40 results on '"Korkmaz, Brice"'
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2. Dipeptidyl peptidase-1 inhibition with brensocatib reduces the activity of all major neutrophil serine proteases in patients with bronchiectasis: results from the WILLOW trial
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Cipolla, David, Zhang, Jimin, Korkmaz, Brice, Chalmers, James D., Basso, Jessica, Lasala, Daniel, Fernandez, Carlos, Teper, Ariel, Mange, Kevin C., Perkins, Walter R., and Sullivan, Eugene J.
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- 2023
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3. Cathepsin C role in inflammatory gastroenterological, renal, rheumatic, and pulmonary disorders
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Aghdassi, Ali A., Pham, Christine, Zierke, Lukas, Mariaule, Vincent, Korkmaz, Brice, and Rhimi, Moez
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- 2024
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4. Neutrophils play a major role in the destruction of the olfactory epithelium during SARS-CoV-2 infection in hamsters
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Bourgon, Clara, Albin, Audrey St, Ando-Grard, Ophélie, Da Costa, Bruno, Domain, Roxane, Korkmaz, Brice, Klonjkowski, Bernard, Le Poder, Sophie, and Meunier, Nicolas
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- 2022
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5. Differences in Staining for Neutrophil Elastase and its Controlling Inhibitor SLPI Reveal Heterogeneity among Neutrophils in Psoriasis
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Skrzeczynska-Moncznik, Joanna, Zabieglo, Katarzyna, Osiecka, Oktawia, Morytko, Agnieszka, Brzoza, Piotr, Drozdz, Lukasz, Kapinska-Mrowiecka, Monika, Korkmaz, Brice, Pastuszczak, Maciej, Kosalka-Wegiel, Joanna, Musial, Jacek, and Cichy, Joanna
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- 2020
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6. Preservation with α1-antitrypsin improves primary graft function of murine lung transplants
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Götzfried, Jessica, Smirnova, Natalia F., Morrone, Carmela, Korkmaz, Brice, Yildirim, Ali Önder, Eickelberg, Oliver, and Jenne, Dieter E.
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- 2018
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7. Substrate-derived triazolo- and azapeptides as inhibitors of cathepsins K and S
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Galibert, Matthieu, Wartenberg, Mylène, Lecaille, Fabien, Saidi, Ahlame, Mavel, Sylvie, Joulin-Giet, Alix, Korkmaz, Brice, Brömme, Dieter, Aucagne, Vincent, Delmas, Agnès F., and Lalmanach, Gilles
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- 2018
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8. Activity-based protein profiling guided identification of urine proteinase 3 activity in subclinical rejection after renal transplantation
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Navarrete, Mario, Korkmaz, Brice, Guarino, Carla, Lesner, Adam, Lao, Ying, Ho, Julie, Nickerson, Peter, and Wilkins, John A.
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- 2020
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9. Premedication with a cathepsin C inhibitor alleviates early primary graft dysfunction in mouse recipients after lung transplantation
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Rehm, Salome R. T., Smirnova, Natalia F., Morrone, Carmela, Götzfried, Jessica, Feuchtinger, Annette, Pedersen, John, Korkmaz, Brice, Yildirim, Ali Önder, and Jenne, Dieter E.
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- 2019
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10. Analysis of urinary cathepsin C for diagnosing Papillon–Lefèvre syndrome
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Hamon, Yveline, Legowska, Monika, Fergelot, Patricia, Dallet-Choisy, Sandrine, Newell, Louise, Vanderlynden, Lise, Valeshabad, Ali Kord, Acrich, Karina, Kord, Hadi, Charalampos, Tsamakis, Morice-Picard, Fanny, Surplice, Ian, Zoidakis, Jerome, David, Karen, Vlahou, Antonia, Ragunatha, Shivanna, Nagy, Nikoletta, Farkas, Katalin, Széll, Márta, Goizet, Cyril, Schacher, Beate, Battino, Maurizio, Al Farraj Aldosari, Abdullah, Wang, Xinwen, Liu, Yang, Marchand-Adam, Sylvain, Lesner, Adam, Kara, Elodie, Korkmaz-Icöz, Sevil, Moss, Celia, Eickholz, Peter, Taieb, Alain, Kavukcu, Salih, Jenne, Dieter E., Gauthier, Francis, and Korkmaz, Brice
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- 2016
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11. Pathogenicity of Proteinase 3-Anti-Neutrophil Cytoplasmic Antibody in Granulomatosis With Polyangiitis: Implications as Biomarker and Future Therapies.
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Granel, Jérôme, Korkmaz, Brice, Nouar, Dalila, Weiss, Stefanie A. I., Jenne, Dieter E., Lemoine, Roxane, and Hoarau, Cyrille
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ANTINEUTROPHIL cytoplasmic antibodies ,PROTEINASES ,FC receptors ,DIAGNOSIS ,BIOMARKERS - Abstract
Granulomatosis with polyangiitis (GPA) is a rare but serious necrotizing auto-immune vasculitis. GPA is mostly associated with the presence of Anti-Neutrophil Cytoplasmic Antibody (ANCA) targeting proteinase 3 (PR3-ANCA), a serine protease contained in neutrophil granules but also exposed at the membrane. PR3-ANCAs have a proven fundamental role in GPA: they bind neutrophils allowing their auto-immune activation responsible for vasculitis lesions. PR3-ANCAs bind neutrophil surface on the one hand by their Fab binding PR3 and on the other by their Fc binding Fc gamma receptors. Despite current therapies, GPA is still a serious disease with an important mortality and a high risk of relapse. Furthermore, although PR3-ANCAs are a consistent biomarker for GPA diagnosis, relapse management currently based on their level is inconsistent. Indeed, PR3-ANCA level is not correlated with disease activity in 25% of patients suggesting that not all PR3-ANCAs are pathogenic. Therefore, the development of new biomarkers to evaluate disease activity and predict relapse and new therapies is necessary. Understanding factors influencing PR3-ANCA pathogenicity, i.e. their potential to induce auto-immune activation of neutrophils, offers interesting perspectives in order to improve GPA management. Most relevant factors influencing PR3-ANCA pathogenicity are involved in their interaction with neutrophils: level of PR3 autoantigen at neutrophil surface, epitope of PR3 recognized by PR3-ANCA, isotype and glycosylation of PR3-ANCA. We detailed in this review the advances in understanding these factors influencing PR3-ANCA pathogenicity in order to use them as biomarkers and develop new therapies in GPA as part of a personalized approach. [ABSTRACT FROM AUTHOR]
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- 2021
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12. Lung Protection by Cathepsin C Inhibition: A New Hope for COVID-19 and ARDS?
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Korkmaz, Brice, Lesner, Adam, Marchand-Adam, Sylvain, Moss, Celia, and Jenne, Dieter E.
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- 2020
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13. Human proteinase 3 resistance to inhibition extends to alpha‐2 macroglobulin.
