Inhibition of Lysine 63 Ubiquitination Prevents the Progression of Renal Fibrosis in Diabetic DBA/2J Mice
Diabetic nephropathy (DN) is the most frequent cause of end-stage renal disease. Tubulointerstitial accumulation of lysine 63 (K63)-ubiquitinated (Ub) proteins is involved in the progression of DN fibrosis and correlates with urinary miR-27b-3p downregulation. We explored the renoprotective effect o...
| Main Authors: | , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-05-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/22/10/5194 |
| _version_ | 1797534121836150784 |
|---|---|
| author | Paola Pontrelli Francesca Conserva Rossella Menghini Michele Rossini Alessandra Stasi Chiara Divella Viviana Casagrande Claudia Cinefra Mariagrazia Barozzino Simona Simone Francesco Pesce Giuseppe Castellano Giovanni Stallone Anna Gallone Francesco Giorgino Massimo Federici Loreto Gesualdo |
| author_facet | Paola Pontrelli Francesca Conserva Rossella Menghini Michele Rossini Alessandra Stasi Chiara Divella Viviana Casagrande Claudia Cinefra Mariagrazia Barozzino Simona Simone Francesco Pesce Giuseppe Castellano Giovanni Stallone Anna Gallone Francesco Giorgino Massimo Federici Loreto Gesualdo |
| author_sort | Paola Pontrelli |
| collection | DOAJ |
| description | Diabetic nephropathy (DN) is the most frequent cause of end-stage renal disease. Tubulointerstitial accumulation of lysine 63 (K63)-ubiquitinated (Ub) proteins is involved in the progression of DN fibrosis and correlates with urinary miR-27b-3p downregulation. We explored the renoprotective effect of an inhibitor of K63-Ub (NSC697923), alone or in combination with the ACE-inhibitor ramipril, in vitro and in vivo. Proximal tubular epithelial cells and diabetic DBA/2J mice were treated with NSC697923 and/or ramipril. K63-Ub protein accumulation along with α-SMA, collagen I and III, FSP-1, vimentin, p16<sup>INK4A</sup> expression, SA-α Gal staining, Sirius Red, and PAS staining were measured. Finally, we measured the urinary albumin to creatinine ratio (uACR), and urinary miR-27b-3p expression in mice. NSC697923, both alone and in association with ramipril, in vitro and in vivo inhibited hyperglycemia-induced epithelial to mesenchymal transition by significantly reducing K63-Ub proteins, α-SMA, collagen I, vimentin, FSP-1 expression, and collagen III along with tubulointerstitial and glomerular fibrosis. Treated mice also showed recovery of urinary miR-27b-3p and restored expression of p16<sup>INK4A</sup>. Moreover, NSC697923 in combination with ramipril demonstrated a trend in the reduction of uACR. In conclusion, we suggest that selective inhibition of K63-Ub, when combined with the conventional treatment with ACE inhibitors, might represent a novel treatment strategy to prevent the progression of fibrosis and proteinuria in diabetic nephropathy and we propose miR-27b-3p as a biomarker of treatment efficacy. |
| first_indexed | 2024-03-10T11:25:10Z |
| format | Article |
| id | doaj.art-c2b63f0514bf44729959384e1b703387 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T11:25:10Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-c2b63f0514bf44729959384e1b7033872023-11-21T19:42:21ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-05-012210519410.3390/ijms22105194Inhibition of Lysine 63 Ubiquitination Prevents the Progression of Renal Fibrosis in Diabetic DBA/2J MicePaola Pontrelli0Francesca Conserva1Rossella Menghini2Michele Rossini3Alessandra Stasi4Chiara Divella5Viviana Casagrande6Claudia Cinefra7Mariagrazia Barozzino8Simona Simone9Francesco Pesce10Giuseppe Castellano11Giovanni Stallone12Anna Gallone13Francesco Giorgino14Massimo Federici15Loreto Gesualdo16Department of Emergency and Organ Transplantation, University of Bari, 70124 Bari, ItalyDepartment of Emergency and Organ Transplantation, University of Bari, 70124 Bari, ItalyDepartment of Systems Medicine, University of Rome Tor Vergata, 00133 Rome, ItalyDepartment of Emergency and Organ Transplantation, University of Bari, 70124 Bari, ItalyDepartment of Emergency and Organ Transplantation, University of Bari, 70124 Bari, ItalyDepartment of Emergency and Organ Transplantation, University of Bari, 70124 Bari, ItalyDepartment of Systems Medicine, University of Rome Tor Vergata, 00133 Rome, ItalyDepartment of Emergency and Organ Transplantation, University of Bari, 70124 Bari, ItalyDepartment of Emergency and Organ Transplantation, University of Bari, 70124 Bari, ItalyDepartment of Emergency and Organ Transplantation, University of Bari, 70124 Bari, ItalyDepartment of Emergency and Organ Transplantation, University of Bari, 70124 Bari, ItalyDepartment of Medical and Surgical Sciences, University of Foggia, 71122 Foggia, ItalyDepartment of Medical and Surgical Sciences, University of Foggia, 71122 Foggia, ItalyDepartment of Basic