BACKGROUND:With life expectancies of patients with chronic myeloid leukemia (CML) now approaching those of the general population under tyrosine kinase inhibitor (TKI) therapy, understanding long-term treatment-related toxicities becomes paramount. Although imatinib's nephrotoxic potential is established, the renal safety profiles of second-generation TKIs-dasatinib, nilotinib, and flumatinib-remain incompletely characterized. METHODS:This single-center retrospective study analyzed renal function trajectories in 348 patients with CML receiving first-line imatinib, dasatinib, nilotinib, or flumatinib. Renal function was assessed using serum creatinine and estimated glomerular filtration rate (eGFR). Mixed-effects models incorporating treatment group, time, and their interaction as fixed effects were used to formally compare eGFR trajectories across treatment groups. Chronic renal adverse events and acute kidney injury were also evaluated. RESULTS:Formal comparison of eGFR trajectories across treatment groups revealed a significant time × treatment interaction, indicating differential renal function changes among the 4 TKIs. Significant serum creatinine elevation and eGFR decline were observed in imatinib and flumatinib groups, contrasting with stable creatinine and divergent eGFR patterns in dasatinib (slight increase) and nilotinib (decline) groups. The incidence of chronic renal adverse events varied substantially across cohorts-12.3% (imatinib), 2.6% (nilotinib), 4.3% (dasatinib), and 9.6% (flumatinib). Notably, among 3 documented acute kidney injury cases, all occurred during flumatinib treatment. Furthermore, treatment-free remission achievement did not confer significant renal function recovery, suggesting potential irreversibility of established renal damage. CONCLUSION:TKIs exhibit differential renal safety profiles, underscoring the critical importance of individualized TKI selection based on baseline renal status and implementation of rigorous renal monitoring protocols throughout treatment duration.

BACKGROUND:Myelodysplastic neoplasms (MDS) represent a group of heterogeneous clonal disorders characterized by immune dysregulation in their pathogenesis. Gut microbiota dysbiosis plays a critical role in immune modulation. METHODS:We collected the fecal samples of 23 newly diagnosed MDS, 10 hypomethylating agents (HMA) treated MDS and 13 age and sex matched healthy controls (HC), and analyzed the gut microbiota compositions and functional pathways using metagenomic next-generation sequencing (mNGS). RESULTS:Distinct microbial compositions were observed between newly diagnosed MDS and HC. Notably, the Veillonellaceae family was significantly enriched in MDS patients. Specific bacteroid species demonstrated significant correlations with lymphocyte subtypes, functional activation status, and serum inflammatory cytokines. Functional profiling revealed altered metabolic pathways in newly diagnosed patients, particularly in amino acid metabolism and ATP synthesis. Notably, glutamine/glutamate and tryptophan metabolism pathways were hyperactive in untreated MDS but downregulated following HMA treatment. CONCLUSIONS:The gut microbiota altered in MDS patients and was associated with immune dysregulation and inflammation, which may contribute to MDS pathogenesis and mediate therapeutic effects of HMA treatment, highlighting the gut microbiota-metabolism axis as a potential therapeutic target for MDS management.

Background:T-lineage acute leukemias are aggressive malignancies for which treatment options are limited. Objectives:This study evaluated the efficacy and safety of a non-cytotoxic regimen combining venetoclax, azacitidine, and dexamethasone (VAD) as first-line induction therapy for T-cell acute lymphoblastic leukemia (T-ALL) and T/myeloid mixed-phenotype acute leukemia (T-Myeloid MPAL). Design:We retrospectively analyzed the clinical parameters and survival data of 11 patients with newly-diagnosed T-ALL or T-Myeloid MPAL who received the VAD regimen. Methods:The complete remission (CR) rate and adverse events were assessed. Kaplan-Meier curves were constructed to evaluate survival outcomes. Results:Among 11 patients (7 with early T-cell precursor [ETP] ALL, 2 with T-Myeloid MPAL, and 2 with non-ETP T-ALL), the overall CR rate after one cycle of VAD therapy was 90.9%, with 45.5% achieving measurable residual disease (MRD) negativity by flow cytometry. The historically poor-prognosis ETP-ALL subgroup showed an 85.7% CR rate (6/7), while both T-Myeloid MPAL cases attained MRD negativity. No tumor lysis syndrome or treatment-related mortality occurred. Over a median follow-up of 358 days, all 5 patients who received transplantations remained relapse-free. Conclusion:The VAD regimen demonstrated a high response rate and favorable safety, enabling effective transplantation bridging with avoidance of conventional cytotoxic therapy.

