Carla S B Viegas1, Lúcia Santos1, Anjos L Macedo1, António A Matos1, Ana P Silva1, Pedro L Neves1, An Staes1, Kris Gevaert1, Rute Morais1, Cees Vermeer1, Leon Schurgers1, Dina C Simes2. 1. From the Centre of Marine Sciences (C.S.B.V., L.S., D.C.S.), GenoGla Diagnostics, Centre of Marine Sciences (C.S.B.V., D.C.S.), and Department of Biomedical Sciences and Medicine (A.P.S., P.L.N.), University of Algarve, Faro, Portugal; UCIBIO-REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, Caparica, Portugal (A.L.M., R.M.); Centro de Investigação Interdisciplinar Egas Moniz, Egas Moniz-Cooperativa de Ensino Superior CRL, Caparica, Portugal (A.A.M.); Nephrology Department, Centro Hospitalar do Algarve, Faro, Portugal (A.P.S., P.L.N.); VIB-UGent Center for Medical Biotechnology Center and UGent Department of Biochemistry, Ghent, Belgium (A.S., K.G.); and R&D Group VitaK (C.V.) and Department of Biochemistry - Vascular Aspects, Faculty of Medicine, Health and Life Science (L.S.), Maastricht University, The Netherlands. 2. From the Centre of Marine Sciences (C.S.B.V., L.S., D.C.S.), GenoGla Diagnostics, Centre of Marine Sciences (C.S.B.V., D.C.S.), and Department of Biomedical Sciences and Medicine (A.P.S., P.L.N.), University of Algarve, Faro, Portugal; UCIBIO-REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, Caparica, Portugal (A.L.M., R.M.); Centro de Investigação Interdisciplinar Egas Moniz, Egas Moniz-Cooperativa de Ensino Superior CRL, Caparica, Portugal (A.A.M.); Nephrology Department, Centro Hospitalar do Algarve, Faro, Portugal (A.P.S., P.L.N.); VIB-UGent Center for Medical Biotechnology Center and UGent Department of Biochemistry, Ghent, Belgium (A.S., K.G.); and R&D Group VitaK (C.V.) and Department of Biochemistry - Vascular Aspects, Faculty of Medicine, Health and Life Science (L.S.), Maastricht University, The Netherlands. dsimes@ualg.pt.
Abstract
OBJECTIVE: Inhibition of mineral crystal formation is a crucial step in ectopic calcification. Serum calciprotein particles (CPPs) have been linked to chronic kidney disease (CKD) calcification propensity, but additional knowledge is required to understand their function, assemblage, and composition. The role of other circulating nanostructures, such as extracellular vesicles (EVs) in vascular calcification is currently unknown. Here, we investigated the association of GRP (Gla-rich protein) with circulating CPP and EVs and the role of CKD CPPs and EVs in vascular calcification. APPROACH AND RESULTS: Biological CPPs and EVs were isolated from healthy and CKD patients and comparatively characterized using ultrastructural, analytic, molecular, and immuno-based techniques. Our results show that GRP is a constitutive component of circulating CPPs and EVs. CKD stage 5 serum CPPs and EVs are characterized by lower levels of fetuin-A and GRP, and CPPs CKD stage 5 have increased mineral maturation, resembling secondary CPP particles. Vascular smooth muscle cell calcification assays reveal that CPPs CKD stage 5 and EVs CKD stage 5 are taken up by vascular smooth muscle cells and induce vascular calcification by promoting cell osteochondrogenic differentiation and inflammation. These effects were rescued by incubation of CPPs CKD stage 5 with γ-carboxylated GRP. In vitro, formation and maturation of basic calcium phosphate crystals was highly reduced in the presence of γ-carboxylated GRP, fetuin-A, and MGP (matrix gla protein), and a similar antimineralization system was identified in vivo. CONCLUSIONS: Uremic CPPs and EVs are important players in the mechanisms of widespread calcification in CKD. We propose a major role for cGRP as inhibitory factor to prevent calcification at systemic and tissue levels.
OBJECTIVE: Inhibition of mineral crystal formation is a crucial step in ectopic calcification. Serum calciprotein particles (CPPs) have been linked to chronic kidney disease (CKD) calcification propensity, but additional knowledge is required to understand their function, assemblage, and composition. The role of other circulating nanostructures, such as extracellular vesicles (EVs) in vascular calcification is currently unknown. Here, we investigated the association of GRP (Gla-rich protein) with circulating CPP and EVs and the role of CKD CPPs and EVs in vascular calcification. APPROACH AND RESULTS: Biological CPPs and EVs were isolated from healthy and CKD patients and comparatively characterized using ultrastructural, analytic, molecular, and immuno-based techniques. Our results show that GRP is a constitutive component of circulating CPPs and EVs. CKD stage 5 serum CPPs and EVs are characterized by lower levels of fetuin-A and GRP, and CPPs CKD stage 5 have increased mineral maturation, resembling secondary CPP particles. Vascular smooth muscle cell calcification assays reveal that CPPs CKD stage 5 and EVs CKD stage 5 are taken up by vascular smooth muscle cells and induce vascular calcification by promoting cell osteochondrogenic differentiation and inflammation. These effects were rescued by incubation of CPPs CKD stage 5 with γ-carboxylated GRP. In vitro, formation and maturation of basic calcium phosphate crystals was highly reduced in the presence of γ-carboxylated GRP, fetuin-A, and MGP (matrix gla protein), and a similar antimineralization system was identified in vivo. CONCLUSIONS: Uremic CPPs and EVs are important players in the mechanisms of widespread calcification in CKD. We propose a major role for cGRP as inhibitory factor to prevent calcification at systemic and tissue levels.
Authors: Hong S Lu; Ann Marie Schmidt; Robert A Hegele; Nigel Mackman; Daniel J Rader; Christian Weber; Alan Daugherty Journal: Arterioscler Thromb Vasc Biol Date: 2019-12-23 Impact factor: 8.311
Authors: Wei Chen; Jessica Fitzpatrick; Jose M Monroy-Trujillo; Stephen M Sozio; Bernard G Jaar; Michelle M Estrella; Jishyra Serrano; Viktoriya Anokhina; Benjamin L Miller; Michal L Melamed; David A Bushinsky; Rulan S Parekh Journal: Am J Kidney Dis Date: 2020-08-13 Impact factor: 8.860
Authors: Miles D Witham; Jennifer S Lees; Myra White; Margaret Band; Samira Bell; Donna J Chantler; Ian Ford; Roberta L Fulton; Gwen Kennedy; Roberta C Littleford; Ian V McCrea; Deborah McGlynn; Maurizio Panarelli; Maximilian R Ralston; Elaine Rutherford; Alison Severn; Nicola Thomson; Jamie P Traynor; Allan D Struthers; Kirsty Wetherall; Patrick B Mark Journal: J Am Soc Nephrol Date: 2020-08-13 Impact factor: 10.121