Cell Lymphoma Leukemia Research Articles

Sodium diethyldithiocarbamate trihydrate: an effective and selective compound for hematological malignancies.

Myeloid leukemias and lymphomas are among the most common and well-studied hematological malignancies. However, due to the aggressiveness and rapid progression of certain subtypes, treating these diseases remains a challenge. Considering the promising results of diethyldithiocarbamates in preclinical and clinical oncology trials, this study aimed to investigate the potential of sodium diethyldithiocarbamate trihydrate (DETC) as a prototype for developing new drugs to treat hematological malignancies. In silico analysis using SwissADME was conducted to evaluate the physicochemical characteristics and pharmacokinetic properties of DETC. An in vitro investigation utilizing the MTT assay assessed the cytotoxic effects of DETC on neoplastic and non-neoplastic cell lines. Selectivity was determined using a selectivity index and a hemolysis assay, while the mechanism of cell death in neoplastic cell lines was examined through flow cytometry analysis of pro-apoptotic and anti-apoptotic protein levels. The results demonstrated that the physicochemical characteristics of DETC are suitable for oral administration. Furthermore, the compound showed promising cytotoxic activity against human myeloid leukemia (K562) and Burkitt's lymphoma (Daudi) cell lines, with high selectivity for neoplastic cells over non-neoplastic cells of the bone marrow microenvironment (HS-5 cell line). Moreover, hemolysis was observed only at very high concentrations. The cytotoxicity mechanism of DETC against both neoplastic cell lines involved cell cycle arrest and the production of reactive oxygen species. In K562 cells, cell death was induced via apoptosis. Additional experiments are needed to confirm the exact mechanism of cell death in Daudi Burkitt's lymphoma cells.

Hemophagocytic syndrome secondary to invasive NK cell leukemia and T-cell lymphoma treated with the modified MINE protocol: report of three cases and literature review

Lymphoma-associated hemophagocytic syndrome is aggressive with rapid progression, particularly in NK/T cell lymphoma. The MINE regimen is a salvage treatment for aggressive non-Hodgkin lymphoma. In our center, the modified MINE regimen was applied to treat three patients with hemophagocytic syndrome secondary to aggressive NK cell leukemia and T-cell lymphoma. The modified MINE regimen showed good efficacy against NK/T cell lymphoma, control of the inflammatory state of secondary hemophagocytic syndrome, and good tolerability.

Long-term survival for lymphoid neoplasms and national health expenditure (EUROCARE-6): a retrospective, population-based study

Management of lymphoid malignancies requires substantial health system resources. Total national health expenditure might influence population-based lymphoid malignancy survival. We studied the long-term survival of patients with 12 lymphoid malignancy types and examined whether different levels of national health expenditure might explain differences in lymphoid malignancy prognosis between European countries and regions. For this observational, retrospective, population-based study, we analysed the EUROCARE-6 dataset of patients aged 15 or older diagnosed between 2001 and 2013 with one of 12 lymphoid malignancies defined according to International Classification of Disease for Oncology (third edition) and WHO classification, and followed up to 2014 (Jan 1, 2001-Dec 31, 2014). Countries were classified according to their mean total national health expenditure quartile in 2001-13. For each lymphoid malignancy, 5-year and 10-year age-standardised relative survival (ASRS) was calculated using the period approach. Generalised linear models indicated the effects of age at diagnosis, gender, and total national health expenditure on the relative excess risk of death (RER). 82 cancer registries (61 regional and 21 national) from 27 European countries provided data eligible for 10-year survival estimates comprising 890 730 lymphoid malignancy cases diagnosed in 2001-13. Median follow-up time was 13 years (IQR 13-14). Of the 12 lymphoid malignancies, the 10-year ASRS in Europe was highest for hairy cell leukaemia (82·6% [95% CI 78·9-86·5) and Hodgkin lymphoma (79·3% [78·6-79·9]) and lowest for plasma cell neoplasms (29·5% [28·9-30·0]). RER increased with age at diagnosis, particularly from 55-64 years to 75 years or older, for all lymphoid malignancies. Women had higher ASRS than men for all lymphoid malignancies, except for precursor B, T, or natural killer cell, or not-otherwise specified lymphoblastic lymphoma or leukaemia. 10-year ASRS for each lymphoid malignancy was higher (and the RER lower) in countries in the highest national health expenditure quartile than in countries in the lowest quartile, with a decreasing pattern through quartiles for many lymphoid malignancies. 10-year ASRS for non-Hodgkin lymphoma, the most representative class for lymphoid malignancies based on the number of incident cases, was 59·3% (95% CI 58·7-60·0) in the first quartile, 57·6% (55·2-58·7) in the second quartile, 55·4% (54·3-56·5) in the third quartile, and 44·7% (43·6-45·8) in the fourth quartile; with reference to the European mean, the RER was 0·80 (95% CI 0·79-0·82) in the first, 0·91 (0·90-0·93) in the second, 0·94 (0·92-0·96) in the third, and 1·45 (1·42-1·48) in the fourth quartiles. Total national health expenditure is associated with geographical inequalities in lymphoid malignancy prognosis. Policy decisions on allocating economic resources and implementing evidence-based models of care are needed to reduce these differences. Italian Ministry of Health, European Commission, Estonian Research Council.