Large granular lymphocyte leukemia (LGLL) is a rare lymphoproliferative disorder where somatic STAT3 mutation is common. Although LGLL has been described as an underlying condition associated with pure red cell aplasia (PRCA), the clinical characteristics and therapeutic response of LGLL − associated PRCA are largely unclear. We evaluated a set of 81 patients with LGLL − associated PRCA. Comparative analysis was performed on the clinical characteristics, responses to immunosuppressive therapy, and survival outcomes in patients with STAT3 mutation. Among the 81 LGLL − associated PRCA patients, 21 cases (26

Purpose:This study evaluated the efficacy and safety of a 14-day blinatumomab-venetoclax (BV) regimen as induction therapy for newly diagnosed Ph-negative B-cell acute lymphoblastic leukemia (B-ALL), focusing on rapid remission and tolerability in unfit patients. Patients and Methods:Thirteen patients received venetoclax (100 mg on day 1, 200 mg on day 2, 300 mg on day 3, and 400 mg from days 4 to 14) with blinatumomab (9 to 28 ug/day) for 14 days. Bone marrow assessments were performed at days 14-21. Primary endpoints were complete remission (CR) rate, minimal residual disease (MRD) negativity by flow cytometry, and adverse events. Results:The CR rate after one cycle of BV regimen was 92.3% (12/13), and all patients achieved MRD-negativity; 91.7% (11/12) achieved MRD clearance by day 21. Grade 1-2 cytokine release syndrome occurred in 46.2% (6/13; 1 grade 3). Hematologic toxicity included grade 3-4 neutropenia (92.3%) and thrombocytopenia (46.2%), with only 30.8% febrile neutropenia. All AEs resolved rapidly with supportive care, allowing therapy to continue without interruption. At median follow-up of 283 days, 1-year relapse-free survival rate and overall survival rate were 60.6% and 83.3%. Conclusion:The 14-day BV regimen induced rapid deep remission (91.7% MRD-negative by day 21) with manageable toxicity in Ph-negative B-ALL. Synergistic T-cell activation by venetoclax may explain enhanced efficacy.

BACKGROUND:Splicing factor 3b subunit 1 (SF3B1) mutations have been implicated in hematologic malignancies, but the clinical significance of distinct SF3B1 mutation variants remain unclear. METHODS:The objective of this study was to evaluate clinicopathologic features, mutational profiles, and outcomes of 1691 patients with hematologic malignancies, including myelodysplastic syndromes (MDS; n = 402), acute myeloid leukemia (AML; n = 758), and chronic lymphocytic leukemia (CLL; n = 531). RESULTS:The frequency of SF3B1-mutated (SF3B1MUT) MDS, AML, and CLL was 70 of 402 patients (17.4%), 23 of 758 patients (3.0%), and 45 of 531 patients (8.5%), respectively. p.K700E was the most prevalent SF3B1MUT variant and was identified in 43 of 70 of patients with MDS (61.4%), in seven of 23 patients with AML (30.4%), and in 19 of 45 patients with CLL (42.2%). In MDS and AML, TET2 was the most frequent co-mutated gene in patients with SF3B1MUT disease (20 of 70 patients with MDS [28.6%]; 11 of 23 patients with AML [47.8%]). For patients with SF3B1MUT CLL, the most common co-mutated genes were ATM (11 of 45; 24.4%) and TP53 (11 of 45; 24.4%). Kaplan-Meier analysis indicated that the SF3B1 p.K700E variant was significantly associated with improved overall survival (OS) and progression-free survival (PFS) in patients who had MDS (p < .001 and p = .016, respectively) but with worse OS and PFS in those who had AML (p = .006 and p = .006, respectively) compared with those who had wild-type SF3B1. In patients who had CLL, p.I704F was associated with reduced OS (p < .001) and p.K700E was associated with a shorter time-to-first treatment (p = .028) compared with those who had wild-type SF3B1. Multivariable analysis identified p.K700E as an independent protective factor for OS in patients with MDS (p = .048) but as an independent risk factor for both OS and PFS in patients with AML (p = .036 and p = .035, respectively) and for the time to first treatment in patients with CLL (p = .033). CONCLUSIONS:Specific SF3B1 variants should be incorporated into prognostic stratification for hematologic malignancies.

