Silvia Neri, Corien L. Eckhardt, Boukje M. Beuger, Folman Folman, Eva Rettenbacher, Hanke L. Matlung, Taco W. Kuijpers, Josephine M. I. Vos, Robin van Bruggen
Introduction
IgA-mediated autoimmune hemolytic anemia (AIHA) is a rare condition associated with severe hemolysis and limited therapeutic response. Bortezomib, a proteasome inhibitor, targets plasma cells responsible for autoantibody production. Here, we describe a case of refractory IgA-mediated AIHA in a 13-year-old boy presenting with severe hemolysis, who was successfully treated with bortezomib.
Methods:
Blood samples were collected at different time points throughout the disease course for immunohematology testing.
Results
The patient showed significant hematologic improvement following four doses of Bortezomib with reduction in hemolysis and recovery of hemoglobin levels. Laboratory tests revealed complement-negative, Coombs-positive blood tests combined with altered RBC morphology. Phagocytosis of patient's RBC was absent at all timepoints. Notably, despite hematologic improvement, IgA-positive RBC remained present, accompanied by compensated hemolysis.
Conclusions
The present case demonstrates the potential of bortezomib as a treatment option for refractory AIHA cases, particularly in children.
Trial Registration: The authors have confirmed clinical trial registration is not needed for this submission
{"title":"Successful Treatment of Refractory IgA-Mediated Autoimmune Hemolytic Anemia With Bortezomib","authors":"Silvia Neri, Corien L. Eckhardt, Boukje M. Beuger, Folman Folman, Eva Rettenbacher, Hanke L. Matlung, Taco W. Kuijpers, Josephine M. I. Vos, Robin van Bruggen","doi":"10.1002/jha2.70162","DOIUrl":"https://doi.org/10.1002/jha2.70162","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Introduction</h3>\u0000 \u0000 <p>IgA-mediated autoimmune hemolytic anemia (AIHA) is a rare condition associated with severe hemolysis and limited therapeutic response. Bortezomib, a proteasome inhibitor, targets plasma cells responsible for autoantibody production. Here, we describe a case of refractory IgA-mediated AIHA in a 13-year-old boy presenting with severe hemolysis, who was successfully treated with bortezomib.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods:</h3>\u0000 \u0000 <p>Blood samples were collected at different time points throughout the disease course for immunohematology testing.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The patient showed significant hematologic improvement following four doses of Bortezomib with reduction in hemolysis and recovery of hemoglobin levels. Laboratory tests revealed complement-negative, Coombs-positive blood tests combined with altered RBC morphology. Phagocytosis of patient's RBC was absent at all timepoints. Notably, despite hematologic improvement, IgA-positive RBC remained present, accompanied by compensated hemolysis.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>The present case demonstrates the potential of bortezomib as a treatment option for refractory AIHA cases, particularly in children.</p>\u0000 \u0000 <p><b>Trial Registration</b>: The authors have confirmed clinical trial registration is not needed for this submission</p>\u0000 </section>\u0000 </div>","PeriodicalId":72883,"journal":{"name":"EJHaem","volume":"6 6","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jha2.70162","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145367156","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Natsumi Kawasaki, Sei Shu, Mayu Tomita, Xiaoxiao Liu, Shigeki Yoshiura, Yoriko Yamashita-Kashima
Background
Aggressive non-Hodgkin lymphoma (aNHL) often relapses after first-line treatment. Clinical data supports the safety and efficacy of the combination of mosunetuzumab, a CD20×CD3 bispecific antibody, and polatuzumab vedotin, an anti-CD79b antibody drug conjugate (Mosun-Pola) in relapsed/refractory aNHL. This study investigated the molecular mechanism behind the combination effect of Mosun-Pola in human diffuse large B-cell lymphoma (DLBCL) cell lines.
Methods
The in vitro Mosun-Pola efficacy in DLBCL cells (SU-DHL-8 and HT) was evaluated by T cell-dependent cellular cytotoxicity (TDCC) assay. CD20-stable-knockdown SU-DHL-8 cells were established using lentiviral short hairpin RNA. Surface and T-cell activation marker proteins expression were determined by flow cytometry. Human T-cell-injected mice or humanized NOD/Shi-scid, IL-2Rγnull (huNOG) mice were used for an in vivo study.
Results
An in vitro TDCC assay showed a synergistic effect in SU-DHL-8 and HT cells. Based on our experimental results of suppressing CD20 expression, it was suggested that this combination effect could be caused by an increase in CD20 expression by polatuzumab vedotin. In addition, examining the effects of CD20 upregulation in tumor cells on T-cell activation demonstrated that the combination of Mosun-Pola enhanced T-cell activation markers in both CD4+ and CD8+ T cells during the TDCC reaction. In vivo studies, using human immune system-reconstituted mouse models confirmed that polatuzumab vedotin enhanced CD20 expression in tumors, and the combination of Mosun-Pola showed significantly improved anti-tumor effects compared with single-drug treatments.
Conclusion
These findings suggest that polatuzumab vedotin-induced CD20 upregulation provides a molecular rationale to explain the synergistic effect of this combination therapy.
Trial Registration
The authors have confirmed clinical trial registration is not needed for this submission.