Abstract CT174: A phase 1 study of JBH492, an anti C-C chemokine receptor 7 antibody-drug conjugate (anti-CCR7 ADC), assessing safety and efficacy in lymphoid malignancies

Abstract Background: CCR7 is overexpressed in lymphoid malignancies. JBH492 is a first-in-class CCR7-targeting ADC, comprising a humanized, Fc-silenced anti-CCR7 IgG1 antibody conjugated to maytansinoid microtubule inhibitor DM4 via a cleavable linker. Preclinical studies support avidity-driven differential binding in CCR7-high tumor cells compared to CCR7-low healthy immune cells, and in vivo animal studies demonstrated robust and durable efficacy in multiple clinically relevant lymphoma models (Dang et al. 2022). Methods: This is an ongoing first-in-human, open-label, phase I/Ib, multi-center study in patients (pts) with relapsed/refractory (r/r) chronic lymphocytic leukemia (CLL) and non-Hodgkin’s lymphoma (NHL) (NCT04240704), which completed enrollment for dose-escalation phase. Five dose levels of JBH492 monotherapy ranging from 0.4 to 3.6 mg/kg (intravenous administration every 3 weeks [q3W]) were tested with 3 - 7 pts enrolled per dose level. Preclinical data from xenografts predicted tumor stasis at 1.6 mg/kg q3W. The objective of dose escalation was to assess safety, tolerability, pharmacokinetics (PK), immunogenicity, and preliminary efficacy of JBH492 and to determine maximum tolerated and recommended doses. Pts with r/r CLL and NHL, who failed standard of care therapy (at least 2 prior regimens) and who were intolerant/ineligible to approved therapies, were included; CCR7 expression confirmation was not required prior enrollment. Results: Total 25 pts with r/r stage II-IV NHL (n = 24, [adult T cell leukemia-lymphoma, 3; cutaneous T cell lymphoma, 4; diffuse large B cell lymphoma, 7; follicular lymphoma, 1; Mantle cell lymphoma, 1; marginal zone lymphoma, 2; peripheral T cell lymphoma, 3; others, 3]) and CLL (n = 1) with median age 69.0 y (43 - 85) were enrolled. Mean duration of exposure was 13.5 wks. One pt had dose-limiting toxicity — grade (G) 3 thrombocytopenia lasting for 13 d. Overall, 23 (92%) pts had at least 1 adverse event (AE) (G ≥ 3, n = 11). Treatment-related AEs (TRAEs) were reported in 15 (60%) pts (G ≥ 3, n = 3). Of 6 (24%) pts who had serious AEs (SAEs) (G ≥ 3, n = 3), 2 (8%) had TRSAE. The most frequent AEs (≥ 20%) included anemia (32%), thrombocytopenia (28%), neutropenia and dry eye (20% each). Two deaths due to underlying disease were reported. PK for total ADC was roughly dose proportional. Four NHL pts (2 T-NHL and 2 B-NHL) had partial response (PR) and 2 had stable disease, with PRs observed at doses between 2.4 and 3.6 mg/kg q3W. Post data-cut-off, 1 pt with T-NHL achieved CR following 4 cycles of treatment. No correlation between CCR7 expression and response could be drawn (CCR7 expression was observed in all samples from 14 pts). Conclusions: The data from this phase I study suggest that targeting CCR7 with JBH492 is a promising therapeutic strategy for lymphoid malignancies with manageable hematological and non-hematological safety profile. [K.I. and S.L. contributed equally to this study.] Citation Format: Pau Abrisqueta, Reinhard Marks, Irit Avivi, Martin Wermke, Francesca Lim, Tae Min Kim, Alba Cabirta Touzon, Kristiina Karihtala, Shinichi Makita, Lillian Werner, Chiaki Tanaka, Anhthu Dang, Anwesha Chaudhury, Seth Rice, Rowshan Chowdhury, Deborah Knee, Belén Gomezcarrillo, Elissa Furutani, Koji Izutsu, Sirpa Leppä. A phase 1 study of JBH492, an anti C-C chemokine receptor 7 antibody-drug conjugate (anti-CCR7 ADC), assessing safety and efficacy in lymphoid malignancies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT174.

