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Cancer Research Apr 2024Impairing the BET family coactivator BRD4 with small-molecule inhibitors (BETi) showed encouraging preclinical activity in treating acute myeloid leukemia (AML)....
UNLABELLED
Impairing the BET family coactivator BRD4 with small-molecule inhibitors (BETi) showed encouraging preclinical activity in treating acute myeloid leukemia (AML). However, dose-limiting toxicities and limited clinical activity dampened the enthusiasm for BETi as a single agent. BETi resistance in AML myeloblasts was found to correlate with maintaining mitochondrial respiration, suggesting that identifying the metabolic pathway sustaining mitochondrial integrity could help develop approaches to improve BETi efficacy. Herein, we demonstrated that mitochondria-associated lactate dehydrogenase allows AML myeloblasts to utilize lactate as a metabolic bypass to fuel mitochondrial respiration and maintain cellular viability. Pharmacologically and genetically impairing lactate utilization rendered resistant myeloblasts susceptible to BET inhibition. Low-dose combinations of BETi and oxamate, a lactate dehydrogenase inhibitor, reduced in vivo expansion of BETi-resistant AML in cell line and patient-derived murine models. These results elucidate how AML myeloblasts metabolically adapt to BETi by consuming lactate and demonstrate that combining BETi with inhibitors of lactate utilization may be useful in AML treatment.
SIGNIFICANCE
Lactate utilization allows AML myeloblasts to maintain metabolic integrity and circumvent antileukemic therapy, which supports testing of lactate utilization inhibitors in clinical settings to overcome BET inhibitor resistance in AML. See related commentary by Boët and Sarry, p. 950.
Topics: Humans; Animals; Mice; Nuclear Proteins; Transcription Factors; Lactic Acid; Cell Line, Tumor; Leukemia, Myeloid, Acute; Lactate Dehydrogenases; Bromodomain Containing Proteins; Cell Cycle Proteins
PubMed: 38285895
DOI: 10.1158/0008-5472.CAN-23-0291 -
Blood Advances Apr 2024Congenital neutropenia (CN) is a genetic disorder characterized by persistent or intermittent low peripheral neutrophil counts, thus increasing susceptibility to...
Congenital neutropenia (CN) is a genetic disorder characterized by persistent or intermittent low peripheral neutrophil counts, thus increasing susceptibility to bacterial and fungal infections. Various forms of CN, caused by distinct genetic mutations, exhibit differential responses to granulocyte colony-stimulating factor (G-CSF) therapy, with the underlying mechanisms not fully understood. This study presents an in-depth comparative analysis of clinical and immunological features in 5 CN patient groups (severe congenital neutropenia [SCN]1, SCN3, cyclic neutropenia [CyN], warts, hypogammaglobulinaemia, infections and myelokathexis [WHIM], and Shwachman-Bodian-Diamond Syndrome [SBDS]) associated with mutations in ELANE, HAX1, CXCR4, and SBDS genes. Our analysis led to the identification of 11 novel mutations in ELANE and 1 each in HAX1, CXCR4, and G6PC3 genes. Investigating bone marrow (BM) granulopoiesis and blood absolute neutrophil count after G-CSF treatment, we found that SCN1 and SCN3 presented with severe early-stage disruption between the promyelocyte and myelocyte, leading to a poor response to G-CSF. In contrast, CyN, affected at the late polymorphonuclear stage of neutrophil development, showed a strong G-CSF response. WHIM, displaying normal neutrophil development, responded robustly to G-CSF, whereas SBDS, with moderate disruption from the early myeloblast stage, exhibited a moderate response. Notably, SCN1 uniquely impeded neutrophil development, whereas SCN3, CyN, WHIM, and SBDS also affected eosinophils and basophils. In addition, SCN1, SCN3, and CyN presented with elevated serum immunoglobulins, increased BM plasma cells, and higher A Proliferation-Inducing Ligand levels. Our study reveals a strong correlation between the stage and severity of granulocyte development disruption and the efficacy of G-CSF therapy.
Topics: Humans; Granulocyte Colony-Stimulating Factor; Mutation; Eosinophils; Adaptor Proteins, Signal Transducing; Congenital Bone Marrow Failure Syndromes; Neutropenia
PubMed: 38286463
DOI: 10.1182/bloodadvances.2023012171 -
Nature Structural & Molecular Biology Jun 2024Despite the importance of citrullination in physiology and disease, global identification of citrullinated proteins, and the precise targeted sites, has remained...
