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Development (Cambridge, England) Sep 2021Neutrophils are the most abundant vertebrate leukocytes and they are essential to host defense. Despite extensive investigation, the molecular network controlling...
Neutrophils are the most abundant vertebrate leukocytes and they are essential to host defense. Despite extensive investigation, the molecular network controlling neutrophil differentiation remains incompletely understood. GFI1 is associated with several myeloid disorders, but its role and the role of its co-regulators in granulopoiesis and pathogenesis are far from clear. Here, we demonstrate that zebrafish gfi1aa deficiency induces excessive neutrophil progenitor proliferation, accumulation of immature neutrophils from the embryonic stage, and some phenotypes similar to myelodysplasia syndrome in adulthood. Both genetic and epigenetic analyses demonstrate that immature neutrophil accumulation in gfi1aa-deficient mutants is due to upregulation of cebpa transcription. Increased transcription was associated with Lsd1-altered H3K4 methylation of the cebpa regulatory region. Taken together, our results demonstrate that Gfi1aa, Lsd1 and cebpa form a regulatory network that controls neutrophil development, providing a disease progression-traceable model for myelodysplasia syndrome. Use of this model could provide new insights into the molecular mechanisms underlying GFI1-related myeloid disorders as well as a means by which to develop targeted therapeutic approaches for treatment.
Topics: Animals; CCAAT-Enhancer-Binding Proteins; Cell Differentiation; Cell Proliferation; DNA-Binding Proteins; Embryo, Nonmammalian; Epigenesis, Genetic; Granulocyte Precursor Cells; Hematopoiesis; Histone Demethylases; Neutrophils; Zebrafish; Zebrafish Proteins
PubMed: 34373913
DOI: 10.1242/dev.199516 -
Blood Mar 2021
Topics: Blood Cell Count; Child; Erythrocytes; Female; Granulocyte Precursor Cells; Humans; Leukemia, Myeloid, Acute
PubMed: 33734334
DOI: 10.1182/blood.2020009744 -
The American Journal of Pathology Nov 2021Myelodysplastic syndromes (MDS) are clonal neoplasms of the hematopoietic stem cell that result in aberrant differentiation of hematopoietic lineages caused by a wide...
Myelodysplastic syndromes (MDS) are clonal neoplasms of the hematopoietic stem cell that result in aberrant differentiation of hematopoietic lineages caused by a wide range of underlying genetic, epigenetic, and other causes. Despite the myriad origins, a recognizable MDS phenotype has been associated with miRNA aberrant expression. A model of aberrant myeloid maturation that mimics MDS was generated using a stable knockdown of miR-378-3p. This model exhibited a transcriptional profile indicating aberrant maturation and function, immunophenotypic and morphologic dysplasia, and aberrant growth that characterizes MDS. Moreover, aberrant signal transduction in response to stimulation specific to the stage of myeloid maturation as indicated by CyTOF mass cytometry was similar to that found in samples from patients with MDS. The aberrant signaling, immunophenotypic changes, cellular growth, and colony formation ability seen in this myeloid model could be reversed with azacytidine, albeit without significant improvement of neutrophil function.
Topics: Adult; Aged; Aged, 80 and over; Female; Gene Knockdown Techniques; HL-60 Cells; Humans; Male; MicroRNAs; Middle Aged; Myelodysplastic Syndromes
PubMed: 34364880
DOI: 10.1016/j.ajpath.2021.07.006 -
Natural Product Research Jul 2022Two novel cytochalasans, armochaetoglasin J () and armochaetoglasin K (), along with 14 known analogues (-) were isolated from Their structures were elucidated by...
Two novel cytochalasans, armochaetoglasin J () and armochaetoglasin K (), along with 14 known analogues (-) were isolated from Their structures were elucidated by HRESIMS, NMR spectroscopy, single-crystal X-ray crystallography, and ECD spectra. Armochaetoglasins J and K were found to be inactive against the HepG2, HT-29, K562, HL-60, and A549 cancer cell lines.
Topics: Chaetomium; Crystallography, X-Ray; Cytochalasins; HL-60 Cells; Humans
PubMed: 33487054
DOI: 10.1080/14786419.2021.1872568 -
The EMBO Journal Feb 2021The ability of cells to polarize and move toward external stimuli plays a crucial role in development, as well as in normal and pathological physiology. Migrating cells...
