RT Journal Article
SR Electronic
T1 Proteome analysis of human neutrophil granulocytes from patients with monogenic disease using data-independent acquisition
JF Molecular & Cellular Proteomics
JO Mol Cell Proteomics
FD American Society for Biochemistry and Molecular Biology
SP mcp.RA118.001141
DO 10.1074/mcp.RA118.001141
A1 Grabowski, Piotr
A1 Hesse, Sebastian
A1 Hollizeck, Sebastian
A1 Rohlfs, Meino
A1 Behrends, Uta
A1 Sherkat, Roya
A1 Tamary, Hannah
A1 Ünal, Ekrem
A1 Somech, Raz
A1 Patıroğlu, Türkan
A1 Canzar, Stefan
A1 van der Werff Ten Bosch, Jutte
A1 Klein, Christoph
A1 Rappsilber, Juri
YR 2019
UL http://www.mcponline.org/content/early/2019/01/10/mcp.RA118.001141.abstract
AB Neutrophil granulocytes are critical mediators of innate immunity and tissue regeneration. Rare diseases of neutrophil granulocytes may affect their differentiation and/or functions. However, there are very few validated diagnostic tests assessing the functions of neutrophil granulocytes in these diseases. Here, we set out to probe omics analysis as a novel diagnostic platform for patients with defective differentiation and function of neutrophil granulocytes. We analyzed highly purified neutrophil granulocytes from 68 healthy individuals and 16 patients with rare monogenic diseases. Cells were isolated from fresh venous blood (purity >99%) and used to create a spectral library covering almost 8000 proteins using strong cation exchange fractionation. Patient neutrophil samples were then analyzed by data-independent acquisition proteomics, quantifying 4154 proteins in each sample. Neutrophils with mutations in the neutrophil elastase gene ELANE, showed large proteome changes that suggest these mutations may affect maturation of neutrophil granulocytes and initiate misfolded protein response and cellular stress mechanisms. In contrast, only few proteins changed in patients with leukocyte adhesion deficiency (LAD) and chronic granulomatous disease (CGD). Strikingly, neutrophil transcriptome analysis showed no correlation with its proteome. In case of two patients with undetermined genetic causes, proteome analysis guided the targeted genetic diagnostics and uncovered the underlying genomic mutations. Data-independent acquisition proteomics may help to define novel pathomechanisms in neutrophil diseases and provide a clinically useful diagnostic dimension.