Despite the importance of citrullination in physiology and disease, global identification of citrullinated proteins, and the precise targeted sites, has remained challenging. Here we employed quantitative-mass-spectrometry-based proteomics to generate a comprehensive atlas of citrullination sites within the HL60 leukemia cell line following differentiation into neutrophil-like cells. We identified 14,056 citrullination sites within 4,008 proteins and quantified their regulation upon inhibition of the citrullinating enzyme PADI4. With this resource, we provide quantitative and site-specific information on thousands of PADI4 substrates, including signature histone marks and transcriptional regulators. Additionally, using peptide microarrays, we demonstrate the potential clinical relevance of certain identified sites, through distinct reactivities of antibodies contained in synovial fluid from anti-CCP-positive and anti-CCP-negative people with rheumatoid arthritis. Collectively, we describe the human citrullinome at a systems-wide level, provide a resource for understanding citrullination at the mechanistic level and link the identified targeted sites to rheumatoid arthritis.
Topics: Humans; Citrullination; Protein-Arginine Deiminase Type 4; Arthritis, Rheumatoid; Citrulline; HL-60 Cells; Proteomics; Protein-Arginine Deiminases; Substrate Specificity; Synovial Fluid
PubMed: 38321148
DOI: 10.1038/s41594-024-01214-9 -
Digital Health 2024Acute leukemia (AL) is a life-threatening malignant disease that occurs in the bone marrow and blood, and is classified as either acute myeloid leukemia (AML) or acute...
OBJECTIVE
Acute leukemia (AL) is a life-threatening malignant disease that occurs in the bone marrow and blood, and is classified as either acute myeloid leukemia (AML) or acute lymphoblastic leukemia (ALL). Diagnosing AL warrants testing methods, such as flow cytometry, which require trained professionals, time, and money. We aimed to develop a model that can classify peripheral blood images of 12 cell types, including pathological cells associated with AL, using artificial intelligence.
METHODS
We acquired 42,386 single-cell images of peripheral blood slides from 282 patients (82 with AML, 40 with ALL, and 160 with immature granulocytes).
RESULTS
The performance of EfficientNet-V2 (B2) using the original image size exhibited the greatest accuracy (accuracy, 0.8779; precision, 0.7221; recall, 0.7225; and F1 score, 0.7210). The next-best accuracy was achieved by EfficientNet-V1 (B1), with a 256 × 256 pixels image. F1 score was the greatest for EfficientNet-V1 (B1) with the original image size. EfficientNet-V1 (B1) and EfficientNet-V2 (B2) were used to develop an ensemble model, and the accuracy (0.8858) and F1 score (0.7361) were improved. The classification performance of the developed ensemble model for the 12 cell types was good, with an area under the receiver operating characteristic curve above 0.9, and F1 scores for myeloblasts and lymphoblasts of 0.8873 and 0.8006, respectively.
CONCLUSIONS
The performance of the developed ensemble model for the 12 cell classifications was satisfactory, particularly for myeloblasts and lymphoblasts. We believe that the application of our model will benefit healthcare settings where the rapid and accurate diagnosis of AL is difficult.
PubMed: 38812848
DOI: 10.1177/20552076241258079 -
Revista Medica Del Instituto Mexicano... Sep 2023Inflammatory indexes can reflect the severity of serious diseases such as acute leukemia (AL), which is why they can predict mortality. (Observational Study)
Observational Study
BACKGROUND
Inflammatory indexes can reflect the severity of serious diseases such as acute leukemia (AL), which is why they can predict mortality.
OBJECTIVE
To evaluate the prognostic value of mortality of inflammatory indexes during the remission induction stage in patients with pediatric AL.
MATERIAL AND METHODS
Observational, longitudinal, analytical and retrolective study. Patients aged 0 to 17 years, with a recent and confirmed diagnosis of AL, who had basal (at diagnosis, before the start of treatment) and final (at the end of remission induction, or, in the cases of death, during the period prior to this outcome) complete blood count were included.
RESULTS
We included 78 patients, 67 with acute lymphoblastic leukemia (ALL), and 11 with acute myeloblastic leukemia (AML), with 11 and 2 deaths, respectively. Regarding ALL, no index showed significant cut-off points to distinguish deaths. Concerning AML, the indices whose cut-off points distinguished the patients who died in the basal measurement, were the monocyte-lymphocyte ratio (MLR) ≥ 3.11 (sensitivity [Se] 100%, specificity [Sp] 66.67%, AUC 0.8333, p 0.03), and, at the final measurement, the neutrophil-lymphocyte ratio (NLR) ≥ 1.30 and MLR ≥ 0.57 (both with Se 100% and Sp 88.89%, AUC 1.0, p < 0.00001) and systemic immune index (SII) ≥ 246612 (Se 100%, Sp 88.89%, AUC 0.9444, p < 0.0001). With bivariate analysis, only the latter demonstrated an increase in the risk of mortality (p = 0.02).
