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Transcriptional Regulation of HL60 Neutrophil Differentiation: Promyelocyte trajectory vs. Neutrophil trajectory

ABSTRACT: Using the HL60 multipotent promyelocytic leukemia cell line, we performed experiments that lead to two different cell fate attractors by treatments of varying dosage and duration of the differentiation agent all-trans-retinoic acid (ATRA). The dosage and duration combinations corresponding to the two treatments were chosen by means of of cytometric measurements of CD11b, a well-known early differentiation marker, such that the two populations are poised at the same stage of differentiation. Using the selected treatment combinations, one population proceeds toward the terminally differentiated neutrophil attractor while the other population reverts back toward the undifferentiated promyelocytic attractor. We monitored the gene expression changes in the two populations after their respective treatments over a period of five days and identified a set of genes that diverged in their expression; a subset of which promotes neutrophil differentiation while the other represses cell cycle progression. By employing promoter based transcription factor binding site analysis, we found enrichment of transcription factors functionally linked to tumor progression, cell cycle, and development. Keywords: Time course, dose response

ORGANISM(S): Homo sapiens

PROVIDER: GSE13673 | GEO | 2008/11/20

SECONDARY ACCESSION(S): PRJNA110417

REPOSITORIES: GEO

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Transcriptional Regulation of HL60 Neutrophil Differentiation: Promyelocyte trajectory vs. Neutrophil trajectory

Project description:Using the HL60 multipotent promyelocytic leukemia cell line, we performed experiments that lead to two different cell fate attractors by treatments of varying dosage and duration of the differentiation agent all-trans-retinoic acid (ATRA). The dosage and duration combinations corresponding to the two treatments were chosen by means of of cytometric measurements of CD11b, a well-known early differentiation marker, such that the two populations are poised at the same stage of differentiation. Using the selected treatment combinations, one population proceeds toward the terminally differentiated neutrophil attractor while the other population reverts back toward the undifferentiated promyelocytic attractor. We monitored the gene expression changes in the two populations after their respective treatments over a period of five days and identified a set of genes that diverged in their expression; a subset of which promotes neutrophil differentiation while the other represses cell cycle progression. By employing promoter based transcription factor binding site analysis, we found enrichment of transcription factors functionally linked to tumor progression, cell cycle, and development. Keywords: Time course, dose response Untreated HL60 cells are hybrized on the channel 1 with Cy3, ATRA-Treated HL60 cells are hybridized on channel 2 with Cy5. HK60 cells were treated with high dosage/short duration and low dosage/long duration to achieve 55% of CD11b+ cells. These positive cells were then isolated and culture in fresh media and total RNA were isolated for comparison. Cells treated with high dosage/short duration treatment proceed toward the neutrophil attractor while cells treated with the low dosage/long duration treatment revert back toward the promyelocyte attractor.

2008-11-19 | E-GEOD-13673 | biostudies-arrayexpress

Exocytic events in neutrophil-mediated inflammation.

Project description:The purpose of the present study was to elucidate the exocytic events in neutrophil-mediated inflammation. Therefore, we first investigated the expression of genes implicated in these exocytic events in a cell model for neutrophils, the promyelocytic HL-60 cell line. By the addition of 1.3% DMSO during 4.5 days, these cells are differentiated into neutrophil-like dHL-60 cells. Non-differentiated cells served as control. Upon cell differentiation, we i) identified differentially expressed genes implicated in the mechanisms of vesicle transport, and ii) found some of them being upregulated. This upregulated gene expression upon DMSO-maturation points towards a functional role in these cells. Secondly, we confirmed the expression of the selected genes in human neutrophils, isolated from venous blood of 4 different donors.

2014-11-25 | E-MTAB-2107 | biostudies-arrayexpress

Dissecting the process of human neutrophil lineage determination by using alpha-lipoic acid inducing neutrophil deficiency model

Project description:Neutrophil deficiency or differentiation disorder might cause serious diseases, such as neutropenia and neutrophilia. To reveal the molecular mechanism of human neutrophil lineage commitment, single cell RNAseq and bulk cells RNAseq were performed to reveal the characteristics and essential genes of neutrophil lineage determination.

