Molecular imaging study on in vivo distribution and pharmacokinetics of modified small interfering RNAs (siRNAs).

Molecular imaging was used to study the biodistribution, pharmacokinetics, and activity of naked small interfering RNAs (siRNAs). siRNAs with riboses chemically modified in the 2' position were compared with unmodified siRNA. In vitro, replacement of the 2'-hydroxyl (2'OH) group of certain nucleotides in an siRNA sequence by a fluorine atom (2'F) on both antisense (AS) and sense (S) strands [2'F(AS/S)], or by a methoxy group (2'OMe) on the S strand [2'OH(AS)/2'OMe(S)], was compatible with RNA interference. Different siRNAs [2'F(AS/S), 2'OH(AS)/2'OMe(S), and 2'OH(AS/S)] were labeled with fluorine-18 (conjugation with [(18)F]FPyBrA), and comparative dynamic and quantitative imaging was performed with positron emission tomography. After intravenous injections of [(18)F]siRNAs in rodents, total radioactivity was rapidly eliminated by the kidneys and the liver. Tissue distribution of the different siRNAs were similar, and their bioavailability (as judged from blood persistence and stability) increased in the order 2'OH(AS/S) = 2'OH(AS)/2'OMe(S) < 2'F(AS/S). However, in our in vivo model, the 2'F(AS/S) siRNA, despite its higher bioavailability, was not able to induce a higher interference effect with respect to the 2'OH(AS/S) siRNA. Molecular imaging approaches, applied in the present work to both natural and chemically modified siRNAs, can contribute to the development of these macromolecules as therapeutic agents.

[1]  R. Boisgard,et al.  Fluorine-18 labelling of small interfering RNAs (siRNAs) for PET imaging† , 2007 .

[2]  M. Stoffel,et al.  Mechanisms and optimization of in vivo delivery of lipophilic siRNAs , 2007, Nature Biotechnology.

[3]  Wolfgang A. Weber,et al.  Impact of tumor-specific targeting on the biodistribution and efficacy of siRNA nanoparticles measured by multimodality in vivo imaging , 2007, Proceedings of the National Academy of Sciences.

[4]  Zhihong Zhu,et al.  Radiolabeling small RNA with technetium-99m for visualizing cellular delivery and mouse biodistribution. , 2007, Nuclear medicine and biology.

[5]  Anna Moore,et al.  In vivo imaging of siRNA delivery and silencing in tumors , 2007, Nature Medicine.

[6]  N Toub,et al.  Innovative nanotechnologies for the delivery of oligonucleotides and siRNA. , 2006, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[7]  O. Boerman,et al.  INTRAVENOUSLY ADMINISTERED SHORT INTERFERING RNA ACCUMULATES IN THE KIDNEY AND SELECTIVELY SUPPRESSES GENE FUNCTION IN RENAL PROXIMAL TUBULES , 2006, Drug Metabolism and Disposition.

[8]  L. Lim,et al.  Position-specific chemical modification of siRNAs reduces "off-target" transcript silencing. , 2006, RNA.

[9]  A. Aigner.  Gene silencing through RNA interference (RNAi) in vivo: strategies based on the direct application of siRNAs. , 2006, Journal of biotechnology.

[10]  Matthias John,et al.  RNAi-mediated gene silencing in non-human primates , 2006, Nature.

[11]  A. Gartel,et al.  RNA interference in cancer. , 2006, Biomolecular engineering.

[12]  A. Lee,et al.  Design of noninflammatory synthetic siRNA mediating potent gene silencing in vivo. , 2006, Molecular therapy : the journal of the American Society of Gene Therapy.

[13]  M. Woodle,et al.  Harnessing in vivo siRNA delivery for drug discovery and therapeutic development , 2006, Drug Discovery Today.

[14]  S. Uprichard.  The therapeutic potential of RNA interference , 2005, FEBS Letters.

[15]  R. Griffey,et al.  Positional effect of chemical modifications on short interference RNA activity in mammalian cells. , 2005, Journal of medicinal chemistry.

[16]  J. Rossi,et al.  Sensing the danger in RNA , 2005, Nature Medicine.

[17]  S. Akira,et al.  Sequence-specific potent induction of IFN-α by short interfering RNA in plasmacytoid dendritic cells through TLR7 , 2005, Nature Medicine.

[18]  J. Votaw,et al.  Silencing of CXCR4 blocks breast cancer metastasis. , 2005, Cancer research.

[19]  M. Manoharan.  RNA interference and chemically modified small interfering RNAs. , 2004, Current opinion in chemical biology.

[20]  Matthias John,et al.  Therapeutic silencing of an endogenous gene by systemic administration of modified siRNAs , 2004, Nature.

[21]  Adriano Duatti,et al.  In vivo imaging of oligonucleotides with nuclear tomography. , 2004, Current drug targets.

[22]  S. Ashley,et al.  Systemic siRNA-Mediated Gene Silencing: A New Approach to Targeted Therapy of Cancer , 2004, Annals of surgery.

