Inhibition of PDIA3 in club cells attenuates osteopontin production and lung fibrosis

Kumar, Amit; Elko, Evan; Bruno, S.; Mark, Z.; Chamberlain, Nicolas; Mihavics, Bethany; Chandrasekaran, R; Walzer, Joseph; Ruban, M.; Gold, Clarissa; Lam, Ying‐Wai; Ghandikota, Sudhir; Jegga, Anil G.; Janssen–Heininger, Yvonne M. W.; Anathy, Vikas

doi:10.1136/thoraxjnl-2021-216882

Cited by 28 publications

(18 citation statements)

References 42 publications

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“…CD44 functioned as the receptor of SPP1 ( 10 ), which showed a similar expression pattern to SPP1 ( Figure 3B ), and their expressions showed a strong positive correlation ( r = 0.7875, p < 0.0001) ( Figure 3C ). SPP1 is considered a major driver for renal fibrosis ( 11 ), the secretion of which is stimulated by inflammatory cytokines ( 12 ) and the suppression of which could alleviate kidney fibrosis ( 13 ). In our study, the expressions of fibronectin and Col1A1 were gradually elevated in the WBCR and BCR groups compared to those in MCD ( Figures 3E, F ).…”

Section: Resultsmentioning

confidence: 99%

Digital Spatial Profiling of Individual Glomeruli From Patients With Anti-Neutrophil Cytoplasmic Autoantibody-Associated Glomerulonephritis

Liu²,

Zhu³

et al. 2022

Front. Immunol.

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We previously showed that the rupture of Bowman’s capsule (BC) promotes the progression of crescentic glomerulonephritis by enhancing the entry of CD8+ T cells into the glomeruli. In the present study, we utilized digital spatial profiling to simultaneously profile the altered abundances of the messenger RNA (mRNA) transcripts and proteins in the glomerular and periglomerular areas of four biopsy samples of anti-neutrophil cytoplasmic autoantibody-associated glomerulonephritis (ANCA-GN) and two biopsy specimens of minimal change disease (MCD) controls. The paraffin-embedded biopsy samples were stained with collagen IV, CD45, and SYTO 13 to distinguish the glomeruli with periglomerular infiltration but intact BC, with focal BC rupture, and with extensive rupture of BC and glomeruli without crescent formation and leukocytic infiltration in ANCA-GN. By assessing multiple discrete glomerular areas, we found that the transcript expression levels of the secreted phosphoprotein-1 and its receptor CD44 were upregulated significantly in the glomeruli with more severe ruptures of BC, and their expression levels correlated positively with the fibrotic markers. We also found that both alternative and classic complement pathways were activated in the glomeruli from patients with ANCA-GN. Furthermore, M1 macrophages were involved mostly in the early stage of BC rupture, while M2 macrophages were involved in the late stage and may contribute to the fibrosis process of the crescents. Finally, loss of glomerular cells in ANCA-GN was likely mediated by apoptosis. Our results show that digital spatial profiling allows the comparative analysis of the mRNA and protein profiles in individual glomeruli affected differently by the disease process and the identification of potential novel mechanisms in ANCA-GN.

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Section: Resultsmentioning

confidence: 99%

Digital Spatial Profiling of Individual Glomeruli From Patients With Anti-Neutrophil Cytoplasmic Autoantibody-Associated Glomerulonephritis

Liu²,

Zhu³

et al. 2022

Front. Immunol.

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“…PDIA3 ablation or inhibition resulted in decreases in allergic airway inflammation, airway hyperresponsiveness, peribronchiolar fibrosis, and pulmonary fibrosis. These phenotype improvements were attributed to deficiencies in growth factors (POSTN, EGF, and SPP1), chemoattractant (Eotaxin), and decreases in death receptor, Fas activity [30][31][32][33]. We also observed that PDIA3 depletion decreases influenza-induced apoptosis of lung epithelial cells and disulfide bonds in HA, leading to decrease in influenza burden and airway hyperresponsiveness [3,30].…”

Section: Discussionmentioning

confidence: 67%

“…PDIA3 catalyzes oxidative modifications of numerous proteins, including proteins that are mediators of respiratory diseases [29][30][31][32][33]. PDIA3 ablation or inhibition resulted in decreases in allergic airway inflammation, airway hyperresponsiveness, peribronchiolar fibrosis, and pulmonary fibrosis.…”

Section: Discussionmentioning

confidence: 99%

“…Another important component of our work is a dose-dependent decrease in many cytokines/and chemokines in the BAL fluid of LOC14-treated mice. Cytokines and chemokines are often stabilized by disulfide bonds [33][34][35][36], and these bonds have been shown to be important for their function [33][34][35][36]. Although PDIs direct role in the immunomodulatory effect needs a thorough examination, given that PDIs are a major disulfide catalyzing enzymes in the cell [8], it is likely to play a major role in immunomodulation by catalyzing the disulfide bond formation in cytokines, chemokines, and growth factors such as osteopontin [31,32].…”

