Spatially Resolved Identification of Transglutaminase Substrates by Proteomics in Pulmonary Fibrosis

Takeuchi, Taishu; Tatsukawa, Hideki; Shinoda, Yoshiki; Kuwata, Keiko; Nishiga, Miyuki; Takahashi, Hiroshi; Hase, Naoki; Hitomi, Kiyotaka

doi:10.1165/rcmb.2021-0012oc

Cited by 9 publications

(5 citation statements)

References 47 publications

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“…We obtained similar results using TG2-deficient mice. Indeed, liver fibrosis induced by bile duct ligation was not inhibited in these mice [152], although these mice presented a significant reduction of fibrosis induction in other fibrosis models, such as kidney fibrosis induced by unilateral ureteral obstruction [158] and lung fibrosis resulting from bleomycin treatment [162]. Interestingly, TG2 or pan-TGase inhibitors, including competitive or reversible/irreversible inhibitors, have been demonstrated to be consistently protective in several fibrosis models, including liver fibrosis induced by both CCl4 and bile duct ligation [98,152,158,[184][185][186][187][188][189].…”

Section: Tg2 Functions In Fibrosismentioning

confidence: 95%

“…The reaction products form an N ε -(γ-glutamyl)lysine isopeptide bond resulting from the crosslinking. This is an important step for the maturation and stabilization of ECM components, such as collagens, exacerbating scarring and fibrosis in various tissues, including the liver [96,[151][152][153], kidney [154][155][156][157][158][159], lung [160][161][162][163][164][165], and heart [166,167]. The other enzymatic crosslinker, lysyl oxidase (LOX), has also been reported to contribute to collagen stabilization.…”

Section: Tg2 Functions In Fibrosismentioning

confidence: 99%

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Role of Transglutaminase 2 in Cell Death, Survival, and Fibrosis

Tatsukawa

Hitomi

2021

Cells

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Transglutaminase 2 (TG2) is a ubiquitously expressed enzyme catalyzing the crosslinking between Gln and Lys residues and involved in various pathophysiological events. Besides this crosslinking activity, TG2 functions as a deamidase, GTPase, isopeptidase, adapter/scaffold, protein disulfide isomerase, and kinase. It also plays a role in the regulation of hypusination and serotonylation. Through these activities, TG2 is involved in cell growth, differentiation, cell death, inflammation, tissue repair, and fibrosis. Depending on the cell type and stimulus, TG2 changes its subcellular localization and biological activity, leading to cell death or survival. In normal unstressed cells, intracellular TG2 exhibits a GTP-bound closed conformation, exerting prosurvival functions. However, upon cell stimulation with Ca2+ or other factors, TG2 adopts a Ca2+-bound open conformation, demonstrating a transamidase activity involved in cell death or survival. These functional discrepancies of TG2 open form might be caused by its multifunctional nature, the existence of splicing variants, the cell type and stimulus, and the genetic backgrounds and variations of the mouse models used. TG2 is also involved in the phagocytosis of dead cells by macrophages and in fibrosis during tissue repair. Here, we summarize and discuss the multifunctional and controversial roles of TG2, focusing on cell death/survival and fibrosis.

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Section: Tg2 Functions In Fibrosismentioning

confidence: 95%

Section: Tg2 Functions In Fibrosismentioning

confidence: 99%

Role of Transglutaminase 2 in Cell Death, Survival, and Fibrosis

Tatsukawa

Hitomi

2021

Cells

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“…7A), suggesting that STAT6 is phosphorylated upstream of TG2. In addition, we have previously identi ed the direct molecular targets of TG2 in various tissues and cells based on the incorporation of a biotin-pentylamine probe into proteins via TG2 crosslinking activity 30,[35][36][37] . However, as there were no signi cant changes in levels of substrate proteins incorporated by the probe in IL-4-treated macrophages (Suppl.…”

Section: Tg2 Regulates M2 Macrophage Polarization Via Alox15 Inductionmentioning

confidence: 99%

Tissue transglutaminase exacerbates renal fibrosis via alternative activation of monocyte-derived macrophages

