Impact of Chromosomal Aberrations Detected by Chromosome Banding Analysis in Symptomatic Waldenström’s Macroglobulinemia

Impact of Chromosomal Aberrations Detected by Chromosome Banding Analysis in Symptomatic Waldenström’s Macroglobulinemia | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Impact of Chromosomal Aberrations Detected by Chromosome Banding Analysis in Symptomatic Waldenström’s Macroglobulinemia Kenichi Ito, Tomoko Kitagawa, Kunihiko Harada, Kazuhiko Hirano, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4736953/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 16 Oct, 2024 Read the published version in Annals of Hematology → Version 1 posted 9 You are reading this latest preprint version Abstract Background The clinicopathologic features and prognostic impact of MYD88 L265P ( MYD L265P ) and CXCR4 mutations ( CXCR4 Mut ) have been well reported, although little is known regarding the impact of chromosomal aberrations (CA) detected by chromosome banding analysis (CBA) in symptomatic Waldenström’s macroglobulinemia (sWM). Thus, we investigated the clinicopathologic features and prognostic impact in sWM with CAs identified by CBA. Methods We retrospectively analyzed the clinicopathologic results and genetic mutations by droplet digital PCR, fluorescence in situ hybridization (FISH), and CBA using the G-banding method from the bone marrow samples of sWM between April 2010 and March 2024 at our institute. The relationship between CAs and clinicopathologic features was evaluated, as well as the time to next treatment (TTNT). Results Thirty-five patients were enrolled. The median age was 71 years, and the median hemoglobin level was 10.1 g/dL. The median serum IgM and M-protein levels were 3,120 mg/dL and 3 g/dL, respectively. MYD L265P was found in 30/35 patients (85.7%), whereas CXCR4 Mut was found in 3/35 patients (8.6%). Deletion 6q identified by FISH in 5/18 patients (28%), and CAs using CBA in 9/34 patients (26%), including 4/34 (12%) complex karyotypes. sWM with CAs had more anemia ( p = 0.04) and hypoalbuminemia ( p = 0.007), in addition to higher serum M-protein and IgM levels ( p = 0.03). With a median follow-up of 73 months, the median TTNT in patients with and without CAs was 27 and 68 months, respectively. Conclusions CAs with CBA may be associated with clinical aggressiveness and shorter TTNT in sWM. Waldenström’s macroglobulinemia chromosomal aberration chromosome banding analysis droplet digital PCR MYD88 CXCR4 Figures Figure 1 Figure 2 1. Introduction Waldenström’s macroglobulinemia (WM) is one of the rare entities of indolent-B-cell lymphoma, characterized by the infiltration of neoplastic B-cells with plasmacytic differentiation into bone marrow (BM), accompanied by immunoglobulin-M (IgM) monoclonal gammopathy [ 1 ]. The invasion of neoplastic cells and the production of IgM cause a range of clinical symptoms, including anemia, lymphadenopathy, B-symptom, and hyperviscosity syndrome (HVS) [ 1 ]. Whole genome sequencing of the neoplastic cells revealed that most WM patients had the MYD88 L265P mutation ( MYD88 L265P ) [ 2 ], followed by the CXCR4 mutation ( CXCR4 Mut ) [ 3 , 4 ]. Detection of these mutations has contributed to the precise diagnosis of other indolent B-cell lymphomas [ 2 , 5 ], and recent advancements in detection techniques, particularly droplet digital polymerase chain reaction (ddPCR), have enabled the highly sensitive detection of these mutations [ 6 – 8 ]. Furthermore, the genetic mutation status was shown to affect the clinicopathological manifestations, including tumor burden, serum IgM levels, the frequency of HVS [ 9 ], and the response and long-term survival after treatment using Bruton’s tyrosine kinase inhibitor (BTKi) [ 10 – 12 ]. Cytogenetic aberration in symptomatic WM (sWM) also has clinical significance. Deletion of the long arm of chromosome 6 (del 6q) was recognized in 30–55% of the sWM patients [ 13 – 16 ], contributing to the disease aggressiveness and poor prognosis [ 16 , 17 ]. WM patients with deletion of the short arm of chromosome 17 (del 17p) have also been reported to have shorter progression-free survival (PFS) and overall survival (OS) [ 18 ]. Del 6q and del 17p are generally detected by fluorescence in situ hybridization (FISH), as recommended in several guidelines [ 19 , 20 ]. In contrast, little is known regarding the role of chromosomal aberrations (CA) identified by chromosome banding analysis (CBA) (e.g., using the G-banding method) in sWM, although their detection may help in differentiating the diagnosis from other B-cell lymphomas and may have some clinical impacts [ 19 , 21 – 22 ]. Thus, the present study was conducted to clarify the impact of CAs identified by CBA on the clinicopathological features in sWM. 2. Methods 2.1. Patients Patients with sWM attending the National Hospital Organization Disaster Medical Center between April 2010 and March 2024 were enrolled in this study. The diagnosis of WM was made according to the revised 5th edition of the World Health Organization (WHO) Classification of Haematolymphoid Tumours: Lymphoid Neoplasms [ 1 ]. Written informed consent was obtained from the patients who participated in this study. Clinical data were obtained from the medical charts, including age, sex, performance status according to the Eastern Cooperative Oncology Group scale [ 23 ], hemoglobin level, platelet count, serum 𝛽2-microglobulin level, serum albumin level, serum lactate dehydrogenase level, serum M-protein level, serum IgM level, B symptoms, splenomegaly, lymphadenopathy, HVS, cold agglutinin disease, international prognostic scoring system for WM (IPSSWM) [ 24 ], revised IPSSWM (R-IPSSWM) [ 25 ], the best response to the primary therapy, the time to next treatment (TTNT), and OS. 2.2. Pathological Review A pathological review confirmed that all cases were diagnosed as WM according to the revised WHO classification [ 1 ]. Multiple myeloma or marginal zone lymphoma with monoclonal IgM protein were carefully excluded from the study. The infiltration percentage of plasma cells, lymphoplasmacytic cells, lymphocytes, and neoplastic cells in the BM was evaluated using BM smears. 2.3. ddPCR Analysis A ddPCR assay was conducted utilizing the QX200 AutoDG Droplet Digital PCR System (Bio-Rad). The detailed method is provided in the Supplementary Information (SI) (Manuscript S1). For the mutation analysis, ddPCR Mutation Detection assays (Bio-Rad) were employed, namely CXCR4 p.T322fs*26 (Assay ID dHsaMDS736661010, Bio-Rad) for detection of the T318 frameshift (FS) mutation, CXCR4 p.S342* c.1025C > A (Assay ID dHsaMDS858629821, Bio-Rad) and CXCR4 p.S342* c.1025C > G (Assay ID dHsaMDS130738061, Bio-Rad) for the S338 nonsense (NS) mutation, CXCR4 p.S342fs*6 (Assay ID dHsaMDS190756596, Bio-Rad) for S338 FS, and MYD88 p.L265P (Assay ID dHsaMDS2516944, Bio-Rad). 2.4. Chromosomal Banding and FISH Analysis CBA (using the G-banding method) and interphase FISH analysis were performed using the BM aspirates at the first diagnosis of sWM. All karyotypes were described according to the International System for Human Cytogenetic Nomenclature (2020) [ 26 ]. Briefly, at least 20 metaphases were examined in the conventional karyotyping analysis; a karyotype was defined as complex if more than 3 clonal abnormalities were found. For interphase FISH analysis, del 17p (Vysis TP53/CEP17 FISH Probe kit) and del 6q (A20/PRDM1/SHGC-79576 DNA-FISH probe; Cancer Genetics Italia S.R.I., Milano, Italy) were evaluated as previously described [ 17 ]. 2.5 Primary Treatment and Response Assessment The primary therapy regimen was performed according to the doctor’s selection and the prevailing treatment recommendation at the time [ 27 – 29 ]. Responses were assessed following the 6th International Workshop for WM response criteria [ 30 ]. 2.6. Statistical Analysis The patients’ characteristics according to the status of MYD88 / CXCR4 mutations, del 6q, and CAs were compared using the Fisher exact test for categorical data and the Mann–Whitney test for numerical data. The TTNT was defined as the time from the induction of the first-line treatment to that of the second-line treatment, including palliative therapy. Survival and TTNT were censored at the time of the last follow-up date or enrollment in a clinical trial. The OS and TTNT were calculated using the Kaplan–Meier method according to the status of MYD88 / CXCR4 mutations, del 6q, and CAs. The differences in survival were tested using the log-rank test. A p -value of < 0.05 was considered significant, and 0.05 < p < 0.10 was considered a trend. The median follow-up was calculated as the time from the diagnosis of sWM to the last follow-up. All statistical analyses were performed using STATA15 software. Results are shown as the median ± standard deviation. 