Precise differential diagnosis between lymphoplasmacytic lymphoma/Waldenström macroglobulinemia (LPL/WM) and marginal zone lymphoma (MZL) remains a challenging issue because of overlapping clinicopathological and immunophenotypic features. In the present study, the differential diagnostic potential of CD180 was assessed by determining its expression patterns in patients with MZL and LPL/WM through flow cytometry. The results indicated that LPL/WM cases exhibited a complete absence of CD180 expression on malignant B cells, whereas MZL cases showed robust CD180 expression (P < .001). Receiver operating characteristic analysis demonstrated that CD180 expression percentage showed optimal diagnostic accuracy in LPL/WM and MZL cases (area under the curve = 0.998, sensitivity = 100%, and specificity = 98.0%), with a further improvement in differentiation potential by the CD180 mean fluorescence intensity ratio (lymphocytes/monocytes) of ≤ 0.47 (area under the curve = 0.937). Moreover, although the MYD88L265P/CXCR4 mutation was not detected by next-generation sequencing in 2 LPL/WM cases, these cases still showed the absence of CD180 expression. Subsequently, droplet digital PCR revealed low-frequency MYD88L265P mutations (0.95% and 1.6% variant allele frequency) in these CD180-negative cases, which confirmed the superior sensitivity of CD180 in identifying LPL/WM with low tumor burden. Overall, our findings establish CD180 as a novel, rapid, and precise flow cytometry-based biomarker with robust ability for the differential diagnosis of LPL/WM and MZL, particularly relevant in resolving diagnostically challenging cases and providing prompt tumor diagnosis in time-constrained situations.

Flumatinib is a novel second-generation tyrosine kinase inhibitor (2G-TKI), which was approved in November 2019 in China. A previous phase III study evaluated the efficacy and safety of flumatinib as a first-line therapy for patients with chronic phase chronic myeloid leukemia (CML-CP). However, randomized trials comparing flumatinib with other 2G-TKIs remain lacking. To assess the efficacy and safety of flumatinib versus nilotinib as a first-line treatment for CML-CP. A multicenter retrospective study. We retrospectively analyzed 101 and 64 patients treated with flumatinib and nilotinib during the same period, respectively. Patients in the flumatinib group were significantly older than patients in the nilotinib group (median age, 44 vs 37 years; p = 0.004). The optimal response and treatment failure rates at 24 months were comparable between the two groups. At 12 months, 85.1% and 88.2% of patients in the flumatinib and nilotinib groups, respectively, achieved a major molecular response (MMR; p = 0.648). By 24 months, 9.9% and 12.5% of patients suffered treatment failure in the flumatinib and nilotinib groups, respectively (p = 0.602). At 9, 12, and 24 months, the rate of MR4 (a BCR::ABL1 transcript level ⩽0.01%) achievement was significantly higher in patients treated with nilotinib than in those treated with flumatinib (26.0% vs 53.7%, p = 0.007; 40.4% vs 60.8%, p = 0.044; and 41.7% vs 80.8%, p = 0.042, respectively). In addition, elevated alanine aminotransferase or aspartate aminotransferase (ALT/AST), glucose, and serum lipid; hyperbilirubinemia; rash; and alopecia were more frequent among patients receiving nilotinib, whereas diarrhea was more frequent in those receiving flumatinib. Flumatinib is a suitable alternative as a first-line treatment for patients with CML-CP to achieve a fast MMR with better tolerability.

Myelodysplastic syndromes (MDS) are heterogeneous stem cell disorders with a 30%-40% risk of transformation to acute myeloid leukaemia (AML). This study characterised the genetic and clinicopathological features of 437 newly diagnosed MDS patients. A predictive model for AML transformation was developed, which identified the bone marrow blast percentage, KRAS, STAG2 and TP53 mutations as significant predictors of disease progression. Novel prognostic models for overall survival (OS) and progression-free survival (PFS) were established and validated. The OS model incorporated age, sex and mutations in TP53, NF1, SRSF2, CSF3R, ETV6, CEBPA, BCOR, TET2, JAK2 and SF3B1. The PFS model incorporated age, sex, bone marrow blast percentage and mutations in STAT3, TP53, NF1, CEBPA, ETV6, CALR, DNMT3A, IDH2, GATA2, BCOR, JAK2, TET2 and SF3B1. These models stratify patients into favourable, intermediate-1, intermediate-2 and adverse risk groups, demonstrating superior risk stratification compared to the Revised/Molecular International Prognostic Scoring System (IPSS-R/M). This work provides the first genomic characterisation of a diagnosed MDS cohort in China and establishes the first risk prediction model for MDS-to-AML transformation, alongside novel prognostic models for OS and PFS. These tools offer improved prognostic prediction and potential guidance for therapeutic strategies in Chinese patients with MDS.