Shinoda

Tatsukawa

Yonaga

et al. 2022

Preprint

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Macrophages are important components in modulating homeostatic and inflammatory responses and are generally categorized into two broad but distinct subsets: classical activated (M1) and alternatively activated (M2) depending on the microenvironment. Fibrosis is a chronic inflammatory disease exacerbated by M2 macrophages, although the detailed mechanism by which M2 macrophage polarization is regulated remains unclear. These polarization mechanisms have little in common between mice and humans, making it difficult to adapt research results obtained in mice to human diseases. Tissue transglutaminase (TG2) is a known marker common to mouse and human M2 macrophages and is a multifunctional enzyme responsible for crosslinking reactions. Here we sought to identify the role of TG2 in macrophage polarization and fibrosis. In IL-4-treated macrophages derived from mouse bone marrow and human monocyte cells, the expression of TG2 was increased with enhancement of M2 macrophage markers, whereas knockout or inhibitor treatment of TG2 markedly suppressed M2 macrophage polarization. In the renal fibrosis model, accumulation of M2 macrophages in fibrotic kidney was significantly reduced in TG2 knockout or inhibitor-administrated mice, along with the resolution of fibrosis. Bone marrow transplantation using TG2-knockout mice revealed that TG2 is involved in M2 polarization of infiltrating macrophages derived from circulating monocytes and exacerbates renal fibrosis. Furthermore, the suppression of renal fibrosis in TG2-knockout mice was abolished by transplantation of wild-type bone marrow or by renal subcapsular injection of IL4-treated macrophages derived from bone marrow of wild-type, but not TG2 knockout. Transcriptome analysis of downstream targets involved in M2 macrophages polarization revealed that ALOX15 expression was enhanced by TG2 activation and promoted M2 macrophage polarization. Furthermore, the increase in the abundance of ALOX15-expressing macrophages in fibrotic kidney was dramatically suppressed in TG2-knockout mice. These findings demonstrated that TG2 activity exacerbates renal fibrosis by polarization of M2 macrophages from monocytes via ALOX15.

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“…Over the past ten years, proteomic analysis through mass spectrometry (MS) has become an extremely sensitive tool for characterizing key proteins involved in the progression of IPF as a chronic disease, elucidating the molecular mechanisms and profile of patients (23)(24)(25)(26). Therefore, the performance of MS proteomics has been maximized with spatially resolved proteomic to precisely delineate the molecular profile characterized by distinct protein compositions in the histologically defined regions of tissue in comparison to whole lung proteome (27)(28)(29). However, given the limitation of fresh tissues from patients with rare and chronic condition, the MS of archived tissues stored as formalin-fixed paraffin-embedded (FFPE) blocks have been widely and more easily used in research (30,31).…”

Section: Introductionmentioning

confidence: 99%

Proteomic profiling of formalin-fixed paraffine-embedded tissue reveals key proteins related to lung dysfunction in idiopathic pulmonary fibrosis

Samarelli,

Tonelli,

Raineri

et al. 2024

Front. Oncol.

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IntroductionIdiopathic pulmonary fibrosis (IPF) severely affects the lung leading to aberrant deposition of extracellular matrix and parenchymal stiffness with progressive functional derangement. The limited availability of fresh tissues represents one of the major limitations to study the molecular profiling of IPF lung tissue. The primary aim of this study was to explore the proteomic profiling yield of archived formalin-fixed paraffin-embedded (FFPE) specimens of IPF lung tissues.MethodsWe further determined the protein expression according to respiratory functional decline at the time of biopsy. The total proteins isolated from 11 FFPE samples of IPF patients compared to 3 FFPE samples from a non-fibrotic lung defined as controls, were subjected to label-free quantitative proteomic analysis by liquid chromatography-mass spectrometry (LC-MS/MS) and resulted in the detection of about 400 proteins.ResultsAfter the pairwise comparison between controls and IPF, functional enrichment analysis identified differentially expressed proteins that were involved in extracellular matrix signaling pathways, focal adhesion and transforming growth factor β (TGF-β) signaling pathways strongly associated with IPF onset and progression. Five proteins were significantly over- expressed in the lung of IPF patients with either advanced disease stage (Stage II) or impaired pulmonary function (FVC<75, DLCO<55) compared to controls; these were lymphocyte cytosolic protein 1 (LCP1), peroxiredoxin-2 (PRDX2), transgelin 2 (TAGLN2), lumican (LUM) and mimecan (OGN) that might play a key role in the fibrogenic processes.DiscussionOur work showed that the analysis of FFPE samples was able to identify key proteins that might be crucial for the IPF pathogenesis. These proteins are correlated with lung carcinogenesis or involved in the immune landscape of lung cancer, thus making possible common mechanisms between lung carcinogenesis and fibrosis progression, two pathological conditions at risk for each other in the real life.

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Spatially Resolved Identification of Transglutaminase Substrates by Proteomics in Pulmonary Fibrosis

Cited by 9 publications

References 47 publications

Role of Transglutaminase 2 in Cell Death, Survival, and Fibrosis

Role of Transglutaminase 2 in Cell Death, Survival, and Fibrosis

Tissue transglutaminase exacerbates renal fibrosis via alternative activation of monocyte-derived macrophages

Proteomic profiling of formalin-fixed paraffine-embedded tissue reveals key proteins related to lung dysfunction in idiopathic pulmonary fibrosis

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