The ability of cells to polarize and move toward external stimuli plays a crucial role in development, as well as in normal and pathological physiology. Migrating cells maintain dynamic complementary distributions of Ras activity and of the phospholipid phosphatidylinositol-3,4-bisphosphate (PI(3,4)P2). Here, we show that lagging-edge component PI(3,4)P2 also localizes to retracting leading-edge protrusions and nascent macropinosomes, even in the absence of phosphatidylinositol 3,4,5-trisphosphate (PIP3). Once internalized, macropinosomes break up into smaller PI(3,4)P2-enriched vesicles, which fuse with the plasma membrane at the rear of the cell. Subsequently, the phosphoinositide diffuses toward the front of the cell, where it is degraded. Computational modeling confirms that this cycle gives rise to stable back-to-front gradient. These results uncover a surprising "reverse-fountain flow" of PI(3,4)P2 that regulates polarity.
Topics: Cell Membrane; Cell Movement; Dictyostelium; HL-60 Cells; Humans; Microtubules; Phosphatidylinositol Phosphates
PubMed: 33586225
DOI: 10.15252/embj.2020105094 -
Journal of Applied Microbiology Aug 2020Utilization of l-asparaginase has been one of the effective strategies for the treatment of lymphoblastic leukaemia. Since the currently used bacterial l-asparaginase...
AIMS
Utilization of l-asparaginase has been one of the effective strategies for the treatment of lymphoblastic leukaemia. Since the currently used bacterial l-asparaginase causes side effects, searching for new enzyme sources has been an active field of research. This study focuses on the characterization of an l-asparaginase-producing fungal strain.
METHODS AND RESULTS
Sarocladium strictum was identified as a potent enzyme-producing strain. For the enhancement of enzyme production, we used two-level factorial design and response surface methodology. The optimization of significant factors showed a 1·84-fold increase in enzyme production. The K and V values of the enzyme were 9·74 mmol l and 8·19 μmol min . The toxicity of the produced l-asparaginase was measured on K562 and HL60 cancer cell lines and L6 as normal cells. The IC values were calculated as 0·4 and 0·5 IU ml for K562 and HL60 respectively and no significant effect was observed in L6. BrdU proliferation and caspase-3 activity assay in l-asparaginase treated HL60 and K562 cells indicated that cell proliferation rates and apoptotic cell death were reduced.
CONCLUSIONS
The cytotoxic properties of the produced fungal enzyme indicated significant growth inhibition in cancer cells while having a little toxic effect on normal cells. The possibility of mass production alongside having suitable cytotoxic and kinetic properties suggest the probable use of the produced l-asparaginase for further researches as a potential chemotherapeutic agent.
SIGNIFICANCE AND IMPACT OF THE STUDY
The lack of significant l-glutaminase activity and promising toxicity properties in S. strictum and the closer evolutionary relativeness of fungi enzymes to human enzymes compared to bacterial enzymes suggest a new source with lower toxicity and anti-cancerous properties, causing less side effect problems.
Topics: Antineoplastic Agents; Apoptosis; Asparaginase; Cell Proliferation; HL-60 Cells; Humans; Hypocreales; K562 Cells; Kinetics; Precursor Cell Lymphoblastic Leukemia-Lymphoma
PubMed: 32119169
DOI: 10.1111/jam.14623 -
Phytochemistry Jan 2022Under OSMAC strategy, seven unreported multioxidized aromatic polyketides, penicanesins A‒G, were discovered from a soil-derived fungus Penicillium canescens along...
Under OSMAC strategy, seven unreported multioxidized aromatic polyketides, penicanesins A‒G, were discovered from a soil-derived fungus Penicillium canescens along with seven known compounds. Their structures were assigned by extensive 1D and 2D NMR spectra in combination with HRESIMS and single crystal X-ray diffraction. Absolute stereochemistry of penicanesins A and D were elucidated by theoretical ECD calculation. (±)-Penicanesins A and B are two pairs of racemic aromatic polyketides with an unusual 6/6/6/6 heterotetracyclic ring core. In bioassay, (-)-penicanesin A shows potential cytotoxicity against human cancer cell lines HL-60 and SW480 with IC values at 13.8 ± 0.6 and 12.5 ± 0.9 μM, respectively, whereas the enantiomer (+)-penicanesin A is less active.
Topics: HL-60 Cells; Humans; Molecular Structure; Penicillium; Polyketides; Soil
PubMed: 34773753
DOI: 10.1016/j.phytochem.2021.113012 -
Chemico-biological Interactions Sep 2022β-lapachone is a 1,2-naphthoquinone of great therapeutic interest that induces cell death by autophagy and apoptosis in tumor cells due to oxidative stress increasing....
