Calreticulin mutations affect its chaperone function and perturb the glycoproteome

Schürch, Patrick M.; Malinovska, Liliana; Hleihil, Mohammad; Losa, Marco; Hofstetter, Mara Carina; Wildschut, Mattheus H.E.; Lysenko, Veronika; Lakkaraju, Asvin K. K.; Maat, Christina A.; Benke, Dietmar; Aguzzi, Adriano; Wollscheid, Bernd; Picotti, Paola; Theocharides, Alexandre

doi:10.1016/j.celrep.2022.111689

Cited by 10 publications

(5 citation statements)

References 53 publications

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“…Thus, reduced adhesion of CALRdel52 mutant neutrophils to E-selectin together with no change in adhesion to VCAM-1 offer novel insights into the pathophysiological mechanisms underlying the less elevated thrombotic risk in CALR mutant patients as compared to JAK2-V617F patients. In their publication, Schürch et al [ 108 ] demonstrated myeloperoxidase deficiency in homozygous CALRdel52 mutations, which was induced by impaired folding of newly synthesized myeloperoxidase due to defective CALR-mediated ER quality control in neutrophils. Considering the central role of CALR in quality control of many glycoproteins, it is tempting to hypothesize that reduced adhesion of CALR +/del neutrophils to E-selectin is the result of altered post-translational modifications of the PSGL-1 receptor.…”

Section: Discussionmentioning

confidence: 99%

Neutrophil-specific expression of JAK2-V617F or CALRmut induces distinct inflammatory profiles in myeloproliferative neoplasia

Haage,

Charakopoulos,

Bhuria

et al. 2024

J Hematol Oncol

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Background Neutrophils play a crucial role in inflammation and in the increased thrombotic risk in myeloproliferative neoplasms (MPNs). We have investigated how neutrophil-specific expression of JAK2-V617F or CALRdel re-programs the functions of neutrophils. Methods Ly6G-Cre JAK2-V617F and Ly6G-Cre CALRdel mice were generated. MPN parameters as blood counts, splenomegaly and bone marrow histology were compared to wild-type mice. Megakaryocyte differentiation was investigated using lineage-negative bone marrow cells upon in vitro incubation with TPO/IL-1β. Cytokine concentrations in serum of mice were determined by Mouse Cytokine Array. IL-1α expression in various hematopoietic cell populations was determined by intracellular FACS analysis. RNA-seq to analyse gene expression of inflammatory cytokines was performed in isolated neutrophils from JAK2-V617F and CALR-mutated mice and patients. Bioenergetics of neutrophils were recorded on a Seahorse extracellular flux analyzer. Cell motility of neutrophils was monitored in vitro (time lapse microscopy), and in vivo (two-photon microscopy) upon creating an inflammatory environment. Cell adhesion to integrins, E-selectin and P-selection was investigated in-vitro. Statistical analysis was carried out using GraphPad Prism. Data are shown as mean ± SEM. Unpaired, two-tailed t-tests were applied. Results Strikingly, neutrophil-specific expression of JAK2-V617F, but not CALRdel, was sufficient to induce pro-inflammatory cytokines including IL-1 in serum of mice. RNA-seq analysis in neutrophils from JAK2-V617F mice and patients revealed a distinct inflammatory chemokine signature which was not expressed in CALR-mutant neutrophils. In addition, IL-1 response genes were significantly enriched in neutrophils of JAK2-V617F patients as compared to CALR-mutant patients. Thus, JAK2-V617F positive neutrophils, but not CALR-mutant neutrophils, are pathogenic drivers of inflammation in MPN. In line with this, expression of JAK2-V617F or CALRdel elicited a significant difference in the metabolic phenotype of neutrophils, suggesting a stronger inflammatory activity of JAK2-V617F cells. Furthermore, JAK2-V617F, but not CALRdel, induced a VLA4 integrin-mediated adhesive phenotype in neutrophils. This resulted in reduced neutrophil migration in vitro and in an inflamed vessel. This mechanism may contribute to the increased thrombotic risk of JAK2-V617F patients compared to CALR-mutant individuals. Conclusions Taken together, our findings highlight genotype-specific differences in MPN-neutrophils that have implications for the differential pathophysiology of JAK2-V617F versus CALR-mutant disease.

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Section: Discussionmentioning

confidence: 99%

Neutrophil-specific expression of JAK2-V617F or CALRmut induces distinct inflammatory profiles in myeloproliferative neoplasia

Haage,

Charakopoulos,

Bhuria

et al. 2024

J Hematol Oncol

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“…These genes act as endoplasmic reticulum chaperones that can perform protein quality control and maintain endoplasmic reticulum protein homeostasis [ 37 ]. Calr gene is a highly conserved molecular chaperone protein that is mainly found in the endoplasmic reticulum [ 38 ] and plays an important role in cell proliferation, apoptosis and immune response [ 39 ]. In this study, we found that calr was involved in a variety of cellular processes, including cell adhesion, cell proliferation.…”

Section: Discussionmentioning

confidence: 99%

Integrated transcriptome and microRNA analysis reveals molecular responses to high-temperature stress in the liver of American shad (Alosa sapidissima)