Novel meriolin derivatives activate the mitochondrial apoptosis pathway in the presence of antiapoptotic Bcl-2

Meriolin derivatives represent a new class of kinase inhibitors with a pronounced cytotoxic potential. Here, we investigated a newly synthesized meriolin derivative (termed meriolin 16) that displayed a strong apoptotic potential in Jurkat leukemia and Ramos lymphoma cells. Meriolin 16 induced apoptosis in rapid kinetics (within 2–3 h) and more potently (IC50: 50 nM) than the previously described derivatives meriolin 31 and 36 [1]. Exposure of Ramos cells to meriolin 16, 31, or 36 for 5 min was sufficient to trigger severe and irreversible cytotoxicity. Apoptosis induction by all three meriolin derivatives was independent of death receptor signaling but required caspase-9 and Apaf-1 as central mediators of the mitochondrial death pathway. Meriolin-induced mitochondrial toxicity was demonstrated by disruption of the mitochondrial membrane potential (ΔΨm), mitochondrial release of proapoptotic Smac, processing of the dynamin-like GTPase OPA1, and subsequent fragmentation of mitochondria. Remarkably, all meriolin derivatives were able to activate the mitochondrial death pathway in Jurkat cells, even in the presence of the antiapoptotic Bcl-2 protein. In addition, meriolins were capable of inducing cell death in imatinib-resistant K562 and KCL22 chronic myeloid leukemia cells as well as in cisplatin-resistant J82 urothelial carcinoma and 2102EP germ cell tumor cells. Given the frequent inactivation of the mitochondrial apoptosis pathway by tumor cells, such as through overexpression of antiapoptotic Bcl-2, meriolin derivatives emerge as promising therapeutic agents for overcoming treatment resistance.

Evaluation of Relationship between Bone Marrow Flow Cytometric Typing and Pathological Immunohistochemical Analysis of Lymphoma Cell Leukemia

Evaluation of Relationship between Bone Marrow Flow Cytometric Typing and Pathological Immunohistochemical Analysis of Lymphoma Cell Leukemia

Recurrence of HBV Reactivation after Allogeneic Hematopoietic Transplantation in Recipients with Resolved HBV Infection: A Nation-Wide Multicenter Retrospective Study

Introduction Reactivation of hepatitis B virus (HBV) is known to cause fulminant hepatitis after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in recipients with resolved HBV infection. Monitoring of HBV-DNA and HBV-DNA-guided preemptive therapy using nucleotide analogue (NA) is recommended to prevent developing hepatitis due to HBV reactivation after allo-HSCT. However, little is known about the adequate duration of NA treatment and the frequency of the recurrence of HBV reactivation (2nd HBV reactivation) after discontinuation of NA. We conducted a nationwide retrospective study of HBV reactivation after allo- HSCT. Patients and Methods We retrospectively reviewed the clinical information of recipients with resolved HBV infection (HBsAg-negative, anti-HBc-positive) before allo-HSCT and were diagnosed as HBV reactivation (HBsAg and/or HBV-DNA positive) after allo-HSCT from January 2010 to December 2020. Patients who received prophylactic NA before HBV reactivation were excluded. This study was approved by the Hokkaido University Hospital institutional review boards and registered as UMIN 000046803. Results A total of 72 patients were enrolled (median age of 60 years, range, 27-73 years; 42 males and 30 females) from 16 institutions. Initial hematological diseases were acute myeloid leukemia 21, myelodysplastic syndrome 14, chronic myeloid leukemia 4, acute lymphoblastic leukemia 12, non-Hodgkin lymphoma 8, adult T cell leukemia-lymphoma 6, mixed phenotype acute leukemia 4, and others 3. HSCT details were as follows: donor source, unrelated bone marrow 33, cord blood 24, related peripheral blood 11, unrelated peripheral blood 4; acute graft versus host disease (GVHD) 48 (66.7%); chronic GVHD 44 (61.1%); systemic steroid usage 41 (56.9%). Initial HBV reactivation after allo-HSCT was observed on day 10-3034 (median 512.5). Immunosuppressants and systemic steroids were administered in 35 (48.6%) and 28 (38.9%) patients respectively at the time of HBV reactivation. The patients who showed anti-HBs below protection levels (<10mIL/mL) were 18 (25%) of 72 patients at pre-HSCT, however, the number increased to 35 (81.4%) of 43 patients who were analyzed at the point of HBV reactivation after allo-HSCT. All patients received preemptive NA (entecavir 65, tenofovir alafenamide 6, tenofovir 1). Elevated aspartate transaminase of more than 200 U/L was observed in 7 cases at HBV reactivation, and all of them showed HBV-DNA more than 4 Log IU/mL. No one developed fulminant hepatitis but 5 patients resulted in continuous HBV carriers. Twenty-four of 72 patients were discontinued NA by each physician's decision. Of 24 patients who discontinued NA, 11 (45.8%) patients developed 2 nd HBV reactivation. The median duration from discontinuation of NA to 2 nd HBV reactivation was 113 days (range, 15 to 434). Duration of initial NA administration was not significantly different between patients with 2 ndHBV reactivation and those without 2 nd HBV reactivation (90-1968 days, median 553 vs 145-3734 days, median 861; p=0.11). In those who analyzed anti-HBs at discontinuation of NA, 4 of 8 cases (50.0%) with anti-HBs <10 mIU/mL developed 2 nd HBV reactivation, whereas only 1 of 6 cases (16.7%) with anti-HBs >10 mIU/mL developed 2 nd HBV reactivation. Four of the 11 cases with 2 nd HBV reactivation were attempted rechallenge of NA discontinuation, however, all 4 cases developed 3 rd HBV reactivation on day 28-127 after discontinuation of NA. These 4 cases further developed 4 th HBV reactivation after the rechallenge of discontinuation of NA. One patient developed protective anti-HBs titer after 2 courses of HB vaccination, and finally successfully discontinue the NA after 4 th HBV reactivation. The patient did not develop further reactivation for more than 3 years. Conclusion 2 nd reactivation of HBV after NA discontinuation was common in patients with HBV reactivation who received allo-HSCT despite the long duration of NA. Acquisition of anti-HBs by HBV reactivation itself or HB vaccine would be necessary to suppress further HBV reactivation. Careful monitoring of HBV-DNA is important even after the discontinuation of NA in the case with HBV reactivation after allo-HSCT because multiple reactivations could occur. A prospective randomized trial of the HB vaccine after allo-HSCT for preventing HBV reactivation is now ongoing (UMIN000034113).