3. Results 3.1. Clinical Features A total of 35 sWM patients were enrolled in this study. Each patient’s clinical features and summary are presented in Table 1 . The median age was 71 years, with a male predominance (29 of the 35 patients). The median hemoglobin level was 10.1 g/dL, the median serum albumin level was 3.2 g/dL, the median serum IgM level was 3,120 mg/dL, and the median amount of serum M-protein was 3 g/dL. The number of patients with very low/low/intermediate/high/very high R-IPSSWM scores was 3/9/9/9/5. Moreover, 8 of the 35 patients exhibited B symptoms, 10 had splenomegaly, and 11 had lymphadenopathies. HVS was observed in 12 of the 35 patients (34.3%). Table 1 Patient characteristics Characteristic N = 35 Median age, years (range) 71 (49–85) Sex (M/F), n 29/6 PS > 1, n (%) 8/35 (23) Median Hb, g/dL (range) 10.1 (5.3–18.5) Median plt count, ×10 9 /L (range) 231 (57–616) Median serum β2MG, mg/L (range) 3.4 (1.4–8.8) Median serum ALB, g/dL (range) 3.2 (1.7–4.5) Median serum LDH, IU/L (range) 137 (65–269) Median serum IgM, mg/dL (range) 3,009 (327-7,682) Median serum M-protein, g/dL (range) 2.95 (1.1–6.6) IPSSWM (low/int/high), n 8/11/16 R-IPSSWM (very low/low/int/high/very high), n 3/9/9/9/5 B-symptom, n (%) 8/35 (23) Splenomegaly, n (%) 10/35 (29) Lymphadenopathy, n (%) 11/35 (31) Hyperviscosity, n (%) 12/35 (34) Cold agglutinin disease, n (%) 5/35 (14) Median plasma cells in BM, % (range) 0.9 (0–7) Median total neoplastic cells in BM, % (range) 44 (20–87) Primary treatment regimen, N = 33 R-chemotherapy 26/33 (79) R-monotherapy 1/33 (3) BTKi 5/33 (15) Fludarabine 1/33 (3) Abbreviations: M, male; F, female; PS, performance status; Hb, hemoglobin; Plt, platelets; β2MG, β2-microglobulin; ALB, albumin; LDH, lactate dehydrogenase; IgM, immunoglobulin M; IPSSWM, international prognostic scoring system for Waldenstrom macroglobulinemia; R-IPSSWM, revised IPSSWM; BM, bone marrow; R, rituximab; BTKi, Bruton's tyrosine kinase inhibitors. 3.2 Mutation, FISH, and Chromosomal Banding Analysis The distributions of the mutations and cytogenetic abnormalities are illustrated in Fig. 1 . MYD88 L265P was observed in 30/35 patients (85.7%), whereas CXCR4 Mut was observed in 3/35 patients (8.6%, 2 FS mutations ( CXCR4 S338 , CXCR4 T318 ) and 1 NS mutation ( CXCR4 S338 )). Del 6q was found in 5/18 patients (28%), and 17p del was found in 2/31 patients (7%). CAs were identified by CBA in 9/34 patients (26%), including 4/34 patients with complex karyotypes (12%). The detailed individual karyotypes are shown in the SI (Table S1 ). 3.3 Correlation Between Clinical Features and Cytogenetic Abnormalities The differences in clinical manifestations according to the MYD88 and CXCR4 mutation status are presented in Table S2 . The MYD88 L265P /CXCR4 Mut group showed a trend toward higher serum M-protein levels ( p = 0.08 ) compared with the MYD88 L265P /CXCR4 WT group. Table S3 presents a comparison of the clinical manifestations associated with the del 6q status. The patients with del 6q had a trend toward lower serum albumin levels (5/5 (100%) vs. 7/13 (54%), p = 0.09). A comparison of the clinicopathological features of sWM between patients with and without CAs detected by CBA is shown in Table 2 . Patients with CAs exhibited higher rates of anemia (9/9 (100%) vs. 16/25 (64%), p = 0.04) and hypoalbuminemia (8/8 (100%) vs. 11/24 (46%), p = 0.007), as well as higher serum IgM (3,620 vs. 2,662 mg/dL, p = 0.03) and serum M-protein levels (3.75 vs. 2.6 g/dL, p = 0.03). Table 2 Patient characteristics according to the chromosomal aberrations identified by chromosomal banding analysis Characteristic Any chromosomal aberration (N = 9) Normal karyotype (N = 25) p Median age, years (range) 71 (49–85) 70 (50–83) 0.25 Sex (M/F), n 8/1 20/5 0.49 PS > 1, n (%) 4/13 (31) 4/25 (16) 0.26 Hb < 11.5 g/dL, n (%) 9/9 (100) 16/25 (64) 0.04 Plt 3 mg/L, n (%) 6/9 (67) 15/23 (65) 0.64 Serum ALB 250 IU/L, n (%) 0/9 (0) 1/24 (4) 0.7 Median serum IgM, mg/dL (range) 3,620 (1,575-6,810) 2,662 (327-7,682) 0.03 Median M-protein, g/dL (range) 3.75 (2.2–6.6) 2.6 (1.1–5.3) 0.03 IPSSWM (low/int/high), n 0/4/5 8/6/11 R-IPSSWM (very low/low/int/high/very high), n 0/0/4/2/3 3/9/4/7/2 MYD88 L265P mutation, n (%) 8/9 (89) 21/25 (84) 0.6 CXCR4 mutation, n (%) 1/9 (11) 2/25 (8) 0.61 Deletion chromosome 6q, n (%) 3/7 (43) 2/11 (18) 0.27 Deletion chromosome 17p, n (%) 1/7 (9) 1/24 (4) 0.41 Complex karyotype, n (%) 4/9 (44) 0/21 (0) Hyperviscosity, n (%) 5/9 (56) 7/25 (24) 0.14 B-symptom, n (%) 2/9 (22) 6/25 (24) 0.65 Splenomegaly, n (%) 2/9 (22) 7/25 (28) 0.55 Lymphadenopathy, n (%) 3/9 (33) 8/25 (32) 0.63 Cold agglutinin disease, n (%) 1/9 (11) 3/25 (12) 0.72 Median plasma cells in BM, % (range) 1 (0–7) 0.9 (0-4.3) 0.96 Median total neoplastic cells in BM, % (range) 55.9 (35.1–86.7) 41.4 (27.9–79.2) 0.1 Abbreviations: PS, performance status; Hb, hemoglobin; Plt, platelets; β2MG, β2-microglobulin; ALB, albumin; LDH, lactate dehydrogenase; IgM, immunoglobulin M; IPSSWM, international prognostic scoring system for Waldenstrom macroglobulinemia; R-IPSSWM, revised IPSSWM; BM, bone marrow. 3.4 Treatment and Survival Outcomes Among the 35 sWM patients, one patient was observed without intervention and one was excluded due to participation in a clinical trial. Thus, 33 patients who received primary therapy were analyzed in terms of treatment and survival (Table 1 ). The median follow-up duration of the 33 patients was 73 months. Overall, 26 patients underwent chemotherapy with rituximab (R), one patient received R monotherapy, 5 patients received BTKi, and one patient received another treatment. Furthermore, 10 of the 12 patients with HVS underwent plasma exchange prior to therapeutic intervention. The initial best responses involved a complete response in 15% (5 patients), very good partial response in 24% (8 patients), partial response in 36% (12 patients), stable disease in 21% (7 patients), and progressive disease in 3% (1 patient). Figure 2 shows the Kaplan–Meier curves of TTNT and OS in the 33 sWM patients, analyzed according to mutation status, del 6q, and CAs using CBA. During the median follow-up of 73 months, no differences in TTNT or OS were correlated with the status of MYD88 / CXCR4 or del 6q. Conversely, the median TTNT was 27 and 68 months with and without CAs ( p = 0.14), and the median TTNT was 27 and 61 months with and without complex karyotypes ( p = 0.42 ), respectively. 4. Discussion MYD88 L265P is recognized in approximately 90% of WM patients and plays a major role in tumor cell survival and proliferation through constitutive NF-kB activation in WM [ 2 , 31 ]. In contrast, CXCR4 Mut is recognized in 25–40% of WM patients [ 1 , 3 – 4 , 12 ] and is generally accepted as the second most common hit, with poor prognostic factors. In particular, the CXCR4 NS mutation has been associated with elevated serum IgM levels, an increased risk of HVS, and an inferior response and survival in WM patients treated with ibrutinib monotherapy [ 9 , 12 ]. In the present study, MYD88 L265P was found in 85.7% of the patients, which is comparable to previous reports [ 1 , 2 ]. The CXCR4 Mut rate was 8.6% in this study, which is equal to or lower than that found in previous reports [ 1 , 3 – 4 , 12 ], although CXCR4 Mut exhibited a trend toward higher serum IgM levels. One reason for this discrepancy may be that only 4 probes were used for detecting CXCR4 Mut : two NS mutations for S338 and an FS mutation for S338 and T318. Another reason may be that all the BM samples were obtained at the first diagnosis of sWM. For reference, regarding East Asia, 4/26 patients (15%) exhibited CXCR4 Mut based on next-generation sequencing from unselected BM in a Japanese phase II study of tirabrutinib for untreated and relapsed/refractory WM (RRWM) [ 32 ]. In a trial of orelabrutinib conducted in China for RRWM, the detection rate of CXCR4 Mut was 4/47 patients (8.8%) based on allele specific-PCR for the S338 mutation [ 33 ]. The detection of del 6q and del 17p by FISH analyses is recommended in several guidelines [ 19 , 20 ]. Del 6q is observed in 30–55% of WM patients [ 13 – 16 ] and is less frequently identified (0–4%) in patients with IgM monoclonal gammopathy of undetermined significance; therefore, it is considered to be the secondary event [ 14 , 16 ]. WM patients with del 6q exhibited higher serum M-protein levels [ 14 ], higher IPSSWM scores, and poor prognosis [ 16 ]. Del 17p has also been associated with unfavorable prognosis [ 18 ]. In the present study, FISH analysis revealed the presence of del 6q in 5/18 patients (28%) and del 17p in 2/31 patients (7%). Although we were unable to fully analyze the significance of del 6q due to missing data, the results demonstrated that serum albumin levels tended to be low, and no difference was