Chronic Lymphoblastic Leukemia Research Articles

Advancing chronic lymphoblastic Leukemia management: A systematic review of preclinical studies on nanotechnology-based diagnostics and therapeutics

Advancing chronic lymphoblastic Leukemia management: A systematic review of preclinical studies on nanotechnology-based diagnostics and therapeutics

Hematological malignancies: Prevalence and hematological characteristics in a single center in southern Saudi Arabia.

To determine the prevalence of leukemia in the Aseer region of Saudi Arabia and the importance of hematological, biochemical and coagulation profiles for leukemic patients in the context of disease management. This retrospective study comprised 210 patients between 2012 and 2022 who had been diagnosed with leukemia at different ages. The multiple unpaired t-test was used to compare leukemic patients with control samples, which consisted of healthy individuals, and p<0.05 was taken as significant. The data was compiled from Aseer Central Hospital in the Aseer region and collected through peripheral blood smear and bone marrow biopsy (2012-2017) or by flow cytometry (2018-2022), according to the hospital information system and registry data. Of the total 210 leukemic patients (61.4% males and 38.6% females), 104 cases (2012-2017) were diagnosed based on peripheral blood smear and bone marrow biopsy, and 106 cases (2018-2022) based on flow cytometry. Fifteen subtypes of leukemia were identified, with chronic myeloid leukemia being the most common (34.2%), followed by acute myeloid leukemia (17.6%), chronic lymphoblastic leukemia (11.9%), and B-cell acute lymphoid leukemia (9.5%). Other rare cases were also found. Of the 210 leukemia cases diagnosed in the Aseer region between 2012-2022, the most common subtype was chronic myeloid leukemia, followed by acute myeloid leukemia. In all leukemia subtypes, distinctive significant changes were observed in hematological parameters, biochemical parameters, and coagulation profiles.

Proportion and Clinical Significance of γδT17/Th17/Tc17 Cells in Peripheral Blood of Uygur Patients with Chronic Lymphoblastic Leukemia

To explore the distribution of γδT17/Th17/Tc17 cells in the peripheral blood of Uygur patients with chronic lymphoblastic leukemia (CLL) and clinical significance. ELISA method was used to detect the levels of IL-17, IL-23, IL-6, and IFN-γ in the peripheral blood serum of 53 newly diagnosed Uygur patients with CLL and 30 healthy controls. Flow cytometry was used to determine the proportion of γδT/γδT17/Th17/Tc17 cells in the peripheral blood of Uygur CLL patients and controls, and the changes of the abover indexes in CLL Binet staging were observed. Compared with the control group, the proportion of γδT cells, γδT17 cells, and Th17 cells in the peripheral blood of Uygur CLL patients increased significantly （P <0.05）. γδT17 cell proportion in total lymphocytes was significantly higher than Th17 and Tc17 cell proportions（P <0.05）, and proportions of γδT, γδT17 cells increased gradually as the disease progressed. The levels of cytokines IL-17, IL-23, and IL-6 in peripheral blood of Uygur patients with CLL were significantly higher than those in the control group（P <0.05）, while the level of cytokine IFN-γ was significantly lower（P <0.05）. The level of IL-17 in peripheral blood decreased gradually as the disease progressed（P <0.05）. γδT and γδT17 are abnormally highly expressed in Uygur CLL , which are related to the stage of disease and participate in the occurrence and development of CLL.

Comparison of bolus administration and short-term infusion versus long-term infusion of doxorubicin in terms of cardiotoxicity and efficacy.

Cardiotoxicity caused by anthracyclines chemotherapy is one of the leading causes of mortality and morbidity in cancer survivors. Continuous infusion (CI) instead of bolus (BOL) injection is one of the methods that seem to be effective in reducing doxorubicin (DOX) cardiotoxicity. Due to the variety of results, we decided to compare these two approaches regarding toxicity and efficacy and report the final results for different cancers. We included 21 studies (four preclinical and seventeen clinical trials) up to May 15, 2023. In children with acute lymphoblastic leukemia (ALL) and adults with chronic lymphoblastic leukemia (CLL) and gastric cancer, results were in favor of BOL injection, without increase in cardiotoxicity. On the other hand, CI showed to be better option in patients with small-cell lung cancer (SCLC) and breast cancer. Various results were also observed in adult patients with sarcoma. Overall, it can be concluded that the benefits of CI, especially in adults, outweigh its disadvantages. However, due to the variety of results and heterogeneity of studies, further clinical trials with a larger sample size and a longer duration of follow-up are needed to make a more accurate comparison between CI and BOL injection.

