TY - JOUR
T1 - Therapeutic Potential of OMe-PS-miR-29b1 for Treating Liver Fibrosis
AU - Kumar, Virender
AU - Kumar, Vinod
AU - Luo, Jiangtao
AU - Mahato, Ram I.
N1 - Funding Information:
The faculty start-up fund from the University of Nebraska Medical Center is duly acknowledged for providing financial support for this work.
PY - 2018/12/5
Y1 - 2018/12/5
N2 - Trans-differentiation of quiescent hepatic stellate cells (HSCs) into active myofibroblasts secretes excess amounts of extracellular matrix (ECM) proteins. miR-29b1 has the potential to treat liver fibrosis, because it targets several profibrotic genes. We previously demonstrated that miR-29b1 and the hedgehog (Hh) pathway inhibitor GDC-0449 could, together, inhibit the activation of HSCs and ECM production in common bile-duct-ligated (CBDL) mice. Herein, we determined the effect of chemical modifications of miR-29b1 on its stability, immunogenicity, and Argonaute-2 (Ago2) loading in vitro, after modifying its antisense strand with phosphorothioate (PS-miR-29b1), 2′-O-methyl-phosphorothioate (OMe-miR-29b1), locked nucleic acid (LNA-miR-29b1), and N,N’-diethyl-4-(4-nitronaphthalen-1-ylazo)-phenylamine (ZEN-miR-29b1). Chemical modifications significantly improved stability of miR-29b1 in 50% FBS. Among all the modified miRNAs tested, OMe-PS-miR-29b1 showed the highest stability with low immunogenicity, without the loss of efficacy in vitro. Therefore, OMe-PS-miR-29b1 was complexed with poly(ethylene glycol)-block-poly(2-methyl-2-carboxyl-propylenecarbonate-graft-dodecanol-graft-tetraethylenepentamine (mPEG-b-PCC-g-DC-g-TEPA) cationic micelles, and anti-fibrotic efficacy was evaluated in CBDL mice. There was a significant improvement in liver histology and decrease in the levels of injury markers. Further, mRNA/protein levels of collagen, α-SMA, and TIMP-1 were significantly lower for the OMe-PS-miR-29b1-loaded micelles compared to miR-29b1-loaded micelles. In conclusion, micellar delivery of OMe-PS-miR-29b1 is a promising strategy to treat liver fibrosis. Kumar et al. demonstrated the improved serum stability, immunogenicity, and efficacy of miR-29b1 when modified with 2′-O-methyl-phosphorothioate. Systemic administration of OMe-PS-miR-29b1 micellar formulation significantly decreased the levels of liver injury markers, collagen, and proinflammatory cytokines. Therefore, this is a promising strategy for treating liver and other fibroses.
AB - Trans-differentiation of quiescent hepatic stellate cells (HSCs) into active myofibroblasts secretes excess amounts of extracellular matrix (ECM) proteins. miR-29b1 has the potential to treat liver fibrosis, because it targets several profibrotic genes. We previously demonstrated that miR-29b1 and the hedgehog (Hh) pathway inhibitor GDC-0449 could, together, inhibit the activation of HSCs and ECM production in common bile-duct-ligated (CBDL) mice. Herein, we determined the effect of chemical modifications of miR-29b1 on its stability, immunogenicity, and Argonaute-2 (Ago2) loading in vitro, after modifying its antisense strand with phosphorothioate (PS-miR-29b1), 2′-O-methyl-phosphorothioate (OMe-miR-29b1), locked nucleic acid (LNA-miR-29b1), and N,N’-diethyl-4-(4-nitronaphthalen-1-ylazo)-phenylamine (ZEN-miR-29b1). Chemical modifications significantly improved stability of miR-29b1 in 50% FBS. Among all the modified miRNAs tested, OMe-PS-miR-29b1 showed the highest stability with low immunogenicity, without the loss of efficacy in vitro. Therefore, OMe-PS-miR-29b1 was complexed with poly(ethylene glycol)-block-poly(2-methyl-2-carboxyl-propylenecarbonate-graft-dodecanol-graft-tetraethylenepentamine (mPEG-b-PCC-g-DC-g-TEPA) cationic micelles, and anti-fibrotic efficacy was evaluated in CBDL mice. There was a significant improvement in liver histology and decrease in the levels of injury markers. Further, mRNA/protein levels of collagen, α-SMA, and TIMP-1 were significantly lower for the OMe-PS-miR-29b1-loaded micelles compared to miR-29b1-loaded micelles. In conclusion, micellar delivery of OMe-PS-miR-29b1 is a promising strategy to treat liver fibrosis. Kumar et al. demonstrated the improved serum stability, immunogenicity, and efficacy of miR-29b1 when modified with 2′-O-methyl-phosphorothioate. Systemic administration of OMe-PS-miR-29b1 micellar formulation significantly decreased the levels of liver injury markers, collagen, and proinflammatory cytokines. Therefore, this is a promising strategy for treating liver and other fibroses.
KW - CBDL
KW - backbone modifications
KW - liver fibrosis
KW - miR-29b1
KW - micelles
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UR - http://www.scopus.com/inward/citedby.url?scp=85054124042&partnerID=8YFLogxK
U2 - 10.1016/j.ymthe.2018.08.022
DO - 10.1016/j.ymthe.2018.08.022
M3 - Article
C2 - 30287074
AN - SCOPUS:85054124042
VL - 26
SP - 2798
EP - 2811
JO - Molecular Therapy
JF - Molecular Therapy
SN - 1525-0016
IS - 12
ER -