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N'Guessan, Koffi, Grzywa, Renata, Seren, Seda, Gabant, Guillaume, Juliano, Maria A., Moniatte, Marc, Dorsselaer, Alain, Bieth, Joseph G., Kellenberger, Christine, Gauthier, Francis, Wysocka, Magdalena, Lesner, Adam, Sienczyk, Marcin, Cadene, Martine, and Korkmaz, Brice
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ELASTASES ,SERINE proteinases ,MASS spectrometry ,LEUCOCYTE elastase ,GRANULOMATOSIS with polyangiitis ,ENDOPEPTIDASES ,PROTEINASES ,PEPTIDASE - Abstract
Polymorphonuclear neutrophils contain at least four serine endopeptidases, namely neutrophil elastase (NE), proteinase 3 (PR3), cathepsin G (CatG), and NSP4, which contribute to the regulation of infection and of inflammatory processes. In physiological conditions, endogenous inhibitors including α2‐macroglobulin (α2‐M), serpins [α1‐proteinase inhibitor (α1‐PI)], monocyte neutrophil elastase inhibitor (MNEI), α1‐antichymotrypsin, and locally produced chelonianins (elafin, SLPI) control excessive proteolytic activity of neutrophilic serine proteinases. In contrast to human NE (hNE), hPR3 is weakly inhibited by α1‐PI and MNEI but not by SLPI. α2‐M is a large spectrum inhibitor that traps a variety of proteinases in response to cleavage(s) in its bait region. We report here that α2‐M was more rapidly processed by hNE than hPR3 or hCatG. This was confirmed by the observation that the association between α2‐M and hPR3 is governed by a kass in the ≤ 105 m−1·s−1 range. Since α2‐M‐trapped proteinases retain peptidase activity, we first predicted the putative cleavage sites within the α2‐M bait region (residues 690–728) using kinetic and molecular modeling approaches. We then identified by mass spectrum analysis the cleavage sites of hPR3 in a synthetic peptide spanning the 39‐residue bait region of α2‐M (39pep‐α2‐M). Since the 39pep‐α2‐M peptide and the corresponding bait area in the whole protein do not contain sequences with a high probability of specific cleavage by hPR3 and were indeed only slowly cleaved by hPR3, it can be concluded that α2‐M is a poor inhibitor of hPR3. The resistance of hPR3 to inhibition by endogenous inhibitors explains at least in part its role in tissue injury during chronic inflammatory diseases and its well‐recognized function of major target autoantigen in granulomatosis with polyangiitis. [ABSTRACT FROM AUTHOR]
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- 2020
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14. Structure-based design and in vivo anti-arthritic activity evaluation of a potent dipeptidyl cyclopropyl nitrile inhibitor of cathepsin C.
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Korkmaz, Brice, Lesner, Adam, Wysocka, Magdalena, Gieldon, Artur, Håkansson, Maria, Gauthier, Francis, Logan, Derek T., Jenne, Dieter E., Lauritzen, Conni, and Pedersen, John
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ELASTASES , *BONE marrow cells , *CYCLOPROPYL compounds , *BONE marrow , *RHEUMATOID arthritis , *AUTOIMMUNE diseases - Abstract
Cathepsin C (CatC) is a dipeptidyl-exopeptidase which activates neutrophil serine protease precursors (elastase, proteinase 3, cathepsin G and NSP4) by removing their N-terminal propeptide in bone marrow cells at the promyelocytic stage of neutrophil differentiation. The resulting active proteases are implicated in chronic inflammatory and autoimmune diseases. Hence, inhibition of CatC represents a therapeutic strategy to suppress excessive protease activities in various neutrophil mediated diseases. We designed and synthesized a series of dipeptidyl cyclopropyl nitrile compounds as putative CatC inhibitors. One compound, IcatC XPZ-01 ((S)-2-amino-N-((1 R ,2 R)-1-cyano-2-(4′-(4-methylpiperazin-1-ylsulfonyl)biphenyl-4-yl)cyclopropyl)butanamide)) was identified as a potent inhibitor of both human and rodent CatC. In mice, pharmacokinetic studies revealed that IcatC XPZ-01 accumulated in the bone marrow reaching levels suitable for CatC inhibition. Subcutaneous administration of IcatC XPZ-01 in a monoclonal anti-collagen antibody induced mouse model of rheumatoid arthritis resulted in statistically significant anti-arthritic activity with persistent decrease in arthritis scores and paw thickness. [ABSTRACT FROM AUTHOR]
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- 2019
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15. Therapeutic targeting of cathepsin C: from pathophysiology to treatment.
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Korkmaz, Brice, Caughey, George H., Chapple, Iain, Gauthier, Francis, Hirschfeld, Josefine, Jenne, Dieter E., Kettritz, Ralph, Lalmanach, Gilles, Lamort, Anne-Sophie, Lauritzen, Conni, Łȩgowska, Monika, Lesner, Adam, Marchand-Adam, Sylvain, McKaig, Sarah J., Moss, Celia, Pedersen, John, Roberts, Helen, Schreiber, Adrian, Seren, Seda, and Thakker, Nalin S.
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CATHEPSINS , *PATHOLOGICAL physiology , *AMINOPEPTIDASES , *SERINE proteinases , *DIPEPTIDES - Abstract
Abstract Cathepsin C (CatC) is a highly conserved tetrameric lysosomal cysteine dipeptidyl aminopeptidase. The best characterized physiological function of CatC is the activation of pro-inflammatory granule-associated serine proteases. These proteases are synthesized as inactive zymogens containing an N-terminal pro-dipeptide, which maintains the zymogen in its inactive conformation and prevents premature activation, which is potentially toxic to the cell. The activation of serine protease zymogens occurs through cleavage of the N-terminal dipeptide by CatC during cell maturation in the bone marrow. In vivo data suggest that pharmacological inhibition of pro-inflammatory serine proteases would suppress or attenuate deleterious effects mediated by these proteases in inflammatory/auto-immune disorders. The pathological deficiency in CatC is associated with Papillon-Lefèvre syndrome (PLS). The patients however do not present marked immunodeficiency despite the absence of active serine proteases in immune defense cells. Hence, the transitory pharmacological blockade of CatC activity in the precursor cells of the bone marrow may represent an attractive therapeutic strategy to regulate activity of serine proteases in inflammatory and immunologic conditions. A variety of CatC inhibitors have been developed both by pharmaceutical companies and academic investigators, some of which are currently being employed and evaluated in preclinical/clinical trials. [ABSTRACT FROM AUTHOR]
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- 2018
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16. Exploiting the S4-S5 Specificity of Human Neutrophil Proteinase 3 to Improve the Potency of Peptidyl Di(chlorophenyl)-phosphonate Ester Inhibitors: A Kinetic and Molecular Modeling Analysis.
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Guarino, Carla, Gruba, Natalia, Grzywa, Renata, Dyguda-Kazimierowicz, Edyta, Hamon, Yveline, Łȩgowska, Monika, Skoreński, Marcin, Dallet-Choisy, Sandrine, Marchand-Adam, Sylvain, Kellenberger, Christine, Jenne#, Dieter E., Sieńczyk, Marcin, Lesner, Adam, Gauthier, Francis, and Korkmaz, Brice
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- 2018
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17. Pseudomonas aeruginosa proteolytically alters the interleukin 22-dependent lung mucosal defense.
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Guillon, Antoine, Brea, Deborah, Morello, Eric, Tang, Aihua, Jouan, Youenn, Ramphal, Reuben, Korkmaz, Brice, Perez-Cruz, Magdiel, Trottein, Francois, O'Callaghan, Richard J., Gosset, Philippe, and Si-Tahar, Mustapha
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PSEUDOMONAS aeruginosa ,INTERLEUKINS ,MICROBIAL peptides ,EPITHELIAL cells ,CELL proliferation - Abstract
The IL-22 signaling pathway is critical for regulating mucosal defense and limiting bacterial dissemination. IL-22 is unusual among interleukins because it does not directly regulate the function of conventional immune cells, but instead targets cells at outer body barriers, such as respiratory epithelial cells. Consequently, IL-22 signaling participates in the maintenance of the lung mucosal barrier by controlling cell proliferation and tissue repair, and enhancing the production of specific chemokines and anti-microbial peptides.Pseudomonas aeruginosais a major pathogen of ventilator-associated pneumonia and causes considerable lung tissue damage. A feature underlying the pathogenicity of this bacterium is its capacity to persist and develop in the host, particularly in the clinical context of nosocomial lung infections. We aimed to investigate the ability ofP. auruginosato disrupt immune-epithelial cells cross-talk. We found thatP. aeruginosaescapes the host mucosal defenses by degrading IL-22, leading to severe inhibition of IL-22-mediated immune responses. We demonstratedin vitrothat, protease IV, a type 2 secretion system-dependent serine protease, is responsible for the degradation of IL-22 byP. aeruginosa. Moreover, the major anti-proteases molecules present in the lungs were unable to inhibit protease IV enzymatic activity. In addition, tracheal aspirates of patients infected byP. aeruginosacontain protease IV activity which further results in IL-22 degradation. This so far undescribed cleavage of IL-22 by a bacterial protease is likely to be an immune-evasion strategy that contributes toP. aeruginosa-triggered respiratory infections. [ABSTRACT FROM AUTHOR]
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- 2017
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18. Bile Acids: Key Players in Inflammatory Bowel Diseases?
