Dataset Information

Neutrophil specific granule and NETosis defect in Gray Platelet Syndrome

ABSTRACT: Gray Platelet Syndrome (GPS) is an autosomal recessive bleeding disorder characterized by a lack of α-granules in platelets and progressive myelofibrosis. Rare loss of function variants in NBEAL2, a member of the family of BEACH genes, are causal of GPS. The gene is involved in fusion, fission and trafficking of vesicles and granules. Whether NBEAL2 controls the ontogeny of granules of myeloid cells remains disputed. We found that neutrophils obtained from the peripheral blood from GPS patients have a normal distribution of azurophilic granules, but show a deficiency of specific granules, as confirmed by immuno-electron microscopy and mass spectrometry proteomics analyses. In cultures from peripheral CD34+ hematopoietic stem cells (HSCs) into mature neutrophils, the time dynamics showed concordance of NBEAL2 and specific granule protein expression at transcriptional and protein level, which were discordant in neutrophils obtained GPS-HSCs. This is indicative of normal granulopoiesis in GPS and identifies NBEAL2 as an important regulator of granule release (similar to platelets) which is suggested to occur upon egress into the blood stream. Patient neutrophil functions, including production of reactive oxygen species, chemotaxis and killing of bacteria and fungi were intact. Since GPS patients do not excessively suffer from infections, the consequence of the reduced specific granule content and lack of NET formation for innate immunity remains to be explored.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Neutrophil, Blood, Cell Line Cell

DISEASE(S): Gray Platelet Syndrome

SUBMITTER: Arie Hoogendijk

LAB HEAD: M. van den Biggelaar

PROVIDER: PXD017784 | Pride | 2021-01-28

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	07072017_CTRL_CD34d17_15_11b_16-.raw	Raw
	07072017_CTRL_CD34d17_15_11b_16_.raw	Raw
	07072017_GPS_CD34d17_15_11b_16-.raw	Raw
	07072017_GPS_CD34d17_15_11b_16_.raw	Raw
	07072017_MPB_CD34d10_15_11b-16-.raw	Raw

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Publications

Neutrophil specific granule and NETosis defects in gray platelet syndrome.

Aarts Cathelijn E M CEM Downes Kate K Hoogendijk Arie J AJ Sprenkeler Evelien G G EGG Gazendam Roel P RP Favier Rémi R Favier Marie M Tool Anton T J ATJ van Hamme John L JL Kostadima Myrto A MA Waller Kate K Zieger Barbara B van Bergen Maaike G J M MGJM Langemeijer Saskia M C SMC van der Reijden Bert A BA Janssen Hans H van den Berg Timo K TK van Bruggen Robin R Meijer Alexander B AB Ouwehand Willem H WH Kuijpers Taco W TW

Blood advances 20210101 2

Gray platelet syndrome (GPS) is an autosomal recessive bleeding disorder characterized by a lack of α-granules in platelets and progressive myelofibrosis. Rare loss-of-function variants in neurobeachin-like 2 (NBEAL2), a member of the family of beige and Chédiak-Higashi (BEACH) genes, are causal of GPS. It is suggested that BEACH domain containing proteins are involved in fusion, fission, and trafficking of vesicles and granules. Studies in knockout mice suggest that NBEAL2 may control the forma ...[more]

PMID: 33496751

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Project description:Neutrophils are short-lived yet vital innate immune cells equipped with bioactive glycoproteins packed in cytosolic granules that can readily be mobilised to elicit an effective and timely response against invading pathogens. Since the dynamic glycosylation processes underpinning the dramatic metamorphosis associated with myeloid progenitor-to-neutrophil differentiation remain unmapped, we here perform a comprehensive spatiotemporal profiling of the complex N-glycoproteome of isolated granule populations from blood-derived neutrophils and during maturation of bone marrow-derived progenitors using glycomics-assisted glycoproteomics. Firstly, glycomics indicated that the granule populations of mature (resting) neutrophils exhibit distinctive glycophenotypes including, most strikingly, unusual paucimannosidic- and monoantennary complex-type N-glycans in azurophilic granules. The granule-specific N-glycosylation features were recapitulated by glycoproteomics, which also uncovered extensive site- and protein-specific N-glycosylation decorating 4,772 N-glycopeptides from 352 N-glycoproteins across the neutrophil granules. Excitingly, comprehensive mining of proteomics and transcriptomics data collected from discrete myeloid progenitor stages for glycopeptide and glyco-enzyme expression revealed that dramatic glycoproteome remodelling underpins the promyelocytic-to-metamyelocyte transition and that remodelling is driven predominantly by proteome expression changes rather than modulation of the glycosylation machinery. Notable exceptions were the oligosaccharyltransferase subunits responsible for initiation of N-glycoprotein biosynthesis that were strongly reduced with myeloid differentiation leading to reduced N-glycosylation efficiency in late-stage neutrophil maturation and in corresponding granules. Our study provides new spatiotemporal insights into the dynamic neutrophil N-glycoproteome, which exhibits intriguingly complex site-, protein- and granule-specific N-glycosylation features and which undergoes strong remodelling during myeloid progenitor-to-neutrophil metamorphosis.