observed in serum M-protein levels or prognosis. In contrast, the impact of CAs using CBA in sWM remains poorly understood. Several studies reported the clinicopathological significance of CAs identified by CBA in WM [ 13 , 21 – 22 ]. Nguyen-Khac et al. found that 66/141 cases (47%) had abnormal karyotypes, including 30% complex karyotypes [ 13 ]. In the report, del 6q was identified with greater frequency in complex karyotypes than in abnormal karyotypes with fewer abnormalities. A group in the French Innovative Leukemia Organization retrospectively analyzed the clinicopathological impact of CAs in 239 WM patients [ 21 ]. In their study, highly complex karyotypes with 5 or more CAs were found in 5% of patients and were associated with poor PFS and OS. In addition, a significant correlation between highly complex karyotypes and del 6q and/or TP53-related abnormalities was observed. Recently, Danesin and colleagues reported that WM patients with CAs were significantly older at the time of diagnosis (median age, 72 vs. 65 years) and had inferior median OS (76.1 vs. 167.7 months) compared with WM patients that had normal karyotypes, across 23 years in the single-center study [ 22 ]. In our study, the presence of CAs detected by CBA was associated with significantly higher rates of anemia and hypoalbuminemia, which are associated with poor prognostic factors in IPSSWM, and higher serum IgM levels. The distribution of age was found to be identical regardless of the presence or absence of CAs. The TTNT in sWM patients with CAs tended to be shorter, although a significant difference was not observed, possibly due to the small sample size. Considering our results and the previous reports, CAs identified by CBA in sWM patients may have some clinical impacts. Regarding the limitations of this work, the present retrospective study was based on 35 patients at a single institute. Second, CXCR4 Mut was not comprehensively examined; only four probes were used, and other types of CXCR4 Mut may have been missed. Third, 17 of the 35 cases lacked del 6q data, providing insufficient analysis. Finally, the outcomes of TTNT and OS may have been influenced by the treatment regimens. In conclusion, our findings suggest that CAs identified by CBA are associated with clinical aggressiveness and shorter TTNT in sWM. Further studies are necessary to clarify and validate the significance of CAs in sWM. Declarations Competing interest Naohiro Sekiguchi receives honoraria from Janssen, Ono, and research funding from Incyte Biosciences Japan, Janssen, Mitsubishi Tanabe Pharma Corporation, MSD, Ono. Ethics approval This study was approved by the institutional review board at our hospital (Disaster Medical Center IRB-2022-33). Consent to participate Written consent was obtained from the patients. Funding This research received no specific grant or funding from agencies in the public, commercial, or not-for-profit sectors. Author Contribution Kenichi Ito analyzed and interpreted the data and drafted the manuscript. Tomoko Kitagawa organized the clinical trial and analyzed the data. Kunihiko Harada acquired the data. Kazuhiko Hirano reviewed pathological diagnoses and provided the pictures for figures. Naohiro Sekiguchi organized the clinical trial, analyzed the data, and revised the manuscript critically for important intellectual content. All authors read and approved the final manuscript. Data Availability The data that support the fndings of this study are available upon request from the corresponding author. References Alaggio R, Amador C, Anagnostopoulos I, Attygalle AD, Araujo IBO, Berti E et al (2022) The 5th edition of the World Health Organization Classification of Haematolymphoid Tumours. Lymphoid Neoplasms Leuk 36(7):1720–1748. https://doi.org/10.1038/s41375-022-01620-2 Treon SP, Xu L, Yang G, Zhou Y, Liu X, Cao Y et al (2012) MYD88 L265P somatic mutation in Waldenström's macroglobulinemia. 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Am J Hematol 96(12):1569–1579. https://doi.org/10.1002/ajh.26339 Danesin N, Bonaldi L, Martines A, Nalio S, Bertorelle R, Compagno S et al (2024) Impact of the presence and number of chromosomal abnormalities on the clinical outcome in Waldenström Macroglobulinemia: a monocentric experience. Ann Hematol. https://doi.org/10.1007/s00277-024-05770-4 Oken MM, Creech RH, Tormey DC, Horton J, Davis TE, McFadden ET, Carbone PP (1982) Toxicity and response criteria of the Eastern Cooperative Oncology Group. Am J Clin Oncol 5(6):649–655 Morel P, Duhamel A, Gobbi P, Dimopoulos MA, Dhodapkar MV, McCoy J et al (2009) International prognostic scoring system for Waldenstrom macroglobulinemia. Blood 113(18):4163–4170. https://doi.org/10.1182/blood-2008-08-174961 Kastritis E, Morel P, Duhamel A, Gavriatopoulou M, Kyrtsonis MC, Durot E et al (2019) A revised international prognostic score system for Waldenström's macroglobulinemia. Leukemia 33(11):2654–2661. https://doi.org/10.1038/s41375-019-0431-y McGowan-Jordan J, Hastings RJ, Moore S, International Standing Committee on Human Cytogenomic Nomenclature (2020) ISCN 2020: an Internatonal System. for Human Cytogenomic Nomenclature. Karger Dimopoulos MA, Kastritis E, Owen RG, Kyle RA, Landgren O, Morra E et al (2014) Treatment recommendations for patients with Waldenström macroglobulinemia (WM) and related disorders: IWWM-7 consensus. Blood 124(9):1404–1411. https://doi.org/10.1182/blood-2014-03-565135 Kobayashi Y (2019) JSH practical guidelines for hematological malignancies, 2018: II. Lymphoma-3. Lymphoplasmacytic lymphoma/Waldenström's macroglobulinemia (LPL/WM). Int J Hematol, 110(5), 524–528. https://doi.org/10.1007/s12185-019-02730-x Castillo JJ, Advani RH, Branagan AR, Buske C, Dimopoulos MA, D'Sa S et al (2020) Consensus treatment recommendations from the tenth International Workshop for Waldenström Macroglobulinaemia. Lancet Haematol, 7(11), e827-e837. https://doi.org/10.1016/s2352-3026(20)30224-6 Owen RG, Kyle RA, Stone MJ, Rawstron AC, Leblond V, Merlini G et al (2013) Response assessment in Waldenström macroglobulinaemia: update from the VIth International Workshop. Br J Haematol, 160(2), 171–176. https://doi.org/10.1111/bjh.12102 Yang G, Zhou Y, Liu X, Xu L, Cao Y, Manning RJ et al (2013) A mutation in MYD88 (L265P) supports the survival of lymphoplasmacytic cells by activation of Bruton tyrosine kinase in Waldenström macroglobulinemia. Blood 122(7):1222–1232. https://doi.org/10.1182/blood-2012-12-475111 Sekiguchi N, Rai S, Munakata W et al (2020) A multicenter, open-label, phase II study of tirabrutinib (ONO/GS-4059) in patients with Waldenström's macroglobulinemia. Cancer Sci 111(9):3327–3337. 10.1111/cas.14561 Cao XX, Jin J, Fu CC, Yi SH, Zhao WL, Sun ZM et al (2022) Evaluation of orelabrutinib monotherapy in patients with relapsed or refractory Waldenström's macroglobulinemia in a single-arm, multicenter, open-label, phase 2 study. EClinicalMedicine 52:101682. https://doi.org/10.1016/j.eclinm.2022.101682 Additional Declarations Competing interest reported. Naohiro Sekiguchi receives honoraria from Janssen, Ono, and research funding from Incyte Biosciences Japan, Janssen, Mitsubishi Tanabe Pharma Corporation, MSD, Ono. Supplementary Files SuppTable1.xlsx SuppTable2.xlsx SuppTable3.xlsx Suppledocument.docx Cite Share Download PDF Status: Published Journal Publication published 16 Oct, 2024 Read the published version in Annals of Hematology → Version 1 posted Editorial decision: Revision requested 12 Sep, 2024 Reviews received at journal 12 Sep, 2024 Reviews received at journal 04 Sep, 2024 Reviewers agreed at journal 03 Sep, 2024 Reviewers agreed at journal 20 Aug, 2024 Reviewers invited by journal 20 Jul, 2024 Editor assigned by journal 17 Jul, 2024 Submission checks completed at journal 17 Jul, 2024 First submitted to journal 13 Jul, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4736953","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":331733750,"identity":"486a2f92-2545-4fb0-99fe-85b47b31c334","order_by":0,"name":"Kenichi Ito","email":"","orcid":"","institution":"National Hospital Organization Disaster Medical Center","correspondingAuthor":false,"prefix":"","firstName":"Kenichi","middleName":"","lastName":"Ito","suffix":""},{"id":331733754,"identity":"03982f04-e6b3-4703-9093-123dc12574ad","order_by":1,"name":"Tomoko Kitagawa","email":"","orcid":"","institution":"National Hospital Organization Disaster Medical Center","correspondingAuthor":false,"prefix":"","firstName":"Tomoko","middleName":"","lastName":"Kitagawa","suffix":""},{"id":331733756,"identity":"391f72c8-4fb8-49cb-95c8-c7e4e8947be3","order_by":2,"name":"Kunihiko Harada","email":"","orcid":"","institution":"National Hospital Organization Tokyo Medical