CONCLUSIONS
The basal MLR and the final NLR, MLR and SII are prognostic inflammatory indices of mortality in patients with AML undergoing remission induction.
Topics: Humans; Child; Retrospective Studies; Prognosis; Lymphocytes; Acute Disease; Leukemia, Myeloid, Acute; Remission Induction; Inflammation
PubMed: 38016441
DOI: No ID Found -
OncoTargets and Therapy 2023We performed sequential molecular analyses of a 75-year-old woman with -ITD positive acute myeloid leukemia (AML) who had received gilteritinib therapy for 43 months. At...
Gilteritinib Affects the Selection of Dominant Clones in Clonal Hematopoiesis: Sequential Genetic Analysis of an -ITD Positive AML Patient with Long-Term Gilteritinib Therapy.
We performed sequential molecular analyses of a 75-year-old woman with -ITD positive acute myeloid leukemia (AML) who had received gilteritinib therapy for 43 months. At the time of diagnosis, her karyotype was normal; however, -ITD, , and mutations were detected. She received induction therapy with daunorubicin and cytarabine and achieved hematological complete remission (HCR). After attaining HCR, she underwent consolidation therapy with azacytidine or cytarabine, aclarubicin, and granulocyte-colony stimulating factor. However, AML relapsed eight months after the first HCR. -ITD and mutations were persistently positive, and the patient received gilteritinib therapy. Although the -ITD clone was not detected during gilteritinib treatment, a clone harboring monosomy 7 and mutations emerged. Bone marrow examinations at 15, 24, and 32 months after gilteritinib treatment revealed multi-lineage blood cell dysplasia without an increase in myeloblasts. After 33 months of treatment, gilteritinib was discontinued for two months because to ileus development, and the -ITD clone was detected again. Gilteritinib treatment was restarted, and -ITD became negative. Our analysis demonstrated that: (1) hematopoiesis derived from gilteritinib-resistant clones was generated by long-term gilteritinib treatment, and (2) -ITD clones regained clonal dominance in the absence of FLT3 inhibition. These findings suggest that gilteritinib affects the selection of dominant clones during clonal hematopoiesis.
PubMed: 37465589
DOI: 10.2147/OTT.S417137 -
European Journal of Cell Biology Jun 2024We analyzed actin cytoskeleton alterations during NET extrusion by neutrophil-like dHL-60 cells and human neutrophils in the absence of DNase1 containing serum to avoid...
We analyzed actin cytoskeleton alterations during NET extrusion by neutrophil-like dHL-60 cells and human neutrophils in the absence of DNase1 containing serum to avoid chromatin degradation and microfilament disassembly. NET-formation by dHL-60 cells and neutrophils was induced by Ionomycin or phorbol-12-myristat-13-acetate (PMA). Subsequent staining with anti-actin and TRITC-phalloidin showed depolymerization of the cortical F-actin at spatially confined areas, the NET extrusion sites, effected by transient activation of the monooxygenase MICAL-1 supported by the G-actin binding proteins cofilin, profilin, thymosin ß4 and probably the F-actin fragmenting activity of gelsolin and/or its fragments, which also decorated the formed NETs. MICAL-1 itself appeared to be proteolyzed by neutrophil elastase possibly to confine its activity to the NET-extrusion area. The F-actin oxidization activity of MICAL-1 is inhibited by Levosimendan leading to reduced NET-formation. Anti-gasdermin-D immunohistochemistry showed a cytoplasmic distribution in non-stimulated cells. After stimulation the NET-extrusion pore displayed reduced anti-gasdermin-D staining but accumulated underneath the plasma membrane of the remaining cell body. A similar distribution was observed for myosin that concentrated together with cortical F-actin along the periphery of the remaining cell body suggesting force production by acto-myosin interactions supporting NET expulsion as indicated by the inhibitory action of the myosin ATPase inhibitor blebbistatin. Isolated human neutrophils displayed differences in their content of certain cytoskeletal proteins. After stimulation neutrophils with high gelsolin content preferentially formed "cloud"-like NETs, whereas those with low or no gelsolin formed long "filamentous" NETs.
Topics: Humans; Extracellular Traps; Neutrophils; Actin Cytoskeleton; HL-60 Cells; Actins; Gelsolin
PubMed: 38555846
DOI: 10.1016/j.ejcb.2024.151407 -
BMJ Case Reports Jan 2024Myeloid sarcoma is a very rare extramedullary malignant tumour, most often associated with acute myeloid leukaemia. We report the case of a man in his early 20s who...