The causal relationship between gut microbiota and leukemia: a two-sample Mendelian randomization study

BackgroundThe association between gut microbiota and leukemia has been established, but the causal relationship between the two remains unclear.MethodsA bidirectional two-sample Mendelian randomization (MR) was used to analyze the causal relationship between gut microbiota and leukemia. Microbiome data (n = 14,306) and leukemia (n = 1,145) data were both sourced from European populations. Single nucleotide polymorphisms (SNPs) were selected as instrumental variables based on several criteria. We employed various MR methods, such as the inverse variance weighted (IVW) method, to evaluate the causal effect between exposure and outcomes and conducted sensitivity analyses to validate the heterogeneity and pleiotropy of the instrumental variables.Results5,742 qualified instrumental variables were included. In the primary MR results, a total of 10 gut microbial taxa were associated with leukemia risk. Genus Blautia and genus Lactococcus are risk factors for acute lymphoblastic leukemia [genus Blautia odds ratio (OR): 1.643, 95% confidence interval (CI): 1.592 ~ 1.695, Adjusted p &amp;lt; 0.001; genus Lactococcus OR: 2.152, 95% CI: 1.447 ~ 3.199, Adjusted p = 0.011]. Genus Rikenellaceae RC9 gut group, genus Anaerostipes, genus Slackia, and genus Lachnospiraceae ND3007 group are risk factors for acute myeloid leukemia [genus Rikenellaceae RC9 gut group OR: 1.964, 95% CI: 1.573 ~ 2.453, Adjusted p &amp;lt; 0.001; genus Anaerostipes OR: 2.515, 95% CI: 1.503 ~ 4.209, Adjusted p = 0.017; genus Slackia OR: 2.553, 95% CI: 1.481 ~ 4.401, Adjusted p = 0.022; genus Lachnospiraceae ND3007 group OR: 3.417, 95% CI: 1.960 ~ 5.959, Adjusted p = 0.001]. Genus Ruminococcaceae UCG011 and genus Ruminococcaceae UCG014 were risk factors for chronic myeloid leukemia (genus Ruminococcaceae UCG011 OR: 2.010, 95% CI: 1.363 ~ 2.963, Adjusted p = 0.044; genus Ruminococcaceae UCG014 OR: 3.101, 95% CI: 1.626 ~ 5.915, Adjusted p = 0.044). Genus Slackia was a protective factor for acute lymphoblastic leukemia (genus Slackia OR: 0.166, 95% CI: 0.062 ~ 0.443, Adjusted p = 0.017). Family Acidaminococcaceae was a protective factor for acute myeloid leukemia (family Acidaminococcaceae OR: 0.208, 95% CI: 0.120 ~ 0.361, Adjusted p &amp;lt; 0.001). Genus Desulfovibrio was a protective factor for chronic lymphoblastic leukemia (genus Desulfovibrio OR: 0.581, 95% CI: 0.440 ~ 0.768, Adjusted p = 0.020). Sensitivity analysis revealed no heterogeneity or pleiotropy between SNPs.ConclusionThis study revealed the causal relationship between the gut microbiota and leukemia, and identified potential pathogenic bacteria and probiotic taxa associated with the onset of leukemia. This research may aid in the early detection of various types of leukemia and offer a new direction for the prevention and treatment of leukemia.

Using Human 'Personalized' Cybrids to Identify Drugs/Agents That Can Regulate Chronic Lymphoblastic Leukemia Mitochondrial Dysfunction.