Possible Concomitant Aggressive NK Cell Leukemia and EBV-positive T-cell lymphoma; Using the online beta version of WHO-HAEM5 and videoconferencing software to make diagnoses accessible in an emerging economy

BackgroundUsing the World Health Organization Classification 5th edition (beta version online; WHO-HAEM5bv) in emerging economies is key to global healthcare equity. Although there may be ongoing updates, hesitancy in accepting and reporting these diagnoses in publication conflicts with the WHO’s commitment to global accessibility. Aggressive NK cell leukemia (ANKL) and systemic EBV-positive T-cell lymphoma of childhood (SEBVTCL) with CD4-positive immunophenotype are both rare entities, are most described in Asians and East Asians, are associated with prior systemic chronic active EBV disease (CAEBV), and presentation with Hemophagocytic Lymphohistiocytosis (HLH). Recognizing and diagnosing any one of these entities requires not only training and experience in hematopathology, but good cooperation between clinical physicians and all areas of the laboratory. We describe a 30-year-old woman who presented to a Vietnam hospital and was rapidly diagnosed with ANKL, SEBVTCL, and HLH using WHO-HAEM5bv essential criteria, aided by expert consultation from a United States (US) board certified hematopathologist in real-time using video conferencing software.MethodsZoom™ videoconferencing software; Immunohistochemistry; flow cytometric immunophenotyping; polymerase chain reaction (PCR), Next Generation Sequencing (NGS).ResultsAt the time of hospital admission, automated complete blood count (CBC) with differential count showed slight anemia, slight lymphocytosis, and moderate thrombocytopenia. HIV serology was negative. Whole blood PCR for EBV was positive showing 98,000 copies/ml. A lymph node biopsy revealed histology and immunohistochemistry consistent with the online beta version WHO-HAEM5 classification of SEBVTCL arising in CAEBV. Blood and bone marrow studies performed for staging revealed no histologic or immunohistochemical evidence of T-cell lymphoma in the bone marrow core, however, atypical blood smear lymphocyte morphology and blood immunophenotyping by flow cytometry were consistent with WHO-HAEM5 classification of ANKL. NGS revealed no evidence of genetic variant(s) associated with HLH in Vietnam. All laboratory studies were performed at Blood Transfusion Hematology Hospital (BTHH) in Ho Chi Minh City Vietnam.ConclusionAlthough Vietnam, an emerging economy, currently lacks the laboratory infrastructure to more rigorously confirm a rare synchronous presentation of two distinct EBV-driven T/NK cell neoplasms, these two concomitant diagnoses were made using only laboratory techniques available in Vietnam with the help of WHO-HAEM5bv and real-time video consultation by a US hematopathologist.

Improving the cytotoxicity of immunotoxins by reducing the affinity of the antibody in acidic pH

BackgroundImmunotoxins are antibody-toxin conjugates that bind to surface antigens and exert effective cytotoxic activity after internalization into tumor cells. Immunotoxins exhibit effective cytotoxicity and have been approved by the FDA to treat multiple hematological malignancies, such as hairy cell leukemia and cutaneous T-cell lymphoma. However, most of the internalized immunotoxin is degraded in lysosomes, and only approximately 5% of free toxin escapes into the cytosol to exert cytotoxicity. Many studies have improved immunotoxins by engineering the toxin fragment to reduce immunogenicity or increase stability, but how the antibody fragment contributes to the activity of immunotoxins has not been well demonstrated.MethodsIn the current study, we used 32A9 and 42A1, two anti-GPC3 antibodies with similar antigen-binding capabilities and internalization rates, to construct scFv-mPE24 immunotoxins and evaluated their in vitro and in vivo antitumor activities. Next, the antigen-binding capacity, trafficking, intracellular protein stability and release of free toxin of 32A9 scFv-mPE24 and 42A1 scFv-mPE24 were compared to elucidate their different antitumor activities. Furthermore, we used a lysosome inhibitor to evaluate the degradation behavior of 32A9 scFv-mPE24 and 42A1 scFv-mPE24. Finally, the antigen-binding patterns of 32A9 and 42A1 were compared under neutral and acidic pH conditions.ResultsAlthough 32A9 and 42A1 had similar antigen binding capacities and internalization rates, 32A9 scFv-mPE24 had superior antitumor activity compared to 42A1 scFv-mPE24. We found that 32A9 scFv-mPE24 exhibited faster degradation and drove efficient free toxin release compared to 42A1 scFv-mPE24. These phenomena were determined by the different degradation behaviors of 32A9 scFv-mPE24 and 42A1 scFv-mPE24 in lysosomes. Moreover, 32A9 was sensitive to the low-pH environment, which made the 32A9 conjugate easily lose antigen binding and undergo degradation in lysosomes, and the free toxin was then efficiently produced to exert cytotoxicity, whereas 42A1 was resistant to the acidic environment, which kept the 42A1 conjugate relatively stable in lysosomes and delayed the release of free toxin.ConclusionsThese results showed that a low pH-sensitive antibody-based immunotoxin degraded faster in lysosomes, caused effective free toxin release, and led to improved cytotoxicity compared to an immunotoxin based on a normal antibody. Our findings suggested that a low pH-sensitive antibody might have an advantage in the design of immunotoxins and other lysosomal degradation-dependent antibody conjugate drugs.