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Kriaa, Aicha, Mariaule, Vincent, Jablaoui, Amin, Rhimi, Soufien, Mkaouar, Hela, Hernandez, Juan, Korkmaz, Brice, Lesner, Adam, Maguin, Emmanuelle, Aghdassi, Ali, and Rhimi, Moez
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INFLAMMATORY bowel diseases ,BILE acids ,GUT microbiome - Abstract
Inflammatory bowel diseases (IBDs) have emerged as a public health problem worldwide with a limited number of efficient therapeutic options despite advances in medical therapy. Although changes in the gut microbiota composition are recognized as key drivers of dysregulated intestinal immunity, alterations in bile acids (BAs) have been shown to influence gut homeostasis and contribute to the pathogenesis of the disease. In this review, we explore the interactions involving BAs and gut microbiota in IBDs, and discuss how the gut microbiota–BA–host axis may influence digestive inflammation. [ABSTRACT FROM AUTHOR]
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- 2022
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19. Cathepsin C inhibition as a potential treatment strategy in cancer.
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Korkmaz, Brice, Lamort, Anne-Sophie, Domain, Roxane, Beauvillain, Céline, Gieldon, Artur, Yildirim, Ali Önder, Stathopoulos, Georgios T., Rhimi, Moez, Jenne, Dieter E., and Kettritz, Ralph
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ELASTASES , *DISEASE risk factors , *CARCINOGENESIS , *SERINE proteinases , *PROTEOLYTIC enzymes , *NEUTROPHILS - Abstract
[Display omitted] Epidemiological studies established an association between chronic inflammation and higher risk of cancer. Inhibition of proteolytic enzymes represents a potential treatment strategy for cancer and prevention of cancer metastasis. Cathepsin C (CatC) is a highly conserved lysosomal cysteine dipeptidyl aminopeptidase required for the activation of pro-inflammatory neutrophil serine proteases (NSPs, elastase, proteinase 3, cathepsin G and NSP-4). NSPs are locally released by activated neutrophils in response to pathogens and non-infectious danger signals. Activated neutrophils also release neutrophil extracellular traps (NETs) that are decorated with several neutrophil proteins, including NSPs. NSPs are not only NETs constituents but also play a role in NET formation and release. Although immune cells harbor large amounts of CatC, additional cell sources for this protease exists. Upregulation of CatC expression was observed in different tissues during carcinogenesis and correlated with metastasis and poor patient survival. Recent mechanistic studies indicated an important interaction of tumor-associated CatC, NSPs, and NETs in cancer development and metastasis and suggested CatC as a therapeutic target in a several cancer types. Cancer cell-derived CatC promotes neutrophil recruitment in the inflammatory tumor microenvironment. Because the clinical consequences of genetic CatC deficiency in humans resulting in the elimination of NSPs are mild, small molecule inhibitors of CatC are assumed as safe drugs to reduce the NSP burden. Brensocatib, a nitrile CatC inhibitor is currently tested in a phase 3 clinical trial as a novel anti-inflammatory therapy for patients with bronchiectasis. However, recently developed CatC inhibitors possibly have protective effects beyond inflammation. In this review, we describe the pathophysiological function of CatC and discuss molecular mechanisms substantiating pharmacological CatC inhibition as a potential strategy for cancer treatment. [ABSTRACT FROM AUTHOR]
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- 2021
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20. Gut Serpinome: Emerging Evidence in IBD.
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Mkaouar, Héla, Mariaule, Vincent, Rhimi, Soufien, Hernandez, Juan, Kriaa, Aicha, Jablaoui, Amin, Akermi, Nizar, Maguin, Emmanuelle, Lesner, Adam, Korkmaz, Brice, and Rhimi, Moez
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INFLAMMATORY bowel diseases ,GUT microbiome ,SERINE proteinases ,SERINE proteinase inhibitors ,PROTEASE inhibitors ,SERPINS - Abstract
Inflammatory bowel diseases (IBD) are incurable disorders whose prevalence and global socioeconomic impact are increasing. While the role of host genetics and immunity is well documented, that of gut microbiota dysbiosis is increasingly being studied. However, the molecular basis of the dialogue between the gut microbiota and the host remains poorly understood. Increased activity of serine proteases is demonstrated in IBD patients and may contribute to the onset and the maintenance of the disease. The intestinal proteolytic balance is the result of an equilibrium between the proteases and their corresponding inhibitors. Interestingly, the serine protease inhibitors (serpins) encoded by the host are well reported; in contrast, those from the gut microbiota remain poorly studied. In this review, we provide a concise analysis of the roles of serine protease in IBD physiopathology and we focus on the serpins from the gut microbiota (gut serpinome) and their relevance as a promising therapeutic approach. [ABSTRACT FROM AUTHOR]
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- 2021
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21. Processing and Maturation of Cathepsin C Zymogen: A Biochemical and Molecular Modeling Analysis.
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Lamort, Anne-Sophie, Hamon, Yveline, Czaplewski, Cezary, Gieldon, Artur, Seren, Seda, Coquet, Laurent, Lecaille, Fabien, Lesner, Adam, Lalmanach, Gilles, Gauthier, Francis, Jenne, Dieter, and Korkmaz, Brice
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ELASTASES ,BIOCHEMICAL models ,SERINE proteinases ,MOLECULAR models ,GRANZYMES ,MISSENSE mutation - Abstract
Cysteine cathepsin C (CatC) is a ubiquitously expressed, lysosomal aminopeptidase involved in the activation of zymogens of immune-cell-associated serine proteinases (elastase, cathepsin G, proteinase 3, neutrophil serine proteinase 4, lymphocyte granzymes, and mast cell chymases). CatC is first synthetized as an inactive zymogen containing an intramolecular chain propeptide, the dimeric form of which is processed into the mature tetrameric form by proteolytic cleavages. A molecular modeling analysis of proCatC indicated that its propeptide displayed a similar fold to those of other lysosomal cysteine cathepsins, and could be involved in dimer formation. Our in vitro experiments revealed that human proCatC was processed and activated by CatF, CatK, and CatV in two consecutive steps of maturation, as reported for CatL and CatS previously. The unique positioning of the propeptide domains in the proCatC dimer complex allows this order of cleavages to be understood. The missense mutation Leu172Pro within the propeptide region associated with the Papillon–Lefèvre and Haim–Munk syndrome altered the proform stability as well as the maturation of the recombinant Leu172Pro proform. [ABSTRACT FROM AUTHOR]
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- 2019
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22. 196. Characterization of cathepsin c as a treatment target in ANCA-associated vasculitis.
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Jerke, Uwe, Eulenberg-Gustavus, Claudia, Rousselle, Anthony, Kreideweiss, Stefan, Grundl, Marc, Eickholz, Peter, Nickles, Katrin, Schreiber, Adrian, Korkmaz, Brice, and Kettriz, Ralph
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VASCULITIS ,CONFERENCES & conventions ,PROTEOLYTIC enzymes ,DIAGNOSIS - Published
- 2019
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23. Proteinase 3 phosphonic inhibitors.
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Grzywa, Renata, Lesner, Adam, Korkmaz, Brice, and Sieńczyk, Marcin
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HEAT shock proteins , *PROTEINASES , *SERINE proteinases , *ARMED Forces , *MILITARY service - Abstract
Neutrophils are one of the most important military services of the armed forces of the immune system, a crucial line of defense against bacterial or fungal onslaughts. One of their mechanisms of action relies on the production of serine proteases. One of these enzymes is proteinase 3 (PR3), which is engaged in the processing of pro-inflammatory cytokines, receptors, heat shock proteins and in the generation of antibacterial peptides. Despite its protective function, uncontrolled activity of PR3 has been associated with the progression of inflammation and tissue injury. Although PR3 was characterized at the beginning of 90's of the last century for the first time, the research on the development of its inhibitors is barely noticeable. Here we present the recent findings on the design, synthesis and the activity of phosphonic PR3 inhibitors together with the historical perspective. Image 1 • Proteinase 3 is considered an interesting drug target since its activity has been associated with different pathophysiological disorders. • Several classes of PR3 inhibitors such as isocoumarins, N -hydroxysuccinimides, thiatriazolidine and phosphonates have been developed. • Diaryl esters of phosphonic analogues of amino acids represent an interesting class of irreversible inhibitors of proteinase 3. • Phosphonic inhibitors can be easily turned into the activity-based probes useful for protease visualization either in vitro or in cell assays. [ABSTRACT FROM AUTHOR]
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- 2019
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24. Constitutive and induced forms of membrane-bound proteinase 3 interact with antineutrophil cytoplasmic antibodies and promote immune activation of neutrophils.