Center","correspondingAuthor":false,"prefix":"","firstName":"Kunihiko","middleName":"","lastName":"Harada","suffix":""},{"id":331733757,"identity":"09f0ef6c-1f90-4d43-bc15-52c830d92074","order_by":3,"name":"Kazuhiko Hirano","email":"","orcid":"","institution":"National Hospital Organization Disaster Medical Center","correspondingAuthor":false,"prefix":"","firstName":"Kazuhiko","middleName":"","lastName":"Hirano","suffix":""},{"id":331733758,"identity":"8f981603-b76f-4c9e-a1d8-7d06b4e6fa5e","order_by":4,"name":"Naohiro Sekiguchi","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA60lEQVRIiWNgGAWjYHACxgcfgCQ/e/MBICUhQ4wWZsMZQFKy51gCSAsPMVrYpDmApMENHwMQj7AWc/bDB6QZd9jJMdzg+fzqRo0FDwP74aMb8Gmx7ElLMC48k2zMOLt3m3XOMaDDeNLSbuDTYnAgxyB5ZhtzYrPM2W3GOWxALRI8Zvi1nH9jcJi3rT6xTSLnmXHOP2K03MgxbOZtO5zYI5HD/Di3jSgtz5IZZ7YdN5bgOWbGnNsnwcNG0C/nk4//+NhWLWd/vPnx55xvdXL87IeP4dWCDNgkwCSxykGA+QMpqkfBKBgFo2DkAAALCUltAvABXwAAAABJRU5ErkJggg==","orcid":"","institution":"National Hospital Organization Disaster Medical Center","correspondingAuthor":true,"prefix":"","firstName":"Naohiro","middleName":"","lastName":"Sekiguchi","suffix":""}],"badges":[],"createdAt":"2024-07-14 04:08:16","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4736953/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4736953/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s00277-024-06041-y","type":"published","date":"2024-10-16T15:57:39+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":62652943,"identity":"e67aa980-cf7a-4792-b761-051bde1e310f","added_by":"auto","created_at":"2024-08-17 01:05:15","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":168252,"visible":true,"origin":"","legend":"\u003cp\u003eThis illustration depicts the prevalence of genetic mutations and chromosomal abnormalities in patients diagnosed with Waldenström’s macroglobulinemia. Each column represents an individual patient (numbers 1–35), and the rows indicate the status of mutations, fluorescence in situ hybridization, and chromosomal abnormalities. The beige and white squares indicate the presence and absence of alterations in each category, respectively. The black squares indicate that the results were unavailable.\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-4736953/v1/1ae32cdfd8308bc3f962e411.jpeg"},{"id":62652944,"identity":"af26ddac-e876-4d8a-8ce0-1fee965c20f8","added_by":"auto","created_at":"2024-08-17 01:05:15","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":160247,"visible":true,"origin":"","legend":"\u003cp\u003eThe time to next treatment (TTNT) and overall survival (OS) for the status of genetic mutations and chromosomal abnormalities: (a, b)\u003cem\u003e MYD88/CXCR4\u003c/em\u003e, (c, d) chromosome 6q deletion, (e, f) chromosomal aberrations detected by chromosomal banding analysis (G-banding), and (g, h) complex karyotypes. Abbreviations: mut, mutated; WT, wild-type; 6q del, chromosome 6q deletion.\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-4736953/v1/f2efe0ba23d09697e4179744.png"},{"id":67148990,"identity":"398c95c5-4092-4549-b358-a0cd7998bb00","added_by":"auto","created_at":"2024-10-21 16:10:41","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":945894,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4736953/v1/796ae891-5de0-4bcd-ab0f-bfe4506f8b90.pdf"},{"id":62654103,"identity":"516b7c4c-c229-4ddd-beab-59ca2051d71d","added_by":"auto","created_at":"2024-08-17 01:21:15","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":14991,"visible":true,"origin":"","legend":"","description":"","filename":"SuppTable1.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4736953/v1/3b4229eb0eec3997b5e89768.xlsx"},{"id":62652948,"identity":"983a951c-a11e-428c-8179-0a47fb4c5a7f","added_by":"auto","created_at":"2024-08-17 01:05:15","extension":"xlsx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":13516,"visible":true,"origin":"","legend":"","description":"","filename":"SuppTable2.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4736953/v1/e570e87eca84c74175441dc6.xlsx"},{"id":62653547,"identity":"111ffa6c-2779-4c30-ae9f-0979d7d07d86","added_by":"auto","created_at":"2024-08-17 01:13:15","extension":"xlsx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":12932,"visible":true,"origin":"","legend":"","description":"","filename":"SuppTable3.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4736953/v1/a7c37f346e404573f9c19253.xlsx"},{"id":62654102,"identity":"ee078553-baeb-434f-8ea0-f7ff99376f7e","added_by":"auto","created_at":"2024-08-17 01:21:15","extension":"docx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":14296,"visible":true,"origin":"","legend":"","description":"","filename":"Suppledocument.docx","url":"https://assets-eu.researchsquare.com/files/rs-4736953/v1/14d84b713971f2bd9aa59e67.docx"}],"financialInterests":"Competing interest reported. Naohiro Sekiguchi receives honoraria from Janssen, Ono, and research funding from Incyte Biosciences Japan, Janssen, Mitsubishi Tanabe Pharma Corporation, MSD, Ono.","formattedTitle":"Impact of Chromosomal Aberrations Detected by Chromosome Banding Analysis in Symptomatic Waldenström’s Macroglobulinemia","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eWaldenstr\u0026ouml;m\u0026rsquo;s macroglobulinemia (WM) is one of the rare entities of indolent-B-cell lymphoma, characterized by the infiltration of neoplastic B-cells with plasmacytic differentiation into bone marrow (BM), accompanied by immunoglobulin-M (IgM) monoclonal gammopathy [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. The invasion of neoplastic cells and the production of IgM cause a range of clinical symptoms, including anemia, lymphadenopathy, B-symptom, and hyperviscosity syndrome (HVS) [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eWhole genome sequencing of the neoplastic cells revealed that most WM patients had the \u003cem\u003eMYD88 L265P\u003c/em\u003e mutation (\u003cem\u003eMYD88\u003c/em\u003e\u003csup\u003e\u003cem\u003eL265P\u003c/em\u003e\u003c/sup\u003e) [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e], followed by the \u003cem\u003eCXCR4\u003c/em\u003e mutation (\u003cem\u003eCXCR4\u003c/em\u003e\u003csup\u003e\u003cem\u003eMut\u003c/em\u003e\u003c/sup\u003e) [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Detection of these mutations has contributed to the precise diagnosis of other indolent B-cell lymphomas [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e], and recent advancements in detection techniques, particularly droplet digital polymerase chain reaction (ddPCR), have enabled the highly sensitive detection of these mutations [\u003cspan additionalcitationids=\"CR7\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Furthermore, the genetic mutation status was shown to affect the clinicopathological manifestations, including tumor burden, serum IgM levels, the frequency of HVS [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e], and the response and long-term survival after treatment using Bruton\u0026rsquo;s tyrosine kinase inhibitor (BTKi) [\u003cspan additionalcitationids=\"CR11\" citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eCytogenetic aberration in symptomatic WM (sWM) also has clinical significance. Deletion of the long arm of chromosome 6 (del 6q) was recognized in 30\u0026ndash;55% of the sWM patients [\u003cspan additionalcitationids=\"CR14 CR15\" citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e], contributing to the disease aggressiveness and poor prognosis [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. WM patients with deletion of the short arm of chromosome 17 (del 17p) have also been reported to have shorter progression-free survival (PFS) and overall survival (OS) [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Del 6q and del 17p are generally detected by fluorescence in situ hybridization (FISH), as recommended in several guidelines [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. In contrast, little is known regarding the role of chromosomal aberrations (CA) identified by chromosome banding analysis (CBA) (e.g., using the G-banding method) in sWM, although their detection may help in differentiating the diagnosis from other B-cell lymphomas and may have some clinical impacts [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThus, the present study was conducted to clarify the impact of CAs identified by CBA on the clinicopathological features in sWM.\u003c/p\u003e"},{"header":"2. Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Patients\u003c/h2\u003e \u003cp\u003ePatients with sWM attending the National Hospital Organization Disaster Medical Center between April 2010 and March 2024 were enrolled in this study. The diagnosis of WM was made according to the revised 5th edition of the World Health Organization (WHO) Classification of Haematolymphoid Tumours: Lymphoid Neoplasms [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Written informed consent was obtained from the patients who participated in this study.