{"title":"Polatuzumab Vedotin Induced CD20 Upregulation Contributes to the Efficacy of Mosunetuzumab in Combination With Polatuzumab Vedotin in Diffuse Large B-Cell Lymphoma Preclinical Models","authors":"Natsumi Kawasaki, Sei Shu, Mayu Tomita, Xiaoxiao Liu, Shigeki Yoshiura, Yoriko Yamashita-Kashima","doi":"10.1002/jha2.70169","DOIUrl":"https://doi.org/10.1002/jha2.70169","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Aggressive non-Hodgkin lymphoma (aNHL) often relapses after first-line treatment. Clinical data supports the safety and efficacy of the combination of mosunetuzumab, a CD20×CD3 bispecific antibody, and polatuzumab vedotin, an anti-CD79b antibody drug conjugate (Mosun-Pola) in relapsed/refractory aNHL. This study investigated the molecular mechanism behind the combination effect of Mosun-Pola in human diffuse large B-cell lymphoma (DLBCL) cell lines.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>The in vitro Mosun-Pola efficacy in DLBCL cells (SU-DHL-8 and HT) was evaluated by T cell-dependent cellular cytotoxicity (TDCC) assay. CD20-stable-knockdown SU-DHL-8 cells were established using lentiviral short hairpin RNA. Surface and T-cell activation marker proteins expression were determined by flow cytometry. Human T-cell-injected mice or humanized NOD/Shi-scid, IL-2Rγnull (huNOG) mice were used for an in vivo study.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>An in vitro TDCC assay showed a synergistic effect in SU-DHL-8 and HT cells. Based on our experimental results of suppressing CD20 expression, it was suggested that this combination effect could be caused by an increase in CD20 expression by polatuzumab vedotin. In addition, examining the effects of CD20 upregulation in tumor cells on T-cell activation demonstrated that the combination of Mosun-Pola enhanced T-cell activation markers in both CD4+ and CD8+ T cells during the TDCC reaction. In vivo studies, using human immune system-reconstituted mouse models confirmed that polatuzumab vedotin enhanced CD20 expression in tumors, and the combination of Mosun-Pola showed significantly improved anti-tumor effects compared with single-drug treatments.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>These findings suggest that polatuzumab vedotin-induced CD20 upregulation provides a molecular rationale to explain the synergistic effect of this combination therapy.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Trial Registration</h3>\u0000 \u0000 <p>The authors have confirmed clinical trial registration is not needed for this submission.</p>\u0000 </section>\u0000 </div>","PeriodicalId":72883,"journal":{"name":"EJHaem","volume":"6 5","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jha2.70169","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145367016","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Alraddadi, J. P. Smalley, W. Alzahrani, et al. “Targeted Degradation of Class 1 HDACs With PROTACs is Highly Effective at Inducing DLBCL Cell Death,” EJHaem 6, no. 4 (2025): e70127, https://doi.org/10.1002/jha2.70127.
Co-author Martin J. S. Dyer was incorrectly listed as Martin Dyer.
We apologize for this error.
[更正文章DOI: 10.1002/jha2.70127.]。
{"title":"Correction to “Targeted Degradation of Class 1 HDACs With PROTACs is Highly Effective at Inducing DLBCL Cell Death”","authors":"","doi":"10.1002/jha2.70159","DOIUrl":"10.1002/jha2.70159","url":null,"abstract":"<p>A. Alraddadi, J. P. Smalley, W. Alzahrani, et al. “Targeted Degradation of Class 1 HDACs With PROTACs is Highly Effective at Inducing DLBCL Cell Death,” <i>EJHaem</i> 6, no. 4 (2025): e70127, https://doi.org/10.1002/jha2.70127.</p><p>Co-author Martin J. S. Dyer was incorrectly listed as Martin Dyer.</p><p>We apologize for this error.</p>","PeriodicalId":72883,"journal":{"name":"EJHaem","volume":"6 5","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12538633/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145350081","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
<p>A 69-year-old man complained of melena. He was diagnosed with gastric primary nasal-type extranodal natural killer/T-cell lymphoma (ENKTL) in October 2017. On <sup>18</sup>F-fluorodeoxyglucose positron emission tomography/computed tomography (<sup>18</sup>F-FDG-PET/CT), accumulations were observed in the stomach and small intestine, indicating stage IV disease. He received combination chemotherapy including alkylating agents, topoisomerase 2 inhibitors, dexamethasone, a nucleoside metabolic inhibitor, and a programmed death-ligand 1 inhibitor, with repeated recurrences and remissions up to September 2022. <sup>18</sup>F-FDG-PET/CT showed a complete metabolic response in November 2022. There was no bone marrow invasion by lymphoma cells during this period. The following March, he contracted COVID-19 and was treated with nirmatrelvir. After recovering from the infection, he presented with pancytopenia without blasts. Although pancytopenia might be due to viral infection, bone marrow examination was performed since he had a long history of chemotherapy including alkylating agents and topoisomerase 2 inhibitors.