Peptide-siRNA nanocomplexes targeting NF-κB subunit p65 suppress nascent experimental arthritis
Hui-fang Zhou, … , Samuel A. Wickline, Christine T.N. Pham
Hui-fang Zhou, … , Samuel A. Wickline, Christine T.N. Pham
Published August 26, 2014
Citation Information: J Clin Invest. 2014;124(10):4363-4374. https://doi.org/10.1172/JCI75673.
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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 3

p5RHH-p65 siRNA nanotherapy suppresses inflammation in CAIA.

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p5RHH-p65 siRNA nanotherapy suppresses inflammation in CAIA. CAIA arthri...
CAIA arthritis was induced as detailed in Methods (day 3 LPS injection is indicated). On day 4, mice were randomly divided into 3 treatment groups: HBSS, p5RHH siRNA nanoparticles, and p5RHH-p65 siRNA nanoparticles. Nanoparticles or HBSS were administered i.v. serially on days 4, 5, and 6 (Tx). (A) Representative photographs of day 10 paws from the 3 treatment groups. (B–D) Change in ankle thickness (B), arthritis score (C), and percent weight loss (D) were chronicled over time. Values are mean ± SEM (n = 6–8 per group). (E) Day 10 paws were harvested, sectioned, and stained with H&E to evaluate for the number of inflammatory cells/erosions and toluidine blue to evaluate for cartilage integrity. Note the intense inflammatory infiltrates in and surrounding the joint space in HBSS- and p5RHH siRNA–treated animals while joint space in p5RHH-p65 siRNA–treated animals was largely devoid of cellular infiltrates (left). In higher-power images, erosions (asterisks) were noted along the bone surface in HBSS- and p5RHH siRNA–treated animals (middle). Extensive proteoglycan depletion (arrowheads) was also noted in HBSS- and p5RHH siRNA–treated animals (right). b, bone; js, joint space. Scale bars: 400 μm (left), 100 μm (middle), 50 μm (right). (F–H) Inflammatory infiltrates per high-power field (HPF; F), erosions (G), and degree of proteoglycan depletion (H) were quantified as detailed in Methods. *P < 0.05, **P < 0.01, ***P < 0.001.