[en] Mesenchymal stem cells (MSCs) are known to migrate to tissue injury sites to participate in immune modulation, tissue remodelling and wound healing, reducing tissue damage. Upon neutrophil activation, there is a release of myeloperoxidase (MPO), an oxidant enzyme. But little is known about the direct role of MSCs on MPO activity. The aim of this study was to investigate the effect of equine mesenchymal stem cells derived from muscle microinvasive biopsy (mdMSC) on the oxidant response of neutrophils and particularly on the activity of the myeloperoxidase released by stimulated equine neutrophils. After specific treatment (trypsin and washings in phosphate buffer saline), the mdMSCs were exposed to isolated neutrophils. The effect of the suspended mdMSCs was studied on the ROS production and the release of total and active MPO by stimulated neutrophils and specifically on the activity of MPO in a neutrophil-free model. Additionally, we developed a model combining adherent mdMSCs with neutrophils to study total and active MPO from the neutrophil extracellular trap (NET). Our results show that mdMSCs inhibited the ROS production, the activity of MPO released by stimulated neutrophils and the activity of MPO bound to the NET. Moreover, the co-incubation of mdMSCs directly with MPO results in a strong inhibition of the peroxidase activity of MPO, probably by affecting the active site of the enzyme. We confirm the strong potential of mdMSCs to lower the oxidant response of neutrophils. The novelty of our study is an evident inhibition of the activity of MPO by MSCs. The results indicated a new potential therapeutic approach of mdMSCs in the inhibition of MPO, which is considered as a pro-oxidant actor in numerous chronic and acute inflammatory pathologies.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Franck, Thierry ; Université de Liège - ULiège > Centre de l'oxygène : Recherche et développement (C.O.R.D.)
Ceusters, Justine ; Université de Liège - ULiège > Département d'Ens. et de Clinique des Equidés (DCE) > Chirurgie des équidés
Regmi, S.; Pathak, S.; Kim, J.O.; Yong, C.S.; Jeong, J.H. Mesenchymal stem cell therapy for the treatment of inflammatory diseases: Challenges, opportunities, and future perspectives. Eur. J. Cell Biol. 2019, 98, 151041. [CrossRef]
Saeedi, P.; Halabian, R.; Imani Fooladi, A.A. A revealing review of mesenchymal stem cells therapy, clinical perspectives and Modification strategies. Stem Cell Investig. 2019, 6, 34. [CrossRef]
Jiang, W.; Xu, J. Immune modulation by mesenchymal stem cells. Cell Prolif. 2020, 53, e12712. [CrossRef] [PubMed]
Maumus, M.; Jorgensen, C.; Noël, D. Mesenchymal stem cells in regenerative medicine applied to rheumatic diseases: Role of secretome and exosomes. Biochimie 2013, 95, 2229–2234. [CrossRef]
Mittal, S.K.; Mashaghi, A.; Amouzegar, A.; Li, M.; Foulsham, W.; Sahu, S.K.; Chauhan, S.K. Mesenchymal stromal cells inhibit neutrophil effector functions in a murine model of ocular inflammation. Investig. Ophthalmol. Vis. Sci. 2018, 59, 1191–1198. [CrossRef] [PubMed]
Jiang, D.; Muschhammer, J.; Qi, Y.; Kügler, A.; de Vries, J.C.; Saffarzadeh, M.; Sindrilaru, A.; Beken, S.V.; Wlaschek, M.; Kluth, M.A.; et al. Suppression of neutrophil-mediated tissue damage—A novel skill of mesenchymal stem cells. Stem Cells 2016, 34, 2393–2406. [CrossRef]
Stavely, R.; Nurgali, K. The emerging antioxidant paradigm of mesenchymal stem cell therapy. Stem Cells Transl. Med. 2020, 9, 985–1006. [CrossRef] [PubMed]
Pedrazza, L.; Cunha, A.A.; Luft, C.; Nunes, N.K.; Schimitz, F.; Gassen, R.B.; Breda, R.V.; Donadio, M.V.F.; de Souza Wyse, A.T.; Pitrez, P.M.C.; et al. Mesenchymal stem cells improves survival in LPS-induced acute lung injury acting through inhibition of NETs formation. J. Cell. Physiol. 2017, 232, 3552–3564. [CrossRef]
Raffaghello, L.; Bianchi, G.; Bertolotto, M.; Montecucco, F.; Busca, A.; Dallegri, F.; Ottonello, L.; Pistoia, V. Human mesenchymal stem cells inhibit neutrophil apoptosis: A model for neutrophil preservation in the bone marrow niche. Stem Cells 2008, 26, 151–162. [CrossRef]