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Guarino, Carla, Seren, Seda, Lemoine, Roxane, Hummel, Amber M., Margotin, Jean-Edouard, El-Benna, Jamel, Hoarau, Cyrille, Specks, Ulrich, Jenne, Dieter E., and Korkmaz, Brice
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ANTINEUTROPHIL cytoplasmic antibodies , *NEUTROPHILS , *PROTEINASES , *ELASTASES - Abstract
Proteinase 3 (PR3) is the main target antigen of antineutrophil cytoplasmic antibodies (ANCAs) in PR3-ANCA-associated vasculitis. A small fraction of PR3 is constitutively exposed on the surface of quiescent blood neutrophils in a proteolytically inactive form. When activated, neutrophils expose an induced form of membrane-bound PR3 (PR3mb) on their surface as well, which is enzymatically less active than unbound PR3 in solution due to its altered conformation. In this work, our objective was to understand the respective role of constitutive and induced PR3mb in the immune activation of neutrophils triggered by murine anti-PR3 mAbs and human PR3-ANCA. We quantified immune activation of neutrophils by the measurement of the production of superoxide anions and secreted protease activity in the cell supernatant before and after treatment of the cells by alpha-1 protease inhibitor that clears induced PR3mb from the cell surface. Incubation of TNFa-primed neutrophils with anti-PR3 antibodies resulted in a significant increase in superoxide anion production, membrane activation marker exposition, and secreted protease activity. When primed neutrophils were first treated with alpha-1 protease inhibitor, we observed a partial reduction in antibody-induced neutrophil activation, suggesting that constitutive PR3mb is sufficient to activate neutrophils. The pretreatment of primed neutrophils with purified antigenbinding fragments used as competitor significantly reduced cell activation by whole antibodies. This led us to the conclusion that PR3mb promoted immune activation of neutrophils. We propose that blocking and/or elimination of PR3mb offers a new therapeutic strategy to attenuate neutrophil activation in patients with PR3-ANCA-associated vasculitis. [ABSTRACT FROM AUTHOR]
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- 2023
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25. Alpha-1-Antitrypsin Protects Vascular Grafts of Brain-Dead Rats Against Ischemia/Reperfusion Injury.
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Ding, Qingwei, Loganathan, Sivakkanan, Zhou, Pengyu, Sayour, Alex Ali, Brlecic, Paige, Radovits, Tamás, Domain, Roxane, Korkmaz, Brice, Karck, Matthias, Szabó, Gábor, and Korkmaz-Icöz, Sevil
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VASCULAR grafts , *REPERFUSION injury , *HEART transplantation , *ISCHEMIA , *BRAIN death - Abstract
Endothelial dysfunction is a potential side effect of brain death (BD). Ischemia/reperfusion (IR) injury during heart transplantation may lead to further endothelial damage. Protective effects of alpha-1-antitrypsin (AAT), a human neutrophil serine protease inhibitor, have been demonstrated against IR injury. We hypothesized that AAT protects brain-dead rats' vascular grafts from IR injury. Donor rats were subjected to BD by inflation of a subdural balloon. After 5.5 h, aortic rings were immediately mounted in organ baths (BD, n = 6 rats) or preserved in saline, supplemented either with vehicle (BD-IR, n = 8 rats) or AAT (BD-IR + AAT, n = 14 rats) for 24 h. During organ bath experiment, rings from both IR groups were exposed to hypochlorite to simulate warm reperfusion-associated endothelial injury. Endothelial function was measured ex vivo. Immunohistochemical staining for caspases was carried out and DNA-strand breaks were evaluated using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling. Data are presented as median (interquartile range). AAT improved IR-induced decreased maximum endothelium-dependent vasorelaxation to acetylcholine in the BD-IR + AAT aortas compared to the BD-IR group (BD: 83 (9-28) % versus BD-IR: 49 (39-60) % versus BD-IR + AAT: 64 (24-42) %, P < 0.05). Additionally, an increase in the rings' sensitivity to acetylcholine was noted after AAT (pD 2 -value: BD-IR + AAT: 7.35 (7.06-7.89) versus BD-IR: 6.96 (6.65-7.21), P < 0.05). Caspase-3, -8, -9, and -12 immunoreactivity and the number of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive cells were significantly decreased by AAT. AAT alleviates endothelial dysfunction, prevents increased caspase-3, -8, -9, and -12 levels, and decreases apoptotic DNA breakage due to BD and IR injury. This suggests that AAT treatment may be therapeutically beneficial to reduce IR-induced vascular damage. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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26. Prolonged pharmacological inhibition of cathepsin C results in elimination of neutrophil serine proteases.
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Guarino, Carla, Hamon, Yveline, Croix, Cécile, Lamort, Anne-Sophie, Dallet-Choisy, Sandrine, Marchand-Adam, Sylvain, Lesner, Adam, Baranek, Thomas, Viaud-Massuard, Marie-Claude, Lauritzen, Conni, Pedersen, John, Heuzé-Vourc'h, Nathalie, Si-Tahar, Mustapha, Fıratlı, Erhan, Jenne, Dieter E., Gauthier, Francis, Horwitz, Marshall S., Borregaard, Niels, and Korkmaz, Brice
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CATHEPSINS , *NEUTROPHILS , *SERINE proteinases , *ZYMOGENS , *GENETIC mutation , *PAPILLON Lefevre syndrome - Abstract
Cathepsin C (CatC) is a tetrameric cysteine dipeptidyl aminopeptidase that plays a key role in activation of pro-inflammatory serine protease zymogens by removal of a N-terminal pro-dipeptide sequence. Loss of function mutations in the CatC gene is associated with lack of immune cell serine protease activities and cause Papillon-Lefèvre syndrome (PLS). Also, only very low levels of elastase-like protease zymogens are detected by proteome analysis of neutrophils from PLS patients. Thus, CatC inhibitors represent new alternatives for the treatment of neutrophil protease-driven inflammatory or autoimmune diseases. We aimed to experimentally inactivate and lower neutrophil elastase-like proteases by pharmacological blocking of CatC-dependent maturation in cell-based assays and in vivo . Isolated, immature bone marrow cells from healthy donors pulse-chased in the presence of a new cell permeable cyclopropyl nitrile CatC inhibitor almost totally lack elastase. We confirmed the elimination of neutrophil elastase-like proteases by prolonged inhibition of CatC in a non-human primate. We also showed that neutrophils lacking elastase-like protease activities were still recruited to inflammatory sites. These preclinical results demonstrate that the disappearance of neutrophil elastase-like proteases as observed in PLS patients can be achieved by pharmacological inhibition of bone marrow CatC. Such a transitory inhibition of CatC might thus help to rebalance the protease load during chronic inflammatory diseases, which opens new perspectives for therapeutic applications in humans. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
27. Development of the first internally-quenched fluorescent substrates of human cathepsin C: The application in the enzyme detection in biological samples.