Background: A phase II trial has shown, first-line treatment with iFCG (ibrutinib, fludarabine, cyclophosphamide, and obinutuzumab) led to a bone marrow (BM) undetectable minimal residual disease (uMRD) rate of 98% (44/45) as best response in treatment-naïve patients with chronic lymphocytic leukemia/small lymphocytic leukemia (CLL/SLL). But this study[1] does not include patients with unmutated immunoglobulin heavy⁃chain variable region gene ( IGHV) and/or del(17p)/ TP53 mutation, and ibrutinib was the first generation BTK (BTKi) which had relatively high off-target adverse effects (AEs). Our study has demonstrated the efficacy and safety of the second generation BTKi orelabrutinib in patients with relapsed/refractory CLL [2]. Therefore, we used orelabrutinib (O), and initiated the phase II OFCG trial in younger, fit patients with previously untreated CLL/SLL unrestricted by IGHV mutation status and del(17p)/ TP53 aberrations (cwCLL-001 study, NCT05322733). Methods：This is a multicenter, open-label, non-randomized phase II study for previously untreated patients with CLL/SLL without restriction by del(17p)/ TP53 aberrations and/or IGHV mutation status. A 7-day lead-in period with single-agent oral orelabrutinib（150 mg daily）was given in all patients, followed by OFCG regimen which consisted of orelabrutinib, intravenous Obinutuzumab, fludarabine and cyclophosphamide for up to three 28-day cycles. MRD was detected by 8 multi-color flow cytometry (FCM) at 10 -4 sensitivity and cloneSEQ using NGS (next generation sequencing) assay with a sensitivity of up to 10 −6. If the patients could achieve undetectable MRD in BM, they will discontinue FC and receive 3 cycles of OG (orelabrutinib and obinutuzumab). Otherwise, the patients will receive another 3 cycles of OFCG. Patients will discontinue obinutuzumab if they achieve BM uMRD after 6 cycles and continue six additional cycles of orelabrutinib; all other patients will receive six cycles of OG. Patients who achieved BM uMRD after 12 cycles discontinue all treatments, otherwise will continue orelabrutinib for up to additional 1 year (Fig. 1). The primary endpoint is the percentage of uMRD in BM after 6 cycles by FCM (at 10 -4 sensitivity). The secondary endpoint is the percentage of uMRD in BM and peripheral blood (PB) at other key time points by FCM, best overall response rate (ORR), complete response rate (CRR), progression-free survival (PFS), 2- and 3-year PFS rate, duration of response (DOR), overall survival (OS), 3-year OS rate as well as safety. The percentage of uMRD by NGS is an exploratory endpoint. Results：From 30 May 2022 to 30 Jun 2023, 25 patients were enrolled. Median age was 49 years old (range, 23-62). Unmutated IGHV, TP53 mutation/del(17p) and del(11q) were detected in 56.5% (13/23), 16.0% (4/25), and 24.0% (6/25) of patients, respectively. At data cut off (30 Jun 2023), 19 patients completed 3 cycles of OFCG and were evaluable. 75% (15/20, 1 patient who off study was assessed at the end of C2 cycle) of patients achieved PB uMRD and 57.9% (11/19) achieved BM uMRD by FCM, ORR and CR/CRi rate was 100% (18/18) and 38.9% (7/18) (Fig. 2). One patient died after cycle 3 due to COVID-19 infection. 16 patients completed 6 cycles, 14 of them were evaluable and achieved deeper response irrespective of different strategies during cycle 4 to 6, PB uMRD and BM uMRD was 100% (14/14) and 85.7% (12/14) by FCM, and the ORR and CRR was 100% (14/14) and 57.1% (8/14), respectively. In IGHV unmutated subgroup, ORR was 100% (10/10) with 50% (5/10) CRR, 87.5% (7/8) patients achieved BM uMRD by FCM after cycle 6. The results of MRD response by NGS assay will be presented in the formal report. 91.7% patients experienced at least one treatment emergent AE at the data cutoff. Most AEs were mild and tolerable. The most frequent AEs (≥10% of any grade) were neutropenia, leukopenia and thrombocytopenia. No bleeding or cardiovascular AEs greater than grade 3 occurred in these patients. Overall, OFCG demonstrated a favorable safety profile. Conclusion：This is the first clinical trial exploring the efficacy and safety of the second generation BTKi plus chemoimmunotherapy in patients with CLL. The OFCG regimen shows a rapid and deep molecular remission with a pleasant safety profile in the TN CLL patients including the ones with unfavorable factors. References: 1. Jain N, et al., Leukemia. 2021 Dec;35(12):3421-3429. 2. Xu W, et al. Am J Hematol. 2023 Apr;98(4):571-579.