β-lapachone is a 1,2-naphthoquinone of great therapeutic interest that induces cell death by autophagy and apoptosis in tumor cells due to oxidative stress increasing. However, its high toxicity in healthy tissues limits its clinical use, which stimulates the planning and synthesis of more selective analogs. The aim of this study was to investigate the cytotoxic activity of three thiosemicarbazones derived from β-lapachone (BV2, BV3 and BV5) in leukemia cells. Cytotoxicity tests were performed on tumor cells (HL-60, K562, K562-Lucena and MOLT-4) and normal peripheral blood mononuclear cells (PBMCs). Subsequently, the mode of action of compounds was accessed by optical microscopy, transmission electron microscopy or fluorescence microscopy. Flow cytometry analysis was performed to investigate apoptosis induction, cell cycle, DNA fragmentation and mitochondrial depolarization. All derivatives inhibited tumor cell growth after 72 h (IC < 10 μM to all cell lines, including the resistant K562-Lucena) with less toxic effects in PBMC cells, being BV3 the most selective compound with selective index (SI) of 275 for HL-60; SI of 40 to K562; SI of 10 for MOLT-4 and SI of 50 to K562-Lucena compared to β-lapachone with SI of 18 to HL-60, SI of 3.7 to K562; SI of 2.4 to MOLT-4 and SI of 0.9 to K562-Lucena. In addition, the K562 or MOLT-4 cells treated with BV3 showed characteristics of both apoptosis and autophagy cell death, mainly by autophagy. These results demonstrate the potent cytotoxic effect of thiosemicarbazones derived from β-lapachone as promising anticancer drugs candidates, encouraging the continuity of in vivo tests.
Topics: Antineoplastic Agents; Apoptosis; HL-60 Cells; Humans; Leukocytes, Mononuclear; Naphthoquinones; Thiosemicarbazones
PubMed: 35934135
DOI: 10.1016/j.cbi.2022.110057 -
Molecular Biology of the Cell Jul 2020The dynamic rearrangement of the actin cytoskeleton is an essential component of many mechanotransduction and cellular force generation pathways. Here we use periodic...
The dynamic rearrangement of the actin cytoskeleton is an essential component of many mechanotransduction and cellular force generation pathways. Here we use periodic surface topographies with feature sizes comparable to those of in vivo collagen fibers to measure and compare actin dynamics for two representative cell types that have markedly different migratory modes and physiological purposes: slowly migrating epithelial MCF10A cells and polarizing, fast-migrating, neutrophil-like HL60 cells. Both cell types exhibit reproducible guidance of actin waves (esotaxis) on these topographies, enabling quantitative comparisons of actin dynamics. We adapt a computer-vision algorithm, optical flow, to measure the directions of actin waves at the submicron scale. Clustering the optical flow into regions that move in similar directions enables micron-scale measurements of actin-wave speed and direction. Although the speed and morphology of actin waves differ between MCF10A and HL60 cells, the underlying actin guidance by nanotopography is similar in both cell types at the micron and submicron scales.
Topics: Actin Cytoskeleton; Actins; Cell Movement; Cytoskeleton; Epithelial Cells; HL-60 Cells; Humans; Image Processing, Computer-Assisted; Mechanical Phenomena; Mechanotransduction, Cellular; Models, Biological
PubMed: 32023172
DOI: 10.1091/mbc.E19-11-0614 -
Journal of Natural Products Jul 2020Five new drimane-type sesquiterpenoids, named proversilins A-E (-), were isolated from the endophytic fungus F210 isolated from the bulbs of . Their structures and...
Five new drimane-type sesquiterpenoids, named proversilins A-E (-), were isolated from the endophytic fungus F210 isolated from the bulbs of . Their structures and absolute configurations were characterized by extensive spectroscopic analysis, including 1D and 2D NMR and HRESIMS data, comparison of experimental and calculated electronic circular dichroism data, and X-ray crystallography. Proversilins B-E (-) represent the first examples of natural products featuring an -acetyl-β-phenylalanine moiety. Compounds and inhibited the growth of HL-60 cells with IC values of 7.3 and 9.9 μM, respectively.
Topics: Aspergillus; Crystallography, X-Ray; HL-60 Cells; Humans; Molecular Structure; Sesquiterpenes; Spectrum Analysis
PubMed: 32628478
DOI: 10.1021/acs.jnatprod.0c00298