title: Rostafuroxin protects from podocyte injury and proteinuria induced by adducin genetic variants and ouabain.
abstract: Glomerulopathies are important causes of morbidity and mortality. Selective therapies that address the underlying mechanisms are still lacking. Recently, two mechanisms, mutant β-adducin and ouabain, have been found to be involved in glomerular podocytopathies and proteinuria through nephrin downregulation. The main purpose of the present study was to investigate whether rostafuroxin, a novel antihypertensive agent developed as a selective inhibitor of Src-SH2 interaction with mutant adducin- and ouabain-activated Na,K-ATPase, may protect podocytes from adducin- and ouabain-induced effects, thus representing a novel pharmacologic approach for the therapy of podocytopathies and proteinuria caused by the aforementioned mechanisms. To study the effect of rostafuroxin on podocyte protein changes and proteinuria, mice carrying mutant β-adducin and ouabain hypertensive rats were orally treated with 100 μg/kg per day rostafuroxin. Primary podocytes from congenic rats carrying mutant α-adducin or β-adducin (NB) from Milan hypertensive rats and normal rat podocytes incubated with 10(-9) M ouabain were cultured with 10(-9) M rostafuroxin. The results indicated that mutant β-adducin and ouabain caused podocyte nephrin loss and proteinuria in animal models. These alterations were reproduced in primary podocytes from NB rats and normal rats incubated with ouabain. Treatment of animals, or incubation of cultured podocytes with rostafuroxin, reverted mutant β-adducin- and ouabain-induced effects on nephrin protein expression and proteinuria. We conclude that rostafuroxin prevented podocyte lesions and proteinuria due to mutant β-adducin and ouabain in animal models. This suggests a potential therapeutic effect of rostafuroxin in patients with glomerular disease progression associated with these two mechanisms.
authors.affiliation: Prassis Sigma-Tau Research Institute, Settimo Milanese, Milan, Italy (M.F., I.M., P.F.); Division of Genetics and Cell Biology, Genomics of Renal Diseases and Hypertension Unit, IRCC San Raffaele Scientific Institute, Milan, Italy (M.F., I.M., P.M.); Renal Research Laboratory, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico & Fondazione D'Amico per la Ricerca sulle Malattie Renali, Milan, Italy (M.P.R.); CVie Therapeutics, Hong Kong, China (P.F., G.B.); and Chair of Nephrology, University Vita-Salute, Milan, Italy (G.B., P.M.) mara.ferrandi@gmail.com.
authors.forename: Mara
authors.initials: M
authors.lastname: Ferrandi
authors.affiliation: Prassis Sigma-Tau Research Institute, Settimo Milanese, Milan, Italy (M.F., I.M., P.F.); Division of Genetics and Cell Biology, Genomics of Renal Diseases and Hypertension Unit, IRCC San Raffaele Scientific Institute, Milan, Italy (M.F., I.M., P.M.); Renal Research Laboratory, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico & Fondazione D'Amico per la Ricerca sulle Malattie Renali, Milan, Italy (M.P.R.); CVie Therapeutics, Hong Kong, China (P.F., G.B.); and Chair of Nephrology, University Vita-Salute, Milan, Italy (G.B., P.M.).
authors.forename: Isabella
authors.initials: I
authors.lastname: Molinari
authors.affiliation: Prassis Sigma-Tau Research Institute, Settimo Milanese, Milan, Italy (M.F., I.M., P.F.); Division of Genetics and Cell Biology, Genomics of Renal Diseases and Hypertension Unit, IRCC San Raffaele Scientific Institute, Milan, Italy (M.F., I.M., P.M.); Renal Research Laboratory, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico & Fondazione D'Amico per la Ricerca sulle Malattie Renali, Milan, Italy (M.P.R.); CVie Therapeutics, Hong Kong, China (P.F., G.B.); and Chair of Nephrology, University Vita-Salute, Milan, Italy (G.B., P.M.).
authors.forename: Maria Pia
authors.initials: MP
authors.lastname: Rastaldi
authors.affiliation: Prassis Sigma-Tau Research Institute, Settimo Milanese, Milan, Italy (M.F., I.M., P.F.); Division of Genetics and Cell Biology, Genomics of Renal Diseases and Hypertension Unit, IRCC San Raffaele Scientific Institute, Milan, Italy (M.F., I.M., P.M.); Renal Research Laboratory, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico & Fondazione D'Amico per la Ricerca sulle Malattie Renali, Milan, Italy (M.P.R.); CVie Therapeutics, Hong Kong, China (P.F., G.B.); and Chair of Nephrology, University Vita-Salute, Milan, Italy (G.B., P.M.).
authors.forename: Patrizia
authors.initials: P
authors.lastname: Ferrari
authors.affiliation: Prassis Sigma-Tau Research Institute, Settimo Milanese, Milan, Italy (M.F., I.M., P.F.); Division of Genetics and Cell Biology, Genomics of Renal Diseases and Hypertension Unit, IRCC San Raffaele Scientific Institute, Milan, Italy (M.F., I.M., P.M.); Renal Research Laboratory, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico & Fondazione D'Amico per la Ricerca sulle Malattie Renali, Milan, Italy (M.P.R.); CVie Therapeutics, Hong Kong, China (P.F., G.B.); and Chair of Nephrology, University Vita-Salute, Milan, Italy (G.B., P.M.).
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Bianchi
authors.affiliation: Prassis Sigma-Tau Research Institute, Settimo Milanese, Milan, Italy (M.F., I.M., P.F.); Division of Genetics and Cell Biology, Genomics of Renal Diseases and Hypertension Unit, IRCC San Raffaele Scientific Institute, Milan, Italy (M.F., I.M., P.M.); Renal Research Laboratory, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico & Fondazione D'Amico per la Ricerca sulle Malattie Renali, Milan, Italy (M.P.R.); CVie Therapeutics, Hong Kong, China (P.F., G.B.); and Chair of Nephrology, University Vita-Salute, Milan, Italy (G.B., P.M.).
authors.forename: Paolo
authors.initials: P
authors.lastname: Manunta
mesh_terms: D000732:Androstanols / Q000494:pharmacology*; D000818:Animals; D000959:Antihypertensive Agents / Q000494:pharmacology; D002148:Calmodulin-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D005260:Female; D014644:Genetic Variation / Q000235:genetics*; D006973:Hypertension / Q000139:chemically induced / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D010042:Ouabain / Q000009:adverse effects*; D050199:Podocytes / Q000187:drug effects* / Q000378:metabolism; D011507:Proteinuria / Q000139:chemically induced / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D000099291:Adducins
chemical_list: D000732:Androstanols; D000959:Antihypertensive Agents; D002148:Calmodulin-Binding Proteins; D010042:Ouabain; D000254:Sodium-Potassium-Exchanging ATPase; D000099291:Adducins; C106263:rostafuroxin