\u003c/p\u003e \u003cp\u003eClinical data were obtained from the medical charts, including age, sex, performance status according to the Eastern Cooperative Oncology Group scale [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e], hemoglobin level, platelet count, serum \u0026#120573;2-microglobulin level, serum albumin level, serum lactate dehydrogenase level, serum M-protein level, serum IgM level, B symptoms, splenomegaly, lymphadenopathy, HVS, cold agglutinin disease, international prognostic scoring system for WM (IPSSWM) [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e], revised IPSSWM (R-IPSSWM) [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e], the best response to the primary therapy, the time to next treatment (TTNT), and OS.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Pathological Review\u003c/h2\u003e \u003cp\u003eA pathological review confirmed that all cases were diagnosed as WM according to the revised WHO classification [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Multiple myeloma or marginal zone lymphoma with monoclonal IgM protein were carefully excluded from the study. The infiltration percentage of plasma cells, lymphoplasmacytic cells, lymphocytes, and neoplastic cells in the BM was evaluated using BM smears.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3. ddPCR Analysis\u003c/h2\u003e \u003cp\u003eA ddPCR assay was conducted utilizing the QX200 AutoDG Droplet Digital PCR System (Bio-Rad). The detailed method is provided in the Supplementary Information (SI) (Manuscript S1). For the mutation analysis, ddPCR Mutation Detection assays (Bio-Rad) were employed, namely \u003cem\u003eCXCR4\u003c/em\u003e p.T322fs*26 (Assay ID dHsaMDS736661010, Bio-Rad) for detection of the \u003cem\u003eT318\u003c/em\u003e frameshift (FS) mutation, \u003cem\u003eCXCR4\u003c/em\u003e p.S342* c.1025C\u0026thinsp;\u0026gt;\u0026thinsp;A (Assay ID dHsaMDS858629821, Bio-Rad) and \u003cem\u003eCXCR4\u003c/em\u003e p.S342* c.1025C\u0026thinsp;\u0026gt;\u0026thinsp;G (Assay ID dHsaMDS130738061, Bio-Rad) for the \u003cem\u003eS338\u003c/em\u003e nonsense (NS) mutation, \u003cem\u003eCXCR4\u003c/em\u003e p.S342fs*6 (Assay ID dHsaMDS190756596, Bio-Rad) for \u003cem\u003eS338\u003c/em\u003e FS, and MYD88 p.L265P (Assay ID dHsaMDS2516944, Bio-Rad).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4. Chromosomal Banding and FISH Analysis\u003c/h2\u003e \u003cp\u003eCBA (using the G-banding method) and interphase FISH analysis were performed using the BM aspirates at the first diagnosis of sWM. All karyotypes were described according to the International System for Human Cytogenetic Nomenclature (2020) [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Briefly, at least 20 metaphases were examined in the conventional karyotyping analysis; a karyotype was defined as complex if more than 3 clonal abnormalities were found.\u003c/p\u003e \u003cp\u003eFor interphase FISH analysis, del 17p (Vysis TP53/CEP17 FISH Probe kit) and del 6q (A20/PRDM1/SHGC-79576 DNA-FISH probe; Cancer Genetics Italia S.R.I., Milano, Italy) were evaluated as previously described [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Primary Treatment and Response Assessment\u003c/h2\u003e \u003cp\u003eThe primary therapy regimen was performed according to the doctor\u0026rsquo;s selection and the prevailing treatment recommendation at the time [\u003cspan additionalcitationids=\"CR28\" citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Responses were assessed following the 6th International Workshop for WM response criteria [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.6. Statistical Analysis\u003c/h2\u003e \u003cp\u003eThe patients\u0026rsquo; characteristics according to the status of \u003cem\u003eMYD88\u003c/em\u003e/\u003cem\u003eCXCR4\u003c/em\u003e mutations, del 6q, and CAs were compared using the Fisher exact test for categorical data and the Mann\u0026ndash;Whitney test for numerical data. The TTNT was defined as the time from the induction of the first-line treatment to that of the second-line treatment, including palliative therapy. Survival and TTNT were censored at the time of the last follow-up date or enrollment in a clinical trial. The OS and TTNT were calculated using the Kaplan\u0026ndash;Meier method according to the status of \u003cem\u003eMYD88\u003c/em\u003e/\u003cem\u003eCXCR4\u003c/em\u003e mutations, del 6q, and CAs. The differences in survival were tested using the log-rank test. A \u003cem\u003ep\u003c/em\u003e-value of \u003cem\u003e\u0026lt;\u0026thinsp;0.05\u003c/em\u003e was considered significant, and \u003cem\u003e0.05\u0026thinsp;\u0026lt;\u0026thinsp;p\u0026thinsp;\u0026lt;\u0026thinsp;0.10\u003c/em\u003e was considered a trend. The median follow-up was calculated as the time from the diagnosis of sWM to the last follow-up. All statistical analyses were performed using STATA15 software. Results are shown as the median\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e3.1. Clinical Features\u003c/h2\u003e \u003cp\u003eA total of 35 sWM patients were enrolled in this study. Each patient\u0026rsquo;s clinical features and summary are presented in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The median age was 71 years, with a male predominance (29 of the 35 patients). The median hemoglobin level was 10.1 g/dL, the median serum albumin level was 3.2 g/dL, the median serum IgM level was 3,120 mg/dL, and the median amount of serum M-protein was 3 g/dL. The number of patients with very low/low/intermediate/high/very high R-IPSSWM scores was 3/9/9/9/5. Moreover, 8 of the 35 patients exhibited B symptoms, 10 had splenomegaly, and 11 had lymphadenopathies. HVS was observed in 12 of the 35 patients (34.3%).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePatient characteristics\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"2\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCharacteristic\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eN\u0026thinsp;=\u0026thinsp;35\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian age, years (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e71 (49\u0026ndash;85)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSex (M/F), n\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e29/6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePS\u0026thinsp;\u0026gt;\u0026thinsp;1, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8/35 (23)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian Hb, g/dL (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10.1 (5.3\u0026ndash;18.5)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian plt count, \u0026times;10\u003csup\u003e9\u003c/sup\u003e /L (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e231 (57\u0026ndash;616)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian serum β2MG, mg/L (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.4 (1.4\u0026ndash;8.8)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian serum ALB, g/dL (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.2 (1.7\u0026ndash;4.5)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian serum LDH, IU/L (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e137 (65\u0026ndash;269)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian serum IgM, mg/dL (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3,009 (327-7,682)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian serum M-protein, g/dL (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.95 (1.1\u0026ndash;6.6)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIPSSWM (low/int/high), n\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8/11/16\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eR-IPSSWM (very low/low/int/high/very high), n\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3/9/9/9/5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eB-symptom, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8/35 (23)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSplenomegaly, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10/35 (29)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLymphadenopathy, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11/35 (31)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHyperviscosity, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e12/35 (34)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCold agglutinin disease, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5/35 (14)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian plasma cells in BM, % (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.9 (0\u0026ndash;7)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian total neoplastic cells in BM, % (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e44 (20\u0026ndash;87)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003ePrimary treatment regimen, N\u0026thinsp;=\u0026thinsp;33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eR-chemotherapy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e26/33 (79)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eR-monotherapy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1/33 (3)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBTKi\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5/33 (15)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFludarabine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1/33 (3)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eAbbreviations: M, male; F, female; PS, performance status; Hb, hemoglobin; Plt, platelets; β2MG, β2-microglobulin; ALB, albumin; LDH, lactate dehydrogenase; IgM, immunoglobulin M; IPSSWM, international prognostic scoring system for Waldenstrom macroglobulinemia; R-IPSSWM, revised IPSSWM; BM, bone marrow; R, rituximab; BTKi, Bruton's tyrosine kinase inhibitors.