</p><p>Bone marrow showed hypocellularity with 9.2% blasts (Figure 1a), and dysplasia was detected in myeloid (pseudo-Pelger-Huët anomaly and giant neutrophil: Figures 1b,c), erythroid (nuclear budding: Figure 1d), and megakaryocytic (micromegakaryocyte, multinucleated megakaryocyte: Figures 1e,f) lineages. Flow cytometry showed that the leukemic blasts were positive for CD13, CD33, CD34, CD56, and myeloperoxidase. Cytogenetic analysis demonstrated a complicated karyotype with deletion of chromosomes 5q and 7. Based on these findings, myeloid neoplasm post cytotoxic therapy (MN-pCT) was diagnosed.</p><p>In bone marrow, there was an increase in eosinophils (14.7%), in addition to blasts, and some eosinophils were dysplastic with large basophilic granules of purple-violet color (Figures 2a,b), which are referred to as harlequin cells [<span>1</span>].</p><p>ENKTL is associated with Epstein–Barr virus infection and rarely involves the gastrointestinal tract. Advances in ENKTL treatment have led to an increase in the number of survivors. Consequently, the risk of patients developing MN-pCT may have increased.</p><p>Two cases of MN-pCT with harlequin cells have been reported. Although the primary cancers were different (thymoma and colon adenocarcinoma), alkylating agents were administered in both cases [<span>2, 3</span>].</p><p>The harlequin cells are classified as Code 9 by the International Working Group on Morphology of MDS [<span>4</span>] and are more frequently observed in de novo acute myeloid leukemia harboring <i>CBFB::MYH11</i> fusion [<span>1, 4</span>]. Harlequin cells are observed not only in hematologic malignancies but also in patients with non-hematologic diseases [<span>1, 4</span>]. However, in AML with the <i>CBFB::MYH11</i> fusion, these eosinophils are considered part of the neoplastic clone [<span>4</span>].</p><p>The app
{"title":"Harlequin Cells in Myeloid Neoplasm Post Cytotoxic Therapy for Gastric Primary Nasal-Type Extranodal Natural Killer/T-Cell Lymphoma","authors":"Sachi Tanaka, Hiromi Kinoshita, Naoki Takahashi, Yasuhiro Ebihara","doi":"10.1002/jha2.70167","DOIUrl":"https://doi.org/10.1002/jha2.70167","url":null,"abstract":"<p>A 69-year-old man complained of melena. He was diagnosed with gastric primary nasal-type extranodal natural killer/T-cell lymphoma (ENKTL) in October 2017. On <sup>18</sup>F-fluorodeoxyglucose positron emission tomography/computed tomography (<sup>18</sup>F-FDG-PET/CT), accumulations were observed in the stomach and small intestine, indicating stage IV disease. He received combination chemotherapy including alkylating agents, topoisomerase 2 inhibitors, dexamethasone, a nucleoside metabolic inhibitor, and a programmed death-ligand 1 inhibitor, with repeated recurrences and remissions up to September 2022. <sup>18</sup>F-FDG-PET/CT showed a complete metabolic response in November 2022. There was no bone marrow invasion by lymphoma cells during this period. The following March, he contracted COVID-19 and was treated with nirmatrelvir. After recovering from the infection, he presented with pancytopenia without blasts. Although pancytopenia might be due to viral infection, bone marrow examination was performed since he had a long history of chemotherapy including alkylating agents and topoisomerase 2 inhibitors.</p><p>Bone marrow showed hypocellularity with 9.2% blasts (Figure 1a), and dysplasia was detected in myeloid (pseudo-Pelger-Huët anomaly and giant neutrophil: Figures 1b,c), erythroid (nuclear budding: Figure 1d), and megakaryocytic (micromegakaryocyte, multinucleated megakaryocyte: Figures 1e,f) lineages. Flow cytometry showed that the leukemic blasts were positive for CD13, CD33, CD34, CD56, and myeloperoxidase. Cytogenetic analysis demonstrated a complicated karyotype with deletion of chromosomes 5q and 7. Based on these findings, myeloid neoplasm post cytotoxic therapy (MN-pCT) was diagnosed.</p><p>In bone marrow, there was an increase in eosinophils (14.7%), in addition to blasts, and some eosinophils were dysplastic with large basophilic granules of purple-violet color (Figures 2a,b), which are referred to as harlequin cells [<span>1</span>].</p><p>ENKTL is associated with Epstein–Barr virus infection and rarely involves the gastrointestinal tract. Advances in ENKTL treatment have led to an increase in the number of survivors. Consequently, the risk of patients developing MN-pCT may have increased.</p><p>Two cases of MN-pCT with harlequin cells have been reported. Although the primary cancers were different (thymoma and colon adenocarcinoma), alkylating agents were administered in both cases [<span>2, 3</span>].</p><p>The harlequin cells are classified as Code 9 by the International Working Group on Morphology of MDS [<span>4</span>] and are more frequently observed in de novo acute myeloid leukemia harboring <i>CBFB::MYH11</i> fusion [<span>1, 4</span>]. Harlequin cells are observed not only in hematologic malignancies but also in patients with non-hematologic diseases [<span>1, 4</span>]. However, in AML with the <i>CBFB::MYH11</i> fusion, these eosinophils are considered part of the neoplastic clone [<span>4</span>].</p><p>The app","PeriodicalId":72883,"journal":{"name":"EJHaem","volume":"6 5","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jha2.70167","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145317712","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Van Tuong Nguyen, Lina Han, Jing Xu, Miguel Cantu, Franklin Fuda, Samuel John, Tamra Slone, Weina Chen
Background
NUP98::BPTF rearranged (r) leukaemia is rare with only five reported cases, including acute myeloid leukemia and T-lymphoblastic leukemia (T-ALL).
