Efficient new cationic liposome formulation for systemic delivery of small interfering RNA silencing tumor necrosis factor α in experimental arthritis

Abstract

Objective. Tumor necrosis factor ␣ (TNF␣) is among the most prominent cytokines in rheumatoid arthritis (RA) and is secreted mainly by macrophages. A direct method for restoring the immunologic balance in RA is use of small interfering RNA (siRNA) for silencing the TNF␣ transcript. The aim of this study was to determine the therapeutic effect of systemic administration of TNF␣ siRNA in an experimental model of RA, optimizing its delivery using new liposome formulations.Methods. Murine macrophages were transfected with siRNA targeting TNF␣, and expression was measured. The therapeutic effect in collagen-induced arthritis (CIA) was assessed after intravenous delivery of TNF␣ siRNA. Delivery was optimized using a carrier DNA for complexation with the cationic liposome RPR209120/DOPE. Levels of TNF␣ and other cytokines were measured in sera and joint tissue-conditioned media. Biodistribution was determined using a fluorescent siRNA.Results. In vitro, TNF␣ siRNA efficiently and specifically modulated the expression of TNF␣ at both the messenger RNA and protein levels. In vivo, complete cure of CIA was observed when TNF␣ siRNA was administered weekly, complexed with the liposome and combined with carrier DNA. Inhibition (50-70%) of articular and systemic TNF␣ secretion was detected in the siRNA-injected groups, which correlated with a decrease in the levels of interleukin-6 and monocyte chemotactic protein 1. The main organs targeted by siRNA were the liver and spleen; the addition of liposome RPR209120 and carrier DNA significantly increased organ uptake.Conclusion. We demonstrated the efficiency of systemic delivery of siRNA designed to silence TNF␣ in CIA, using a liposome carrier system as a way to address the methodologic limitations in vivo.

In vivo RNAi-mediated silencing of TAK1 decreases inflammatory Th1 and Th17 cells through targeting of myeloid cells

“…39 To address in vivo the effect of TAK1 silencing in CIA as a prototype model of RA, we took advantage of a simple, efficient RNAi-based lipoplex formulation that we previously developed. 24 Although RNAi has emerged as a potent technology for silencing gene expression, the use of RNAi-based therapeutics has to overcome major obstacles, mainly effective delivery to target cells. We have previously demonstrated that the cationic liposome, RPR209120/DOPE, in combination with a carrier DNA, and siRNAs, provides high nucleic acid stability and effective lipofection rates at low amounts of formulated siRNAs.…”

“…capacities, we have previously shown that no type I IFN response could be observed upon intravenous injection of 0.5 mg/kg of unmodified siRNA duplexes formulated with our delivery system. 24,40,45 Different types of cells or tissues have been reported to be sensitive for TAK1 deletion and undergone apoptotic cell death. 19 Here, we show that targeted organs of TAK1-siRNA lipoplex-injected mice exhibited similar numbers of mononuclear cells, indicating that TAK1 RNAimediated knockdown does not induce cell death.…”

“…24 In vivo delivery experiment using fluorescently labeled Cy3 siRNA (Dharmacon) was carried out by administering formulated siRNA intravenously (0.5 mg/kg). Mice were killed at defined time points, and fluorescence uptake was examined by flow cytometry for indicated phenotypic markers.…”

Prevention of experimental autoimmune uveoretinitis by blockade of osteopontin with small interfering RNA

“…In particular, small interfering RNAs (siRNAs), 21-23 nucleotide-length fragments (Elbashir et al 2001;Hannon 2002;Xie et al 2006), have been shown to be of great value in decreasing the expression of the target gene by in vivo administration (Song et al 2003;Nakamura et al 2004;Schiffelers et al 2005;Khoury et al 2006) In the present study, we applied siRNA targeting to an OPN coding sequence (OPN-siRNA) to inhibit OPN function by reducing OPN expression. We demonstrate the remarkable efficacy of OPN-siRNA to prevent EAU in mice.…”

Amphoteric liposomes enable systemic antigen‐presenting cell–directed delivery of CD40 antisense and are therapeutically effective in experimental arthritis

“…However, their in vivo application requires effective delivery to immune cells and/or sites of inflammation (28). Recent efforts to facilitate systemic oligonucleotide delivery to inflammation sites in vivo have included the use of neutral liposomes (17) or atelocollagen (29) in experimental allergic dermatitis, cationic liposomes in experimental arthritis (30), and antibody-targeted stabilized nanoparticles in experimental colitis (31).…”