Deceptive Presentation of Low-Grade Lymphoma with Grade 3 Marrow Fibrosis and Aplasia: Diagnostic and Clinical Considerations

AbstractBone marrow fibrosis with lymphoproliferative disorders is rare with the exception of hairy cell leukemia and nodular sclerosis Hodgkin's lymphoma. We report the case of a 63-year-old gentleman with indolent B-cell lymphoma presenting with myelofibrosis and aplasia. He was evaluated for pancytopenia with no organomegaly or lymphadenopathy. Bone marrow aspiration was a dry tap and biopsy revealed a hypocellular marrow with a cellularity of 10 to 20% with absent megakaryocytes and grade 2 to 3 reticulin fibrosis. Myeloproliferative neoplasms were ruled out based on morphology and absence of myeloid mutations on next-generation sequencing. Further sections revealed interstitial infiltrates of lymphoid cells with round, clumped chromatin and inconspicuous nucleoli, which on immunohistochemistry (IHC) were positive for CD20 and BCL2, and negative for CD5, CD10, BCL6, annexin A1, cyclin D1, and TdT. The final diagnosis was thus confirmed as CD5-negative low-grade B-cell lymphoma and he was initiated on therapy with a combination of Bendamustine and Rituximab. He had resolution of symptoms and cytopenia after six cycles of the same. Presence of significant myelofibrosis on the background of a hypocellular marrow can mimic several subtypes of myeloproliferative neoplasms or myelodysplastic syndromes, providing a diagnostic challenge. IHC is essential in determining the exact subtype to decide further therapy. Based on literature search, only a handful of patients with this presentation have been described and this case will be a valuable addition to the same.

Cytokines induced memory-like NK cells engineered to express CD19 CAR exhibit enhanced responses against B cell malignancies.

CD19 chimeric antigen receptor (CAR) engineered NK cells have been used for treating patients with relapsed and/or refractory B cell malignancies and show encouraging outcomes and safety profile. However, the poor persistence of NK cells remains a major challenge for CAR NK cell therapy. Memory-like NK cells (MLNK) induced by IL-12, IL-15, and IL-18 have shown enhanced and prolonged responses to tumor re-stimulation, making them an attractive candidate for adoptive cellular immunotherapy. Here, we show efficient and stable gene delivery of CD19 CAR to memory-like NK cells using retroviral vectors with transduction efficiency comparable to those achieved with conventional NK cells. Analysis of surface molecules revealed a distinct phenotypic profile in CAR engineered memory-like NK cells (CAR MLNK), as evidenced by increased expression of CD94 and downregulation of NKp30 as well as KIR2DL1. Compared to conventional CAR NK cells, CAR MLNK cells exhibited significantly increased IFN-γ production and degranulation in response to CD19+ target cells, resulting in enhanced cytotoxic activity against CD19+ leukemia cells and lymphoma cells. Furthermore, memory properties induced by IL-12/-15/-18 improved the in vivo persistence of CAR MLNK cells and significantly suppressed tumor growth in a exnograft mouse model of lymphoma, leading to prolonged survival of CD19+ tumor-bearing mouse. Altogether, our data indicate that CD19 CAR engineered memory-like NK cells exhibited superior persistence and antitumor activity against CD19+ tumors, which might be an attractive approach for treating patient with relapse or refractory B cell malignancies.

Transformations of marginal zone lymphomas and lymphoplasmacytic lymphomas: Report from the 2021 SH/EAHP Workshop.