Medical Sciences, Neurosciences and Sense Organs, University of Bari, 70124 Bari, ItalyDepartment of Emergency and Organ Transplantation, University of Bari, 70124 Bari, ItalyDepartment of Systems Medicine, University of Rome Tor Vergata, 00133 Rome, ItalyDepartment of Emergency and Organ Transplantation, University of Bari, 70124 Bari, ItalyDiabetic nephropathy (DN) is the most frequent cause of end-stage renal disease. Tubulointerstitial accumulation of lysine 63 (K63)-ubiquitinated (Ub) proteins is involved in the progression of DN fibrosis and correlates with urinary miR-27b-3p downregulation. We explored the renoprotective effect of an inhibitor of K63-Ub (NSC697923), alone or in combination with the ACE-inhibitor ramipril, in vitro and in vivo. Proximal tubular epithelial cells and diabetic DBA/2J mice were treated with NSC697923 and/or ramipril. K63-Ub protein accumulation along with α-SMA, collagen I and III, FSP-1, vimentin, p16<sup>INK4A</sup> expression, SA-α Gal staining, Sirius Red, and PAS staining were measured. Finally, we measured the urinary albumin to creatinine ratio (uACR), and urinary miR-27b-3p expression in mice. NSC697923, both alone and in association with ramipril, in vitro and in vivo inhibited hyperglycemia-induced epithelial to mesenchymal transition by significantly reducing K63-Ub proteins, α-SMA, collagen I, vimentin, FSP-1 expression, and collagen III along with tubulointerstitial and glomerular fibrosis. Treated mice also showed recovery of urinary miR-27b-3p and restored expression of p16<sup>INK4A</sup>. Moreover, NSC697923 in combination with ramipril demonstrated a trend in the reduction of uACR. In conclusion, we suggest that selective inhibition of K63-Ub, when combined with the conventional treatment with ACE inhibitors, might represent a novel treatment strategy to prevent the progression of fibrosis and proteinuria in diabetic nephropathy and we propose miR-27b-3p as a biomarker of treatment efficacy.https://www.mdpi.com/1422-0067/22/10/5194DBA2/J micelysine 63 ubiquitinationmicroRNArenal fibrosissenescencediabetes |
| spellingShingle | Paola Pontrelli Francesca Conserva Rossella Menghini Michele Rossini Alessandra Stasi Chiara Divella Viviana Casagrande Claudia Cinefra Mariagrazia Barozzino Simona Simone Francesco Pesce Giuseppe Castellano Giovanni Stallone Anna Gallone Francesco Giorgino Massimo Federici Loreto Gesualdo Inhibition of Lysine 63 Ubiquitination Prevents the Progression of Renal Fibrosis in Diabetic DBA/2J Mice International Journal of Molecular Sciences DBA2/J mice lysine 63 ubiquitination microRNA renal fibrosis senescence diabetes |
| title | Inhibition of Lysine 63 Ubiquitination Prevents the Progression of Renal Fibrosis in Diabetic DBA/2J Mice |
| title_full | Inhibition of Lysine 63 Ubiquitination Prevents the Progression of Renal Fibrosis in Diabetic DBA/2J Mice |
| title_fullStr | Inhibition of Lysine 63 Ubiquitination Prevents the Progression of Renal Fibrosis in Diabetic DBA/2J Mice |
| title_full_unstemmed | Inhibition of Lysine 63 Ubiquitination Prevents the Progression of Renal Fibrosis in Diabetic DBA/2J Mice |
| title_short | Inhibition of Lysine 63 Ubiquitination Prevents the Progression of Renal Fibrosis in Diabetic DBA/2J Mice |
| title_sort | inhibition of lysine 63 ubiquitination prevents the progression of renal fibrosis in diabetic dba 2j mice |
| topic | DBA2/J mice lysine 63 ubiquitination microRNA renal fibrosis senescence diabetes |
| url | https://www.mdpi.com/1422-0067/22/10/5194 |
| work_keys_str_mv | AT paolapontrelli inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT francescaconserva inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT rossellamenghini inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT michelerossini inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT alessandrastasi inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT chiaradivella inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT vivianacasagrande inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT claudiacinefra inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT mariagraziabarozzino inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT simonasimone inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT francescopesce inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT giuseppecastellano inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT giovannistallone inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT annagallone inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT francescogiorgino inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT massimofederici inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice AT loretogesualdo inhibitionoflysine63ubiquitinationpreventstheprogressionofrenalfibrosisindiabeticdba2jmice |