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e3.2 Mutation, FISH, and Chromosomal Banding Analysis\u003c/h2\u003e \u003cp\u003eThe distributions of the mutations and cytogenetic abnormalities are illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. \u003cem\u003eMYD88\u003c/em\u003e\u003csup\u003e\u003cem\u003eL265P\u003c/em\u003e\u003c/sup\u003e was observed in 30/35 patients (85.7%), whereas \u003cem\u003eCXCR4\u003c/em\u003e\u003csup\u003e\u003cem\u003eMut\u003c/em\u003e\u003c/sup\u003e was observed in 3/35 patients (8.6%, 2 FS mutations (\u003cem\u003eCXCR4 S338\u003c/em\u003e, \u003cem\u003eCXCR4 T318\u003c/em\u003e) and 1 NS mutation (\u003cem\u003eCXCR4 S338\u003c/em\u003e)). Del 6q was found in 5/18 patients (28%), and 17p del was found in 2/31 patients (7%). CAs were identified by CBA in 9/34 patients (26%), including 4/34 patients with complex karyotypes (12%). The detailed individual karyotypes are shown in the SI (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e3.3 Correlation Between Clinical Features and Cytogenetic Abnormalities\u003c/h2\u003e \u003cp\u003eThe differences in clinical manifestations according to the \u003cem\u003eMYD88\u003c/em\u003e and \u003cem\u003eCXCR4\u003c/em\u003e mutation status are presented in Table \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e. The \u003cem\u003eMYD88\u003c/em\u003e\u003csup\u003e\u003cem\u003eL265P\u003c/em\u003e\u003c/sup\u003e\u003cem\u003e/CXCR4\u003c/em\u003e\u003csup\u003e\u003cem\u003eMut\u003c/em\u003e\u003c/sup\u003e group showed a trend toward higher serum M-protein levels (\u003cem\u003ep\u0026thinsp;=\u0026thinsp;0.08\u003c/em\u003e) compared with the \u003cem\u003eMYD88\u003c/em\u003e\u003csup\u003e\u003cem\u003eL265P\u003c/em\u003e\u003c/sup\u003e\u003cem\u003e/CXCR4\u003c/em\u003e\u003csup\u003e\u003cem\u003eWT\u003c/em\u003e\u003c/sup\u003e group.\u003c/p\u003e \u003cp\u003eTable \u003cspan refid=\"MOESM3\" class=\"InternalRef\"\u003eS3\u003c/span\u003e presents a comparison of the clinical manifestations associated with the del 6q status. The patients with del 6q had a trend toward lower serum albumin levels (5/5 (100%) vs. 7/13 (54%), \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.09).\u003c/p\u003e \u003cp\u003eA comparison of the clinicopathological features of sWM between patients with and without CAs detected by CBA is shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. Patients with CAs exhibited higher rates of anemia (9/9 (100%) vs. 16/25 (64%), \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.04) and hypoalbuminemia (8/8 (100%) vs. 11/24 (46%), \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.007), as well as higher serum IgM (3,620 vs. 2,662 mg/dL, \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.03) and serum M-protein levels (3.75 vs. 2.6 g/dL, \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.03).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePatient characteristics according to the chromosomal aberrations identified by chromosomal banding analysis\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCharacteristic\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAny chromosomal aberration\u003c/p\u003e \u003cp\u003e(N\u0026thinsp;=\u0026thinsp;9)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNormal karyotype\u003c/p\u003e \u003cp\u003e(N\u0026thinsp;=\u0026thinsp;25)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian age, years (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e71 (49\u0026ndash;85)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e70 (50\u0026ndash;83)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.25\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSex (M/F), n\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8/1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e20/5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.49\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePS\u0026thinsp;\u0026gt;\u0026thinsp;1, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4/13 (31)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4/25 (16)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.26\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHb\u0026thinsp;\u0026lt;\u0026thinsp;11.5 g/dL, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9/9 (100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16/25 (64)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.04\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePlt\u0026thinsp;\u0026lt;\u0026thinsp;100 \u0026times; 109 /L, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2/9 (22)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1/25 (4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.16\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eβ2MG\u0026thinsp;\u0026gt;\u0026thinsp;3 mg/L, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6/9 (67)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15/23 (65)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.64\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSerum ALB\u0026thinsp;\u0026lt;\u0026thinsp;3.5 g/dL, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8/8 (100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e11/24 (46)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.007\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLDH\u0026thinsp;\u0026gt;\u0026thinsp;250 IU/L, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0/9 (0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1/24 (4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian serum IgM, mg/dL (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3,620 (1,575-6,810)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2,662 (327-7,682)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.03\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian M-protein, g/dL (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.75 (2.2\u0026ndash;6.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.6 (1.1\u0026ndash;5.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.03\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIPSSWM (low/int/high), n\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0/4/5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8/6/11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eR-IPSSWM (very low/low/int/high/very high), n\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0/0/4/2/3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3/9/4/7/2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMYD88 L265P mutation, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8/9 (89)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e21/25 (84)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCXCR4 mutation, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1/9 (11)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2/25 (8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.61\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDeletion chromosome 6q, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3/7 (43)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2/11 (18)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.27\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDeletion chromosome 17p, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1/7 (9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1/24 (4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.41\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eComplex karyotype, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4/9 (44)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0/21 (0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHyperviscosity, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5/9 (56)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7/25 (24)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.14\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eB-symptom, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2/9 (22)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6/25 (24)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.65\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSplenomegaly, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2/9 (22)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7/25 (28)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.55\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLymphadenopathy, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3/9 (33)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8/25 (32)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.63\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCold agglutinin disease, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1/9 (11)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3/25 (12)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.72\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian plasma cells in BM, % (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1 (0\u0026ndash;7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.9 (0-4.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.96\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedian total neoplastic cells in BM, % (range)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e55.9 (35.1\u0026ndash;86.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e41.4 (27.9\u0026ndash;79.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"4\" nameend=\"c4\" namest=\"c1\"\u003e \u003cp\u003eAbbreviations: PS, performance status; Hb, hemoglobin; Plt, platelets; β2MG, β2-microglobulin; ALB, albumin; LDH, lactate dehydrogenase; IgM, immunoglobulin M; IPSSWM, international prognostic scoring system for Waldenstrom macroglobulinemia; R-IPSSWM, revised IPSSWM; BM, bone marrow.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e3.4 Treatment and Survival Outcomes\u003c/h2\u003e \u003cp\u003eAmong the 35 sWM patients, one patient was observed without intervention and one was excluded due to participation in a clinical trial. Thus, 33 patients who received primary therapy were analyzed in terms of treatment and survival (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The median follow-up duration of the 33 patients was 73 months.\u003c/p\u003e \u003cp\u003eOverall, 26 patients underwent chemotherapy with rituximab (R), one patient received R monotherapy, 5 patients received BTKi, and one patient received another treatment. Furthermore, 10 of the 12 patients with HVS underwent plasma exchange prior to therapeutic intervention. The initial best responses involved a complete response in 15% (5 patients), very good partial response in 24% (8 patients), partial response in 36% (12 patients), stable disease in 21% (7 patients), and progressive disease in 3% (1 patient). Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e shows the Kaplan\u0026ndash;Meier curves of TTNT and OS in the 33 sWM patients, analyzed according to mutation status, del 6q, and CAs using CBA. During the median follow-up of 73 months, no differences in TTNT or OS were correlated with the status of \u003cem\u003eMYD88\u003c/em\u003e/\u003cem\u003eCXCR4\u003c/em\u003e or del 6q. Conversely, the median TTNT was 27 and 68 months with and without CAs (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.14), and the median TTNT was 27 and 61 months with and without complex karyotypes (\u003cem\u003ep\u0026thinsp;=\u0026thinsp;0.42\u003c/em\u003e), respectively.\u003c/p\u003e \u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003e \u003cem\u003eMYD88\u003c/em\u003e \u003csup\u003e \u003cem\u003eL265P\u003c/em\u003e \u003c/sup\u003e is recognized in approximately 90% of WM patients and plays a major role in tumor cell survival and proliferation through constitutive NF-kB activation in WM [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. In contrast, \u003cem\u003eCXCR4\u003c/em\u003e\u003csup\u003e\u003cem\u003eMut\u003c/em\u003e\u003c/sup\u003e is recognized in 25\u0026ndash;40% of WM patients [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e] and is generally accepted as the second most common hit, with poor prognostic factors. In particular, the \u003cem\u003eCXCR4\u003c/em\u003e NS mutation has been associated with elevated serum IgM levels, an increased risk of HVS, and an inferior response and survival in WM patients treated with ibrutinib monotherapy [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. In the present study, \u003cem\u003eMYD88\u003c/em\u003e \u003csup\u003e\u003cem\u003eL265P\u003c/em\u003e\u003c/sup\u003e was found in 85.7% of the patients, which is comparable to previous reports [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. The \u003cem\u003eCXCR4\u003c/em\u003e\u003csup\u003e\u003cem\u003eMut\u003c/em\u003e\u003c/sup\u003e rate was 8.6% in this study, which is equal to or lower than that found in previous reports [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e], although \u003cem\u003eCXCR4\u003c/em\u003e\u003csup\u003e\u003cem\u003eMut\u003c/em\u003e\u003c/sup\u003e exhibited a trend toward higher serum IgM levels. One reason for this discrepancy may be that only 4 probes were used for detecting \u003cem\u003eCXCR4\u003c/em\u003e\u003csup\u003e\u003cem\u003eMut\u003c/em\u003e\u003c/sup\u003e: two NS mutations for S338 and an FS mutation for S338 and T318. Another reason may be that all the BM samples were obtained at the first diagnosis of sWM. For reference, regarding East Asia, 4/26 patients (15%) exhibited \u003cem\u003eCXCR4\u003c/em\u003e\u003csup\u003e\u003cem\u003eMut\u003c/em\u003e\u003c/sup\u003e based on next-generation sequencing from unselected BM in a Japanese phase II study of tirabrutinib for untreated and relapsed/refractory WM (RRWM) [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. In a trial of orelabrutinib conducted in China for RRWM, the detection rate of \u003cem\u003eCXCR4\u003c/em\u003e\u003csup\u003e\u003cem\u003eMut\u003c/em\u003e\u003c/sup\u003e was 4/47 patients (8.8%) based on allele specific-PCR for the \u003cem\u003eS338\u003c/em\u003e mutation [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe detection of del 6q and del 17p by FISH analyses is recommended in several guidelines [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Del 6q is observed in 30\u0026ndash;55% of WM patients [\u003cspan additionalcitationids=\"CR14 CR15\" citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e] and is less frequently identified (0\u0026ndash;4%) in patients with IgM monoclonal gammopathy of undetermined significance; therefore, it is considered to be the secondary event [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. WM patients with del 6q exhibited higher serum M-protein levels [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], higher IPSSWM scores, and poor prognosis [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Del 17p has also been associated with unfavorable prognosis [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. In the present study, FISH analysis revealed the presence of del 6q in 5/18 patients (28%) and del 17p in 2/31 patients (7%). Although we were unable to fully analyze the significance of del 6q due to missing data, the results demonstrated that serum albumin levels tended to be low, and no difference was observed in serum M-protein levels or prognosis.\u003c/p\u003e \u003cp\u003eIn contrast, the impact of CAs using CBA in sWM remains poorly understood. Several studies reported the clinicopathological significance of CAs identified by CBA in WM [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Nguyen-Khac et al. found that 66/141 cases (47%) had abnormal karyotypes, including 30% complex karyotypes [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. In the report, del 6q was identified with greater frequency in complex karyotypes than in abnormal karyotypes with fewer abnormalities. A group in the French Innovative Leukemia Organization retrospectively analyzed the clinicopathological impact of CAs in 239 WM patients [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. In their study, highly complex karyotypes with 5 or more CAs were found in 5% of patients and were associated with poor PFS and OS. In addition, a significant correlation between highly complex karyotypes and del 6q and/or TP53-related abnormalities was observed. Recently, Danesin and colleagues reported that WM patients with CAs were significantly older at the time of diagnosis (median age, 72 vs. 65 years) and had inferior median OS (76.1 vs. 167.7 months) compared with WM patients that had normal karyotypes, across 23 years in the single-center study [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. In our study, the presence of CAs detected by CBA was associated with significantly higher rates of anemia and hypoalbuminemia, which are associated with poor prognostic factors in IPSSWM, and higher serum IgM levels. The distribution of age was found to be identical regardless of the presence or absence of CAs. The TTNT in sWM patients with CAs tended to be shorter, although a significant difference was not observed, possibly due to the small sample size. Considering our results and the previous reports, CAs identified by CBA in sWM patients may have some clinical impacts.