Biochemical and Biophysical Research Communications
Electro-transfer of small interfering RNA ameliorated arthritis in rats
Section snippets
Materials and methods
siRNA duplexes. siRNA duplexes targeting rat TNF-α gene were synthesized (A: 5′-GCCCGUAGCCCACGUCGUAd(TT)-3′ and 5′-UACGACGUGGGCUACGGGCd(TT)-3′; B: 5′-UGGGCUCCCUCUCAUCAGUd(TT)-3′ and 5′-ACUGAUGAGAGGGAGCCCAd(TT)-3′; C: 5′-GGAGGAGAAGUUCCCAAAUd(TT)-3′, and 5′-AUUUGGGAACUUCUCCUCCd(TT)-3′; and D: 5′-AGACAACCAACUGGUGGUAd(TT), and 5′-UACCACCAGUUGGUUGUCUd(TT)-3′). Two nucleotide mismatches were introduced into the TNF-α-siRNA-A to generate the mismatched TNF-α-siRNA (5′-GCCCGUAGAACACGUCGUAd(TT)-3′ and
Electro-transfer of siRNA into the synovium of the rat knee joint
GAPDH-specific siRNA duplex was labeled with 6-carboxyfluorescein (FAM) and injected into the left knee joint of DA rats, to which electric field was applied subsequently at the joint. Twenty-four hours later, the labeled siRNA was present at the intra-articular synovial tissue as revealed by fluorescence stereomicroscopic observation (data not shown). The siRNA was more effectively transduced when it was conjugated with a polyamine (siPORT Amine) before electro-transfer (Figs. 1A–D). The
Discussion
Therapeutic application of RNAi has been performed against a variety of disorders including hepatic [8], respiratory [10], [11], ocular [12], neuronal [13], [14], renal [15], and malignant [16], [17], [18], [19] diseases. To obtain therapeutic gene silencing in the liver, synthetic siRNA was intravenously administered to the animals [8], while RNAi in other organs was induced by siRNA [11], [12], [14], [16], [17], [19], [20] or a siRNA-expression plasmid [10], [18] that were administered into
Acknowledgments
We thank Dr. M. Okabe for supplying the EGFP transgenic rat. This study was supported by a Grant-in-Aid for Scientific Research (No. 15790799) from the Ministry of Education, Culture, Sports, Science and Technology of Japan and JSPS Fujita Memorial Fund for Medical Research.
References (36)
- et al.
Gene silencing by systemic delivery of synthetic siRNAs in adult mice
J. Mol. Biol.
(2003) - et al.
Inhibitory effect of TNF alpha antibodies on synovial cell interleukin-1 production in rheumatoid arthritis
Lancet
(1989) - et al.
Successful genetic transduction in vivo into synovium by means of electroporation
Biochem. Biophys. Res. Commun.
(2002) - et al.
‘Green mice’ as a source of ubiquitous green cells
FEBS Lett.
(1997) - et al.
Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans
Nature
(1998) RNA interference
Nature
(2002)- et al.
Gene silencing as an adaptive defence against viruses
Nature
(2001) - et al.
Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells
Nature
(2001) - et al.
Efficient delivery of siRNA for inhibition of gene expression in postnatal mice
Nat. Genet.
(2002) - et al.
RNA interference in adult mice
Nature
(2002)
Sequence-specific gene silencing in murine muscle induced by electroporation-mediated transfer of short interfering RNA
J. Gene Med.
RNA interference targeting Fas protects mice from fulminant hepatitis
Nat. Med.
Inhibition of respiratory syncytial virus infection with intranasal siRNA nanoparticles targeting the viral NS1 gene
Nat. Med.
In vivo gene silencing (with siRNA) of pulmonary expression of MIP-2 versus KC results in divergent effects on hemorrhage-induced, neutrophil-mediated septic acute lung injury
J. Leukoc. Biol.
RNA interference targeting transforming growth factor-beta type II receptor suppresses ocular inflammation and fibrosis
Mol. Vis.
RNAi suppresses polyglutamine-induced neurodegeneration in a model of spinocerebellar ataxia
Nat. Med.
Neurochemical and behavioral consequences of widespread gene knockdown in the adult mouse brain by using nonviral RNA interference
Proc. Natl. Acad. Sci. USA
Exploring RNA interference as a therapeutic strategy for renal disease
Gene Ther.
Cited by (65)
Cyclin B1 knockdown mediated by clinically approved pulsed electric fields siRNA delivery induces tumor regression in murine melanoma
2020, International Journal of PharmaceuticsTargeted electro-delivery of oligonucleotides for RNA interference: SiRNA and antimiR
2015, Advanced Drug Delivery ReviewsCitation Excerpt :As a result, as cells must be permeabilized to initiate oligonucleotide uptake, silencing is obtained only on a sub-population. EP has been used successfully for in vivo siRNA electrotransfer in a wide variety of tissues such as muscle [74,75], eyes [76], joint tissue [77], the kidney [78] and the brain [79]. EP has also proven its efficacy in siRNA tumor electrotransfer [37,61,80] (Table 1).
CCR5 small interfering RNA ameliorated joint inflammation in rats with adjuvant-induced arthritis
2014, Immunology LettersCitation Excerpt :Now, the therapeutic potential of RNAi was highlighted in experiments indicating the efficacy of the approach against viral infection, such as human immunodeficiency virus [12,13], hepatitis B virus [14] and simplex virus 2 [15]. In addition, the siRNA-dependent strategy also be used in RA model [16–18], neurodegenerative diseases [19,20] and cancers [21,22]. Moreover, RNAi-based therapy for age-related macular degeneration has already reached clinical trials in 2004 [23].
PLGA microspheres encapsulating siRNA anti-TNFalpha: Efficient RNAi-mediated treatment of arthritic joints
2012, European Journal of Pharmaceutics and BiopharmaceuticsCitation Excerpt :The use of short interfering (si)RNA has been recently proposed as new approach to efficiently inhibit the expression of TNF-α in the treatment of RA [5–8]. Direct intra-articular electro-transfer of anti-TNF siRNAs has been reported to reduce joint inflammation in the pre-clinical model of collagen-induced arthritis (CIA) and supports the use of RNAi-based therapies [5,6]. Long term in vivo application of RNAi, however, needs to face several hurdles and requires the development of specifically tailored vehicles.
Alternative for anti-TNF antibodies for arthritis treatment
2011, Molecular TherapyCitation Excerpt :Recent studies have proven the success of siRNA in targeting specific protein in vitro6,8 and in vivo.17,18 Recently, siRNA targeting TNF-α was shown to modulate disease severity and inhibit gene expression in electrotransferred knee joints of arthritic animals.7,19 On the other hand, TFO molecules are 12–28 long oligonucleotides20 which recognize the double-helical DNA by sequence-specific binding, due to Hoogsteen or reverse-Hoogsteen hydrogen bounding.21,22