Results
Herein, we report a case of NUP98::BPTF-r mixed phenotype acute leukemia (MPAL), B/T with T-lineage-predominance in a 15-year-old female. Cytogenetic and molecular studies revealed a t(11;17)(p15;q23)/NUP98::BPTF fusion and cooperative gene alterations characteristic of T-ALL (NOTCH1, FBXW7, and PHF6). Our patient had a primary induction failure with T-ALL-directed chemotherapy and achieved complete remission following consolidation.
Conclusion
This is the first case study characterizing the clinicopathological and genomic features of B/T MPAL harboring NUP98::BPTF fusion and providing insights into molecular pathogenesis.
Trial Registration
The authors have confirmed clinical trial registration is not needed for this submission.
背景：BPTF重排(r)白血病是罕见的，仅有5例报道，包括急性髓系白血病和t淋巴母细胞白血病（T-ALL）。结果：在此，我们报告了一例15岁女性的NUP98::BPTF-r混合表型急性白血病（MPAL）， B/T伴T谱系优势。细胞遗传学和分子生物学研究显示t(11;17)(p15;q23)/NUP98::BPTF融合和t - all （NOTCH1、FBXW7和PHF6）的协同基因改变特征。我们的患者在接受t - all定向化疗时出现了原发性诱导失败，并在巩固后完全缓解。结论：这是第一个描述含有NUP98::BPTF融合的B/T MPAL的临床病理和基因组特征的病例研究，并为分子发病机制提供了新的见解。试验注册：作者已确认本次提交不需要临床试验注册。
{"title":"B/T Mixed Phenotype Acute Leukaemia Harbouring NUP98::BPTF Fusion","authors":"Van Tuong Nguyen, Lina Han, Jing Xu, Miguel Cantu, Franklin Fuda, Samuel John, Tamra Slone, Weina Chen","doi":"10.1002/jha2.70166","DOIUrl":"10.1002/jha2.70166","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p><i>NUP98</i>::<i>BPTF</i> rearranged (r) leukaemia is rare with only five reported cases, including acute myeloid leukemia and T-lymphoblastic leukemia (T-ALL).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Herein, we report a case of <i>NUP98::BPTF</i>-r mixed phenotype acute leukemia (MPAL), B/T with T-lineage-predominance in a 15-year-old female. Cytogenetic and molecular studies revealed a t(11;17)(p15;q23)/<i>NUP98::BPTF</i> fusion and cooperative gene alterations characteristic of T-ALL (<i>NOTCH1</i>, <i>FBXW7</i>, and <i>PHF6</i>). Our patient had a primary induction failure with T-ALL-directed chemotherapy and achieved complete remission following consolidation.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>This is the first case study characterizing the clinicopathological and genomic features of B/T MPAL harboring <i>NUP98::BPTF</i> fusion and providing insights into molecular pathogenesis.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Trial Registration</h3>\u0000 \u0000 <p>The authors have confirmed clinical trial registration is not needed for this submission.</p>\u0000 </section>\u0000 </div>","PeriodicalId":72883,"journal":{"name":"EJHaem","volume":"6 5","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12519876/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145304804","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mark Anthony Turingan, Ehsan Bahrami Hezaveh, Cuihong Wei, Verna Cheung, Dawn Maze, Hong Chang
Background
The emergence of JAK2-mutated myeloproliferative neoplasms (MPNs) after remission from acute myeloid leukaemia (AML) is exceedingly rare.
Case Description
This case series describes two patients who developed JAK2-mutated MPNs years after achieving AML remission, each retaining persistent TET2 and SRSF2 mutations while losing AML-defining mutations.
Pathogenetic Insight
Findings support clonal persistence from a shared haematopoietic progenitor with divergent progression into distinct myeloid neoplasms. A two-pathway model is proposed to explain this trajectory.
Implications
These observations highlight the biological relevance of CHIP-associated mutations and underscore the value of post-remission molecular surveillance to detect emerging secondary neoplasms in AML survivors.
Trial Registration: The authors have confirmed clinical trial registration is not needed for this submission.
