Neutrophils are important innate immune cells that tackle invading pathogens with different effector mechanisms. They acquire this antimicrobial potential during their maturation in the bone marrow, where they differentiate from hematopoietic stem cells in a process called granulopoiesis. Mature neutrophils are terminally differentiated and short-lived with a high turnover rate. Here, we show a critical role for linker histone H1 on the differentiation and function of neutrophils using a genome-wide CRISPR/Cas9 screen in the human cell line PLB-985. We systematically disrupted expression of somatic H1 subtypes to show that individual H1 subtypes affect PLB-985 maturation in opposite ways. Loss of H1.2 and H1.4 induced an eosinophil-like transcriptional program, thereby negatively regulating the differentiation into the neutrophil lineage. Importantly, H1 subtypes also affect neutrophil differentiation and the eosinophil-directed bias of murine bone marrow stem cells, demonstrating an unexpected subtype-specific role for H1 in granulopoiesis.
RNA sequencing data have been deposited in ArrayExpress - accession no. E-MTAB-8459All data generated or analysed during this study are included in the manuscript and supplemental files. Source data files are provided for Figure 1 and Figure 4. A supplementary table with all used qPCR primers, sgRNA sequences, antibodies and other reagents is provided.
BP_August_2016_RNA-Seq_band_form_neutrophil_on_GRCh38 - samplesEGAD00001002446.
neutrophilic myelocyte from bone marrow of donor: BM230614 ,BM220513EGAX00001244028.
- Gabriel Sollberger
- Gabriel Sollberger
- Robert Streeck
- Brian Edward Caffrey
- Arturo Zychlinsky
- Arthur I Skoultchi
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Animal experimentation: Breeding of mice and isolation of blood and bone marrow were approved by the Berlin state authority Landesamt für Gesundheit und Soziales. All mice were bred at the Max Planck Insitute for Infection Biology under specific pathogen-free conditions. Animals were maintained on a 12-hour light/12-hour dark cycle and fed ad libitum
Human subjects: Human primary neutrophils, PBMCs and monocytes were isolated from blood samples of healthy volunteers according to the declaration of Helsinki. All donors provided written informed consent and all blood samples were collected with approval from the local ethics committee.
- Florent Ginhoux, Agency for Science Technology and Research, Singapore
© 2020, Sollberger et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.