2022-07-20 | GSE184864 | GEO

Functional evaluation of neutrophil of patients at high risk for infection treated with monocyte/ neutrophil apheresis

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Functional evaluation of neutrophil of patients at high risk for infection treated with multiple monocyte/ neutrophil aphaereses

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Single-cell transcriptome profiling reveals neutrophil heterogeneity in homeostasis and infection

Project description:The full neutrophil heterogeneity and differentiation landscape remains incompletely characterized. Here we profiled more than 25,000 mouse differentiating and mature neutrophils using single-cell RNA sequencing to provide a comprehensive transcriptional landscape of neutrophil maturation, function, and fate decision in their steady state and during bacterial infection. Eight neutrophil populations (including the GMP population) were defined by distinct molecular signatures, including a new circulating mature neutrophil population highly expressing interferon-stimulated genes. The three mature peripheral blood neutrophil subsets arise from distinct maturing bone marrow neutrophil subsets, a novel mechanism that highlights the complex and precise regulation of neutrophil production. Neutrophil heterogeneity and differentiation are driven by both known and uncharacterized transcription factors. Neutrophils gradually acquire microbicidal capability as cells traverse the transcriptional landscape, representing an evolved mechanism for fine-tuned regulation of an effective but balanced neutrophil response. Bacterial infection reprograms the genetic architecture of neutrophil population, alters dynamic transition between each subpopulation, and primes neutrophils for augmented functionality without affecting overall heterogeneity. Bacterial infection-induced emergency granulopoiesis is mediated by augmented proliferation of early-stage neutrophil progenitors and accelerated post-mitotic maturation. In summary, single-cell transcriptomics enabled the reconstruction of neutrophil differentiation and maturation trajectories and uncovered neutrophil subpopulations, gene pathways, and regulators of neutrophil function and fate decisions. These data establish a reference model and general framework for studying neutrophil-related disease mechanisms, biomarkers, and therapeutic targets at single-cell resolution.

2020-05-27 | GSE137539 | GEO

A map of gene expression in neutrophil-like cell lines

Project description:We report gene expression data for the human cell lines HL-60 and PLB-985, which serve as models for human neutrophils. We measured gene expression using RNA-Seq for these cell lines both prior and after differentiation into a neutrophil-like state using two differentiation protocols (treatment with DMSO or treatment with DMSO and replacement of serum with Nutridoma).

2018-08-03 | GSE103706 | GEO

Identification and characterization of human CD34+ and CD34dim/- neutrophil-committed progenitors [Bulk RNA-seq]

Project description:Here we report the identification of human CD66b-CD64dimCD115- neutrophil-committed progenitor cells (NCPs) within the SSCloCD45dimCD34+ and CD34dim/- subsets in the bone marrow. NCPs were either CD45RA+ or CD45RA-, and in vitro experiments showed that CD45RA acquisition was not mandatory for their maturation process. NCPs exclusively generated human CD66b+ neutrophils in both in vitro differentiation and in vivo adoptive transfer experiments. Single-cell RNA-sequencing analysis indicated NCPs fell into four clusters, characterized by different maturation stages and distributed along two differentiation routes. One of the clusters was characterized by an interferon-stimulated gene signature, consistent with the reported expansion of peripheral mature neutrophil subsets that express interferon-stimulated genes in diseased individuals. Finally, comparison of transcriptomic and phenotypic profiles indicated NCPs represented earlier neutrophil precursors than the previously described early neutrophil progenitors (eNePs), proNeus and COVID-19 proNeus. Altogether, our data shed light on the very early phases of neutrophil ontogeny.

2022-03-11 | GSE175880 | GEO

Tissue neutrophil heterogeneity

Project description:Tissue neutrophil heterogeneity

| PRJNA594618 | ENA