This study uses personalized chronic lymphoblastic leukemia (CLL) cybrid cells to test various drugs/agents designed to improve mitochondrial function and cell longevity. Age-matched control (NL) and CLL cybrids were created. The NL and CLL cybrids were treated with ibrutinib (Ibr-10 μM), mitochondrial-targeted nutraceuticals such as alpha lipoic acid (ALA-1 mM), amla (Aml-300 μg), melatonin (Mel-1 mM), resveratrol (Res-100 μM) alone, or a combination of ibrutinib with nutraceuticals (Ibr + ALA, Ibr + Aml, Ibr + Mel, or Ibr + Res) for 48 h. MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliumbromide), H2DCFDA(2',7' Dichlorodihydrofluorescein diacetate), and JC1 assays were used to measure the cellular metabolism, intracellular ROS levels, and mitochondrial membrane potential (∆ψm), respectively. The expression levels of genes associated with antioxidant enzymes (SOD2, GPX3, and NOX4), apoptosis (BAX and CASP3), and inflammation (IL6, IL-1β, TNFα, and TGFβ) were measured using quantitative real-time PCR (qRT-PCR). CLL cybrids treated with Ibr + ALA, Ibr + Aml, Ibr + Mel, and Ibr + Res had (a) reduced cell survivability, (b) increased ROS production, (c) increased ∆ψm levels, (d) decreased antioxidant gene expression levels, and (e) increased apoptotic and inflammatory genes in CLL cybrids when compared with ibrutinib-alone-treated CLL cybrids. Our findings show that the addition of nutraceuticals makes the CLL cybrids more pro-apoptotic with decreased cell survival compared with CLL cybrids exposed to ibrutinib alone.

γδ T cells in immunotherapies for B-cell malignancies.

Despite the advancements in therapy for B cell malignancies and the increase in long-term survival of patients, almost half of them lead to relapse. Combinations of chemotherapy and monoclonal antibodies such as anti-CD20 leads to mixed outcomes. Recent developments in immune cell-based therapies are showing many encouraging results. γδ T cells, with their potential of functional plasticity and their anti-tumoral properties, emerged as good candidates for cancer immunotherapies. The representation and the diversity of γδ T cells in tissues and in the blood, in physiological conditions or in B-cell malignancies such as B cell lymphoma, chronic lymphoblastic leukemia or multiple myeloma, provides the possibility to manipulate them with immunotherapeutic approaches for these patients. In this review, we summarized several strategies based on the activation and tumor-targeting of γδ T cells, optimization of expansion protocols, and development of gene-modified γδ T cells, using combinations of antibodies and therapeutic drugs and adoptive cell therapy with autologous or allogenic γδ T cells following potential genetic modifications.

Synthesis and Biological Evaluation of α-Tocopherol Derivatives as Potential Anticancer Agents

α-Tocopheryl succinate (α-TS) and α-tocopheryloxyacetic acid (α-TEA) are potent inducers of apoptosis in cancer cells and efficient suppressors of tumors in experimental model cancer cell lines. They exhibit selective cytotoxicity against tumor cells and very limited or no toxicity toward nonmalignant cells. In the present work, a series of new α-tocopherol derivatives were synthesized as analogs of α-TS and α-TEA. The cytotoxic activity of obtained compounds was tested using three human cancer cell lines, including chronic lymphoblastic leukemia (CEM), breast adenocarcinoma (MCF7), cervical adenocarcinoma (HeLa), and normal human fibroblasts (BJ). The introduction of an alkyl substituent into the ether-linked acetic acid moiety in α-TEA increased anticancer activity. α-Tocopheryloxy-2-methylpropanoic acid with two additional geminal methyl groups was more active against CEM cells compared to α-TEA and non-toxic to normal cells. In order to acquire a deeper understanding of the biological activity of synthesized compounds, a molecular docking study was also conducted. Our research confirmed that vitamin E derivatives are interesting and valuable compounds in terms of their potential therapeutic use as anticancer agents.

Standardize and enhance the diagnosis and treatment capabilities of chronic lymphoblastic leukemia in China

Standardize and enhance the diagnosis and treatment capabilities of chronic lymphoblastic leukemia in China

The Impact of Improved Treatments on Survival of Adult U.S. Leukemia Patients: 1990-2018.