{"title":"Uncommon Evolution From Acute Myeloid Leukaemia to JAK2-Mutated Myeloproliferative Neoplasm: Evidence of Clonal Persistence and Divergence From TET2/SRSF2-Mutated Haematopoietic Progenitors","authors":"Mark Anthony Turingan, Ehsan Bahrami Hezaveh, Cuihong Wei, Verna Cheung, Dawn Maze, Hong Chang","doi":"10.1002/jha2.70137","DOIUrl":"10.1002/jha2.70137","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>The emergence of <i>JAK2</i>-mutated myeloproliferative neoplasms (MPNs) after remission from acute myeloid leukaemia (AML) is exceedingly rare.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Case Description</h3>\u0000 \u0000 <p>This case series describes two patients who developed <i>JAK2</i>-mutated MPNs years after achieving AML remission, each retaining persistent <i>TET2</i> and <i>SRSF2</i> mutations while losing AML-defining mutations.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Pathogenetic Insight</h3>\u0000 \u0000 <p>Findings support clonal persistence from a shared haematopoietic progenitor with divergent progression into distinct myeloid neoplasms. A two-pathway model is proposed to explain this trajectory.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Implications</h3>\u0000 \u0000 <p>These observations highlight the biological relevance of CHIP-associated mutations and underscore the value of post-remission molecular surveillance to detect emerging secondary neoplasms in AML survivors.</p>\u0000 \u0000 <p><b>Trial Registration</b>: The authors have confirmed clinical trial registration is not needed for this submission.</p>\u0000 </section>\u0000 </div>","PeriodicalId":72883,"journal":{"name":"EJHaem","volume":"6 5","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12519878/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145304846","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Maura Rosane Valerio Ikoma-Colturato, Felipe Magalhães Furtado, Camila Marques Bertolucci, Alef Rafael Severino, Elizabeth Xisto Souto, Tharsis Cardoso Ferreira dos Santos, Ana Paula Fortunato Dametto, Miriam Perlingeiro Beltrame, Nydia Strachman Bacal, Danielle Marchetti Vitelli Avelar, Raquel Arrieche Fernandes, Bernadete Evangelho Gomes, Elaine Sobral da Costa, Berta Elisa Fonseca da Silva Santos, Robéria Mendonça de Pontes, Maria Daniela Holthausen Périco, Fabíola Gevert, Patrícia Fernanda Rosa de Siqueira, Fernanda Marquezotti, Andressa Oliveira Martin Wagner, Ana Claudia Carramaschi Villela Soares, Fernando Barroso Duarte, Mary Evelyn Flowers, Afonso Celso Vigorito
Introduction
Measurable residual disease (MRD) is a strong predictor of the risk of relapse of acute myeloid leukemia (AML). Therefore, for use in clinical decision-making, methods for MRD assessment must achieve adequate accuracy, sensitivity, specificity, and reproducibility. Multiparametric flow cytometry (MFC) is the most widely used method for assessing AML-MRD, but its sensitivity varies considerably due to the differing approaches used across centers, in addition to the different experiences of flow cytometrists, especially during clonal evolution. This study aimed to standardize AML-MRD by MFC in a multicenter project involving 16 Brazilian laboratories.
Methods
In the first phase, specialists were trained in pre-analytical standard operating procedures (SOPs) and analysis strategies of pre-validated 8- and 10-color protocols, followed by a data-only, that is, a Dry Phase of flow cytometry standard (FCS) file exchange by the coordinating laboratory in a comparability assessment. In the second or Wet Phase, laboratories prepared and analyzed their samples, and the FCS files were submitted for central analysis.
Results
The agreement of MRD results was 81% and 80% between laboratories and central analysts in the Dry and Wet Phases, respectively. However, non-suitable application of pre-analytical SOPs hampered MRD interpretation for 30% of the laboratories in the Wet Phase.
Conclusions
This study demonstrated that standardized flow cytometry protocols are reproducible as long as rigorous SOPs are implemented. The project's results underscore that continuous education and external quality control are essential to build expertise and ensure reliable AML-MRD results in clinical practice.
Trial Registration:The authors have confirmed clinical trial registration is not needed for this submission.
{"title":"Standardization of Measurable Residual Disease in Acute Myeloid Leukemia by Flow Cytometry: A Multicenter Study","authors":"Maura Rosane Valerio Ikoma-Colturato, Felipe Magalhães Furtado, Camila Marques Bertolucci, Alef Rafael Severino, Elizabeth Xisto Souto, Tharsis Cardoso Ferreira dos Santos, Ana Paula Fortunato Dametto, Miriam Perlingeiro Beltrame, Nydia Strachman Bacal, Danielle Marchetti Vitelli Avelar, Raquel Arrieche Fernandes, Bernadete Evangelho Gomes, Elaine Sobral da Costa, Berta Elisa Fonseca da Silva Santos, Robéria Mendonça de Pontes, Maria Daniela Holthausen Périco, Fabíola Gevert, Patrícia Fernanda Rosa de Siqueira, Fernanda Marquezotti, Andressa Oliveira Martin Wagner, Ana Claudia Carramaschi Villela Soares, Fernando Barroso Duarte, Mary Evelyn