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Transcriptomic analysis of emergency granulopoiesis-generated neutrophils compared to steady state (control) neutrophils

ABSTRACT: Functional analysis demonstrates that neutrophils generated by emergency granulopoiesis have elevated bactericidal activity when compared to control (steady state) neutrophils. To determine if transcriptomic changes support these functional adaptations, we performed gene expression analysis on emergency granulopoiesis and steady state neutrophils before and after infection.

ORGANISM(S): Danio rerio

PROVIDER: GSE217064 | GEO | 2023/05/01

REPOSITORIES: GEO

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Changes in gene expression during G-CSF-induced emergency granulopoiesis in humans

Project description:Emergency granulopoiesis refers to the increased production of neutrophils in bone marrow and their release into circulation induced by severe infection. Several studies point to a critical role for granulocyte colony-stimulating factor (G-CSF) as the main mediator of emergency granulopoiesis. However, the consequences of G-CSF stimulation on the transcriptome of neutrophils and their precursors have not yet been investigated in humans. Here, we examine the changes in mRNA expression induced by administration of G-CSF in vivo, as a model of emergency granulopoiesis in humans. Blood samples were collected from healthy individuals after five days of G-CSF administration. Neutrophil precursors were sorted into discrete stages of maturation by flow cytometry, and RNA was subjected to microarray analysis. mRNA levels were compared to previously published expression levels in corresponding populations of neutrophil precursors isolated from bone marrow of untreated, healthy individuals. 1110 mRNAs were differentially expressed more than 2-fold throughout terminal granulopoiesis. Major changes were seen in pathways involved in apoptosis, cytokine signaling, and Toll-like receptor pathways. In addition, G-CSF treatment reduced the levels of four out of five measured granule proteins in mature neutrophils including the proantibacterial protein hCAP-18, which was completely deficient in neutrophils from G-CSF-treated donors. These results indicate that multiple biological processes are altered in order to satisfy the increased demand for neutrophils during G-CSF-induced emergency granulopoiesis in humans.

2017-04-04 | E-MTAB-4668 | biostudies-arrayexpress

Age-induced alterations of granulopoiesis generate atypical neutrophils that aggravate stroke pathology [scRNA-Seq 1]

Project description:Stroke in aging leads to a dysregulated emergency granulopoiesis impacting neurological outcome.

2023-05-16 | GSE174439 | GEO

Age-induced alterations of granulopoiesis generate atypical neutrophils that aggravate stroke pathology. [scRNA-Seq 2]

Project description:Stroke in aging leads to a dysregulated emergency granulopoiesis impacting neurological outcome.

2023-05-16 | GSE231822 | GEO

Mitochondrial pyruvate metabolism and glutaminolysis toggle steady-state and emergency myelopoiesis

Project description:RNA-sequencing was performed to look for transcriptional differences between wild-type and Mpc2-deficient granulocyte-macrophage progenitors immediately following the deletion of Mpc2. Paper abstract: Hematopoietic stem and progenitor cells likely engage specific metabolic pathways to promote homeostasis of downstream mature lineages. We examined hematopoiesis in mice conditionally deficient in genes required for long-chain fatty acid oxidation (Cpt2), glutaminolysis (Gls), or mitochondrial pyruvate import (Mpc2). Genetic ablation of Cpt2 or Gls minimally impacted most blood lineages. Deletion of Mpc2 led to a sharp decline in mature myeloid cells and a slower reduction in T cells, whereas other hematopoietic lineages were unaffected. Yet MPC2-deficient monocytes and neutrophils rapidly recovered due to a transient increase in proliferation of myeloid but not other progenitors. Competitive bone marrow chimera and stable isotope tracing experiments demonstrated that this proliferative burst was progenitor-intrinsic and accompanied by a metabolic switch to glutaminolysis. Myeloid recovery after loss of MPC2 or cyclophosphamide treatment was delayed in the absence of GLS. Reciprocally, MPC2 was not required for myeloid recovery after cyclophosphamide treatment. Thus, whereas mitochondrial pyruvate metabolism maintains myelopoiesis under steady-state conditions, rapid recovery from acute loss of myeloid cells requires a switch to glutaminolysis in progenitors.