\u003c/p\u003e \u003cp\u003eRegarding the limitations of this work, the present retrospective study was based on 35 patients at a single institute. Second, \u003cem\u003eCXCR4\u003c/em\u003e\u003csup\u003e\u003cem\u003eMut\u003c/em\u003e\u003c/sup\u003e was not comprehensively examined; only four probes were used, and other types of \u003cem\u003eCXCR4\u003c/em\u003e\u003csup\u003e\u003cem\u003eMut\u003c/em\u003e\u003c/sup\u003e may have been missed. Third, 17 of the 35 cases lacked del 6q data, providing insufficient analysis. Finally, the outcomes of TTNT and OS may have been influenced by the treatment regimens.\u003c/p\u003e \u003cp\u003eIn conclusion, our findings suggest that CAs identified by CBA are associated with clinical aggressiveness and shorter TTNT in sWM. Further studies are necessary to clarify and validate the significance of CAs in sWM.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eCompeting interest\u003c/h2\u003e \u003cp\u003eNaohiro Sekiguchi receives honoraria from Janssen, Ono, and research funding from Incyte Biosciences Japan, Janssen, Mitsubishi Tanabe Pharma Corporation, MSD, Ono.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eEthics approval\u003c/strong\u003e \u003cp\u003e This study was approved by the institutional review board at our hospital (Disaster Medical Center IRB-2022-33).\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eConsent to participate\u003c/strong\u003e \u003cp\u003eWritten consent was obtained from the patients.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThis research received no specific grant or funding from agencies in the public, commercial, or not-for-profit sectors.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eKenichi Ito analyzed and interpreted the data and drafted the manuscript. Tomoko Kitagawa organized the clinical trial and analyzed the data. Kunihiko Harada acquired the data. Kazuhiko Hirano reviewed pathological diagnoses and provided the pictures for figures. Naohiro Sekiguchi organized the clinical trial, analyzed the data, and revised the manuscript critically for important intellectual content. All authors read and approved the final manuscript.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe data that support the fndings of this study are available upon request from the corresponding author.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAlaggio R, Amador C, Anagnostopoulos I, Attygalle AD, Araujo IBO, Berti E et al (2022) The 5th edition of the World Health Organization Classification of Haematolymphoid Tumours. 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EClinicalMedicine 52:101682. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.eclinm.2022.101682\u003c/span\u003e\u003cspan address=\"10.1016/j.eclinm.2022.101682\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"annals-of-hematology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"aohe","sideBox":"Learn more about [Annals of Hematology](http://link.springer.com/journal/277)","snPcode":"277","submissionUrl":"https://submission.nature.com/new-submission/277/3","title":"Annals of Hematology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Waldenström’s macroglobulinemia, chromosomal aberration, chromosome banding analysis, droplet digital PCR, MYD88, CXCR4","lastPublishedDoi":"10.21203/rs.3.rs-4736953/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4736953/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eThe clinicopathologic features and prognostic impact of \u003cem\u003eMYD88 L265P\u003c/em\u003e (\u003cem\u003eMYD\u003c/em\u003e\u003csup\u003e\u003cem\u003eL265P\u003c/em\u003e\u003c/sup\u003e) and \u003cem\u003eCXCR4\u003c/em\u003e mutations (\u003cem\u003eCXCR4\u003c/em\u003e\u003csup\u003e\u003cem\u003eMut\u003c/em\u003e\u003c/sup\u003e) have been well reported, although little is known regarding the impact of chromosomal aberrations (CA) detected by chromosome banding analysis (CBA) in symptomatic Waldenstr\u0026ouml;m\u0026rsquo;s macroglobulinemia (sWM). Thus, we investigated the clinicopathologic features and prognostic impact in sWM with CAs identified by CBA.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eWe retrospectively analyzed the clinicopathologic results and genetic mutations by droplet digital PCR, fluorescence in situ hybridization (FISH), and CBA using the G-banding method from the bone marrow samples of sWM between April 2010 and March 2024 at our institute. The relationship between CAs and clinicopathologic features was evaluated, as well as the time to next treatment (TTNT).\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eThirty-five patients were enrolled. The median age was 71 years, and the median hemoglobin level was 10.1 g/dL. The median serum IgM and M-protein levels were 3,120 mg/dL and 3 g/dL, respectively. \u003cem\u003eMYD\u003c/em\u003e\u003csup\u003e\u003cem\u003eL265P\u003c/em\u003e\u003c/sup\u003e was found in 30/35 patients (85.7%), whereas \u003cem\u003eCXCR4\u003c/em\u003e\u003csup\u003e\u003cem\u003eMut\u003c/em\u003e\u003c/sup\u003e was found in 3/35 patients (8.6%). Deletion 6q identified by FISH in 5/18 patients (28%), and CAs using CBA in 9/34 patients (26%), including 4/34 (12%) complex karyotypes. sWM with CAs had more anemia (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.04) and hypoalbuminemia (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.007), in addition to higher serum M-protein and IgM levels (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.03). With a median follow-up of 73 months, the median TTNT in patients with and without CAs was 27 and 68 months, respectively.\u003c/p\u003e\u003ch2\u003eConclusions\u003c/h2\u003e \u003cp\u003eCAs with CBA may be associated with clinical aggressiveness and shorter TTNT in sWM.\u003c/p\u003e","manuscriptTitle":"Impact of Chromosomal Aberrations Detected by Chromosome Banding Analysis in Symptomatic Waldenström’s Macroglobulinemia","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-08-17 01:05:10","doi":"10.21203/rs.3.rs-4736953/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-09-12T13:46:27+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-09-12T10:28:20+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-09-04T16:41:58+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"97616998672997866441751769498481830140","date":"2024-09-03T10:15:56+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"313403019500027344246605772515405938354","date":"2024-08-20T11:53:32+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-07-20T13:05:09+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-07-17T13:46:44+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-07-17T13:46:33+00:00","index":"","fulltext":""},{"type":"submitted","content":"Annals of Hematology","date":"2024-07-14T03:55:23+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"annals-of-hematology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"aohe","sideBox":"Learn more about [Annals of Hematology](http://link.springer.com/journal/277)","snPcode":"277","submissionUrl":"https://submission.nature.com/new-submission/277/3","title":"Annals of Hematology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"6a000f58-2421-4d2d-bdf1-a0c6b4fccda6","owner":[],"postedDate":"August 17th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-10-21T16:02:28+00:00","versionOfRecord":{"articleIdentity":"rs-4736953","link":"https://doi.org/10.1007/s00277-024-06041-y","journal":{"identity":"annals-of-hematology","isVorOnly":false,"title":"Annals of Hematology"},"publishedOn":"2024-10-16 15:57:39","publishedOnDateReadable":"October 16th, 2024"},"versionCreatedAt":"2024-08-17 01:05:10","video":"","vorDoi":"10.1007/s00277-024-06041-y","vorDoiUrl":"https://doi.org/10.1007/s00277-024-06041-y","workflowStages":[]},"version":"v1","identity":"rs-4736953","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4736953","identity":"rs-4736953","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}