Myeloid sarcoma is a very rare extramedullary malignant tumour, most often associated with acute myeloid leukaemia. We report the case of a man in his early 20s who presented with chronic headache, raised intracranial pressure and progressive vision loss of 2 years duration with no systemic manifestations. He had a history of myeloid sarcoma of the left thigh 15 years ago, treated with external beam radiotherapy and in complete remission for more than 13 years. However, the progressive blindness remained unexplained for 2 years, and he was eventually diagnosed with isolated meningeal relapse without marrow or systemic involvement. Imaging revealed subarachnoid haemorrhage, diffuse leptomeningeal enhancement and involvement of lower dorsal cord and conus, and cerebrospinal fluid cytology showed myeloid blasts. He was managed with intrathecal chemotherapy and craniospinal irradiation, after which he had mild improvement in vision.
Topics: Male; Humans; Sarcoma, Myeloid; Neoplasm Recurrence, Local; Meningeal Neoplasms; Blindness; Granulocyte Precursor Cells
PubMed: 38191225
DOI: 10.1136/bcr-2023-256821 -
American Journal of Cancer Research 2023Acute myeloblastic leukemia (AML) is the most prevalent form of AML in adults. Despite the availability of various treatment options, including radiotherapy and...
Acute myeloblastic leukemia (AML) is the most prevalent form of AML in adults. Despite the availability of various treatment options, including radiotherapy and chemotherapy, many patients fail to respond to treatment or relapse. Copper is a necessary cofactor for all organisms; however, it turns toxic when concentrations reach a certain threshold maintained by homeostatic systems that have been conserved through evolution. However, the mechanism through which excess copper triggers cell death remains unknown. In this study, data on long non-coding RNAs (lncRNAs) related to cuproptosis were retrieved from publicly available databases. LASSO and univariate and multivariate Cox regression analyses were performed to establish an lncRNA model associated with cuproptosis specific to AML. To investigate the risk model, the Kaplan-Meier curve, principal component analysis, functional enrichment analysis, and nomographs were employed. The underlying clinicopathological characteristics were determined, and drug sensitivity predictions against the model were identified. Six cuproptosis-related lncRNA-based risk models were identified as the independent prognostic factors. By regrouping patients using a model-based method, we were able to more accurately differentiate patients according to their responses to immunotherapy. In addition, prospective compounds targeting AML subtypes have been identified. Using qRT-PCR, we examined the expression levels of six cuproptosis-associated lncRNAs in 30 clinical specimens. The cuproptosis-associated lncRNA risk-scoring model developed herein has implications in monitoring AML prognosis and in the clinical prediction of the response to immunotherapy. Furthermore, we identified and verified the ceRNA of the cuproptosis-related lncRNA HAGLR/miR-326/CDKN2A regulatory axis using bioinformatic tools. HAGLR is highly expressed in AML and AML cell lines. HAGLR inhibition significantly reduced the proliferation of AML cells and promoted apoptosis. Elesclomol promotes the degradation of CDKN2A and inhibits the proliferation of AML cells. Elesclomol combined with si-HAGLR inhibited the AML progression of AML both and .
PubMed: 37818049
DOI: No ID Found -
European Journal of Haematology Jun 2024Acute myeloid leukemia (AML) is distinguished by clonal growth of myeloid precursor cells, which impairs normal hematopoiesis. Minimal residual disease (MRD) refers to... (Review)
Review
Acute myeloid leukemia (AML) is distinguished by clonal growth of myeloid precursor cells, which impairs normal hematopoiesis. Minimal residual disease (MRD) refers to the residual leukemia cells that persist after chemotherapy. Patients who test positive for MRD have a higher likelihood of experiencing a recurrence, regardless of the specific chemotherapy approach used. Multi-parameter flow cytometry (MFC), polymerase chain reaction (PCR), and next-generation sequencing (NGS) are commonly employed techniques for identifying MRD. In the context of AML, patients are frequently monitored for measurable residual disease via multi-parameter flow cytometry (MFC-MRD). In order to explore recent advancements in AML and MRD diagnosis, an extensive search of the PubMed database was conducted, focusing on relevant research in the past 20 years. This review aims to examine various MRD monitoring methods, the optimal time points for assessment, as well as different specimen types used. Additionally, it underscores the significance of MFC-MRD assessment in guiding the treatment of elderly AML.
Topics: Neoplasm, Residual; Humans; Leukemia, Myeloid, Acute; Flow Cytometry; Aged; Disease Management; High-Throughput Nucleotide Sequencing; Prognosis; Immunophenotyping; Age Factors
PubMed: 38342613
DOI: 10.1111/ejh.14187