TY - JOUR
T1 - Human genetic defects in SRP19 and SRPRA cause severe congenital neutropenia with distinctive proteome changes
AU - Linder, Monika I.
AU - Mizoguchi, Yoko
AU - Hesse, Sebastian
AU - Csaba, Gergely
AU - Tatematsu, Megumi
AU - Łyszkiewicz, Marcin
AU - Ziȩtara, Natalia
AU - Jeske, Tim
AU - Hastreiter, Maximilian
AU - Rohlfs, Meino
AU - Liu, Yanshan
AU - Grabowski, Piotr
AU - Ahomaa, Kaarin
AU - Maier-Begandt, Daniela
AU - Schwestka, Marko
AU - Pazhakh, Vahid
AU - Isiaku, Abdulsalam I.
AU - Briones Miranda, Brenda
AU - Blombery, Piers
AU - Saito, Megumu K.
AU - Rusha, Ejona
AU - Alizadeh, Zahra
AU - Pourpak, Zahra
AU - Kobayashi, Masao
AU - Rezaei, Nima
AU - Unal, Ekrem
AU - Hauck, Fabian
AU - Drukker, Micha
AU - Walzog, Barbara
AU - Rappsilber, Juri
AU - Zimmer, Ralf
AU - Lieschke, Graham J.
AU - Klein, Christoph
N1 - Funding Information:
The authors thank all the patients and their families for allowing us to study their cells and all clinical staff for taking excellent care. The authors are particularly indebted to Sorin Iurian (Romania) for referring family A. The authors acknowledge Sebastian Hollizeck and Isabelle Plonner for outstanding expertise in the C4R sequencing and computational facility. The authors are very grateful to Thomas U. Mayer for providing the HeLa-Flp-In/T-Rex cell line. The authors thank Tanja Vlaovic for isolating neutrophils from healthy volunteers and Diana Laverde for her guidance in performing the phagocytosis assay. The authors acknowledge Manfred Rohde (Department of Medical Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany) for the acquisition and processing of electron microscopy studies. The authors are very grateful to the Pediatric Radiology team at Ludwig-Maximilians-Universität for acquisition of the computed tomography scan. The authors thank Lucy Fox and Ella Thompson for the help with the zebrafish next-generation sequencing analysis. This work was supported by grants from the DFG, Gottfried Wilhelm Leibniz Program CRC914 (project A08 and A02); the TRR332 (project B1); the BMBF (PIDNET); and the Care-for-Rare Foundation. The Peter MacCallum Cancer Centre Molecular Hematology Laboratory is supported by funding from Wilson Centre for Lymphoma Genomics through the Snowdome Foundation. The Australian Regenerative Medicine Institute is supported by grants from the State Government of Victoria and the Australian Government. This work was supported by grants from the National Health and Medical Research Council (1044754, 1086020, and 1159278) (G.J.L.) and Maddie Riewoldt's Vision (ARMI-MRV-2018G) (G.J.L. C.K. and P.B.). Members of the Lieschke Laboratory are supported by grants of the Monash University (Graduate Scholarship and International Tuition Scholarship). Whole-genome sequencing on family A was supported by the Junior Researcher Fund of Ludwig-Maximilians-Universität Excellence Initiative (N.Z.). Contribution: C.K. conceived the original idea; C.K. M.I.L. Y.M. S.H. and R.Z. conceptualized and developed the methodology; C.K. S.H. Z.A. M.K. N.R. E.U. and F.H. performed clinical and patient-related investigations and performed the experiments; S.H. G.C. K.A. T.J. M.H. and R.Z. performed computational analysis; M.S. V.P. A.I.I. B.B.M. P.B. and G.J.L. conceptualized and performed the zebrafish experiments; M.D. B.W. J.R. R.Z. C.K.; P.B. and G.J.L. provided resources; C.G. M.R. R.Z. and M.S. curated data; C.K. M.I.L. S.H. Y.M. and G.J.L. wrote the original draft; C.K. B.W. V.P. R.Z. J.R. and G.J.L. provided supervision; and all authors wrote, reviewed and edited the draft and read and approved the manuscript before submission.
Funding Information:
This work was supported by grants from the DFG , Gottfried Wilhelm Leibniz Program CRC914 (project A08 and A02); the TRR332 (project B1); the BMBF (PIDNET); and the Care-for-Rare Foundation . The Peter MacCallum Cancer Centre Molecular Hematology Laboratory is supported by funding from Wilson Centre for Lymphoma Genomics through the Snowdome Foundation. The Australian Regenerative Medicine Institute is supported by grants from the State Government of Victoria and the Australian Government . This work was supported by grants from the National Health and Medical Research Council (1044754, 1086020, and 1159278) (G.J.L.) and Maddie Riewoldt’s Vision (ARMI-MRV-2018G) (G.J.L., C.K., and P.B.). Members of the Lieschke Laboratory are supported by grants of the Monash University (Graduate Scholarship and International Tuition Scholarship). Whole-genome sequencing on family A was supported by the Junior Researcher Fund of Ludwig-Maximilians-Universität Excellence Initiative (N.Z.).
Publisher Copyright:
© 2023 The American Society of Hematology
PY - 2023/2/9
Y1 - 2023/2/9
N2 - The mechanisms of coordinated changes in proteome composition and their relevance for the differentiation of neutrophil granulocytes are not well studied. Here, we discover 2 novel human genetic defects in signal recognition particle receptor alpha (SRPRA) and SRP19, constituents of the mammalian cotranslational targeting machinery, and characterize their roles in neutrophil granulocyte differentiation. We systematically study the proteome of neutrophil granulocytes from patients with variants in the SRP genes, HAX1, and ELANE, and identify global as well as specific proteome aberrations. Using in vitro differentiation of human induced pluripotent stem cells and in vivo zebrafish models, we study the effects of SRP deficiency on neutrophil granulocyte development. In a heterologous cell–based inducible protein expression system, we validate the effects conferred by SRP dysfunction for selected proteins that we identified in our proteome screen. Thus, SRP-dependent protein processing, intracellular trafficking, and homeostasis are critically important for the differentiation of neutrophil granulocytes.
AB - The mechanisms of coordinated changes in proteome composition and their relevance for the differentiation of neutrophil granulocytes are not well studied. Here, we discover 2 novel human genetic defects in signal recognition particle receptor alpha (SRPRA) and SRP19, constituents of the mammalian cotranslational targeting machinery, and characterize their roles in neutrophil granulocyte differentiation. We systematically study the proteome of neutrophil granulocytes from patients with variants in the SRP genes, HAX1, and ELANE, and identify global as well as specific proteome aberrations. Using in vitro differentiation of human induced pluripotent stem cells and in vivo zebrafish models, we study the effects of SRP deficiency on neutrophil granulocyte development. In a heterologous cell–based inducible protein expression system, we validate the effects conferred by SRP dysfunction for selected proteins that we identified in our proteome screen. Thus, SRP-dependent protein processing, intracellular trafficking, and homeostasis are critically important for the differentiation of neutrophil granulocytes.
UR - http://www.scopus.com/inward/record.url?scp=85145182715&partnerID=8YFLogxK
U2 - 10.1182/blood.2022016783
DO - 10.1182/blood.2022016783
M3 - Article
C2 - 36223592
AN - SCOPUS:85145182715
SN - 0006-4971
VL - 141
SP - 645
EP - 658
JO - Blood
JF - Blood
IS - 6
ER -