Molecularly targeted therapies such as tyrosine kinase inhibitors (TKI) are effective treatments for B-cell receptor (BCR)-ABL-bearing leukemias. We evaluated the impact of TKIs on historical chronic myeloid leukemia (CML) mortality trends compared with acute lymphoblastic leukemia (ALL) and chronic lymphoblastic leukemia (CLL). Because mortality trends reflect combined effects of leukemia incidence and survival, we also evaluated the contribution of incidence and survival trends to mortality trends by subtypes. We used data from 13 U.S. (SEER) registries (1992-2017) among U.S. adults. We utilized histology codes to identify cases of CML, ALL, and CLL and death certificate data to calculate mortality. We used Joinpoint to characterize incidence (1992-2017) and mortality (1992-2018) trends by subtype and diagnosis year. For CML, mortality rates started declining in 1998 at an average rate of 12% annually. Imatinib was approved by the FDA for treating CML and ALL in 2001, leading to clear benefits for patients with CML. Five-year CML survival increased dramatically over time, especially between 1996 to 2011, 2.3% per year on average. ALL incidence increased 1.5% annually from 1992 to 2017. ALL mortality decreased 0.6% annually during 1992 to 2012 and then stopped declining. CLL incidence fluctuated during 1992 to 2017 while mortality decreased 1.1% annually during 1992 to 2011 and at a faster rate of 3.6% per year from 2011. Five-year survival increased 0.7% per year on average during 1992 to 2016. Survival benefit from TKIs and other novel therapies for treating leukemia subtypes has been demonstrated in clinical trials. Our study highlights the impact of molecularly targeted therapies at the population level.

The association of BTLA gene polymorphisms with non-small lung cancer risk in smokers and never-smokers.

Lung cancer is the predominant cause of death among cancer patients and non-small cell lung cancer (NSCLC) is the most common type. Cigarette smoking is the prevailing risk factor for NSCLC, nevertheless, this cancer is also diagnosed in never-smokers. B and T lymphocyte attenuator (BTLA) belongs to immunological checkpoints which are key regulatory molecules of the immune response. A growing body of evidence highlights the important role of BTLA in cancer. In our previous studies, we showed a significant association between BTLA gene variants and susceptibility to chronic lymphoblastic leukemia and renal cell carcinoma in the Polish population. The present study aimed to analyze the impact of BTLA polymorphic variants on the susceptibility to NSCLC and NSCLC patients' overall survival (OS). Using TaqMan probes we genotyped seven BTLA single-nucleotide polymorphisms (SNPs): rs2705511, rs1982809, rs9288952, rs9288953, rs1844089, rs11921669 and rs2633582 with the use of ViiA 7 Real-Time PCR System. We found that rs1982809 within BTLA is associated with NSCLC risk, where carriers of rs1982809G allele (AG+GG genotypes) were more frequent in patients compared to controls. In subgroup analyses, we also noticed that rs1982809G carriers are significantly overrepresented in never-smokers, but not in smokers compared to controls. Additionally, the global distribution of the haplotypes differed between the never-smokers and smokers, where haplotypes A G G C A, C G A C G, and C G A T G were more frequent in never-smoking patients. Furthermore, the presence rs1982809G (AG+GG genotypes) allele as well as the presence of rs9288953T allele (CT+TT genotypes) increased NSCLC risk in females' patients. After stratification by histological type, we noticed that rs1982809G and rs2705511C carriers were more frequent among adenocarcinoma patients. Moreover, rs1982809G and rs2705511C correlated with the more advanced stages of NSCLC (stage II and III), but not with stage IV. Furthermore, we showed that rs2705511 and rs1982809 significantly modified OS, while rs9288952 tend to be associated with patients' survival. Our results indicate that BTLA polymorphic variants may be considered low penetrating risk factors for NSCLC especially in never-smokers, and in females, and are associated with OS of NSCLC patients.

Emerging roles of circRNAs in leukemia and the clinical prospects: An update.

