Immune activation and target organ damage are consequences of hydrodynamic treatment but not delivery of naked siRNAs in mice.

Short-interfering RNAs (siRNAs), key mediators of RNA interference comprise a promising therapeutic tool, although side effects such as interferon (IFN) response are still not perfectly understood. Further, delivery to target organs is a major challenge, possibly associated with side effects including immune activation or organ damage. We investigated whether immune activation as a consequence of double-stranded RNA induced IFN response (Jak/STAT pathway activation or cytokine production) or target organ damage is induced by in vivo low-volume (LV) or high-volume (HV) hydrodynamic delivery or treatment with naked siRNA. NMRI mice were injected with naked siRNAs or saline by hydrodynamic injection (HDI) and positive control mice received polyinosinic-polycytidilic acid (poly I:C). LV (1 mL/mouse) and HV (10% of body weight) HDI were compared. After LV HDI, STAT1 and OAS1 gene expression inflammatory cytokine plasma levels and target organ injury were assessed. LV HDI induced slight alanine aminotransferase elevation and mild hepatocyte injury, whereas HV HDI resulted in high ALAT level and extensive hepatocyte necrosis. STAT1 or OAS1 was not induced by LV siRNA; however, HV saline led to a time-dependent slight increase in gene expression. Inflammatory cytokine plasma level and organ histology and functional parameters demonstrated no damage following LV HDI with or without siRNA. Our data demonstrate that naked siRNAs may be harnessed, without the induction of IFN response or immune activation, and that LV HDI is preferable, because HV HDI may cause organ damage.

Post-transcriptional gene-expression regulation by micro RNA (miRNA) network in renal disease.

Micro RNAs (miRNAs) are a recently discovered class of small, non-coding RNAs with the function of post-transcriptional gene expression regulation. MiRNAs may function in networks, forming a complex relationship with diseases. Alterations of specific miRNA levels have significant correlation with diseases of divergent origin, such as diabetic or ischemic organ injury including nephropathy, and malignant diseases including renal tumors. After identification of disease-associated miRNAs, there are two options of influencing their tissue expression. The function of miRNAs can be inhibited by antisense oligonucleotides (ASOs), which have been shown to silence specific miRNAs in vivo. Moreover, miRNA activity can be also mimicked or enhanced by delivering chemically synthesized miRNAs. Thus, modifying the expression of miRNAs is a potential future gene-therapeutic tool to influence posttranscriptional regulation of multiple genes in a single therapy. In this review we focus on key renal miRNAs with the aim of revealing the pathomechanisms of renal diseases. Nucleic acid therapy with oligonucleotides and short interfering RNA (siRNA) are under clinical evaluation presently. Similar therapeutic strategies, to influence miRNA function is also already under clinical investigation in RNA interference trials. We summarize here studies specifically aimed at the modification of miRNA expression. Research on the post-transcriptional regulation of gene expression by miRNA may reshape our understanding of renal pathophysiology and consequently may bring new diagnostic markers and therapeutic agents.

Application of siRNA in targeting protein expression in kidney disease.

Although it is one of the major targeted organs by systemically administered siRNA, when compared to other tissues the kidney receives only moderate interest regarding therapeutic siRNA delivery. Here we review recent approaches to target renal protein expression under normal and pathological conditions. Experimental evidence to support the clinical relevance of siRNA administration in the treatment of renal disease is discussed. High-throughput screening using recently available genome-wide RNA interference libraries provides a new, powerful tool that can be applied to conventional and 3D in vitro culture models for lead finding or the identification of signal pathway involvement in renal disease.

[SiRNA technology, the gene therapy of the future?]. / SiRNS-technológia, a jövo génterápiája?

A new era in genetics started 17 years ago, when co-suppression in petunia was discovered. Later, co-suppression was identified as RNA interference (RNAi) in many plant and lower eukaryote animals. Although an ancient antiviral host defense mechanism in plants, the physiologic role of RNAi in mammals is still not completely understood. RNAi is directed by short interfering RNAs (siRNAs), one subtype of short double stranded RNAs. In this review we summarize the history and mechanisms of RNAi. We also aim to highlight the correlation between structure and efficacy of siRNAs. Delivery is the most important obstacle for siRNA based gene therapy. Viral and nonviral deliveries are discussed. In vivo delivery is the next obstacle to clinical trials with siRNAs. Although hydrodynamic treatment is effective in animals, it cannot be used in human therapy. One possibility is organ selective catheterization. The known side effects of synthesized siRNAs are also discussed. Although there are many problems to face in this new field of gene therapy, successful in vitro and in vivo experiments raise hope for treating human disease with siRNA.