[23]  B. Mahon,et al.  Efficient delivery of small interfering RNA for inhibition of IL-12p40 expression in vivo , 2004, Journal of Inflammation.

[24]  G. Lucignani,et al.  Techniques: reporter mice - a new way to look at drug action. , 2004, Trends in pharmacological sciences.

[25]  Anton P. McCaffrey,et al.  In vivo activity of nuclease-resistant siRNAs. , 2004, RNA.

[26]  B. Tavitian,et al.  Design and synthesis of a new [18F]fluoropyridine-based haloacetamide reagent for the labeling of oligonucleotides: 2-bromo-N-[3-(2-[18F]fluoropyridin-3-yloxy)propyl]acetamide. , 2004, Bioconjugate chemistry.

[27]  Z. Paroo,et al.  Biodistribution of phosphodiester and phosphorothioate siRNA. , 2004, Bioorganic & medicinal chemistry letters.

[28]  S. Ashley,et al.  RNA interference targeting the M2 subunit of ribonucleotide reductase enhances pancreatic adenocarcinoma chemosensitivity to gemcitabine , 2004, Oncogene.

[29]  R. Boisgard,et al.  Fluorine‐18‐ and iodine‐125‐labelling of spiegelmers , 2003 .

[30]  T. Rana,et al.  siRNA function in RNAi: a chemical modification analysis. , 2003, RNA.

[31]  Robert H. Silverman,et al.  Activation of the interferon system by short-interfering RNAs , 2003, Nature Cell Biology.

[32]  A. Harel-Bellan,et al.  SiRNA-mediated inhibition of vascular endothelial growth factor severely limits tumor resistance to antiangiogenic thrombospondin-1 and slows tumor vascularization and growth. , 2003, Cancer research.

[33]  B. Tavitian.  In vivo imaging with oligonucleotides for diagnosis and drug development , 2003, Gut.

[34]  A. Klippel,et al.  Structural variations and stabilising modifications of synthetic siRNAs in mammalian cells. , 2003, Nucleic acids research.

[35]  B. Li,et al.  Expression profiling reveals off-target gene regulation by RNAi , 2003, Nature Biotechnology.

[36]  Mouldy Sioud,et al.  Gene silencing by systemic delivery of synthetic siRNAs in adult mice. , 2003, Journal of molecular biology.

[37]  M. Amarzguioui,et al.  Tolerance for mutations and chemical modifications in a siRNA. , 2003, Nucleic acids research.

[38]  T. Tuschl,et al.  Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells , 2001, Nature.

[39]  C. Crouzel,et al.  General method to label antisense oligonucleotides with radioactive halogens for pharmacological and imaging studies. , 2000, Bioconjugate chemistry.

[40]  S. Gambhir,et al.  Rapid synthesis of a 5'-fluorinated oligodeoxy-nucleotide: a model antisense probe for use in imaging with positron emission tomography (PET). , 1998, Bioorganic & medicinal chemistry letters.

[41]  Bernard Bendriem,et al.  In vivo imaging of oligonucleotides with positron emission tomography , 1998, Nature Medicine.

[42]  A. Fire,et al.  Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans , 1998, Nature.

[43]  S. Agrawal,et al.  Pharmacokinetics and metabolism of an oligodeoxynucleotide phosphorothioate (GEM91) in cynomolgus monkeys following intravenous infusion. , 1998, Antisense & nucleic acid drug development.

[44]  Richard A. Jorgensen,et al.  Chalcone synthase cosuppression phenotypes in petunia flowers: comparison of sense vs. antisense constructs and single-copy vs. complex T-DNA sequences , 1996, Plant Molecular Biology.

[45]  C. Napoli,et al.  Introduction of a Chimeric Chalcone Synthase Gene into Petunia Results in Reversible Co-Suppression of Homologous Genes in trans. , 1990, The Plant cell.

[46]  V. Wacheck,et al.  Short interfering RNA (siRNA): tool or therapeutic? , 2006, Clinical science.

[47]  M. Kitajima,et al.  Selective sentinel lymphadenectomy for human solid cancer , 2005 .

[48]  S. Fesik,et al.  siRNA-mediated gene silencing: a global genome view. , 2004, Nucleic acids research.

[49]  Michael R. Green,et al.  Nonspecific, concentration-dependent stimulation and repression of mammalian gene expression by small interfering RNAs (siRNAs). , 2004, RNA.

[50]  Volker A Erdmann,et al.  Application of locked nucleic acids to improve aptamer in vivo stability and targeting function. , 2004, Nucleic acids research.

[51]  Phillip A Sharp,et al.  siRNAs can function as miRNAs , 2003 .

[52]  C. Cantor,et al.  Comparative properties of a technetium-99m-labeled single-stranded natural DNA and a phosphorothioate derivative in vitro and in mice. , 1996, The Journal of pharmacology and experimental therapeutics.

[53]  S. Agrawal,et al.  Pharmacokinetics of Antisense Oligonucleotides , 1995, Clinical pharmacokinetics.