Circular RNAs (circRNAs) are a new category of endogenous non-protein coding RNAs (ncRNAs), and show the characteristics of high conservation, stability, and tissue specificity. Due to rapid advances in next-generation sequencing and transcriptome profiling technologies, circRNAs have been widely discovered in many organisms and participated in the development and progress of a variety of diseases. As a type of molecular sponge, circRNAs mainly absorb micro RNAs competitively and interplay with RNA-binding proteins to modulate the splicing as well as transcription of target genes. This review is based on a literature search using the Medline database. Search terms used were "circular RNAs and leukemia," "circRNAs and leukemia," "circRNAs and acute lymphoblastic leukemia," "circRNAs and chronic lymphoblastic leukemia," "circRNAs and acute myeloid leukemia," "circRNAs and chronic myeloid leukemia," and "circRNAs, biomarker, and hematological system." CircRNAs have been proven as potential biomarkers and therapeutic targets in a variety of tumors. Recent research has found that circRNAs aberrantly exist in hematological cancers, especially leukemia, and are significantly associated with the incidence, progress, and metastasis of diseases as well as the prognosis of patients. The current work summarizes the latest findings on circRNAs in various types of leukemia, aiming to propose prospective therapies and new drug screening methods for the treatment of leukemia.

A survival prediction model and nomogram based on immune-related gene expression in chronic lymphocytic leukemia cells.

There are many different chronic lymphoblastic leukemia (CLL) survival prediction models and scores. But none provide information on expression of immune-related genes in the CLL cells. We interrogated data from the Gene Expression Omnibus database (GEO, GSE22762; Number = 151; training) and International Cancer Genome Consortium database (ICGC, CLLE-ES; Number = 491; validation) to develop an immune risk score (IRS) using Least absolute shrinkage and selection operator (LASSO) Cox regression analyses based on expression of immune-related genes in CLL cells. The accuracy of the predicted nomogram we developed using the IRS, Binet stage, and del(17p) cytogenetic data was subsequently assessed using calibration curves. A survival model based on expression of 5 immune-related genes was constructed. Areas under the curve (AUC) for 1-year survivals were 0.90 (95% confidence interval, 0.78, 0.99) and 0.75 (0.54, 0.87) in the training and validation datasets, respectively. 5-year survivals of low- and high-risk subjects were 89% (83, 95%) vs. 6% (0, 17%; p < 0.001) and 98% (95, 100%) vs. 92% (88, 96%; p < 0.001) in two datasets. The IRS was an independent survival predictor of both datasets. A calibration curve showed good performance of the nomogram. In vitro, the high expression of CDKN2A and SREBF2 in the bone marrow of patients with CLL was verified by immunohistochemistry analysis (IHC), which were associated with poor prognosis and may play an important role in the complex bone marrow immune environment. The IRS is an accurate independent survival predictor with a high C-statistic. A combined nomogram had good survival prediction accuracy in calibration curves. These data demonstrate the potential impact of immune related genes on survival in CLL.

A Human iPSC Model Reveals Specific Upregulation of Linker Histone H1-0 in ETV6::RUNX1+ Preleukemia and B Cell Precursor Acute Lymphoblastic Leukemia

