Extracellular DNA traps released by acute promyelocytic leukemia cells through autophagy

Ma, Ruishuang; Li, T; Cao, Mingming; Yu, Shaohua; Wu, Xiaoming; Zhao, Lu; Yao, Zhipeng; Zhang, Y; Fang, Shaohong; Deng, Ruijuan; Novakovic, Valerie A.; Bi, Yang; Kou, Junjie; Yu, Bo; Yang, Sichang; Wang, J; Zhou, Jin; Shi, Jialan

doi:10.1038/cddis.2016.186

Cited by 33 publications

(34 citation statements)

References 59 publications

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“…Therefore, we aimed to test whether these NETs‐related processes are activated in differentiated HL‐60 cells post stimulation with PMA or CI. It has been previously suggested that autophagy is necessary for PMA‐induced NETosis and that diminished NETs formation by HL‐60 cells results from defective autophagy . We were able to confirm that PMA stimulates formation of autophagosomes by PBN, which we also observed in DMF‐dHL‐60 cells.…”

Section: Discussionsupporting

confidence: 86%

“…It has been previously suggested that autophagy is necessary for PMA-induced NETosis and that diminished NETs formation by HL-60 cells results from defective autophagy. 4,39 We were able to confirm that PMA stimulates formation of autophagosomes by PBN, which we also observed in DMF-dHL-60 cells. Moreover, we observed increased citH3 levels in unstimulated cells differentiated with DMF or DMSO, but not with ATRA, when compared to control HL-60 cells.…”

Section: Discussionsupporting

confidence: 78%

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The influence of agents differentiating HL‐60 cells toward granulocyte‐like cells on their ability to release neutrophil extracellular traps

et al. 2018

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Studies on neutrophil extracellular traps (NETs) are challenging as neutrophils live shortly and easily become activated. Thus, availability of a cell line model closely resembling the functions of peripheral blood neutrophils would be advantageous. Our purpose was to find a compound that most effectively differentiates human promyelocytic leukemia (HL-60) cells toward granulocyte-like cells able to release NETs. HL-60 cells were differentiated with all-trans retinoic acid (ATRA), dimethyl sulfoxide (DMSO) or dimethylformamide (DMF) and stimulated with phorbol 12-myristate 13-acetate (PMA) or calcium ionophore A23187 (CI). Cell differentiation, phagocytosis and calcium influx were analyzed by flow cytometry. Reactive oxygen species production and NETs release were measured fluorometrically and analyzed microscopically. LC3-II accumulation and histone 3 citrullination were analyzed by western blot. ATRA most effectively differentiated HL-60 cells toward granulocyte-like cells. ATRA-dHL-60 cells released NETs only upon PMA stimulation, DMSO-dHL-60 cells only post CI stimulation, while DMF-dHL-60 cells formed NETs in response to both stimuli. Oxidative burst was induced in ATRA-, DMSO- and DMF-dHL-60 cells post PMA stimulation and only in DMF-dHL-60 cells post CI stimulation. Increased histone 3 citrullination was observed in stimulated DMSO- and DMF-, but not in ATRA-dHL-60 cells. The calcium influx was diminished in ATRA-dHL-60 cells. Significant increase in autophagosomes formation was observed only in PMA-stimulated DMF-dHL-60 cells. Phagocytic index was higher in ATRA-dHL-60 cells than in control, DMSO- and DMF-dHL-60 cells. We conclude that ATRA, DMSO and DMF differentiate HL-60 in different mechanisms. DMF is the best stimulus for HL-60 cell differentiation for NETs studies.

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Section: Discussionsupporting

confidence: 86%

Section: Discussionsupporting

confidence: 78%

The influence of agents differentiating HL‐60 cells toward granulocyte‐like cells on their ability to release neutrophil extracellular traps

et al. 2018

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“…Interesting, in vitro NET generation is impaired in older adults in response to LPS and IL-8 ( Hazeldine et al, 2014 ), and reduced ATG5 gene expression ( Vieira da Silva Pellegrina et al, 2015 ; Xu et al, 2017 ). Consistent with this, inhibition of autophagy by pharmacological inhibitors or by small interfering RNA against ATG7 attenuated LC3 autophagy formation and significantly decreased NET generation in promyelocytes ( Ma et al, 2016 ). Furthermore, knockdown of the inhibitor of PI3K/AKT/mTOR pathway, autophagy inducer, PTEN in HL-60 differentiated neutrophil-like cells resulted in diminished generation of NETs upon stimulation with PMA ( Teimourian and Moghanloo, 2015 ).…”

Section: Autophagy and Net Formationsupporting

confidence: 56%

Autophagy in Neutrophils: From Granulopoiesis to Neutrophil Extracellular Traps

Skendros

Mitroulis

Ritis

2018

Front. Cell Dev. Biol.

