Anti–microRNA-21 oligonucleotides prevent Alport nephropathy progression by stimulating metabolic pathways
Ivan G. Gomez, Deidre A. MacKenna, Bryce G. Johnson, Vivek Kaimal, Allie M. Roach, Shuyu Ren, Naoki Nakagawa, Cuiyan Xin, Rick Newitt, Shweta Pandya, Tai-He Xia, Xueqing Liu, Dorin-Bogdan Borza, Monica Grafals, Stuart J. Shankland, Jonathan Himmelfarb, Didier Portilla, Shiguang Liu, B. Nelson Chau, Jeremy S. Duffield
Ivan G. Gomez, Deidre A. MacKenna, Bryce G. Johnson, Vivek Kaimal, Allie M. Roach, Shuyu Ren, Naoki Nakagawa, Cuiyan Xin, Rick Newitt, Shweta Pandya, Tai-He Xia, Xueqing Liu, Dorin-Bogdan Borza, Monica Grafals, Stuart J. Shankland, Jonathan Himmelfarb, Didier Portilla, Shiguang Liu, B. Nelson Chau, Jeremy S. Duffield
View: Text | PDF
Research Article Nephrology

Anti–microRNA-21 oligonucleotides prevent Alport nephropathy progression by stimulating metabolic pathways

Abstract

MicroRNA-21 (miR-21) contributes to the pathogenesis of fibrogenic diseases in multiple organs, including the kidneys, potentially by silencing metabolic pathways that are critical for cellular ATP generation, ROS production, and inflammatory signaling. Here, we developed highly specific oligonucleotides that distribute to the kidney and inhibit miR-21 function when administered subcutaneously and evaluated the therapeutic potential of these anti–miR-21 oligonucleotides in chronic kidney disease. In a murine model of Alport nephropathy, miR-21 silencing did not produce any adverse effects and resulted in substantially milder kidney disease, with minimal albuminuria and dysfunction, compared with vehicle-treated mice. miR-21 silencing dramatically improved survival of Alport mice and reduced histological end points, including glomerulosclerosis, interstitial fibrosis, tubular injury, and inflammation. Anti–miR-21 enhanced PPARα/retinoid X receptor (PPARα/RXR) activity and downstream signaling pathways in glomerular, tubular, and interstitial cells. Moreover, miR-21 silencing enhanced mitochondrial function, which reduced mitochondrial ROS production and thus preserved tubular functions. Inhibition of miR-21 was protective against TGF-β–induced fibrogenesis and inflammation in glomerular and interstitial cells, likely as the result of enhanced PPARα/RXR activity and improved mitochondrial function. Together, these results demonstrate that inhibition of miR-21 represents a potential therapeutic strategy for chronic kidney diseases including Alport nephropathy.

Authors

Ivan G. Gomez, Deidre A. MacKenna, Bryce G. Johnson, Vivek Kaimal, Allie M. Roach, Shuyu Ren, Naoki Nakagawa, Cuiyan Xin, Rick Newitt, Shweta Pandya, Tai-He Xia, Xueqing Liu, Dorin-Bogdan Borza, Monica Grafals, Stuart J. Shankland, Jonathan Himmelfarb, Didier Portilla, Shiguang Liu, B. Nelson Chau, Jeremy S. Duffield

×

Figure 3

Anti–miR-21 prevents glomerulosclerosis tubular atrophy and interstitial fibrosis.

Options: View larger image (or click on image) Download as PowerPoint
Anti–miR-21 prevents glomerulosclerosis tubular atrophy and interstitial...
(A) Silver methenamine–stained images highlighting glomerulosclerosis (arrowheads); Sirius red–stained images of whole kidney showing interstitial fibrosis; PAS-stained images showing tubule injury; WT-1 immunofluorescence showing podocytes within glomeruli (dotted lines), all at 9 weeks. EM images of glomerular capillary loops that show severe basement membrane thickening, duplication, and podocyte effacement in vehicle-treated Col4a3–/– compared with anti–miR-21–treated Col4a3–/– mice at 9 weeks, which show preserved basement membrane in many glomeruli, with minimal foot-process effacement. At 16 weeks (lowest panel), the GBM in anti–miR-21–treated mice shows small but classical basement membrane humps, whereas foot processes remain partially preserved. U, urinary space; P, podocyte; Pp, podocyte processes; L, capillary lumen; EC, endothelial cell. Scale bars: 100 μm (conventional); 500 nm (EM). (B) Graph of percentage of glomeruli with glomerulosclerosis at 9 weeks, scored from 0 to 4 by extent of sclerosis, where 0 is without sclerosis. (C) Graph showing percentage of glomeruli with glomerular epithelial cell hyperplasia (known as crescent). (D) Podocyte number per glomerular cross section. (E) Tubule injury score. (F) Morphometry of interstitial fibrosis. *P < 0.05; **P < 0.01, 1-way ANOVA or Mann-Whitney U test. n = 12/group.