- Author
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Łęgowska, Monika, Hamon, Yveline, Wojtysiak, Anna, Grzywa, Renata, Sieńczyk, Marcin, Burster, Timo, Korkmaz, Brice, and Lesner, Adam
- Subjects
- *
CATHEPSINS , *CYSTEINE , *PEPTIDASE , *PROTEOLYTIC enzymes , *NEUTROPHILS - Abstract
Cathepsin C is a widely expressed cysteine exopeptidase that is mostly recognized for the activation of the granule-associated proinflammatory serine proteases in neutrophils, cytotoxic T lymphocytes and mast cells. It has been shown that the enzyme can be secreted extracellularly; however, its occurrence in human bodily fluids/physiological samples has not been thoroughly studied. In the course of this study, the first fluorescence resonance energy transfer peptides for the measurement of the activity of human cathepsin C were designed and synthesized. Two series of tetra- and pentapeptide substrates enabled the detailed S′ specificity study of cathepsin C, which has been examined for the first time. The extensive enzymatic studies of the obtained compounds resulted in the selection of the highly specific and selective substrate Thi-Ala(Mca)-Ser - Gly-Tyr(3-NO 2 )-NH 2 , which was successfully employed for the detection of cathepsin C activity in complex biological samples such as cell lysates, urine and bronchoalveolar lavage fluids. Molecular docking of the selected substrate was performed in order to better understand the binding mode of the substrates in the active site of cathepsin C. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
28. Neutrophilic Cathepsin C Is Maturated by a Multistep Proteolytic Process and Secreted by Activated Cells during Inflammatory Lung Diseases.
- Author
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Hamon, Yveline, Legowska, Monika, Hervé, Virginie, Dallet-Choisy, Sandrine, Marchand-Adam, Sylvain, Vanderlynden, Lise, Demonte, Michèle, Williams, Rich, Scott, Christopher J., Si-Tahar, Mustapha, Heuzé-Vourc'h, Nathalie, Lalmanach, Gilles, Jenne, Dieter E., Lesner, Adam, Gauthier, Francis, and Korkmaz, Brice
- Subjects
- *
CYSTEINE proteinase inhibitors , *CATHEPSINS , *PROTEOLYTIC enzymes , *LUNG disease treatment , *SERINE proteinases - Abstract
The cysteine protease cathepsin C (CatC) activates granuleassociated proinflammatory serine proteases in hematopoietic precursor cells. Its early inhibition in the bone marrow is regarded as a new therapeutic strategy for treating proteolysisdriven chronic inflammatory diseases, but its complete inhibition is elusive in vivo. Controlling the activity of CatC may be achieved by directly inhibiting its activity with a specific inhibitor or/and by preventing its maturation. We have investigated immunochemically and kinetically the occurrence of CatC and its proform in human hematopoietic precursor cells and in differentiated mature immune cells in lung secretions. The maturation of proCatC obeys a multistep mechanism that can be entirely managed by CatS in neutrophilic precursor cells. CatS inhibition by a cell-permeable inhibitor abrogated the release of the heavy and light chains from proCatC and blocked ∼80% of CatC activity. Under these conditions the activity of neutrophil serine proteases, however, was not abolished in precursor cell cultures. In patients with neutrophilic lung inflammation, mature CatC is found in large amounts in sputa. It is secreted by activated neutrophils as confirmed through lipopolysaccharide administration in a nonhuman primate model. CatS inhibitors currently in clinical trials are expected to decrease the activity of neutrophilic CatC without affecting those of elastase-like serine proteases. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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- View/download PDF
29. Neutrophil serine proteases exert proteolytic activity on endothelial cells.
- Author
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Jerke, Uwe, Hernandez, Daniel Perez, Beaudette, Patrick, Korkmaz, Brice, Dittmar, Gunnar, and Kettritz, Ralph
- Subjects
- *
APOPTOSIS , *BIOCHEMISTRY , *CELL lines , *CELL physiology , *CELLULAR signal transduction , *CYTOPLASM , *DOSE-effect relationship in pharmacology , *EPITHELIAL cells , *IMMUNITY , *PHENOMENOLOGY , *METABOLISM , *NEUTROPHILS , *PROTEOLYTIC enzymes , *TIME , *ALBUMINS - Abstract
Neutrophil serine proteases (NSPs) are released from activated neutrophils during inflammation. Here we studied the transfer of the three major NSPs, namely proteinase 3, human neutrophil elastase, and cathepsin G, from neutrophils to endothelial cells and used an unbiased approach to identify novel endothelial NSP substrates. Enzymatically active NSPs were released from stimulated neutrophils and internalized by endothelial cells in a dose- and time-dependent manner as shown by immunoblotting, flow cytometry, and the Boc-Ala substrate assay. Using terminal-amine isotopic labeling of substrates in endothelial cells, we identified 121 peptides from 82 different proteins consisting of 36 substrates for proteinase 3, 30 for neutrophil elastase, and 28 for cathepsin G, respectively. We characterized the extended cleavage pattern and provide corresponding IceLogos. Gene ontology analysis showed significant cytoskeletal substrate enrichment and confirmed several cytoskeletal protein substrates by immunoblotting. Finally, ANCA-stimulated neutrophils released all three active NSPs into the supernatant. Supernatants increased endothelial albumin flux and disturbed the endothelial cell cytoskeletal architecture. Serine protease inhibition abrogated this effect. Longer exposure to NSPs reduced endothelial cell viability and increased apoptosis. Thus, we identified novel NSP substrates and suggest NSP inhibition as a therapeutic measure to inhibit neutrophil-mediated inflammatory vascular diseases. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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30. New Selective Peptidyl Di(chlorophenyl) Phosphonate Esters for Visualizing and Blocking Neutrophil Proteinase 3 in Human Diseases.
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Guarino, Carla, Legowska, Monika, Epinette, Christophe, Kellenberger, Christine, Dallet-Choisy, Sandrine, Sieńczyk, Marcin, Gabant, Guillaume, Cadene, Martine, Zoidakis, Jérôme, Vlahou, Antonia, Wysocka, Magdalena, Marchand-Adam, Sylvain, Jenne, Dieter E., Lesner, Adam, Gauthier, Francis, and Korkmaz, Brice
- Subjects
- *
NEUTROPHILS , *AUTOIMMUNE diseases , *ESTERS , *ELASTASES , *BLADDER cancer , *PHYSIOLOGY - Abstract
The function of neutrophil protease 3 (PR3) is poorly understood despite of its role in autoimmune vasculitides and its possible involvement in cell apoptosis. This makes it different from its structural homologue neutrophil elastase (HNE). Endogenous inhibitors of human neutrophil serine proteases preferentially inhibit HNE and to a lesser extent, PR3. We constructed a single-residue mutant PR3 (I217R) to investigate the S4 subsite preferences of PR3 andHNEand used the best peptide substrate sequences to develop selective phosphonate inhibitors with the structure Ac-peptidylP(O-C6H4-4-Cl)2. The combination of a prolyl residue at P4 and an aspartyl residue at P2 was totally selective for PR3. We then synthesized N-terminally biotinylated peptidyl phosphonates to identify the PR3 in complex biological samples. These inhibitors resisted proteolytic degradation and rapidly inactivated PR3 in biological fluids such as inflammatory lung secretions and the urine of patients with bladder cancer. One of these inhibitors revealed intracellular PR3 in permeabilized neutrophils and on the surface of activated cells. They hardly inhibited PR3 bound to the surface of stimulated neutrophils despite their low molecular mass, suggesting that the conformation and reactivity of membrane-bound PR3 is altered. This finding is relevant for autoantibody binding and the subsequent activation of neutrophils in granulomatosis with polyangiitis (formerly Wegener disease). These are the first inhibitors that can be used as probes to monitor, detect, and control PR3 activity in a variety of inflammatory diseases. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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31. Dipeptidyl peptidase 1 inhibition as a potential therapeutic approach in neutrophil-mediated inflammatory disease.