To summarize the conclusions of the 2021 Society for Hematopathology/European Association for Haematopathology workshop regarding transformations of marginal zone lymphoma (MZL) and lymphoplasmacytic lymphoma (LPL). Nineteen cases were submitted to this portion of the workshop. Additional studies were performed in cases with sufficient material. Cases included splenic MZL (n = 4), splenic diffuse red pulp small B-cell lymphoma (n = 2), nodal MZL (n = 4), extranodal MZL (n = 1), and LPL (n = 8). The most common transformation was to diffuse large B-cell lymphoma (DLBCL), but others included classic Hodgkin lymphoma, high-grade B-cell lymphomas with MYC and BCL6 rearrangements, plasmablastic lymphoma, and plasma cell leukemia. Two splenic MZLs with transformation to DLBCL contained t(14;19)(q32;q13.3) IGH::BCL3 rearrangements in both samples. Paired sequencing studies in 5 MZLs with transformation to clonally related DLBCL identified a variety of mutations and gene fusions at the time of transformation, including CARD11, IGH::MYC, NOTCH2, P2RY8, TBLX1X1, and IGH::CD274. Marginal zone lymphoma and LPL may undergo a variety of transformation events, most commonly to DLBCL, which is usually, although not always, directly clonally related to the underlying low-grade lymphoma. Multiparameter analysis including broad-based sequencing studies can assist in the diagnosis and classification of these uncommon cases.

Ferroptosis in hematological malignant tumors.

Ferroptosis is a kind of iron-dependent programmed cell death discovered in recent years. Its main feature is the accumulation of lipid reactive oxygen species in cells, eventually leading to oxidative stress and cell death. It plays a pivotal role in normal physical conditions and the occurrence and development of various diseases. Studies have shown that tumor cells of the blood system, such as leukemia cells and lymphoma cells, are sensitive to the response to ferroptosis. Regulators that modulate the Ferroptosis pathway can accelerate or inhibit tumor disease progression. This article reviews the mechanism of ferroptosis and its research status in hematological malignancies. Understanding the mechanisms of ferroptosis could provide practical guidance for treating and preventing these dreaded diseases.

Abstract CT136: An open-label phase I/IIa study to evaluate the safety and efficacy of CCS1477 as monotherapy and in combination in patients with advanced hematological malignancies

Abstract Background Inobrodib (CCS1477) is a first in class potent, selective, and orally bioavailable inhibitor of the bromodomains of p300 and CBP, two closely related histone acetyl transferases with oncogenic roles in hematological malignancies. In pre-clinical studies inobrodib potently inhibited cell proliferation in acute myeloid leukemia (AML), multiple myeloma (MM) and non-Hodgkin lymphoma (NHL) cell lines. Inobrodib demonstrated dose-dependent efficacy in corresponding xenograft models, eliciting tumor regression at the highest doses. These effects were accompanied by significant reductions in expression of MYC, MYB and IRF4. Low doses of inobrodib also demonstrated good efficacy when added to standard-of-care treatments, providing a strong rationale for combination treatment in patients. Inobrodib represents a novel and differentiated approach and offers a potential new therapeutic option for patients who have relapsed or are refractory to current standard of care therapies in AML, higher risk MDS, MM or NHL. Study Design and Methods NCT04068597 is an adaptive multi-arm/multi-stage trial allowing exploration of p300/CBP inhibition as monotherapy or in combination with different agents across multiple indications including AML/higher risk MDS, MM, and NHL. The trial was initiated as a dose escalation rolling 6 design separately in AML/MDS and MM/NHL to ensure tolerability is explored separately across groups of indications, with the possibility to expand in monotherapy across different indications. Key inclusion criteria include patients with confirmed relapsed or refractory disease, with patients having received standard therapy (typically at least two prior lines of therapy). Response criteria are assessed using standard methodologies for the different indications. Blood and tumor samples are collected for exploratory biomarker analysis to understand mechanisms of response to treatment or disease progression. The recommended phase 2 dose/schedule has been determined, and monotherapy expansion arms are currently open in selected indications. Cohorts investigating dose escalation with standard of care agents pomalidomide/dexamethasone in MM as well azacytidine with or without venetoclax in AML or MDS are currently enrolling, with the possibility of expansion for promising combinations. An additional ‘rescue’ cohort for MM patient who failed inobrodib monotherapy will be used to determine whether addition of pomalidomide with or without dexamethasone may reverse resistance to inobrodib. This cohort will also explore tolerability and clinical activity of steroid-sparing combination regimen. Citation Format: Tomasz Knurowski, Emma Searle, Karen Clegg, Neil Pegg, William West, Debbie Haynes, Kristopher Frese, Tim C. Somervaille. An open-label phase I/IIa study to evaluate the safety and efficacy of CCS1477 as monotherapy and in combination in patients with advanced hematological malignancies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr CT136.

Acquired L1196M ALK mutation in anaplastic lymphoma kinase-positive anaplastic large cell lymphoma during alectinib administration.

Acquired L1196M ALK mutation in anaplastic lymphoma kinase-positive anaplastic large cell lymphoma during alectinib administration.

Comprehensive analysis of clinicalf eatures, treatment options, overall survival, and prognostic factors in lymphoma cell leukemia patients: A retrospective study.

