The Transcriptional Network That Controls Growth Arrest and Macrophage Differentiation in the Human Myeloid Leukemia Cell Line THP-1

Gažová, Iveta; Lefèvre, Lucas; Bush, Stephen J.; Clohisey, Sara; Arner, Peter; Hoon, Michiel de; Severin, Jessica; Duin, Lucas van; Andersson, Robin; Lengeling, Andreas; Hume, David; Summers, Kim M.

doi:10.3389/fcell.2020.00498

Cited by 26 publications

(39 citation statements)

References 90 publications

(172 reference statements)

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“…The aim of this study was to investigate the role of LPS heptose-mediated signaling by H. pylori in human monocyte-like cells, which can be differentiated to a macrophage-like phenotype, exemplified by the model cell line Thp-1 ( 47 , 53 , 67 ). The recently discovered heptose-dependent innate recognition and signaling process ( 68 ) had not been studied specifically in human phagocytic cells.…”

Section: Discussionmentioning

confidence: 99%

Contribution of Heptose Metabolites and the cag Pathogenicity Island to the Activation of Monocytes/Macrophages by Helicobacter pylori

et al. 2021

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The human gastric pathogen Helicobacter pylori activates human epithelial cells by a particular combination of mechanisms, including NOD1 and ALPK1-TIFA activation. These mechanisms are characterized by a strong participation of the bacterial cag pathogenicity island, which forms a type IV secretion system (CagT4SS) that enables the bacteria to transport proteins and diverse bacterial metabolites, including DNA, glycans, and cell wall components, into human host cells. Building on previous findings, we sought to determine the contribution of lipopolysaccharide inner core heptose metabolites (ADP-heptose) in the activation of human phagocytic cells by H. pylori. Using human monocyte/macrophage-like Thp-1 cells and human primary monocytes and macrophages, we were able to determine that a substantial part of early phagocytic cell activation, including NF-κB activation and IL-8 production, by live H. pylori is triggered by bacterial heptose metabolites. This effect was very pronounced in Thp-1 cells exposed to bacterial purified lysates or pure ADP-heptose, in the absence of other bacterial MAMPs, and was significantly reduced upon TIFA knock-down. Pure ADP-heptose on its own was able to strongly activate Thp-1 cells and human primary monocytes/macrophages. Comprehensive transcriptome analysis of Thp-1 cells co-incubated with live H. pylori or pure ADP-heptose confirmed a signature of ADP-heptose-dependent transcript activation in monocyte/macrophages. Bacterial enzyme-treated lysates (ETL) and pure ADP-heptose–dependent activation differentiated monocytes into macrophages of predominantly M1 type. In Thp-1 cells, the active CagT4SS was less required for the heptose-induced proinflammatory response than in epithelial cells, while active heptose biosynthesis or pure ADP-heptose was required and sufficient for their early innate response and NF-κB activation. The present data suggest that early activation and maturation of incoming and resident phagocytic cells (monocytes, macrophages) in the H. pylori–colonized stomach strongly depend on bacterial LPS inner core heptose metabolites, also with a significant contribution of an active CagT4SS.

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Section: Discussionmentioning

confidence: 99%

Contribution of Heptose Metabolites and the cag Pathogenicity Island to the Activation of Monocytes/Macrophages by Helicobacter pylori

et al. 2021

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“…The observed differentiation effect can at least partially explain the decrease in cell cycle progression, as THP-1 cells treated with phorbol myristate acetate for macrophage differentiation only exit G1 phase to a little extent [ 49 ]. CDK4 mRNA and protein levels were unchanged, and the decrease in cell cycle progression would appear to be the result of a downstream target.…”

Section: Discussionmentioning

confidence: 99%

“…CDK4 mRNA and protein levels were unchanged, and the decrease in cell cycle progression would appear to be the result of a downstream target. However, notable downregulation of the CDK4 gene is known to be a later event in macrophage differentiation of THP-1 cells [ 49 ]. We have not identified the exact substances that initiate differentiation or lead to the inhibition of cell cycle progression.…”

Section: Discussionmentioning

confidence: 99%

Platelet Microparticles Protect Acute Myelogenous Leukemia Cells against Daunorubicin-Induced Apoptosis