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Autophagy is an evolutionarily conserved intracellular degradation system aiming to maintain cell homeostasis in response to cellular stress. At physiological states, basal or constitutive level of autophagy activity is usually low; however, it is markedly up-regulated in response to oxidative stress, nutrient starvation, and various immunological stimuli including pathogens. Many studies over the last years have indicated the implication of autophagy in a plethora of cell populations and functions. In this review, we focus on the role of autophagy in the biology of neutrophils. Early studies provided a link between autophagy and neutrophil cell death, a process essential for resolution of inflammation. Since then, several lines of evidence both in the human system and in murine models propose a critical role for autophagy in neutrophil-driven inflammation and defense against pathogens. Autophagy is essential for major neutrophil functions, including degranulation, reactive oxygen species production, and release of neutrophil extracellular traps. Going back to neutrophil generation in the bone marrow, autophagy plays a critical role in myelopoiesis, driving the differentiation of progenitor cells of the myeloid lineage toward neutrophils. Taken together, in this review we discuss the functional role of autophagy in neutrophils throughout their life, from their production in the bone marrow to inflammatory responses and NETotic cell death.

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“…To distinguish the effect of ATO on ETosis and apoptosis, lactadherin and propidium iodide (PI) were used to stain NB4 cells 24 , 25 . In ETotic cells, the chromatin expands while the cytoplasmic membrane remains intact.…”

Section: Resultsmentioning

confidence: 99%

“…Formed mainly by immune cells, ETs can also be released by human leukemia cells when exposed to microorganisms, reactive oxygen species (ROS) or tunicamycin 22 , 23 . Studies from our laboratory have shown that APL cells from patients can also undergo this novel cell death process, producing ETs through autophagy 24 , 25 , that has been linked to the mechanisms of ATO. More interestingly, ATRA promotes ETosis leading to procoagulant promyelocytic extracellular chromatin 25 .…”

Section: Introductionmentioning

confidence: 99%

Arsenic trioxide promoting ETosis in acute promyelocytic leukemia through mTOR-regulated autophagy

Zhang

et al. 2018

Cell Death Dis

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Despite the high efficacy and safety of arsenic trioxide (ATO) in treating acute promyelocytic leukemia (APL) and eradicating APL leukemia-initiating cells (LICs), the mechanism underlying its selective cytotoxicity remains elusive. We have recently demonstrated that APL cells undergo a novel cell death program, termed ETosis, through autophagy. However, the role of ETosis in ATO-induced APL LIC eradication remains unclear. For this study, we evaluated the effects of ATO on ETosis and the contributions of drug-induced ETosis to APL LIC eradication. In NB4 cells, ATO primarily increased ETosis at moderate concentrations (0.5–0.75 μM) and stimulated apoptosis at higher doses (1.0–2.0 μM). Furthermore, ATO induced ETosis through mammalian target of rapamycin (mTOR)-dependent autophagy, which was partially regulated by reactive oxygen species. Additionally, rapamycin-enhanced ATO-induced ETosis in NB4 cells and APL cells from newly diagnosed and relapsed patients. In contrast, rapamycin had no effect on apoptosis in these cells. We also noted that PML/RARA oncoprotein was effectively cleared with this combination. Intriguingly, activation of autophagy with rapamycin-enhanced APL LIC eradication clearance by ATO in vitro and in a xenograft APL model, while inhibition of autophagy spared clonogenic cells. Our current results show that ATO exerts antileukemic effects at least partially through ETosis and targets LICs primarily through ETosis. Addition of drugs that target the ETotic pathway could be a promising therapeutic strategy to further eradicate LICs and reduce relapse.

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Extracellular DNA traps released by acute promyelocytic leukemia cells through autophagy

Cited by 33 publications

References 59 publications

The influence of agents differentiating HL‐60 cells toward granulocyte‐like cells on their ability to release neutrophil extracellular traps

The influence of agents differentiating HL‐60 cells toward granulocyte‐like cells on their ability to release neutrophil extracellular traps

Autophagy in Neutrophils: From Granulopoiesis to Neutrophil Extracellular Traps

Arsenic trioxide promoting ETosis in acute promyelocytic leukemia through mTOR-regulated autophagy

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