Mumaw, J.L.; Schmiedt, C.W.; Breidling, S.; Sigmund, A.; Norton, N.A.; Thoreson, M.; Peroni, J.F.; Hurley, D.J. Feline mesenchymal stem cells and supernatant inhibit reactive oxygen species production in cultured feline neutrophils. Res. Vet. Sci. 2015, 103, 60–69. [CrossRef] [PubMed]
Taghavi-Farahabadi, M.; Mahmoudi, M.; Hashemi, S.M.; Rezaei, N. Evaluation of the effects of mesenchymal stem cells on neutrophils isolated from severe congenital neutropenia patients. Int. Immunopharmacol. 2020, 83, 106463. [CrossRef] [PubMed]
Espinosa, G.; Plaza, A.; Schenffeldt, A.; Alarcón, P.; Gajardo, G.; Uberti, B.; Morán, G.; Henríquez, C. Equine bone marrow-derived mesenchymal stromal cells inhibit reactive oxygen species production by neutrophils. Vet. Immunol. Immunopathol. 2020, 221, 109975. [CrossRef] [PubMed]
Deby-Dupont, G.; Deby, C.; Lamy, M. Neutrophil myeloperoxidase revisited: It’s role in health and disease. Intensivmed. Notf. 1999, 36, 500–513. [CrossRef]
Davies, M.J.; Hawkins, C.L. The role of myeloperoxidase in biomolecule modification, chronic inflammation, and disease. Antioxid. Redox Signal. 2020, 32, 957–981. [CrossRef]
Sarmah, D.; Kaur, H.; Vats, K.; Kalia, K.; Yavagal, D.R.; Bhattacharya, P. Abstract 429: Inflammasome Mediated reduction of myeloperoxidase in ischemic stroke by intra-arterial mesenchymal stem cell therapy. Arterioscler. Thromb. Vasc. Biol. 2019, 39, A429. [CrossRef]
Guillén, M.I.; Platas, J.; del Caz, M.D.; Mirabet, V.; Alcaraz, M.J. Paracrine anti-inflammatory effects of adipose tissue-derived mesenchymal stem cells in human monocytes. Front. Physiol. 2018, 9, 661. [CrossRef]
Čamernik, K.; Mihelič, A.; Mihalič, R.; Marolt Presen, D.; Janež, A.; Trebše, R.; Marc, J.; Zupan, J. Skeletal-muscle-derived mesenchymal stem/stromal cells from patients with osteoarthritis show superior biological properties compared to bone-derived cells. Stem Cell Res. 2019, 38, 101465. [CrossRef]
Ceusters, J.; Lejeune, J.P.; Sandersen, C.; Niesten, A.; Lagneaux, L.; Serteyn, D. From skeletal muscle to stem cells: An innovative and minimally-invasive process for multiple species. Sci. Rep. 2017, 7, 696. [CrossRef]
Serteyn, D.; Ceusters, J. Mammalian Muscle-Derived Stem Cells. Patent WO2015091210A1, 25 June 2015.
Franck, T.; Grulke, S.; Deby-Dupont, G.; Deby, C.; Duvivier, H.; Peters, F.; Serteyn, D. Development of an enzyme-linked immunosorbent assay for specific equine neutrophil myeloperoxidase measurement in blood. J. Vet. Diagn. Investig. 2005, 17, 412–419. [CrossRef] [PubMed]
Strober, W. Trypan blue exclusion test of cell viability. Curr. Protoc. Immunol. 2001, 21, A.3B.1–A.3B.2. [CrossRef]
Pycock, J.F.; Allen, W.E.; Morris, T.H. Rapid, single-step isolation of equine neutrophils on a discontinuous Percoll density gradient. Res. Vet. Sci. 1987, 42, 411–412. [CrossRef]
Franck, T.; Aldib, I.; Zouaoui Boudjeltia, K.; Furtmüller, P.G.; Obinger, C.; Neven, P.; Prévost, M.; Soubhye, J.; Van Antwerpen, P.; Mouithys-Mickalad, A.; et al. The soluble curcumin derivative NDS27 inhibits superoxide anion production by neutrophils and acts as substrate and reversible inhibitor of myeloperoxidase. Chem. Interact. 2018, 297, 34–43. [CrossRef] [PubMed]
Zielonka, J.; Lambeth, J.D.; Kalyanaraman, B. On the use of L-012, a luminol-based chemiluminescent probe, for detecting superoxide and identifying inhibitors of NADPH oxidase: A reevaluation. Free Radic. Biol. Med. 2013, 65, 1310–1314. [CrossRef] [PubMed]
Goiffon, R.J.; Martinez, S.C.; Piwnica-Worms, D. A rapid bioluminescence assay for measuring myeloperoxidase activity in human plasma. Nat. Commun. 2015, 6, 6271. [CrossRef]
Franck, T.; Kohnen, S.; Deby-Dupont, G.; Grulke, S.; Deby, C.; Serteyn, D. A specific method for measurement of equine active myeloperoxidase in biological samples and in in vitro tests. J. Vet. Diagn. Investig. 2006, 18, 326–334. [CrossRef] [PubMed]