The ETV6::RUNX1 translocation is the most common chromosomal rearrangement associated with pediatric B cell precursor acute lymphoblastic leukemia (BCP-ALL). As previously shown, ETV6::RUNX1 leads to a distinct clinically silent preleukemic state in utero that manifests in impaired early B cell differentiation. Acquisition of secondary oncogenic mutations gives rise to overt leukemia in 0.2% of children carrying ETV6::RUNX1. In-depth characterization of ETV6::RUNX1+ preleukemia is essential to gain an understanding of leukemic transformation and develop preventive strategies. Here, we identify dysregulation of the epigenetic regulator H1-0 in ETV6::RUNX1+ preleukemia and BCP-ALL. Utilizing CRISPR/Cas9 genome editing, we generated monoclonal ETV6::RUNX1+ human induced pluripotent stem cell (hiPSC) lines with stable ETV6::RUNX1 expression driven by the endogenous ETV6 promoter. To assess the preleukemic setting mediated by ETV6::RUNX1, we performed RNA sequencing (RNA-seq) analysis of ETV6::RUNX1+ and matching wild-type (WT) hiPSCs of two donors. We identified 20 differentially expressed genes in ETV6::RUNX1+ hiPSCs of both donors compared to the respective WT hiPSCs (up: 15, down: 5, absolute fold change ≥ 2, p ≤ 0.05). Interestingly, our RNA-seq analysis revealed consistent upregulation of H1-0 (mean: 2.43-fold), a linker histone involved in chromatin remodeling, cell proliferation and differentiation. Elevated H1-0 levels in the ETV6::RUNX1+ hiPSC lines were confirmed by quantitative real-time PCR and immunoblot (Figure 1A). To determine H1-0 levels in ETV6::RUNX1+ B cell precursors, we analyzed published gene expression microarray and RNA-seq data sets. Both ETV6::RUNX1+ CD34+ cells (n = 10) derived from umbilical cord blood (Polak et al. Haematologica 2019) and ETV6::RUNX1+ CD34+CD19+ cells (n = 4) generated by hematopoietic differentiation of hiPSCs (Böiers et al. Developmental Cell 2018) showed significant upregulation of H1-0 compared to the respective control cells lacking ETV6::RUNX1 (unpaired t-test, p = 0.0087 and p = 0.0076, respectively). Additionally, we differentiated ETV6::RUNX1+ and WT hiPSCs into CD34+CD45+ hematopoietic progenitor cells and performed single-cell RNA-seq analysis. We identified coexpression of H1-0 with cyclin-dependent kinase inhibitor genes CDKN1A and CDKN1B, indicating a slow or non-cycling phenotype of H1-0+ cells. Next, we examined H1-0 mRNA levels in gene expression data sets of pediatric and adult leukemia patients (St. Jude PeCan database (n = 876), Jerchel et al. Haematologica 2018 (n = 654), MILE study (n = 1299)). H1-0 expression was found to be significantly elevated in ETV6::RUNX1+ BCP-ALL patient samples compared to other leukemia entities, especially TCF3::PBX1+ ALL (ordinary one-way ANOVA, p ≤ 0.0001), KMT2A rearranged ALL (p ≤ 0.0001) and chronic lymphoblastic leukemia (CLL, p ≤ 0.0001). Strikingly, H1-0 expression was 22.37-fold lower in CLL compared to ETV6::RUNX1+ BCP-ALL (Figure 1B). This observation prompted us to investigate H1-0 expression in B cell developmental stages using published data sets. Interestingly, we found that H1-0 levels decrease during early B cell development in both normal bone marrow (Black et al. Nucleic Acids Research 2018) and umbilical cord blood-derived cells (Novershtern et al. Cell 2011). Additionally, analysis of single-cell RNA-seq data of early B cell precursors derived from healthy bone marrow (Mehtonen et al. Genome Medicine 2020) and fetal liver (Developmental Cell Atlas Newcastle University) revealed that relative numbers of H1-0 expressing cells per differentiation state decreased along the pseudotime B cell development trajectory. Taken together, our study identifies H1-0 upregulation as a specific alteration present already at the ETV6::RUNX1+ preleukemic state. High H1-0 expression was associated with slower proliferation and increased stemness. This phenotype might explain the persistence of in utero generated preleukemic ETV6::RUNX1+ clones residing in the bone marrow of approximately 5% of healthy children for years. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Anti-CD70 Monoclonal Antibodies Show Efficacy in Preclinical Models for Chronic Lymphoblastic Leukemia