- Author
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Chalmers JD, Kettritz R, and Korkmaz B
- Subjects
- Humans, Protease Inhibitors, Cathepsin C, Inflammation drug therapy, Inflammation pathology, Neutrophils pathology, Serine Proteases
- Abstract
Neutrophils have a critical role in the innate immune response to infection and the control of inflammation. A key component of this process is the release of neutrophil serine proteases (NSPs), primarily neutrophil elastase, proteinase 3, cathepsin G, and NSP4, which have essential functions in immune modulation and tissue repair following injury. Normally, NSP activity is controlled and modulated by endogenous antiproteases. However, disruption of this homeostatic relationship can cause diseases in which neutrophilic inflammation is central to the pathology, such as chronic obstructive pulmonary disease (COPD), alpha-1 antitrypsin deficiency, bronchiectasis, and cystic fibrosis, as well as many non-pulmonary pathologies. Although the pathobiology of these diseases varies, evidence indicates that excessive NSP activity is common and a principal mediator of tissue damage and clinical decline. NSPs are synthesized as inactive zymogens and activated primarily by the ubiquitous enzyme dipeptidyl peptidase 1, also known as cathepsin C. Preclinical data confirm that inactivation of this protease reduces activation of NSPs. Thus, pharmacological inhibition of dipeptidyl peptidase 1 potentially reduces the contribution of aberrant NSP activity to the severity and/or progression of multiple inflammatory diseases. Initial clinical data support this view. Ongoing research continues to explore the role of NSP activation by dipeptidyl peptidase 1 in different disease states and the potential clinical benefits of dipeptidyl peptidase 1 inhibition., Competing Interests: JDC has received grants and personal fees from AstraZeneca, Boehringer-Ingelheim, GSK, Zambon, and Insmed Incorporated, a grant from Gilead, and personal fees from Novartis and Chiesi. RK has received a grant from Boehringer-Ingelheim and personal fees from Insmed Incorporated. BK has been paid for the time spent as a committee member for advisory boards (Brensocatib Advisory Board [BRAB] Insmed Incorporated, USA), other forms of consulting (Boehringer Ingelheim [Germany], Neuprozyme Therapeutics Aps [Denmark], Santhera Pharmaceuticals [Switzerland], Chiesi [Italy], Gerson Lehrman Group [GLG] [USA]), symposium organisation (Insmed Incorporated) and travel support, and lectures or presentations, outside of the submitted work., (Copyright © 2023 Chalmers, Kettritz and Korkmaz.)
- Published
- 2023
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32. Preservation solution Custodiol containing human alpha-1-antitrypsin improves graft recovery after prolonged cold ischemic storage in a rat model of heart transplantation.
- Author
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Korkmaz-Icöz S, Abulizi S, Li K, Korkmaz B, Georgevici AI, Sayour AA, Loganathan S, Canoglu H, Karck M, and Szabó G
- Subjects
- Animals, Humans, Rats, Heart, Ischemia, Rats, Inbred Lew, Tissue Donors, Heart Transplantation, Organ Preservation Solutions
- Abstract
Introduction: The shortage of available donor hearts and the risk of ischemia/reperfusion injury restrict heart transplantation (HTX). Alpha-1-antitrypsin (AAT), a well-characterized inhibitor of neutrophil serine protease, is used in augmentation therapy to treat emphysema due to severe AAT deficiency. Evidence demonstrates its additional anti-inflammatory and tissue-protective effects. We hypothesized that adding human AAT in a preservation solution reduces graft dysfunction in a rat model of HTX following extended cold ischemic storage., Methods: The hearts from isogenic Lewis donor rats were explanted, stored for either 1h or 5h in cold Custodiol supplemented with either vehicle (1h ischemia, n=7 or 5h ischemia, n=7 groups) or 1 mg/ml AAT (1h ischemia+AAT, n=7 or 5h ischemia+AAT, n=9 groups) before heterotopic HTX. Left-ventricular (LV) graft function was evaluated in vivo 1.5h after HTX. Immunohistochemical detection of myeloperoxydase (MPO) was performed in myocardial tissue and expression of 88 gene quantified with PCR was analyzed both statistical and with machine-learning methods., Results: After HTX, LV systolic function (dP/dt
max 1h ischemia+AAT 4197 ± 256 vs 1h ischemia 3123 ± 110; 5h ischemia+AAT 2858 ± 154 vs 5h ischemia 1843 ± 104mmHg/s, p <0.05) and diastolic function (dP/dtmin 5h ischemia+AAT 1516 ± 68 vs 5h ischemia 1095 ± 67mmHg/s, p <0.05) at an intraventricular volume of 90µl were improved in the AAT groups compared with the corresponding vehicle groups. In addition, the rate pressure product (1h ischemia+AAT 53 ± 4 vs 1h ischemia 26 ± 1; 5h ischemia+AAT 37 ± 3 vs 5h ischemia 21 ± 1mmHg*beats/min at an intraventricular volume of 90µl; p <0.05) was increased in the AAT groups compared with the corresponding vehicle groups. Moreover, the 5h ischemia+AAT hearts exhibited a significant reduction in MPO-positive cell infiltration in comparison to the 5h ischemia group. Our computational analysis shows that ischemia+AAT network displays higher homogeneity, more positive and fewer negative gene correlations than the ischemia+placebo network., Discussion: We provided experimental evidence that AAT protects cardiac grafts from prolonged cold ischemia during HTX in rats., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Korkmaz-Icöz, Abulizi, Li, Korkmaz, Georgevici, Sayour, Loganathan, Canoglu, Karck and Szabó.)- Published
- 2023
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33. Constitutive and induced forms of membrane-bound proteinase 3 interact with antineutrophil cytoplasmic antibodies and promote immune activation of neutrophils.
- Author
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Carla Guarino, Seren S, Lemoine R, Hummel AM, Margotin JE, El-Benna J, Hoarau C, Specks U, Jenne DE, and Korkmaz B
- Subjects
- Animals, Humans, Mice, Neutrophils metabolism, Protease Inhibitors metabolism, Superoxides metabolism, Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis metabolism, Antibodies, Antineutrophil Cytoplasmic, Myeloblastin immunology, Myeloblastin metabolism
- Abstract
Proteinase 3 (PR3) is the main target antigen of antineutrophil cytoplasmic antibodies (ANCAs) in PR3-ANCA-associated vasculitis. A small fraction of PR3 is constitutively exposed on the surface of quiescent blood neutrophils in a proteolytically inactive form. When activated, neutrophils expose an induced form of membrane-bound PR3 (PR3
mb ) on their surface as well, which is enzymatically less active than unbound PR3 in solution due to its altered conformation. In this work, our objective was to understand the respective role of constitutive and induced PR3mb in the immune activation of neutrophils triggered by murine anti-PR3 mAbs and human PR3-ANCA. We quantified immune activation of neutrophils by the measurement of the production of superoxide anions and secreted protease activity in the cell supernatant before and after treatment of the cells by alpha-1 protease inhibitor that clears induced PR3mb from the cell surface. Incubation of TNFα-primed neutrophils with anti-PR3 antibodies resulted in a significant increase in superoxide anion production, membrane activation marker exposition, and secreted protease activity. When primed neutrophils were first treated with alpha-1 protease inhibitor, we observed a partial reduction in antibody-induced neutrophil activation, suggesting that constitutive PR3mb is sufficient to activate neutrophils. The pretreatment of primed neutrophils with purified antigen-binding fragments used as competitor significantly reduced cell activation by whole antibodies. This led us to the conclusion that PR3mb promoted immune activation of neutrophils. We propose that blocking and/or elimination of PR3mb offers a new therapeutic strategy to attenuate neutrophil activation in patients with PR3-ANCA-associated vasculitis., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)- Published