Lymphoma cell leukemia (LCL) is regarded as patients presenting a high extensive lymphoma cell ratio in bone marrow (BM), which is recognized as lymphoma of stage IV by invading into BM. This study aimed to investigate the clinical characteristics, treatment options, survival profiles, and prognostic factors in patients with LCL. Clinical data of 42 patients with LCL were retrospectively reviewed, and baseline characteristics and treatment records were extracted. In addition, overall survival (OS) was calculated, and the causes of death were analyzed. Out of the 42 patients with LCL, 9 (21.4%) had primary BMLCL, 20 (47.6%) had Non-Hodgkin lymphoma (NHL) complicated with LCL, and 13 (31.0%) had NHL evolving into LCL. Common clinical characteristics included B syndromes (n = 21, 50.0%), abnormal white blood count (n = 28, 66.5%), decreased hemoglobin (n = 28, 66.7%), and platelet (n = 30, 71.4%). Additionally, elevated Eastern Cooperative Oncology Group (ECOG) with a score greater than one occurred in 26 patients (61.9%), and elevated lactate dehydrogenase (LDH) occurred in 25 patients (59.5%). For treatments, chemotherapy was the most common therapy (n = 35, 83.2%), followed by symptomatic treatment and radiotherapy plus chemotherapy. Additionally, the mean OS of the patients was 16.9 (95% CI: 12.8-20.9) months, among which primary patients with BMLCL showed shorter OS than those with NHL complicated with LCL and NHL evolving into patients with LCL. A total of 9 (21.4%) patients with LCL died during follow-up, among which the central nervous system (CNS) invasion was the most common cause of death. Furthermore, primary BMLCL, higher ECOG, and higher LDH were potential predictive factors for worse OS in patients with LCL. This study gives an overview of the treatment and prognosis of LCL, which provides additional information for the management of LCL.

A Comprehensive, Multiomics Analysis of Natural Killer-Cell Malignancies

Natural killer/T cell lymphoma (NKTCL) and aggressive NK cell leukemia (ANKL) are rare mature NK cell malignancies which show a geographic predilection for Asia and Central/South America and are strongly associated with EBV infection. Despite recent advances in chemotherapeutical agents, the prognosis (especially for ANKL) remain poor. On the contrary, NK cell proliferations of a more indolent nature also exist. NK-large granular lymphocytic leukemia (NK-LGLL) is a disorder of NK cells with indolent clinical manifestation. Additionally, chronic active Epstein-Barr virus infection (CAEBV) affecting NK cells can have variable presentation, between indolent and aggressive. Due to their rarity, a comprehensive understanding of the events differentiating the wide spectrum of NK malignancies remains elusive. Here, we use a multi-omics approach to investigate the genetic differences underlying the different clinical manifestations of NK malignancies. To discover the pathogenesis behind indolent and aggressive NK malignancies, we studied whole-genome/exome sequencing and whole transcriptome sequencing (RNA-seq) data from over 200 NK-cases collected from 11 previous publications. We investigated driver genes from 145 WES and WGS samples, and transcriptomes from 91 samples. Single-cell RNA-seq analysis was performed to identify and characterize distinct NK-cell subsets. We used pooled CRISPR screens with a single-cell RNA sequencing readout (CROP-seq) to investigate the effect of knocking out the most frequently mutated genes on the transcriptome of NKTCL cells under the effect of commonly used chemotherapeutic agents. We also aim to construct a comprehensive host-EBV interaction map to elucidate the role of EBV in the pathogenesis of NK malignancies We validated previously found driver genes STAT3 and DDX3X as the most frequently mutated genes in NK malignancies, being mutated in 15% and 14% of samples, respectively. STAT3 mutations were found only in NKTCL and ANKL samples. Additionally, our combinatorial analysis allowed for identification of previously unappreciated putative driver genes, MSN and HLA-A, which were mutated in 8% and 4% of samples, respectively. We also discovered associations between important genes, such as significant co-occurrence of mutations in STAT3 and novel driver gene MSN (p <0.05). Samples originating from different diseases were intermingled in non-negative matrix factorization (NMF) analyses, highlighting the effect of mutational profile, rather than disease type, on sample clustering. Using RNA-seq, differentially expressed genes (Q-value < 0.01) were identified between ANKL, NKTCL and healthy samples. Gene set enrichment analysis revealed differential enrichment and depletion of biological processes in ANKL and NKTCL. Upregulation of chemokine signalling and macrophage-associated processes in NKTCL reflected the inflammatory microenvironment of the disease. Of 17 putative driver genes, CRISPR knockout of STAT3 and TP53 led to most transcriptional changes. Knockout of STAT3 caused upregulation of HLA I and II molecules and downregulation of IL10 signaling, thus promoting tumor cell immune evasion. Our preliminary results indicate that NK malignancies are, contrary to previous knowledge, a spectrum of manifestations rather than distinct diseases. Our results concerning the heterogeneous genetic landscape of NK malignancies can lead to better and faster diagnosis as well as improved treatment stratification in the future. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

The mycotoxin viriditoxin induces leukemia- and lymphoma-specific apoptosis by targeting mitochondrial metabolism

