Peptide-siRNA nanocomplexes targeting NF-κB subunit p65 suppress nascent experimental arthritis
Hui-fang Zhou, Huimin Yan, Hua Pan, Kirk K. Hou, Antonina Akk, Luke E. Springer, Ying Hu, J. Stacy Allen, Samuel A. Wickline, Christine T.N. Pham
Hui-fang Zhou, Huimin Yan, Hua Pan, Kirk K. Hou, Antonina Akk, Luke E. Springer, Ying Hu, J. Stacy Allen, Samuel A. Wickline, Christine T.N. Pham
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Technical Advance Inflammation

Peptide-siRNA nanocomplexes targeting NF-κB subunit p65 suppress nascent experimental arthritis

Abstract

The NF-κB signaling pathway is implicated in various inflammatory diseases, including rheumatoid arthritis (RA); therefore, inhibition of this pathway has the potential to ameliorate an array of inflammatory diseases. Given that NF-κB signaling is critical for many immune cell functions, systemic blockade of this pathway may lead to detrimental side effects. siRNAs coupled with a safe and effective delivery nanoplatform may afford the specificity lacking in systemic administration of small-molecule inhibitors. Here we demonstrated that a melittin-derived cationic amphipathic peptide combined with siRNA targeting the p65 subunit of NF-κB (p5RHH-p65) noncovalently self-assemble into stable nanocomplexes that home to the inflamed joints in a murine model of RA. Specifically, administration of p5RHH-p65 siRNA nanocomplexes abrogated inflammatory cytokine expression and cellular influx into the joints, protected against bone erosions, and preserved cartilage integrity. The p5RHH-p65 siRNA nanocomplexes potently suppressed early inflammatory arthritis without affecting p65 expression in off-target organs or eliciting a humoral response after serial injections. These data suggest that this self-assembling, largely nontoxic platform may have broad utility for the specific delivery of siRNA to target and limit inflammatory processes for the treatment of a variety of diseases.

Authors

Hui-fang Zhou, Huimin Yan, Hua Pan, Kirk K. Hou, Antonina Akk, Luke E. Springer, Ying Hu, J. Stacy Allen, Samuel A. Wickline, Christine T.N. Pham

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Figure 1

p5RHH siRNA nanoparticles accumulate in inflamed paws.

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p5RHH siRNA nanoparticles accumulate in inflamed paws. Day 4 arthritic m...
Day 4 arthritic mice received i.v. injection of HBSS, free Cy5.5-labeled scrambled siRNA (Free siRNA), p5RHH-Cy5.5–labeled scrambled siRNA nanoparticles (p5RHH NP), and in vivo fluorescent images were acquired over time. (A) Fluorescent signals were readily detected in the inflamed paws of arthritic mice injected with free Cy5.5 siRNA and p5RHH siRNA nanoparticles, but not HBSS. Note the substantially higher level of fluorescence in the p5RHH siRNA nanoparticle–injected animal at 7 hours. (B) Organs and blood were collected 2 hours after injection for fluorescence imaging. The highest signals were detected mainly in kidneys, while uptake in the reticuloendothelial system (liver and spleen) was markedly lower. Note that p5RHH siRNA nanoparticles were largely cleared from the circulatory system (blood) 2 hours after injection (far right). Pseudocolor “efficiency image” was used to illustrate the organ accumulation of Cy5.5-labeled siRNA.