Thålin, C.; Daleskog, M.; Göransson, S.P.; Schatzberg, D.; Lasselin, J.; Laska, A.-C.; Kallner, A.; Helleday, T.; Wallén, H.; Demers, M. Validation of an enzyme-linked immunosorbent assay for the quantification of citrullinated histone H3 as a marker for neutrophil extracellular traps in human plasma. Immunol. Res. 2017, 65, 706–712. [CrossRef]
Kim, W.S.; Park, B.S.; Kim, H.K.; Park, J.S.; Kim, K.J.; Choi, J.S.; Chung, S.J.; Kim, D.D.; Sung, J.H. Evidence supporting antioxidant action of adipose-derived stem cells: Protection of human dermal fibroblasts from oxidative stress. J. Dermatol. Sci. 2008, 49, 133–142. [CrossRef]
Yao, J.; Zheng, J.; Cai, J.; Zeng, K.; Zhou, C.; Zhang, J.; Li, S.; Li, H.; Chen, L.; He, L.; et al. Extracellular vesicles derived from human umbilical cord mesenchymal stem cells alleviate rat hepatic ischemia-reperfusion injury by suppressing oxidative stress and neutrophil inflammatory response. FASEB J. 2019, 33, 1695–1710. [CrossRef]
Sharma, A.K.; Salmon, M.D.; Lu, G.; Su, G.; Pope, N.H.; Smith, J.R.; Weiss, M.L.; Upchurch, G.R. Mesenchymal stem cells attenuate NADPH oxidase-dependent high mobility group box 1 production and inhibit abdominal aortic aneurysms. Arterioscler. Thromb. Vasc. Biol. 2016, 36, 908–918. [CrossRef]
King, C.C.; Jefferson, M.M.; Thomas, E.L. Secretion and inactivation of myeloperoxidase by isolated neutrophils. J. Leukoc. Biol. 1997, 61, 293–302. [CrossRef]
Franck, T.; Kohnen, S.; de la Rebière, G.; Deby-Dupont, G.; Deby, C.; Niesten, A.; Serteyn, D. Activation of equine neutrophils by phorbol myristate acetate or N-formyl-methionyl-leucyl-phenylalanine induces a different response in reactive oxygen species production and release of active myeloperoxidase. Vet. Immunol. Immunopathol. 2009, 130, 243–250. [CrossRef] [PubMed]
Bei, L.; Hu, T.; Qian, Z.M.; Shen, X. Extracellular Ca2+ regulates the respiratory burst of human neutrophils. Biochim. Biophys. Acta 1998, 1404, 475–483. [CrossRef]
Franck, T.; Minguet, G.; Delporte, C.; Derochette, S.; Zouaoui Boudjeltia, K.; Van Antwerpen, P.; Gach, O.; Deby-Dupont, G.; Mouithys-Mickalad, A.; Serteyn, D. An immunological method to combine the measurement of active and total myeloperoxidase on the same biological fluid, and its application in finding inhibitors which interact directly with the enzyme. Free Radic. Res. 2015, 49, 790–799. [CrossRef] [PubMed]
Bodega, G.; Alique, M.; Puebla, L.; Carracedo, J.; Ramírez, R.M. Microvesicles: ROS scavengers and ROS producers. J. Extracell. Vesicles 2019, 8, 1626654. [CrossRef] [PubMed]
De Bont, C.M.; Koopman, W.J.H.; Boelens, W.C.; Pruijn, G.J.M. Stimulus-dependent chromatin dynamics, citrullination, calcium signalling and ROS production during NET formation. Biochim. Biophys. Acta Mol. Cell Res. 2018, 1865, 1621–1629. [CrossRef]
Mikacenic, C.; Moore, R.; Dmyterko, V.; West, T.E.; Altemeier, W.A.; Liles, W.C.; Lood, C. Neutrophil extracellular traps (NETs) are increased in the alveolar spaces of patients with ventilator-associated pneumonia. Crit. Care 2018, 22, 358. [CrossRef]
Donkel, S.J.; Wolters, F.J.; Ikram, M.A.; de Maat, M.P.M. Circulating Myeloperoxidase (MPO)-DNA complexes as marker for Neutrophil Extracellular Traps (NETs) levels and the association with cardiovascular risk factors in the general population. PLoS ONE 2021, 16, e0253698. [CrossRef]
Jiang, D.; Saffarzadeh, M.; Scharffetter-Kochanek, K. In vitro demonstration and quantification of neutrophil extracellular trap formation. Bio Protoc. 2017, 7. [CrossRef] [PubMed]
Thålin, C.; Aguilera, K.; Hall, N.W.; Marunde, M.R.; Burg, J.M.; Rosell, A.; Daleskog, M.; Månsson, M.; Hisada, Y.; Meiners, M.J.; et al. Quantification of citrullinated histones: Development of an improved assay to reliably quantify nucleosomal H3Cit in human plasma. J. Thromb. Haemost. 2020, 18, 2732–2743. [CrossRef] [PubMed]
Björnsdottir, H.; Welin, A.; Michaëlsson, E.; Osla, V.; Berg, S.; Christenson, K.; Sundqvist, M.; Dahlgren, C.; Karlsson, A.; Bylund, J. Neutrophil NET formation is regulated from the inside by myeloperoxidase-processed reactive oxygen species. Free Radic. Biol. Med. 2015, 89, 1024–1035. [CrossRef] [PubMed]
