Immune Suppressors","volume":"129","author":"Hughes","year":"2017","journal-title":"Blood"},{"doi-asserted-by":"crossref","unstructured":"Kwa\u015bnik, P., and Giannopoulos, K. (2021). Treatment-Free Remission-A New Aim in the Treatment of Chronic Myeloid Leukemia. J. Pers. Med., 11.","key":"ref_31","DOI":"10.3390\/jpm11080697"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"7534","DOI":"10.18632\/oncotarget.23691","article-title":"Prognostic Factors Associated with a Stable MR4.5 Achievement in Chronic Myeloid Leukemia Patients Treated with Imatinib","volume":"9","author":"Breccia","year":"2017","journal-title":"Oncotarget"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1182\/blood.V126.23.597.597","article-title":"Predictive Factors of Stable Deep Molecular Response in Chronic Myeloid Leukemia Patients Treated with Imatinib Standard Dose: A Study from the Gruppo Triveneto LMC","volume":"126","author":"Bonifacio","year":"2015","journal-title":"Blood"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1441","DOI":"10.3324\/haematol.2013.096537","article-title":"Distinct Characteristics of E13a2 versus E14a2 BCR-ABL1 Driven Chronic Myeloid Leukemia under First-Line Therapy with Imatinib","volume":"99","author":"Hanfstein","year":"2014","journal-title":"Haematologica"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2358","DOI":"10.1038\/s41375-019-0562-1","article-title":"Chronic Myeloid Leukemia: The Concepts of Resistance and Persistence and the Relationship with the BCR-ABL1 Transcript Type","volume":"33","author":"Baccarani","year":"2019","journal-title":"Leukemia"},{"doi-asserted-by":"crossref","unstructured":"Marc\u00e9, S., M\u00e9ndez, A., Xicoy, B., Estrada, N., Cabez\u00f3n, M., Sturla, A.L., Garc\u00eda, M.R., Angona, A., Amat, P., and Escribano Serrat, S. (2024). E14a2 Transcript Favors Treatment-Free Remission in Chronic Myeloid Leukemia When Associated with Longer Treatment with Tyrosine Kinase Inhibitors and Sustained Deep Molecular Response. J. Clin. Med., 13.","key":"ref_36","DOI":"10.3390\/jcm13030779"},{"doi-asserted-by":"crossref","unstructured":"E\u015fkazan, A.E., Kok, C.H., Yeung, D., Dalal, J., and Tiribelli, M. (2023). Editorial: Advances in the Treatment of Chronic Myeloid Leukemia. Front. Oncol., 13.","key":"ref_37","DOI":"10.3389\/fonc.2023.1166588"},{"key":"ref_38","first-page":"1596","article-title":"E14a2 and e14a2+e13a2 BCR-ABL Transcripts Are Associated with Earlier Cytogenetic and Molecular Response Rates in CML-CP Treated with Imatinib","volume":"130","author":"Pagnano","year":"2017","journal-title":"Blood"},{"unstructured":"Garofalaki, M., Tziotziou, E., Nikolou, E., Tsonis, I., Dellatola, M., Mellios, Z., Psyllaki, M., Grispou, E., Delimpassi, S., and Baltadakis, I. (2024, July 03). E13A2, E14A2 Transcripts: Do They Have a Different Impact on Outcome In Patients with Chronic Myeloid Leukemia? EHA Library. 9 June 2021. Available online: https:\/\/library.ehaweb.org\/eha\/2021\/eha2021-virtual-congress\/324194\/maria.garofalaki.e13a2.e14a2.transcripts.do.they.have.a.different.impact.on.html.","key":"ref_39"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1879","DOI":"10.1038\/s41375-022-01612-2","article-title":"Impact of BCR::ABL1 Transcript Type on RT-QPCR Amplification Performance and Molecular Response to Therapy","volume":"36","author":"Salmon","year":"2022","journal-title":"Leukemia"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1111\/bjh.18026","article-title":"Assessment of Quantitative Polymerase Chain Reaction for BCR-ABL1 Transcripts in Chronic Myeloid Leukaemia: Are Improved Outcomes in Patients with E14a2 Transcripts an Artefact of Technology?","volume":"197","author":"Dominy","year":"2022","journal-title":"Br. J. Haematol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"843","DOI":"10.1007\/s00432-016-2321-2","article-title":"No Influence of BCR-ABL1 Transcript Types E13a2 and E14a2 on Long-Term Survival: Results in 1494 Patients with Chronic Myeloid Leukemia Treated with Imatinib","volume":"143","author":"Pfirrmann","year":"2017","journal-title":"J. Cancer Res. Clin. Oncol."}],"container-title":["Journal of Clinical Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2077-0383\/14\/7\/2304\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:03:48Z","timestamp":1760029428000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2077-0383\/14\/7\/2304"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,27]]},"references-count":42,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2025,4]]}},"alternative-id":["jcm14072304"],"URL":"https:\/\/doi.org\/10.3390\/jcm14072304","relation":{},"ISSN":["2077-0383"],"issn-type":[{"type":"electronic","value":"2077-0383"}],"subject":[],"published":{"date-parts":[[2025,3,27]]}}}