Flowers, Afonso Celso Vigorito","doi":"10.1002/jha2.70163","DOIUrl":"10.1002/jha2.70163","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Introduction</h3>\u0000 \u0000 <p>Measurable residual disease (MRD) is a strong predictor of the risk of relapse of acute myeloid leukemia (AML). Therefore, for use in clinical decision-making, methods for MRD assessment must achieve adequate accuracy, sensitivity, specificity, and reproducibility. Multiparametric flow cytometry (MFC) is the most widely used method for assessing AML-MRD, but its sensitivity varies considerably due to the differing approaches used across centers, in addition to the different experiences of flow cytometrists, especially during clonal evolution. This study aimed to standardize AML-MRD by MFC in a multicenter project involving 16 Brazilian laboratories.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>In the first phase, specialists were trained in pre-analytical standard operating procedures (SOPs) and analysis strategies of pre-validated 8- and 10-color protocols, followed by a data-only, that is, a Dry Phase of flow cytometry standard (FCS) file exchange by the coordinating laboratory in a comparability assessment. In the second or Wet Phase, laboratories prepared and analyzed their samples, and the FCS files were submitted for central analysis.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The agreement of MRD results was 81% and 80% between laboratories and central analysts in the Dry and Wet Phases, respectively. However, non-suitable application of pre-analytical SOPs hampered MRD interpretation for 30% of the laboratories in the Wet Phase.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>This study demonstrated that standardized flow cytometry protocols are reproducible as long as rigorous SOPs are implemented. The project's results underscore that continuous education and external quality control are essential to build expertise and ensure reliable AML-MRD results in clinical practice.</p>\u0000 \u0000 <p><b>Trial Registration</b>:The authors have confirmed clinical trial registration is not needed for this submission.</p>\u0000 </section>\u0000 </div>","PeriodicalId":72883,"journal":{"name":"EJHaem","volume":"6 5","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12519880/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145304774","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A 21-year-old male diagnosed with B-lymphoblastic leukaemia, who received one cycle of chemotherapy, presented for a follow-up bone marrow biopsy for evaluation of stem cell transplant. A complete blood count revealed haemoglobin levels of 14.7 g/dL, a leucocyte count of 3.2 × 109/L, and a platelet count of 93 × 109/L. The peripheral blood smear was unremarkable apart from mild leucopenia and moderate thrombocytopenia. Bone marrow aspirate smears demonstrated significant myeloid predominance and severe erythroid hypoplasia with maturation arrest; notably, rare giant proerythroblasts with intranuclear inclusions and subtle vacuolated cytoplasm were identified (Figure 1A, 100× objective). The trephine biopsy exhibited a few giant proerythroblasts among maturing myeloid and megakaryocytic precursors (Figure 1B, 100× objective). Immunohistochemistry revealed large CD71-positive cells (Figure 1C, 100× objective) and Parvovirus B19-positive cells (Figure 1D, 100× objective), indicating infected giant proerythroblasts, also known as'lantern cells.' Parvovirus B19 quantitative polymerase chain reaction (qPCR) was positive, and serologies demonstrated positive IgM and negative IgG, confirming an acute parvovirus infection.
Parvovirus B19 is a single-stranded DNA virus that infects precursor red blood cells via the P antigen or globoside (Gb4) present on the cell surface [1]. The diagnosis is typically achieved through serological testing, qPCR, clinical symptomatology, and analysis of red blood cell indices. Immunocompromised individuals have been reported to present with normocytic anaemia, a normal leucocyte count, and either normal or elevated platelet counts [2]. These findings are discordant with the current case. Since mature red blood cells remain in circulation for approximately 120 days, early marrow suppression may not immediately result in a decrease in haemoglobin levels. Eighteen days after bone marrow biopsy, repeated complete blood count revealed haemoglobin levels of 8.2 g/dL, a leucocyte count of 5.0 × 109/L, and a platelet count of 228 × 109/L.
The present case highlights that parvovirus infection can manifest in atypical forms and may go unnoticed in immunocompromised patients lacking characteristic laboratory findings. When encountering severe erythroid hypoplasia with maturation arrest, it is advisable to conduct testing for parvovirus B19 infection, even in the absence of anaemia.
The authors have nothing to report.
The author has confirmed that a patient consent statement is not required for this submission, as no patient-identifying data were used.
The authors declare that they have no relationships with, or financial interests in, any commercial companies related to this article.