Cacic

Reikvam

Nordgård

et al. 2021

Cancers

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The role of platelets in cancer development and progression is increasingly evident, and several platelet–cancer interactions have been discovered, including the uptake of platelet microparticles (PMPs) by cancer cells. PMPs inherit a myriad of proteins and small RNAs from the parental platelets, which in turn can be transferred to cancer cells following internalization. However, the exact effect this may have in acute myelogenous leukemia (AML) is unknown. In this study, we sought to investigate whether PMPs could transfer their contents to the THP-1 cell line and if this could change the biological behavior of the recipient cells. Using acridine orange stained PMPs, we demonstrated that PMPs were internalized by THP-1 cells, which resulted in increased levels of miR-125a, miR-125b, and miR-199. In addition, co-incubation with PMPs protected THP-1 and primary AML cells against daunorubicin-induced cell death. We also showed that PMPs impaired cell growth, partially inhibited cell cycle progression, decreased mitochondrial membrane potential, and induced differentiation toward macrophages in THP-1 cells. Our results suggest that this altering of cell phenotype, in combination with decrease in cell activity may offer resistance to daunorubicin-induced apoptosis, as serum starvation also yielded a lower frequency of dead and apoptotic cells when treated with daunorubicin.

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“…THP-1 cells (high differentiation clone 5, from FANTOM4 consortium, passage number 8, provided by Dr. Mark Barnett, The Roslin Institute, United Kingdom) were cultured as described previously (Gazova et al, 2020). The day before the start of the differentiation assay, cells were counted by hemocytometer and between 3 × 10 6 and 5 × 10 6 cells were pelleted and resuspended in 10 ml fresh medium.…”

Section: Thp-1 Cell Line and Differentiation Assaymentioning

confidence: 99%

“…In the parent THP-1 line available from the American Type Culture Collection (ATCC R TIB-202 TM ), around 50% of cells become adherent in response to PMA, indicating that there is considerable phenotypic heterogeneity (see Supplementary Figure 1 in Suzuki et al, 2009). To enable a detailed study of the transcriptomic response to PMA, the FANTOM consortium isolated a clonal line that was highly responsive to PMA and uniformly differentiated into a macrophage like state (Suzuki et al, 2009;Gazova et al, 2020). The karyotype of the clonal line was established by microarray-based comparative genomic hybridization to be 46XY, with minimal chromosomal aberrations (deletions at 6p, 12p, 17p, and on the X chromosome) (Adati et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

CRISPR-Cas9 Editing of Human Histone Deubiquitinase Gene USP16 in Human Monocytic Leukemia Cell Line THP-1

Gažová

Lefèvre

Bush

et al. 2021

Front. Cell Dev. Biol.

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USP16 is a histone deubiquitinase which facilitates G2/M transition during the cell cycle, regulates DNA damage repair and contributes to inducible gene expression. We mutated the USP16 gene in a high differentiation clone of the acute monocytic leukemia cell line THP-1 using the CRISPR-Cas9 system and generated four homozygous knockout clones. All were able to proliferate and to differentiate in response to phorbol ester (PMA) treatment. One line was highly proliferative prior to PMA treatment and shut down proliferation upon differentiation, like wild type. Three clones showed sustained expression of the progenitor cell marker MYB, indicating that differentiation had not completely blocked proliferation in these clones. Network analysis of transcriptomic differences among wild type, heterozygotes and homozygotes showed clusters of genes that were up- or down-regulated after differentiation in all cell lines. Prior to PMA treatment, the homozygous clones had lower levels than wild type of genes relating to metabolism and mitochondria, including SRPRB, encoding an interaction partner of USP16. There was also apparent loss of interferon signaling. In contrast, a number of genes were up-regulated in the homozygous cells compared to wild type at baseline, including other deubiquitinases (USP12, BAP1, and MYSM1). However, three homozygotes failed to fully induce USP3 during differentiation. Other network clusters showed effects prior to or after differentiation in the homozygous clones. Thus the removal of USP16 affected the transcriptome of the cells, although all these lines were able to survive, which suggests that the functions attributed to USP16 may be redundant. Our analysis indicates that the leukemic line can adapt to the extreme selection pressure applied by the loss of USP16, and the harsh conditions of the gene editing and selection protocol, through different compensatory pathways. Similar selection pressures occur during the evolution of a cancer in vivo, and our results can be seen as a case study in leukemic cell adaptation. USP16 has been considered a target for cancer chemotherapy, but our results suggest that treatment would select for escape mutants that are resistant to USP16 inhibitors.

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The Transcriptional Network That Controls Growth Arrest and Macrophage Differentiation in the Human Myeloid Leukemia Cell Line THP-1

Cited by 26 publications

References 90 publications

Contribution of Heptose Metabolites and the cag Pathogenicity Island to the Activation of Monocytes/Macrophages by Helicobacter pylori

Contribution of Heptose Metabolites and the cag Pathogenicity Island to the Activation of Monocytes/Macrophages by Helicobacter pylori

Platelet Microparticles Protect Acute Myelogenous Leukemia Cells against Daunorubicin-Induced Apoptosis

CRISPR-Cas9 Editing of Human Histone Deubiquitinase Gene USP16 in Human Monocytic Leukemia Cell Line THP-1

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