Liu,

Liang,

et al. 2024

BMC Genomics

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Background Fish reproduction, development and growth are directly affected by temperature, investigating the regulatory mechanisms behind high temperature stress is helpful to construct a finer molecular network. In this study, we systematically analyzed the transcriptome and miRNA information of American shad (Alosa sapidissima) liver tissues at different cultivation temperatures of 24 ℃ (Low), 27 ℃ (Mid) and 30 ℃ (High) based on a high-throughput sequencing platform. Results The results showed that there were 1594 differentially expressed genes (DEGs) and 660 differentially expressed miRNAs (DEMs) in the LowLi vs. MidLi comparison group, 473 DEGs and 84 DEMs in the MidLi vs. HighLi group, 914 DEGs and 442 DEMs in the LowLi vs. HighLi group. These included some important genes and miRNAs such as calr, hsp90b1, hsp70, ssa-miR-125a-3p, ssa-miR-92b-5p, dre-miR-15a-3p and novel-m1018-5p. The DEGs were mainly enriched in the protein folding, processing and export pathways of the endoplasmic reticulum; the target genes of the DEMs were mainly enriched in the focal adhesion pathway. Furthermore, the association analysis revealed that the key genes were mainly enriched in the metabolic pathway. Interestingly, we found a significant increase in the number of genes and miRNAs involved in the regulation of heat stress during the temperature change from 24 °C to 27 °C. In addition, we examined the tissue expression characteristics of some key genes and miRNAs by qPCR, and found that calr, hsp90b1 and dre-miR-125b-2-3p were significantly highly expressed in the liver at 27 ℃, while novel-m0481-5p, ssa-miR-125a-3p, ssa-miR-92b-5p, dre-miR-15a-3p and novel-m1018-5p had the highest expression in the heart at 30℃. Finally, the quantitative expression trends of 10 randomly selected DEGs and 10 DEMs were consistent with the sequencing data, indicating the reliability of the results. Conclusions In summary, this study provides some fundamental data for subsequent in-depth research into the molecular regulatory mechanisms of A. sapidissima response to heat stress, and for the selective breeding of high temperature tolerant varieties.

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“…Hence the presence of ASC specks, but not of BSA, accelerates SAA fibril formation. To confirm direct interaction and to map the protein domains of interaction between ASC and SAA, we employed the limited proteolysis-coupled mass spectrometry (LiP-MS) technology [31][32][33][34][35][36] which detects changes in peptide cleavage exerted by limited proteinase K (PK)-based proteolysis by mass spectroscopy (MS) 37 . If an interaction between proteins exists, certain epitopes become inaccessible to PK and the peptides detected by MS will display an altered profile (Fig.…”

Section: Asc Specks Interact With Saa Through the Pyrin Domain To Pro...mentioning

confidence: 99%

“…LiP-MS was conducted as shown earlier [31][32][33][34][35] . Briefly, we incubated purified recombinants ASC protein or ExpiHEK cell native lysates containing ASC specks with human recombinant SAA1 proteins for 15 min at 37 °C in LiP buffer (100 mM HEPES pH 7.4, 150 mM KCl, 1mM MgCl2).…”

Section: Limited Proteolysis-coupled Mass Spectrometry (Lip-ms)mentioning

confidence: 99%

The ASC inflammasome adapter controls the extent of peripheral protein aggregate deposition in inflammation-associated amyloidosis

Losa

Emmenegger

Rossi³

et al. 2021

Preprint

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The apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) protein is a component of inflammasomes, inflammation-mediating complexes that include Nucleotide-Binding Oligomerization Domain, Leucine Rich Repeat And Pyrin Domain Containing 3 (NLRP3). ASC and NLRP3 components aggregate to form ASC specks, which can cross-seed Aβ aggregation in Alzheimer’s disease (AD). Here we asked whether ASC is involved in additional protein misfolding diseases (PMDs). AA amyloidosis is a severe complication of chronic inflammatory conditions caused by the aggregation of Serum Amyloid A (SAA) protein. Since AA and Aβ aggregates share similar biochemical properties, such as β-sheet structures, we hypothesized that ASC inflammasomes may be involved in SAA/AA recruitment, aggregation, processing and removal. We therefore investigated the role of ASC in AA amyloidosis in vivo, employing a murine AA amyloidosis model in Asc-ablated mice. We show that ASC exerts a crucial role in SAA recruitment in the presence of AA fibrils, that splenic AA amyloid load was decreased in Asc-/- mice, that the presence of ASC accelerates SAA fibril formation and that ASC is important for SAA-activated phagocytosis of macrophages. We conclude that ASC modulates both SAA serum levels and the severity of AA amyloidosis in mice. ASC may represent a therapeutic target in AA as well as other amyloidosis entities.

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Calreticulin mutations affect its chaperone function and perturb the glycoproteome

Cited by 10 publications

References 53 publications

Neutrophil-specific expression of JAK2-V617F or CALRmut induces distinct inflammatory profiles in myeloproliferative neoplasia

Neutrophil-specific expression of JAK2-V617F or CALRmut induces distinct inflammatory profiles in myeloproliferative neoplasia

Integrated transcriptome and microRNA analysis reveals molecular responses to high-temperature stress in the liver of American shad (Alosa sapidissima)

The ASC inflammasome adapter controls the extent of peripheral protein aggregate deposition in inflammation-associated amyloidosis

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