{"title":"Before the Blood Drops: Early Clues of Parvovirus B19","authors":"Joseph Stenberg, Daniel Babu, Nicole Deshmukh","doi":"10.1002/jha2.70165","DOIUrl":"10.1002/jha2.70165","url":null,"abstract":"<p>A 21-year-old male diagnosed with B-lymphoblastic leukaemia, who received one cycle of chemotherapy, presented for a follow-up bone marrow biopsy for evaluation of stem cell transplant. A complete blood count revealed haemoglobin levels of 14.7 g/dL, a leucocyte count of 3.2 × 10<sup>9</sup>/L, and a platelet count of 93 × 10<sup>9</sup>/L. The peripheral blood smear was unremarkable apart from mild leucopenia and moderate thrombocytopenia. Bone marrow aspirate smears demonstrated significant myeloid predominance and severe erythroid hypoplasia with maturation arrest; notably, rare giant proerythroblasts with intranuclear inclusions and subtle vacuolated cytoplasm were identified (Figure 1A, 100× objective). The trephine biopsy exhibited a few giant proerythroblasts among maturing myeloid and megakaryocytic precursors (Figure 1B, 100× objective). Immunohistochemistry revealed large CD71-positive cells (Figure 1C, 100× objective) and Parvovirus B19-positive cells (Figure 1D, 100× objective), indicating infected giant proerythroblasts, also known as'lantern cells.' Parvovirus B19 quantitative polymerase chain reaction (qPCR) was positive, and serologies demonstrated positive IgM and negative IgG, confirming an acute parvovirus infection.</p><p>Parvovirus B19 is a single-stranded DNA virus that infects precursor red blood cells via the P antigen or globoside (Gb4) present on the cell surface [<span>1</span>]. The diagnosis is typically achieved through serological testing, qPCR, clinical symptomatology, and analysis of red blood cell indices. Immunocompromised individuals have been reported to present with normocytic anaemia, a normal leucocyte count, and either normal or elevated platelet counts [<span>2</span>]. These findings are discordant with the current case. Since mature red blood cells remain in circulation for approximately 120 days, early marrow suppression may not immediately result in a decrease in haemoglobin levels. Eighteen days after bone marrow biopsy, repeated complete blood count revealed haemoglobin levels of 8.2 g/dL, a leucocyte count of 5.0 × 10<sup>9</sup>/L, and a platelet count of 228 × 10<sup>9</sup>/L.</p><p>The present case highlights that parvovirus infection can manifest in atypical forms and may go unnoticed in immunocompromised patients lacking characteristic laboratory findings. When encountering severe erythroid hypoplasia with maturation arrest, it is advisable to conduct testing for parvovirus B19 infection, even in the absence of anaemia.</p><p>The authors have nothing to report.</p><p>The author has confirmed that a patient consent statement is not required for this submission, as no patient-identifying data were used.</p><p>The authors declare that they have no relationships with, or financial interests in, any commercial companies related to this article.</p>","PeriodicalId":72883,"journal":{"name":"EJHaem","volume":"6 5","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12515047/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145281976","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kathleen De John, Jan-Edward Koen, Helder De Quintal, Robert Lohlun
<p>A 4-year-old boy presented with pallor and acute respiratory distress requiring nasal prong oxygen. Vital signs showed tachycardia (150 bpm), tachypnoea (38/min) and temperature of 37.8°C. Examination revealed cervical lymphadenopathy, congestive cardiac failure and hepatosplenomegaly with no dysmorphic features. Chest radiography demonstrated right upper lobe consolidation without a mediastinal mass. Laboratory investigations showed severe pancytopenia (Hb 39 g/L, WCC 13.2 × 10⁹/L, ANC 0.13 × 10⁹/L, platelets 29 × 10⁹/L). A leucoerythroblastic blood film revealed 11% abnormal promyelocytes—intermediate size, moderate lightly basophilic granular cytoplasm, occasional vacuoles, folded nuclei, immature chromatin and conspicuous nucleoli (Figure 1A,B). Auer rods and faggot cells were absent. Immunophenotyping by flow cytometry confirmed acute myeloid leukaemia (AML) by identifying a myeloid blast population (35%) with moderate to high complexity expressing CD45, bright cMPO, CD13, CD33, CD15, CD64 and dim HLA-DR, without CD11b, CD14, CD34, CD117 or lymphoid markers. The bone marrow aspirate was markedly hypercellular with 65% abnormal promyelocytes, exhibiting a maturation block (Figure 1C). Megakaryopoiesis and erythropoiesis were reduced with no significant dysplasia. The trephine biopsy was inadequate for assessment.</p><p>Fluorescence in situ hybridization (FISH) detected an extra RARA signal without t(15;17) PML::RARA fusion (Figure 1D). FISH for 17q21 rearrangement was negative, however, 38% of the cells showed three fusion signals, indicating an extra copy of the RARA gene (Figure 1E). Karyotyping revealed an isochromosome 17: 46, XY, i(17)(q10) [10]/46, XY [3] (Figure 1F). Induction chemotherapy with ATRA and dexamethasone was initiated without signs of differentiation syndrome, followed by daunorubicin and intrathecal cytarabine. Bone marrow examination at Day 35 and post-consolidation were consistent with morphological remission.</p><p>Isochromosome 17q [i(17)(q10)] is a cytogenetic abnormality involving the deletion of chromosome 17's short arm and duplication of its long arm, leading to an obligatory loss of a single TP53 allele. It is primarily associated with chronic myeloid leukaemia, myelodysplastic syndromes/myeloproliferative disorders and AML. According to the WHO 5th edition [<span>1</span>], this patient is classified as AML—myelodysplasia-related (isochromosome 17q). To the authors' knowledge, paediatric AML with i(17)(q10) mimicking acute promyelocytic leukaemia is rare, having only been described twice in literature, with these cases generally showing an ineffective response to ATRA and a poor prognosis [<span>2, 3</span>].</p><p>J.E.K. and R.L. performed morphological assessment. R.L. made the diagnosis. H.D.Q. provided clinical data. K.D.J. and R.L. wrote the manuscript which was approved by all authors.</p><p>This study was approved by the University of Cape Town–Human Research Ethics Committee, approval number: 997/202