2023-02-22 | GSE225578 | GEO

The proteome of neutrophils in sickle cell disease

Project description:Polymorphonuclear neutrophils are key actors in the pathophysiology of sickle cell disease, but specific factors underlying their activation and sustained inflammation are not well documented. In the present study, we investigated the proteome of neutrophils by a label-free global comparative approach between 4 non-treated sickle patients (SS genotype) at steady state and 4 healthy donors. We identified 101 proteins differentially expressed in SS and normal neutrophils. We found overexpression of CD64 and under-expression of CD62L suggestive of an activated and aged neutrophil profile in SS patients. Comparison of the two proteomes revealed a strong involvement of the type 1 interferon (IFN) response pathway with a 3- to 84-fold increase of type 1 IFN-induced proteins in SS neutrophils, and overexpression of STAT1 and STAT2. Thus, we next determined the plasmatic concentration of type 1 IFNs (IFNα and IFNβ) using the digital-ELISA technology and found a significant higher concentration of IFNα in the plasma from half of our SS patients compared to controls. Overall, a dramatic high-level expression of IFNα signaling proteins in neutrophils from SS patients suggests auto-inflammatory-like phenotype in sickle cell disease at steady state. This finding could open the way to new anti-inflammatory therapies.

2020-11-11 | PXD014457 | Pride

Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish

Project description:The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins, and which exert non-overlapping functions. However, there is little information about ontogenetically distinct neutrophil populations. In this work, using a photoconvertible transgenic zebrafish line; Tg(mpx:Dendra2), we selectively label rostral blood island-derived and caudal hematopoietic tissue-derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single-cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury, both populations share a core transcriptional profile as well as subset-specific transcriptional signatures. Accordingly, both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly, caudal neutrophils respond heterogeneously. Collectively, our results reveal that co-existing neutrophils populations with ontogenically distinct origin display key functional differences.

2023-08-07 | GSE239880 | GEO

scRNA-seq of mouse hematopoietic stem cells after 4 hours of LPS/PBS treatment in vivo.

Project description:To understand whether hematopoietic stem cells (HSCs) take part in emergency granulopoiesis, WT C57BL6 mice were treated intraperitoneally with 35 ug of LPS (lipopolysacharide) to induce emergency granulopoiesis or PBS control. Mice were sacrificed 4 hours after the treatment andHSCs (Lin-c-Kit+Sca-1+CD48-CD150+) were sorted and subjected to scRNA-seq.

2023-10-16 | E-MTAB-12324 | biostudies-arrayexpress

ATACseq of mouse hematopoietic stem cells after 4 hours of LPS/PBS treatment in vivo

Project description:To understand whether hematopoietic stem cells (HSCs) take part in emergency granulopoiesis, WT C57BL6 mice were treated intraperitoneally with 35 ug of lipopolysacharide (LPS) to induce emergency granulopoiesis, or PBS control. Mice were sacrificed 4 hours after the treatment and HSCs (Lin-c-Kit+Sca-1+CD48-CD150+) were sorted and subjected to bulk ATACseq.

2023-10-16 | E-MTAB-12310 | biostudies-arrayexpress

Expression data from control, aged and activated neutrophils in mouse blood

Project description:Neutrophils provide immune protection against pathogens but also may promote tissue injury in inflammatory diseases. Although neutrophils are generally considered as a relatively homogeneous population, evidence for heterogeneity is emerging. Under steady-state conditions, neutrophil heterogeneity may arise from ageing and the replenishment by newly released neutrophils from the bone marrow. We used microarray to globally analyze gene expression in aged neutrophils and characterize the inflammatory programs that are activated during the aging process in the circulation. Control, aged and activated neutrophils were sorted directly from mouse blood for RNA extraction and hybridization on Affymetrix microarrays. We transfused whole blood and harvested donor neutrophils marked by the CD45.1 allele 6h later to derive neutrophils that had truly aged in vivo. Sorted neutrophils were compared to control donor neutrophils that had been transferred for only 10 min. Additionally, we harvested circulating neutrophils from TNF-? treated mice for comparison with neutrophils activated by systemic inflammation.

2015-09-24 | E-GEOD-69886 | biostudies-arrayexpress

The Pu.1 target gene Zbtb11 regulates neutrophil development through its integrase-like HHCC zinc finger

Project description:From a forward genetic screen in zebrafish, we identified the transcription factor, ZBTB11, as critical for basal and emergency granulopoiesis and showed that ZBTB11 sits in a pathway directly downstream of master myeloid regulators including PU.1, GFI1 and CEBPa. To better understand target genes regulated by Zbtb11, RNAseq profiling was performed in neutrophils from WT and Zbtb11 mutant zebrafish.

2023-08-08 | GSE239949 | GEO

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