- 2023
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- View/download PDF
34. Targeting Cathepsin C in PR3-ANCA Vasculitis.
- Author
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Jerke U, Eulenberg-Gustavus C, Rousselle A, Nicklin P, Kreideweiss S, Grundl MA, Eickholz P, Nickles K, Schreiber A, Korkmaz B, and Kettritz R
- Subjects
- Antibodies, Antineutrophil Cytoplasmic, Cathepsin C metabolism, Endothelial Cells metabolism, Enzyme Precursors metabolism, Humans, Myeloblastin genetics, Neutrophils metabolism, Peroxidase, Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis, Papillon-Lefevre Disease metabolism
- Abstract
Background: The ANCA autoantigens proteinase 3 (PR3) and myeloperoxidase (MPO) are exclusively expressed by neutrophils and monocytes. ANCA-mediated activation of these cells is the key driver of the vascular injury process in ANCA-associated vasculitis (AAV), and neutrophil serine proteases (NSPs) are disease mediators. Cathepsin C (CatC) from zymogens activates the proteolytic function of NSPs, including PR3. Lack of NSP zymogen activation results in neutrophils with strongly reduced NSP proteins., Methods: To explore AAV-relevant consequences of blocking NSP zymogen activation by CatC, we used myeloid cells from patients with Papillon-Lefèvre syndrome, a genetic deficiency of CatC, to assess NSPs and NSP-mediated endothelial cell injury. We also examined pharmacologic CatC inhibition in neutrophil-differentiated human hematopoietic stem cells, primary human umbilical vein cells, and primary glomerular microvascular endothelial cells., Results: Patients with Papillon-Lefèvre syndrome showed strongly reduced NSPs in neutrophils and monocytes. Neutrophils from these patients produced a negative PR3-ANCA test, presented less PR3 on the surface of viable and apoptotic cells, and caused significantly less damage in human umbilical vein cells. These findings were recapitulated in human stem cells, in which a highly specific CatC inhibitor, but not prednisolone, reduced NSPs without affecting neutrophil differentiation, reduced membrane PR3, and diminished neutrophil activation upon PR3-ANCA but not MPO-ANCA stimulation. Compared with healthy controls, neutrophils from patients with Papillon-Lefèvre syndrome transferred less proteolytically active NSPs to glomerular microvascular endothelial cells, the cell type targeted in ANCA-induced necrotizing crescentic glomerulonephritis. Finally, both genetic CatC deficiency and pharmacologic inhibition, but not prednisolone, reduced neutrophil-induced glomerular microvascular endothelial cell damage., Conclusions: These findings may offer encouragement for clinical studies of adjunctive CatC inhibitor in patients with PR3-AAV., (Copyright © 2022 by the American Society of Nephrology.)
- Published
- 2022
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- View/download PDF
35. Monitoring Human Neutrophil Activation by a Proteinase 3 Near-Infrared Fluorescence Substrate-Based Probe.
- Author
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Saidi A, Wartenberg M, Madinier JB, Ilango G, Seren S, Korkmaz B, Lecaille F, Aucagne V, and Lalmanach G
- Subjects
- Cell Survival, Fluorescent Dyes, Gene Expression Regulation, Enzymologic drug effects, Humans, Ionomycin, Microscopy, Confocal, Molecular Structure, Myeloblastin chemistry, Neutrophils drug effects, Spectrophotometry, Infrared, Myeloblastin metabolism, Neutrophil Activation physiology, Neutrophils physiology
- Abstract
A near-infrared fluorescent (NIRF) substrate-based probe (SBP) was conceived to monitor secreted human proteinase 3 (hPR3) activity. This probe, called pro3-SBP, is shaped by a fused peptide hairpin loop structure, which associates a hPR3 recognition domain (Val-Ala-Asp-Nva-Ala-Asp-Tyr-Gln, where Nva is norvaline) and an electrostatic zipper (consisting of complementary polyanionic (d-Glu)
5 and polycationic (d-Arg)5 sequences) in close vicinity of the N- and C-terminal FRET couple (fluorescent donor, sulfoCy5.5; dark quencher, QSY21). Besides its subsequent stability, no intermolecular fluorescence quenching was detected following its complete hydrolysis by hPR3, advocating that pro3-SBP could further afford unbiased imaging. Pro3-SBP was specifically hydrolyzed by hPR3 ( kcat / Km = 440 000 ± 5500 M-1 ·s-1 ) and displayed a sensitive detection threshold for hPR3 (subnanomolar concentration range), while neutrophil elastase showed a weaker potency. Conversely, pro3-SBP was not cleaved by cathepsin G. Pro3-SBP was successfully hydrolyzed by conditioned media of activated human neutrophils but not by quiescent neutrophils. Moreover, unlike unstimulated neutrophils, a strong NIRF signal was specifically detected by confocal microscopy following neutrophil ionomycin-induced degranulation. Fluorescence release was abolished in the presence of a selective hPR3 inhibitor, indicating that pro3-SBP is selectively cleaved by extracellular hPR3. Taken together, the present data support that pro3-SBP could be a convenient tool, allowing straightforward monitoring of human neutrophil activation.- Published
- 2021
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36. Proteinase release from activated neutrophils in mechanically ventilated patients with non-COVID-19 and COVID-19 pneumonia.
- Author
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Seren S, Derian L, Keleş I, Guillon A, Lesner A, Gonzalez L, Baranek T, Si-Tahar M, Marchand-Adam S, Jenne DE, Paget C, Jouan Y, and Korkmaz B
- Subjects
- Humans, Neutrophils, Peptide Hydrolases, Respiration, Artificial, SARS-CoV-2, COVID-19
- Abstract
Competing Interests: Conflict of interest: S. Seren has nothing to disclose. Conflict of interest: L. Derian has nothing to disclose. Conflict of interest: I. Keleş has nothing to disclose. Conflict of interest: A. Guillon has nothing to disclose. Conflict of interest: A. Lesner has nothing to disclose. Conflict of interest: L. Gonzalez has nothing to disclose. Conflict of interest: T. Baranek has nothing to disclose. Conflict of interest: M. Si-Tahar has nothing to disclose. Conflict of interest: S. Marchand-Adam has nothing to disclose. Conflict of interest: D.E. Jenne has nothing to disclose. Conflict of interest: C. Paget has nothing to disclose. Conflict of interest: Y. Jouan has nothing to disclose. Conflict of interest: B. Korkmaz has been paid for the time spent as a committee member for advisory boards (INSMED), other forms of consulting (Neuprozyme Therapeutics Aps (Denmark), Santhera Pharmaceuticals (Switzerland)), symposium organisation (INSMED) and travel support, lectures or presentations, outside the submitted work.
- Published
- 2021
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37. 4C3 Human Monoclonal Antibody: A Proof of Concept for Non-pathogenic Proteinase 3 Anti-neutrophil Cytoplasmic Antibodies in Granulomatosis With Polyangiitis.
- Author
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Granel J, Lemoine R, Morello E, Gallais Y, Mariot J, Drapeau M, Musnier A, Poupon A, Pugnière M, Seren S, Nouar D, Gouilleux-Gruart V, Watier H, Korkmaz B, and Hoarau C
- Subjects
- Aged, Antibodies, Antineutrophil Cytoplasmic metabolism, Antibodies, Monoclonal metabolism, Antibody Affinity, Antibody Specificity, B-Lymphocytes enzymology, Binding Sites, Antibody, Biomarkers metabolism, Case-Control Studies, Cell Line, Epitope Mapping, Epitopes, Female, Glycosylation, Granulomatosis with Polyangiitis diagnosis, Granulomatosis with Polyangiitis enzymology, Humans, Male, Middle Aged, Neutrophil Activation, Proof of Concept Study, Antibodies, Antineutrophil Cytoplasmic immunology, Antibodies, Monoclonal immunology, B-Lymphocytes immunology, Granulomatosis with Polyangiitis immunology, Myeloblastin immunology
- Abstract
Granulomatosis with polyangiitis (GPA) is a severe autoimmune vasculitis associated with the presence of anti-neutrophil cytoplasmic antibodies (ANCA) mainly targeting proteinase 3 (PR3), a neutrophilic serine proteinase. PR3-ANCA binding to membrane-bound PR3 on neutrophils induce their auto-immune activation responsible for vascular lesions. However, the correlation between PR3-ANCA level and disease activity remains inconsistent, suggesting the existence of non-pathogenic PR3-ANCA. In order to prove their existence, we immortalized B lymphocytes from blood samples of GPA patients in remission having persistent PR3-ANCA to isolate non-activating PR3-ANCA. We obtained for the first time a non-activating human IgG1κ anti-PR3 monoclonal antibody (mAb) named 4C3. This new mAb binds soluble PR3 with a high affinity and membrane-bound PR3 on an epitope close to the PR3 hydrophobic patch and in the vicinity of the active site. 4C3 is able to bind FcγRIIA and FcγRIIIB and has a G2F glycosylation profile on asparagine 297. 4C3 did not induce activation of neutrophils and could inhibit human polyclonal PR3-ANCA-induced activation suggesting that 4C3 is non-pathogenic. This characteristic relies on the recognized epitope on PR3 rather than to the Fc portion properties. The existence of non-pathogenic PR3-ANCA, which do not activate neutrophils, could explain the persistence of high PR3-ANCA levels in some GPA patients in remission and why PR3-ANCA would not predict relapse. Finally, these results offer promising perspectives particularly regarding the understanding of PR3-ANCA pathogenicity and the development of new diagnostic and therapeutic strategies in GPA., (Copyright © 2020 Granel, Lemoine, Morello, Gallais, Mariot, Drapeau, Musnier, Poupon, Pugnière, Seren, Nouar, Gouilleux-Gruart, Watier, Korkmaz and Hoarau.)