Objective:To analyze the clinical and genetic characteristics of acute myeloid leukemia,myelodysplasia-related(AML-MR)patients and evaluate their prognostic risk stratification,to guide clinical treatment decisions and improve understanding of the biological characteristics and disease progression.Methods:The study analyzed cellular and molecular genetic information of 307 AML-MR patients,diagnosed based on clinical history,bone marrow morphology,cytogenetics,and molecular genetic abnormalities.The risk stratification followed the 2022 ELN guidelines.Results:57 cases(18.6％)met the AML-MR diagnostic criteria based on morphology and clinical history,110 cases(37.2％)met the AML-MR diagnostic criteria based on cytogenetic results,and 210 cases(74.5％)met the AML-MR diagnostic criteria based on molecular testing results.Among different type of mutations,ASXL1 mutation was the most frequent,followed by SRSF2 and BCOR mutations.Except for 2 cases with incomplete data that could not be classified.263(86.2％)of the 305 patients were classified as poor prognosis,20(6.6％)were classified as good prognosis group,and 22(7.2％)were classified as intermediate prognosis group.Conclusion:Molecular genetic information plays a crucial role in diagnosing AML-MR,highlighting the importance of genetics in diagnosis and prognosis.Most AML-MR patients fall into poor prognosis categories,necessitating early intensive and targeted therapy for better survival outcomes.

Background: Acute myeloid leukemia (AML) patients with various nucleophosmin 1 (NPM1) mutations are controversial in the prognosis. This study aimed to investigate the prognosis of patients according to types of NPM1 mutations (NPM1mut). Methods: Bone marrow samples of 528 patients newly diagnosed with AML, were collected for morphology, immunology, cytogenetics, and molecular biology examinations. Gene mutations were detected by next-generation sequencing (NGS) technology. Results: About 25.2

Philadelphia chromosome-positive (Ph + ) leukemia is a fatal hematological malignancy. Although standard treatments with tyrosine kinase inhibitors (TKIs) have achieved remarkable success in prolonging patient survival, intolerance, relapse, and TKI resistance remain serious issues for patients with Ph + leukemia. Here, we report a new leukemogenic process in which RAPSYN and BCR-ABL co-occur in Ph + leukemia, and RAPSYN mediates the neddylation of BCR-ABL. Consequently, neddylated BCR-ABL enhances the stability by competing its c-CBL-mediated degradation. Furthermore, SRC phosphorylates RAPSYN to activate its NEDD8 E3 ligase activity, promoting BCR-ABL stabilization and disease progression. Moreover, in contrast to in vivo ineffectiveness of PROTAC-based degraders, depletion of RAPSYN expression, or its ligase activity decreased BCR-ABL stability and, in turn, inhibited tumor formation and growth. Collectively, these findings represent an alternative to tyrosine kinase activity for the oncoprotein and leukemogenic cells and generate a rationale of targeting RAPSYN-mediated BCR-ABL neddylation for the treatment of Ph + leukemia.