Inhibition of the mitochondrial metabolism offers a promising therapeutic approach for the treatment of cancer. Here, we identify the mycotoxin viriditoxin (VDT), derived from the endophytic fungus Cladosporium cladosporioides, as an interesting candidate for leukemia and lymphoma treatment. VDT displayed a high cytotoxic potential and rapid kinetics of caspase activation in Jurkat leukemia and Ramos lymphoma cells in contrast to solid tumor cells that were affected to a much lesser extent. Most remarkably, human hematopoietic stem and progenitor cells and peripheral blood mononuclear cells derived from healthy donors were profoundly resilient to VDT-induced cytotoxicity. Likewise, the colony-forming capacity was affected only at very high concentrations, which provides a therapeutic window for cancer treatment. Intriguingly, VDT could directly activate the mitochondrial apoptosis pathway in leukemia cells in the presence of antiapoptotic Bcl-2 proteins. The mitochondrial toxicity of VDT was further confirmed by inhibition of mitochondrial respiration, breakdown of the mitochondrial membrane potential (ΔΨm), the release of mitochondrial cytochrome c, generation of reactive oxygen species (ROS), processing of the dynamin-like GTPase OPA1 and subsequent fission of mitochondria. Thus, VDT-mediated targeting of mitochondrial oxidative phosphorylation (OXPHOS) might represent a promising therapeutic approach for the treatment of leukemia and lymphoma without affecting hematopoietic stem and progenitor cells.

Hodgkin Lymphoma and Hairy Cell Leukemia Arising from Chronic Lymphocytic Leukemia: Case Reports and Literature Review

Richter’s syndrome represents the progression of chronic lymphocytic leukemia (CLL) to more aggressive diseases, most frequently diffuse large B-cell lymphoma, while Hodgkin’s lymphoma (HL) and hairy cell leukemia (HCL) are rarely described. The first case involved a 67-year-old man with a diagnosis of a high-risk stage-II CLL treated with rituximab and ibrutinib, developed a HL nodular sclerosis variant after three months of therapy for CLL. After achieving a complete remission for HL and ibrutinib cessation because of drug-related cardiotoxicity, the patient relapsed after five months off-therapy and died due to disease progression after two cycles of brentuximab-vedotin. The second case involved an 83-year-old female with a diagnosis of stage-IV CLL treated with rituximab plus bendamustine who developed a HCL eight years later. Pentostatin was unsuccessfully employed as upfront HCL therapy, and the patient was then switched to rituximab while in remission for CLL. In conclusion, Richter’s transformation risk rate might be higher in patients treated with novel targeted therapies, and multiparametric flow cytometry and lymph node biopsy at relapse could help in early identifying small clones. The treatment of predominant neoplasia is mandatory, and disease-specific drugs are administered; however, clinical efficacy might be lower in these patients.

Clinical characteristics and prognosis of lymphoma-associated hemophagocytic syndrome

Subjective To investigate clinical characteristics, treatment, and prognosis of lymphoma-associated hemophagocytic syndrome (LAHS) patients. Methods: The clinical data of patients diagnosed with LAHS from January 2010 to October 2021 in West China Hospital were retrospectively analyzed. Clinical characteristics, treatment, overall response rate (ORR), and overall survival (OS) were investigated. Univariate and multivariate analysis of potential factors were conducted. Results: Of all 94 patients included, 59 were male and 35 were female. The age at hemophagocytic lymphohistiocytosis (HLH) diagnosis was (40.5±17.3) years. Seventy-four cases were T/NK cell lymphoma; 15 were B cell lymphoma; 5 were Hodgkin lymphoma. The age at HLH diagnosis of T/NK cell LAHS patients was (37.9±16.2) years, while that of B cell LAHS patients was (55.9±14.0) years. T/NK cell LAHS patients were significantly younger than B cell LAHS patients (P<0.001). Baseline fibrinogen of T/NK cell LAHS patients was 1.34 (0.86, 2.44) g/L, while that of B cell LAHS patients was 2.20 (1.75, 4.25) g/L. T/NK cell LAHS patients showed significantly lower fibrinogen levels than B cell LAHS patients (P=0.008). Combined treatment of anti-HLH and anti-lymphoma treatment was conducted in 35 patients; anti-HLH treatment was conducted in 31 patients; anti-lymphoma treatment was conducted in 8 patients; glucocorticoid treatment was conducted in 7 patients. ORR was 49.4%, and the median OS was 61 days for overall patients. Patients who received anti-HLH treatment and turned to anti-lymphoma treatment early displayed the best ORR and OS, significantly higher than those of anti-HLH patients (69.0 vs 38.7%, P=0.019, and 192.0 vs 24.5 days, P=0.028, respectively), which were also insignificantly higher than those of anti-lymphoma patients. Extranodal NK/T-cell lymphoma or aggressive natural killer cell leukemia was the risk factor of LAHS prognosis (HR=0.113, 95%CI: 0.018-0.728, P=0.022). Conclusions: Prognosis of LAHS patients is poor. Anti-lymphoma treatment should be initiated as soon as HLH is rapidly controlled.