Section: Discussionmentioning

confidence: 99%

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Protein Disulfide Isomerase A3 Regulates Influenza Neuraminidase Activity and Influenza Burden in the Lung

Chamberlain

Ruban

Mark

et al. 2022

IJMS

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Influenza (IAV) neuraminidase (NA) is a glycoprotein required for the viral exit from the cell. NA requires disulfide bonds for proper function. We have recently demonstrated that protein disulfide isomerase (PDI)A3 is required for oxidative folding of IAV hemagglutinin (HA), and viral propagation. However, it not known whether PDIs are required for NA maturation or if these interactions represent a putative target for the treatment of influenza infection. We sought to determine whether PDIA3 is required for disulfide bonds of NA, its activity, and propagation of the virus. Requirement of disulfides for NA oligomerization and activity were determined using biotin switch and redox assays in WT and PDIA3−/− in A549 cells. A PDI specific inhibitor (LOC14) was utilized to determine the requirement of PDIs in NA activity, IAV burden, and inflammatory response in A549 and primary mouse tracheal epithelial cells. Mice were treated with the inhibitor LOC14 and subsequently examined for IAV burden, NA activity, cytokine, and immune response. IAV-NA interacts with PDIA3 and this interaction is required for NA activity. PDIA3 ablation or inhibition decreased NA activity, viral burden, and inflammatory response in lung epithelial cells. LOC14 treatment significantly attenuated the influenza-induced inflammatory response in mice including the overall viral burden. These results provide evidence for PDIA3 inhibition suppressing NA activity, potentially providing a novel platform for host-targeted antiviral therapies.

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“…Specific knockout of PDIA3 in B cells inhibits recruitment of MHC class I molecules onto the peptide-loading complex for antigen presentation ( 67 ). PDIA3 is upregulated and secreted by kidney fibroblasts in response to TGFβ1 ( 34 ), is released on platelet activation ( 68 ), and its upregulation has been correlated with pulmonary fibrosis ( 69 ). Cell-surface PDIA3 has also been shown to act with glycosylated calnexin to reduce extracellular disulfide bonds and make ECM more susceptible to degradation by cancer cells ( 39 ).…”

Section: Discussionmentioning

confidence: 99%

PDIA3/ERp57 promotes a matrix-rich secretome that stimulates fibroblast adhesion through CCN2

Hellewell

Heesom

Jepson

et al. 2022

American Journal of Physiology-Cell Physiology

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The matricellular glycoprotein thrombospondin1 (TSP1) has complex roles in the extracellular matrix and at cell surfaces, but relatively little is known about its intracellular associations prior to secretion. To search for novel intracellular interactions of TSP1 in situ, we carried out a biotin ligase-based TSP1 interactome screen and identified protein disulphide isomerase A3 (PDIA3/ERp57) as a novel candidate binding protein. In validation, TSP1 and PDIA3 were established to bind in vitro and to colocalise in the endoplasmic reticulum of human dermal fibroblasts (HDF). Loss of PDIA3 function, either by pharmacological inhibition in HDF or in Pdia3-/- mouse embryo fibroblasts (Pdia3-/- MEF), led to alterations in the composition of cell-derived ECM, involving changed abundance of fibronectin and TSP1, and was correlated with reduced cell spreading, altered organisation of F-actin and reduced focal adhesions. These cellular phenotypes of Pdia3-/- MEF were normalised by exposure to conditioned medium (WTCM) or extracellular matrix (WTECM) from wild-type (WT)-MEF. Rescue depended on PDIA3 activity in WT-MEF, and was not prevented by immunodepletion of fibronectin. Heparin-binding proteins in WTCM were found to be necessary for rescue. Comparative quantitative tandem-mass-tag proteomics and functional assays on the heparin-binding secretomes of WT-MEF and Pdia3-/- MEF identified multiple ECM and growth factor proteins to be down-regulated in the CM of Pdia3-/- MEF. Of these, CCN2 was identified to be necessary for the adhesion-promoting activity of WTCM on Pdia3-/- MEF and to bind TSP1. Thus, PDIA3 coordinates fibroblast production of an ECM-rich, pro-adhesive microenvironment, with implications for PDIA3 as a translational target.

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Inhibition of PDIA3 in club cells attenuates osteopontin production and lung fibrosis

Cited by 28 publications

References 42 publications

Digital Spatial Profiling of Individual Glomeruli From Patients With Anti-Neutrophil Cytoplasmic Autoantibody-Associated Glomerulonephritis

Digital Spatial Profiling of Individual Glomeruli From Patients With Anti-Neutrophil Cytoplasmic Autoantibody-Associated Glomerulonephritis

Protein Disulfide Isomerase A3 Regulates Influenza Neuraminidase Activity and Influenza Burden in the Lung

PDIA3/ERp57 promotes a matrix-rich secretome that stimulates fibroblast adhesion through CCN2

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