Background: Measurable residual disease (MRD) is an important prognostic indicator of chronic lymphocytic leukemia (CLL). Different flow cytometric panels have been developed for the MRD assessment of CLL in Western countries; however, the application of these panels in China remains largely unexplored. Methods: Owing to the requirements for high accuracy, reproducibility, and comparability of MRD assessment in China, we investigated the performance of a flow cytometric approach (CD45-ROR1 panel) to assess MRD in patients with CLL. The European Research Initiative on CLL (ERIC) eight-color panel was used as the "gold standard." Results: The sensitivity, specificity, and concordance rate of the CD45-ROR1 panel in the MRD assessment of CLL were 100% (87/87), 88.5% (23/26), and 97.3% (110/113), respectively. Two of the three inconsistent samples were further verified using next-generation sequencing. In addition, the MRD results obtained from the CD45-ROR1 panel were positively associated with the ERIC eight-color panel results for MRD assessment (R = 0.98, p < 0.0001). MRD detection at low levels (<= 1.0%) demonstrated a smaller difference between the two methods (bias, -0.11; 95% CI, -0.90 to 0.68) than that at high levels (>1%). In the reproducibility assessment, the bias was smaller at three data points (within 24, 48, and 72 h) in the CD45-ROR1 panel than in the ERIC eight-color panel. Moreover, MRD levels detected using the CD45-ROR1 panel for the same samples from different laboratories showed a strong statistical correlation (R = 0.99, p < 0.0001) with trivial interlaboratory variation (bias, 0.135; 95% CI, -0.439 to 0.709). In addition, the positivity rate of MRD in the bone marrow samples was higher than that in the peripheral blood samples. Conclusions: Collectively, this study demonstrated that the CD45-ROR1 panel is a reliable method for MRD assessment of CLL with high sensitivity, reproducibility, and reliability.

Objective:To analyze the distribution of different SF3B1 genotypes in patients with myelodysplastic syndromes (MDS) and its prognostic value.Methods:Totally, 377MDS patients who were initially diagnosed in the First Affiliated Hospital of Nanjing Medical University from January 2014 to January 2022 were included in the retrospective analysis.The patients were divided into three different groups according to mutation stcote of SF3B1, including 317 patients with SF3B1 wild type (SF3B1 WT) (214 males and 103 females, 63(49, 71) years old),39 patients with SF3B1 K700E mutation(SF3B1 K700E(17 males and 22 females, 65(52, 73)years old)) and 21 patients with SF3B1 non-K700E mutation(SF3B1 non-K700E)(13 males and 8 females, 67(63, 73) years old). MDS-related 20 gene mutations were detected using targeted sequencing technology; Survival curves were constructed by the Kaplan-Meier method; Cox proportional hazards model was established to evaluate different factors at diagnosis on survival by univariate and multivariate analyses.. Results:Compared with SF3B1 non-K700E patients, SF3B1 K700E patients had a higher median absolute neutrophil count ( P=0.002) and were likely to be in the low/int-1 International Prognostic Scoring System (IPSS) categories ( P=0.023). A 20-gene targeted sequencing analysis showed that, compared with SF3B1 WT patients, SF3B1 K700E patients were associated with lower frequency of ASXL1 and U2AF1 mutations ( P=0.018 and P=0.003); while compared with SF3B1 non-K700E patients, the frequency of ASXL1 mutation was significantly lower in SF3B1 K700E cases ( P=0.029). Patients with SF3B1 K700E had better overall survival (OS) in comparison with SF3B1 WT and SF3B1 non-K700E in MDS patients ( P<0.001 and P=0.045, respectively). In comparison with SF3B1 WT patients, SF3B1 MUT patients had more favorable OS and progression-free survival (PFS) in MDS without excess blasts ( P<0.001 and P<0.001, respectively), but no significant difference was found in MDS with excess blasts ( P>0.05). Compared with SF3B1 WT patients, SF3B1 K700E patients had superior OS and PFS in the int-1 IPSS category ( P=0.010 and P=0.013, respectively). By multivariable analysis, the presence of SF3B1 K700Ewas an independent predictor of superior OS ( HR=0.461,95% CI 0.262-0.811, P=0.007). Conclusion:SF3B1 K700E and SF3B1 non-K700E patients had significantly improved OS in comparison with SF3B1 WT MDS patients. Furthermore, SF3B1 K700E patients were associated with a better OS compared with SF3B1 non-K700E MDS patients. SF3B1 mutation could not overcome the poor prognostic effect of excess blasts, which highlights the importance of the SF3B1 mutation subtype in risk assessment of MDS without excess blasts.

