1. In vivo silencing of amphiregulin by a novel effective Self-Assembled-Micelle inhibitory RNA ameliorates renal fibrosis via inhibition of EGFR signals
- Author
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Young Ho Ko, Sung-Il Yun, Jun Hong Park, Eun Young Lee, Soohyun Hwang, Ji-Hye Lee, Beomseok Son, Tae-Rim Kim, Han Oh Park, and Seung Seob Son
- Subjects
Male ,Science ,Down-Regulation ,urologic and male genital diseases ,Amphiregulin ,Article ,Kidney Tubules, Proximal ,Transforming Growth Factor beta1 ,Fibrosis ,Renal fibrosis ,Chronic kidney disease ,Pulmonary fibrosis ,medicine ,Gene silencing ,Animals ,Tissue Distribution ,Gene Silencing ,RNA, Messenger ,Phosphorylation ,Micelles ,Kidney ,Multidisciplinary ,biology ,Chemistry ,Cell adhesion molecule ,Adenine ,Fibroblasts ,medicine.disease ,Diet ,Fibronectin ,ErbB Receptors ,Mice, Inbred C57BL ,Disease Models, Animal ,Kinetics ,medicine.anatomical_structure ,biology.protein ,Cancer research ,Medicine ,Cytokines ,RNA ,Cell Adhesion Molecules ,Signal Transduction ,Ureteral Obstruction - Abstract
Amphiregulin (AREG) is a transmembrane glycoprotein recently implicated in kidney fibrosis. Previously, we reported that the AREG-targeting Self-Assembled-Micelle inhibitory RNA (SAMiRNA-AREG) alleviated fibrosis by stably silencing the AREG gene, and reduced the side effects of conventional siRNA treatment of pulmonary fibrosis. However, the therapeutic effect of SAMiRNA-AREG in renal fibrosis has not been studied until now. We used two animal models of renal fibrosis generated by a unilateral ureteral obstruction (UUO) and an adenine diet (AD) to investigate whether SAMiRNA-AREG inhibited renal fibrosis. To investigate the delivery of SAMiRNA-AREG to the kidney, Cy5-labeled SAMiRNA-AREG was injected into UUO- and AD-induced renal fibrosis models. In both kidney disease models, SAMiRNA-AREG was delivered primarily to the damaged kidney. We also confirmed the protective effect of SAMiRNA-AREG in renal fibrosis models. SAMiRNA-AREG markedly decreased the UUO- and AD-induced AREG mRNA expression. Furthermore, the mRNA expression of fibrosis markers, including α-smooth muscle actin, fibronectin, α1(I) collagen, and α1(III) collagen in the UUO and AD-induced kidneys, was diminished in the SAMiRNA-AREG-treated mice. The transcription of inflammatory markers (tumor necrosis factor-α and monocyte chemoattractant protein-1) and adhesion markers (vascular cell adhesion molecule 1 and intercellular adhesion molecule 1) was attenuated. The hematoxylin and eosin, Masson’s trichrome, and immunohistochemical staining results showed that SAMiRNA-AREG decreased renal fibrosis, AREG expression, and epidermal growth factor receptor (EGFR) phosphorylation in the UUO- and AD-induced models. Moreover, we studied the effects of SAMiRNA-AREG in response to TGF-β1 in mouse and human proximal tubule cells, and mouse fibroblasts. TGF-β1-induced extracellular matrix production and myofibroblast differentiation were attenuated by SAMiRNA-AREG. Finally, we confirmed that upregulated AREG in the UUO or AD models was mainly localized in the distal tubules. In conclusion, SAMiRNA-AREG represents a novel siRNA therapeutic for renal fibrosis by suppressing EGFR signals.
- Published
- 2020