Anti–microRNA-21 oligonucleotides prevent Alport nephropathy progression by stimulating metabolic pathways
Ivan G. Gomez, … , B. Nelson Chau, Jeremy S. Duffield
Ivan G. Gomez, … , B. Nelson Chau, Jeremy S. Duffield
Published November 21, 2014
Citation Information: J Clin Invest. 2015;125(1):141-156. https://doi.org/10.1172/JCI75852.
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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

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Figure 2

Anti–miR-21 is widely distributed to kidney cells, increases life span, and protects Col4a3–/– mice from kidney disease progression.

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Anti–miR-21 is widely distributed to kidney cells, increases life span, ...
(A) Experimental schema indicating anti–miR-21 delivery from 3.5 weeks after birth and analysis at 63 days (9 weeks) or continued to 110 days (16 weeks). (B) Split panel confocal images of kidney cortex showing distribution of a single s.c. injection of Cy3-conjugated anti–miR-21 oligos (red) or vehicle 48 hours previously in 8-week-old Col4a3–/– mice. Sections were colabeled with antibodies against specific cellular markers of myofibroblasts, fibroblasts/pericytes and mesangial cells, endothelium, or podocytes to highlight uptake in particular cell types (arrowheads). Scale bars: 50 μm. g, glomerulus. (C) Confocal images showing vehicle control. Scale bars: 50 μm. (D) Graph of plasma BUN levels at 9 weeks. A-miR-21, anti–miR-21. (E) Graph of time course of urine albumin concentration normalized to urine creatinine. (F) Curves showing body weight changes with time. (G) Kaplan-Meier survival curve. Arrow indicates last delivery of anti–miR-21. **P < 0.01; ***P < 0.001; ****P < 0.0001, Gehan-Breslow-Wilcoxon test for survival; 1-way ANOVA or Mann-Whitney U test for others. n = 12/group.