Abstract Background: Chronic lymphocytic leukemia (CLL) is a common lymphoproliferative disease involving B cells, and small lymphocytic lymphoma (SLL) is regarded as the solid tumor counterpart of CLL. Both of them share the same morphology, which appear as small, mature-appearing lymphocytes with a scanty amount of cytoplasm and a clumped nuclear chromatin. While some cases have reported the presence of binucleated lymphocytes on peripheral blood smears of a small part of CLL patients. However, the prognostic value of this atypical morphologyin CLL patients has not been well acknowledged. Method: We retrospectively reviewed 325 newly diagnosed CLL/SLL patients. Blood smears were performed by Wright-Giemsa stain and used for morphometric assessment. Cytogenetic aberrations were assessed by chromosome banding analysis. Genetic characterization was analyzed by direct Sanger sequencingor FISH. Result: Binucleated lymphocytes observed on peripheral blood smear presented with two similar size and relatively symmetrical nuclei, but varied in size and forms. 308 patients (94.8%) were defined as binucleated lymphocytes negative (BLN) and 17 patients (5.2%) as binucleated lymphocytes positive (BLP). In particular, multinucleated lymphocytes were found in 2 cases. Binucleated lymphocytes were highly correlated with prolymphocytes and dividing lymphocytes. BLP had a higher proportion of complex karyotypes, unmutated IGHV, TP53 mutation or depletion, and a lower proportion of 13q deletion. In addition, there was a significant association between the presence of binucleated lymphocytes and overall survival. In cases withmutated IGHV or without TP53 mutation or deletion, overall survival of BLP was significantly shorter. Conclusion: The observation of binucleated lymphocytes should be considered as a negative prognostic parameter for CLL, especially in lower genetic-risk CLL population. The morphological examination of blood smear in CLL is still worthy, for its advantages of saving of time, laboratory technique, and simplicity and convenience.

Abstract Background The status of minimal residual disease (MRD) has been established as an important prognostic indicator in chronic lymphocytic leukemia (CLL). Methods Owing to the requirements of high accuracy, reproducibility and comparability of MRD, this study investigated the performance of a flow cytometric approach (CD45-ROR1 panel) in the MRD detection of CLL patients, with European Research Initiative on CLL (ERIC) 8-color panel as the “gold standard”. Results The sensitivity, specificity and concordance rate of CD45-ROR1 panel in the MRD assessment of CLL were 100% (87/87), 88.5% (23/26) and 97.3% (110/113), respectively. Two of the 3 non-consistent samples were further verified by the next-generation sequencing. In addition, the MRD results obtained from the CD45-ROR1 panel were positively associated with the ERIC 8-color results for MRD assessment (R = 0.98, p < 0.0001). MRD detection at low levels (≤ 1.0%) demonstrated a smaller difference between the two methods (bias, -0.11; 95% CI, -0.90-0.68) as compared with that at high levels (>0.1%). For the reproducibility assessment, the bias was smaller at three datapoints in the CD45-ROR1 panel as compared with that of the ERIC 8-color panel. Moreover, MRD level detected using the CD45-ROR1 panel for the same samples between different laboratories showed a strong statistical correlation (R = 0.99, p < 0.0001) with a trivial inter-laboratory variation (bias, 0.135; 95% CI, -0.439-0.709). Interesingly, the MRD level detected in the lymph nodes samples were significantly higher than that of the peripheral blood and bone marrow samples (p = 0.029). Conclusions Collectively, this study demonstrates that the CD45-ROR1 panel is a reliable method for the MRD assessment of CLL, with higher sensitivity, reproducibility, and reliability.

BACKGROUND:Real-time monitoring is essential for the management of chronic lymphocytic leukemia (CLL) patients. Utilizing peripheral blood is advantageous due to its affordability and convenience. Existing methods of assessing peripheral blood films have limitations that include lack of automation, dependence on personal experience, and low repeatability and reproducibility. To overcome these challenges, we have designed an artificial intelligence-driven system that provides a clinical perspective to objectively evaluate morphologic features in CLL patients' blood cells.METHODS:Based on our center's CLL dataset, we developed an automated algorithm using a deep convolutional neural network to precisely identify regions of interest on blood films and used the well-established Visual Geometry Group-16 as the encoder to segment cells and extract morphological features. This tool enabled us to extract morphological features of all lymphocytes for subsequent analysis.RESULTS:Our study's lymphocyte identification had a recall of 0.96 and an F1 score of 0.97. Cluster analysis identified three clear, morphological groups of lymphocytes that reflect distinct stages of disease development to some extent. To investigate the longitudinal evolution of lymphocyte, we extracted cellular morphology parameters at various time points from the same patient. The results showed some similar trends to those observed in the aforementioned cluster analysis. Correlation analysis further supports the prognostic potential of cell morphology-based parameters.CONCLUSION:Our study provides valuable insights and potential avenues for further exploration of lymphocyte dynamics in CLL. Investigating morphological changes may help in determining the optimal timing for intervening with CLL patients, but further research is needed.