Although therapy for chronic lymphocytic leukemia (CLL) has improved significantly with combination therapies that include ibrutinib and/or anti-B cell mAbs, resistance can develop, and patients can discontinue therapy for various reasons. Therefore, effective, alternative therapies are needed. CD70, a TNF family member, and its receptor, CD27, are mainly expressed on B and T lymphocytes. Different from normal B cells that only transiently express CD70 upon activation, CLL B cells stably express the molecule at relatively high levels, suggesting a fundamental biologic abnormality and adding another example of the dysregulation of the immune system in CLL. Here we investigated the role of CD70 in CLL using blocking anti-CD70 mAbs in preclinical mouse models of the disease. We first confirmed stable expression of CD27 and CD70 in patient samples as well as on murine TCL1 CLL B cells, including TCL1-192 cells, a set of primary murine leukemic cells within which > 95% cells react to autoantigen stimulation via the B-cell receptor (BCR). Comparing with healthy BM, we found that primary CLL patient B cells express significantly higher levels of the CD27 - CD70 receptor-ligand pair at both mRNA and protein levels, as do TCL-1 mice. We next investigated the effects of an anti-mouse CD70 effector function silenced/blocking mAb, FcD-FR70, on TCL1-192 CLL cells in vitro and in vivo in preclinical models. In vitro, treating TCL1 CLL cells with FcD-FR70 significantly inhibited CD40L-induced CLL cell growth. In vivo, twice weekly 10 mg/kg treatments with FcD-FR70 antibody significantly reduced the number of murine CLL B cells in spleens, lymph nodes and blood. The levels of reduction were comparable to those observed in anti-CD19 mAb or ibrutinib treated mice. There was a significantly better treatment outcome when therapy was initiated earlier. Moreover, the level of soluble mouse CD27, the product of CD27 cleavage after interaction with CD70, was significantly reduced in sera of animals treated with FcD-FR70. Notably, FcD-FR70, but not anti-CD19 mAb or ibrutinib, significantly reduced CD70 membrane levels. CD27 expression levels were not altered by all treatments. As CD27 and CD70 are inversely expressed in B cells, due to tight control at the transcriptional and posttranslational levels, this result strongly suggests preferential depletion of malignant cells with the high CD70 expression, which also affects levels of soluble CD27. We then performed bulk RNAseq analysis in CD19+CD5+ splenic cells from mice treated with control or FcD-FR70 antibody. Genes differing by >1.5 fold (P < 0.01) by FcD-FR70 treatment between the two groups included upregulated cell cycle regulator cyren and interferon gamma inducible protein IfI211, as well as downregulated IL10, CCR7, PIM3, PIM1, TGFb3, TNFaip3, JunB and CD69. Importantly, IPA analysis of the altered genes suggested inhibition of TLR7, TLR9, TNF, NFKB, IL4, BCR, BTK, and CD40L signaling pathways. Thus, FcD-FR70 treatment blocked key survival pathways, inducing death of CLL B cells. Finally, we studied the effect of anti-CD70 antibodies on CLL B-T cell interactions. Adding the FcD-FR70 mAb to an allo-MLR significantly blocked T cell proliferation and survival induced by TCL1-192 leukemic B cells (P < 0.01). We then tested the effects of 41D12-FcD, a blocking mAb in human IgG1 format that binds with picomolar affinity to human CD70, in a primary CLL patient derived xenograft model. This revealed a significant reduction of malignant cells in the spleen, BM, and peritoneum (P < 0.001). Interestingly, the efficacy of the blocking 41D12-FcD antibody was associated with a significant decrease of human T cells, suggesting that the antibody directly blocked the proliferation of malignant B cells and also inhibited T cell-CLL cell interactions. Notably, no significant changes were seen in the differentiation of T cells, expression of CTLA4, PD1, PD-L1, and levels of Th1/Th2 cytokines after anti-CD70 treatment. Overall, data from our preclinical models suggest targeting CD70 in CLL alters BCR, CD40L, IL4, and TNF receptor signaling and delays CLL disease progression. Although the involvement of CD70 and the relationships between CLL B cells and non-malignant cells needs further study, the availability of clinical stage therapeutic anti-CD70 antibodies supports the use anti-CD70 immunotherapy in combination therapies in CLL.

Efficacy of tixagevimab-cilgavimab in preventing SARS-CoV-2 for patients with B-cell malignancies

COVID-19 still represents a major issue for patients with lymphoid malignancies, especially those on therapy, because of immune suppression and suboptimal responses to vaccination. Davis et al report on their experience with double dose tixagevimab-cilgavimab preexposure prophylaxis in a cohort of 251 patients with chronic lymphocytic leukemia, B-cell lymphomas, multiple myeloma, or B-cell acute lymphoblastic leukemia, 63% of whom had received 3 doses of the SARS-CoV-2 vaccine. Breakthrough infections within 3 months occur despite passive immunization, affecting 11% in this series; however, hospitalization rates are low, and mortality was avoided, suggesting benefit from this strategy.

Loop CD20/CD19 CAR-T cells eradicate B-cell malignancies efficiently.