{"title":"Paediatric Acute Myeloid Leukaemia–Myelodysplasia Related With i(17)(q10) Morphologically Mimicking Acute Promyelocytic Leukaemia","authors":"Kathleen De John, Jan-Edward Koen, Helder De Quintal, Robert Lohlun","doi":"10.1002/jha2.70160","DOIUrl":"10.1002/jha2.70160","url":null,"abstract":"<p>A 4-year-old boy presented with pallor and acute respiratory distress requiring nasal prong oxygen. Vital signs showed tachycardia (150 bpm), tachypnoea (38/min) and temperature of 37.8°C. Examination revealed cervical lymphadenopathy, congestive cardiac failure and hepatosplenomegaly with no dysmorphic features. Chest radiography demonstrated right upper lobe consolidation without a mediastinal mass. Laboratory investigations showed severe pancytopenia (Hb 39 g/L, WCC 13.2 × 10⁹/L, ANC 0.13 × 10⁹/L, platelets 29 × 10⁹/L). A leucoerythroblastic blood film revealed 11% abnormal promyelocytes—intermediate size, moderate lightly basophilic granular cytoplasm, occasional vacuoles, folded nuclei, immature chromatin and conspicuous nucleoli (Figure 1A,B). Auer rods and faggot cells were absent. Immunophenotyping by flow cytometry confirmed acute myeloid leukaemia (AML) by identifying a myeloid blast population (35%) with moderate to high complexity expressing CD45, bright cMPO, CD13, CD33, CD15, CD64 and dim HLA-DR, without CD11b, CD14, CD34, CD117 or lymphoid markers. The bone marrow aspirate was markedly hypercellular with 65% abnormal promyelocytes, exhibiting a maturation block (Figure 1C). Megakaryopoiesis and erythropoiesis were reduced with no significant dysplasia. The trephine biopsy was inadequate for assessment.</p><p>Fluorescence in situ hybridization (FISH) detected an extra RARA signal without t(15;17) PML::RARA fusion (Figure 1D). FISH for 17q21 rearrangement was negative, however, 38% of the cells showed three fusion signals, indicating an extra copy of the RARA gene (Figure 1E). Karyotyping revealed an isochromosome 17: 46, XY, i(17)(q10) [10]/46, XY [3] (Figure 1F). Induction chemotherapy with ATRA and dexamethasone was initiated without signs of differentiation syndrome, followed by daunorubicin and intrathecal cytarabine. Bone marrow examination at Day 35 and post-consolidation were consistent with morphological remission.</p><p>Isochromosome 17q [i(17)(q10)] is a cytogenetic abnormality involving the deletion of chromosome 17's short arm and duplication of its long arm, leading to an obligatory loss of a single TP53 allele. It is primarily associated with chronic myeloid leukaemia, myelodysplastic syndromes/myeloproliferative disorders and AML. According to the WHO 5th edition [<span>1</span>], this patient is classified as AML—myelodysplasia-related (isochromosome 17q). To the authors' knowledge, paediatric AML with i(17)(q10) mimicking acute promyelocytic leukaemia is rare, having only been described twice in literature, with these cases generally showing an ineffective response to ATRA and a poor prognosis [<span>2, 3</span>].</p><p>J.E.K. and R.L. performed morphological assessment. R.L. made the diagnosis. H.D.Q. provided clinical data. K.D.J. and R.L. wrote the manuscript which was approved by all authors.</p><p>This study was approved by the University of Cape Town–Human Research Ethics Committee, approval number: 997/202","PeriodicalId":72883,"journal":{"name":"EJHaem","volume":"6 5","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12515045/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145281951","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Gonzalo de Luna, Isabelle Genty, Vincent Parinet, Florence Beckerich, Rabah Redjoul, Anoosha Habibi, Sihem Iles, Geneviève Derumeaux, Nicolas Lellouche, Thibaut Moulin, Laurent Boyer, Sébastien Maury, Pablo Bartolucci, Thomas d'Humières
A Sickle Cell Anemia patient with severe ventricular arrhythmia and left ventricular dysfunction achieved rapid, complete cardiac recovery after haplo-identical stem cell transplantation, raising questions about SCA cardiac pathophysiology and highlighting the potential for reversible cardiac complications following curative treatment.
Trial Registration: The authors have confirmed clinical trial registration is not needed for this submission
{"title":"Haplo-Identical Stem Cell Transplant Resolves Ventricular Arrhythmias and Dysfunction in a Sickle Cell Anemia Patient: Case Report and Discussion","authors":"Gonzalo de Luna, Isabelle Genty, Vincent Parinet, Florence Beckerich, Rabah Redjoul, Anoosha Habibi, Sihem Iles, Geneviève Derumeaux, Nicolas Lellouche, Thibaut Moulin, Laurent Boyer, Sébastien Maury, Pablo Bartolucci, Thomas d'Humières","doi":"10.1002/jha2.70161","DOIUrl":"10.1002/jha2.70161","url":null,"abstract":"<p>A Sickle Cell Anemia patient with severe ventricular arrhythmia and left ventricular dysfunction achieved rapid, complete cardiac recovery after haplo-identical stem cell transplantation, raising questions about SCA cardiac pathophysiology and highlighting the potential for reversible cardiac complications following curative treatment.</p><p><b>Trial Registration</b>: The authors have confirmed clinical trial registration is not needed for this submission</p>","PeriodicalId":72883,"journal":{"name":"EJHaem","volume":"6 5","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12498570/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145245873","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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