- Published
- 2020
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38. Consequences of cathepsin C inactivation for membrane exposure of proteinase 3, the target antigen in autoimmune vasculitis.
- Author
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Seren S, Rashed Abouzaid M, Eulenberg-Gustavus C, Hirschfeld J, Nasr Soliman H, Jerke U, N'Guessan K, Dallet-Choisy S, Lesner A, Lauritzen C, Schacher B, Eickholz P, Nagy N, Szell M, Croix C, Viaud-Massuard MC, Al Farraj Aldosari A, Ragunatha S, Ibrahim Mostafa M, Giampieri F, Battino M, Cornillier H, Lorette G, Stephan JL, Goizet C, Pedersen J, Gauthier F, Jenne DE, Marchand-Adam S, Chapple IL, Kettritz R, and Korkmaz B
- Subjects
- Adolescent, Adult, Case-Control Studies, Child, Child, Preschool, Female, Granulomatosis with Polyangiitis drug therapy, Granulomatosis with Polyangiitis genetics, Granulomatosis with Polyangiitis metabolism, Humans, Male, Myeloblastin genetics, Neutrophils enzymology, Proteolysis, Young Adult, Cathepsin C antagonists & inhibitors, Cell Membrane metabolism, Cysteine Proteinase Inhibitors pharmacology, Granulomatosis with Polyangiitis pathology, Myeloblastin metabolism
- Abstract
Membrane-bound proteinase 3 (PR3
m ) is the main target antigen of anti-neutrophil cytoplasmic autoantibodies (ANCA) in granulomatosis with polyangiitis, a systemic small-vessel vasculitis. Binding of ANCA to PR3m triggers neutrophil activation with the secretion of enzymatically active PR3 and related neutrophil serine proteases, thereby contributing to vascular damage. PR3 and related proteases are activated from pro-forms by the lysosomal cysteine protease cathepsin C (CatC) during neutrophil maturation. We hypothesized that pharmacological inhibition of CatC provides an effective measure to reduce PR3m and therefore has implications as a novel therapeutic approach in granulomatosis with polyangiitis. We first studied neutrophilic PR3 from 24 patients with Papillon-Lefèvre syndrome (PLS), a genetic form of CatC deficiency. PLS neutrophil lysates showed a largely reduced but still detectable (0.5-4%) PR3 activity when compared with healthy control cells. Despite extremely low levels of cellular PR3, the amount of constitutive PR3m expressed on the surface of quiescent neutrophils and the typical bimodal membrane distribution pattern were similar to what was observed in healthy neutrophils. However, following cell activation, there was no significant increase in the total amount of PR3m on PLS neutrophils, whereas the total amount of PR3m on healthy neutrophils was significantly increased. We then explored the effect of pharmacological CatC inhibition on PR3 stability in normal neutrophils using a potent cell-permeable CatC inhibitor and a CD34+ hematopoietic stem cell model. Human CD34+ hematopoietic stem cells were treated with the inhibitor during neutrophil differentiation over 10 days. We observed strong reductions in PR3m , cellular PR3 protein, and proteolytic PR3 activity, whereas neutrophil differentiation was not compromised., (© 2018 Seren et al.)- Published
- 2018
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39. Neutrophils can disarm NK cell response through cleavage of NKp46.
- Author
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Valayer A, Brea D, Lajoie L, Avezard L, Combes-Soia L, Labas V, Korkmaz B, Thibault G, Baranek T, and Si-Tahar M
- Subjects
- Cathepsin G metabolism, Cell Membrane metabolism, Down-Regulation, Humans, K562 Cells, Natural Cytotoxicity Triggering Receptor 1 chemistry, Neutrophil Activation, Killer Cells, Natural metabolism, Natural Cytotoxicity Triggering Receptor 1 metabolism, Neutrophils metabolism
- Abstract
Polymorphonuclear neutrophils (PMNs) can contribute to the regulation of the host immune response by crosstalk with innate and adaptive leukocytes, including NK cells. Mechanisms by which this immunoregulation process occurs remain incompletely understood. Here, we focused on the effect of human neutrophil-derived serine proteases on NKp46, a crucial activating receptor expressed on NK cells. We used flow cytometry, Western blotting, and mass spectrometry (MS) analysis to reveal that cathepsin G [CG; and not elastase or proteinase 3 (PR3)] induces a time- and concentration-dependent, down-regulatory effect on NKp46 expression through a restricted proteolytic mechanism. We also used a functional assay to demonstrate that NKp46 cleavage by CG severely impairs NKp46-mediated responses of NK cells, including IFN-γ production and cell degranulation. Importantly, sputa of cystic fibrosis (CF) patients, which have high concentrations of CG, also alter NKp46 on NK cells. Hence, we have identified a new immunoregulatory mechanism of neutrophils that proteolytically disarms NK cell responses., (© Society for Leukocyte Biology.)
- Published
- 2017
- Full Text
- View/download PDF
40. Inhibitors and Antibody Fragments as Potential Anti-Inflammatory Therapeutics Targeting Neutrophil Proteinase 3 in Human Disease.
- Author
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Korkmaz B, Lesner A, Guarino C, Wysocka M, Kellenberger C, Watier H, Specks U, Gauthier F, and Jenne DE
- Subjects
- Animals, Anti-Inflammatory Agents therapeutic use, Apoptosis drug effects, Humans, Inflammation drug therapy, Inflammation enzymology, Molecular Targeted Therapy, Myeloblastin metabolism, Neutrophils drug effects, Neutrophils physiology, Anti-Inflammatory Agents pharmacology, Immunoglobulin Fragments pharmacology, Immunoglobulin Fragments therapeutic use, Myeloblastin antagonists & inhibitors, Neutrophils enzymology, Protease Inhibitors pharmacology, Protease Inhibitors therapeutic use
- Abstract
Proteinase 3 (PR3) has received great scientific attention after its identification as the essential antigenic target of antineutrophil cytoplasm antibodies in Wegener's granulomatosis (now called granulomatosis with polyangiitis). Despite many structural and functional similarities between neutrophil elastase (NE) and PR3 during biosynthesis, storage, and extracellular release, unique properties and pathobiological functions have emerged from detailed studies in recent years. The development of highly sensitive substrates and inhibitors of human PR3 and the creation of PR3-selective single knockout mice led to the identification of nonredundant roles of PR3 in cell death induction via procaspase-3 activation in cell cultures and in mouse models. According to a study in knockout mice, PR3 shortens the lifespan of infiltrating neutrophils in tissues and accelerates the clearance of aged neutrophils in mice. Membrane exposure of active human PR3 on apoptotic neutrophils reprograms the response of macrophages to phagocytosed neutrophils, triggers secretion of proinflammatory cytokines, and undermines immune silencing and tissue regeneration. PR3-induced disruption of the anti-inflammatory effect of efferocytosis may be relevant for not only granulomatosis with polyangiitis but also for other autoimmune diseases with high neutrophil turnover. Inhibition of membrane-bound PR3 by endogenous inhibitors such as the α-1-protease inhibitor is comparatively weaker than that of NE, suggesting that the adverse effects of unopposed PR3 activity resurface earlier than those of NE in individuals with α-1-protease inhibitor deficiency. Effective coverage of PR3 by anti-inflammatory tools and simultaneous inhibition of both PR3 and NE should be most promising in the future., (Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics.)
- Published
- 2016
- Full Text
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