CD19 chimeric antigen receptor (CAR) T cells have shown robust efficacy in relapsed and refractory acute lymphoblastic leukemia (R/R ALL), but compromising result in chronic lymphoblastic leukemia (CLL) and non-Hodgkin's lymphoma (NHL). CD19 relapse and the lack of CAR-T cell persistence which result in treatment failure are considerable obstacles to overcome. CAR-T targeting CD20 is an option for salvaging CD19 CAR-T failure. Previous studies have established variant structures of bispecific CAR-T which could avoid antigen-loss and immune escape. Here, we constructed tandem and loop CAR structures targeting both CD19 and CD20 antigen. Bispecific CAR-T cells could eliminate either CD19 or CD20 negative lymphoma cells, suggesting they exhibited dual antigen targeting of CD19 and CD20. By comparing the efficiency of four bispecific CAR modified T cells, it was found that loop2019 CAR was the best structure among them to eradicate lymphoma cell lines and patients' primary lymphoma or CLL cells in a very low dose in vitro and prolong the survival time dramatically in lymphoma xenograft mice model. These data highlighted the potential of loop2019 CAR-T in clinical treatment.

Leukaemia Identification based on Texture Analysis of Microscopic Peripheral Blood Images using Feed-Forward Neural Network

ABSTRACT&#x0D; Leukaemia is very dangerous because it includes liquid tumour that it cannot be seen physically and is difficult to detect. Alternative detection of Leukaemia using microscopy can be processed using a computing system. Leukemia disease can be detected by microscopic examination. Microscopic test results can be processed using machine learning for classification systems. The classification system can be obtained using Feed-Forward Neural Network. Extreme Learning Machine (ELM) is a neural network that has a feedforward structure with a single hidden layer. ELM chooses the input weight and hidden neuron bias at random to minimize training time based on the Moore Penrose Pseudoinverse theory. The classification of Leukaemia is based on microscopic peripheral blood images using ELM. The classification stages consist of pre-processing, feature extraction using GLRLM, and classification using ELM. This system is used to classify Leukaemia into three classes, that is acute lymphoblastic Leukaemia, chronic lymphoblastic Leukaemia, and not Leukaemia. The best results were obtained in ten hidden nodes with an accuracy of 100%, a precision of 100%, a withdrawal of 100%.

Acute subdural hematoma due to acute myeloid leukemia and B-cell lymphoma

Chloroma, is an rare malignant tumor characterized by the extramedullary blast proliferation of myeloid lineages that subsequently change the normal architecture of surrounding tissues. Because this is very rare disease, primary central nervous system myeloid sarcoma has not been widely reported. Myeloid sarcoma, as a group of heterogenetic diseases, presents with differential clinical and histological pictures depending upon the primary affected site. We are presenting a 77-year-old gentleman, with past medical history of chronic lymphoblastic leukemia that was not on any treatment and who presented with complaints of lethargy, disorientation, and weakness. CT scan of the head showed left-sided subdural hematoma with significant mass effect with left-to-right shift. Craniotomy and hematoma evacuation was performed and hematoma and subdural lesion was sent for pathological evaluation. Histologic examination of the hematoma revealed solid sheets of small-medium sized atypical hematopoietic cells that was imbedded in an acute and chronic hematoma. To confirm the final diagnosis, flow cytometry was performed and showed two neoplastic clones, consistent with acute myeloid leukemia with monocytic differentiation and a separate mature B-cell lymphoma. To our knowledge we are presenting the first case in the literature of the Intracranial acute myeloid leukemia/myeloid sarcoma and B-cell lymphoma that was found in the same brain lesion.

Features of diagnosis and treatment of multiple myeloma

Multiple myeloma is a variant of chronic lymphoblastic leukemia in which there is an uncontrolled formation of plasma cells due to DNA mutation of B lymphocytes with their subsequent colonization in bone tissue. Increased production of immunoglobulins by plasma cells leads to the disruption of the immune system, and damage to bone tissue causes persistent bone pain and frequent bone destruction. Since the produced pathological immunoglobulins are metabolized by the kidneys, one of the clinical manifestations of the disease is renal amyloidosis. Nonspecific clinical symptoms of multiple myeloma also include peripheral neuropathy, spinal cord compression syndrome, and anemia. The disease is quite rare; it is detected mainly in elderly age, and there is some prevalence of this pathology in people of the Negroid race. The origin of the disease remains not fully understood; a number of authors tend to believe that it is based on a genetic predisposition and possible influence of radiation. Diagnosis of multiple myeloma is based on the detection of an increased content of plasma cells and the detection of pathological immunoglobulins. The disease is considered incurable; symptomatic treatment is aimed at decreasing the production of plasma cells and reducing their production of protein substances.