an-orthotopic-bladder-tumor-model-evaluation-intravesical-sarna

An Orthotopic Bladder Tumor Model and the Evaluation of Intravesical saRNA Treatment

We present a novel method for treating bladder cancer with intravesically delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder ...

Proprotein convertase subtilisin/kexin type 9 (PCSK9) regulates low density lipoprotein receptor (LDLR) protein levels and function. Loss of PCSK9 increases LDLR levels in liver and reduces plasma LDL cholesterol (LDLc), whereas excess PCSK9 activity decreases liver LDLR levels and increases plasma LDLc. Here, we have developed active, cross-species, small interfering RNAs (siRNAs) capable of targeting murine, rat, nonhuman primate (NHP), and human PCSK9. For in vivo studies, PCSK9 and control siRNAs were formulated in a lipidoid nanoparticle (LNP). Liver-specific siRNA silencing of PCSK9 in mice and rats reduced PCSK9 mRNA levels by 50-70%. The reduction in PCSK9 transcript was associated with up to a 60% reduction in plasma cholesterol concentrations. These effects were shown to be mediated by an RNAi mechanism, using 5'-RACE. In transgenic mice expressing human PCSK9, siRNAs silenced the human PCSK9 transcript by >70% and significantly reduced PCSK9 plasma protein levels. In NHP, a single dose of siRNA targeting PCSK9 resulted in a rapid, durable, and reversible lowering of plasma PCSK9, apolipoprotein B, and LDLc, without measurable effects on either HDL cholesterol (HDLc) or triglycerides (TGs). The effects of PCSK9 silencing lasted for 3 weeks after a single bolus i.v. administration. These results validate PCSK9 targeting with RNAi therapeutics as an approach to specifically lower LDLc, paving the way for the development of PCSK9-lowering agents as a future strategy for treatment of hypercholesterolemia.

Therapeutic RNAi targeting PCSK9 acutely lowers plasma cholesterol in rodents and LDL cholesterol in nonhuman primates.

Proprotein-Konvertase Subtilisin/Kexin Typ 9 (PCSK9) regelt low-Density-Lipoprotein-Rezeptor (LDLR)-Protein-Ebene und Funktion. Verlust des PCSK9 erh&ouml;ht LDLR-Konzentrationen in Leber und Plasma LDL-Cholesterin (LDLc), verringert, w&auml;hrend &uuml;bersch&uuml;ssiges PCSK9 Aktivit&auml;t Leber sinkt LDLR Ebenen und Plasma LDLc erh&ouml;ht. Hier haben wir aktive, Kreuz-Arten, kleine st&ouml;rende RNAs (SiRNAs) f&auml;hig Murine, Ratte, nichtmenschlichen Primaten (NHP) und menschliche PCSK9 entwickelt. F&uuml;r in-vivo-Untersuchungen wurden PCSK9 und Kontrolle SiRNAs formuliert, in einem Lipidoid-Nanopartikel (LNP). Leber-spezifischen SiRNA Unterdr&uuml;ckender des PCSK9 in M&auml;usen und Ratten reduziert PCSK9 mRNA-Stufen um 50-70 %. Der R&uuml;ckgang der PCSK9 Abschrift wurde mit bis zu 60 % Erm&auml;&szlig;igung im Plasma-Cholesterin-Konzentrationen. Diese Effekte wurden gezeigt, durch einen Mechanismus zur RNAi, 5&#39;-RACE mit vermittelt werden. Bei transgenen M&auml;usen Ausdruck menschlichen PCSK9 SiRNAs zum Schweigen gebracht der menschlichen PCSK9-Abschrift von &gt; 70 % und deutlich reduzierten PCSK9-Plasma-Protein-Ebene. NHP f&uuml;hrte eine Einzeldosis SiRNA PCSK9 Ausrichtung einer raschen, langlebig und reversible Senkung des Plasma PCSK9, Apolipoprotein B und LDLc, ohne messbare Auswirkungen auf HDL-Cholesterin (HDLc) oder Triglyceride (TGs). Die Auswirkungen des PCSK9 zum Schweigen dauerte 3 Wochen nach einer einzigen Bolus i.v. Verabreichung. Diese Ergebnisse best&auml;tigen PCSK9 targeting mit RNAi-Therapeutika als Ansatz, speziell LDLc, ebnet den Weg f&uuml;r die Entwicklung des PCSK9-senkende Mittel als k&uuml;nftige Strategie zur Behandlung der Hypercholesterin&auml;mie zu senken.

Therapeutischen RNAi gezielt PCSK9 akut senkt Plasma-Cholesterin bei Nagern und LDL-Cholesterin in nicht-menschlichen Primaten.

Nanoparticles are used for delivering therapeutics into cells. However, size, shape, surface chemistry and the presentation of targeting ligands on the surface of nanoparticles can affect circulation half-life and biodistribution, cell-specific internalization, excretion, toxicity and efficacy. A variety of materials have been explored for delivering small interfering RNAs (siRNAs)--a therapeutic agent that suppresses the expression of targeted genes. However, conventional delivery nanoparticles such as liposomes and polymeric systems are heterogeneous in size, composition and surface chemistry, and this can lead to suboptimal performance, a lack of tissue specificity and potential toxicity. Here, we show that self-assembled DNA tetrahedral nanoparticles with a well-defined size can deliver siRNAs into cells and silence target genes in tumours. Monodisperse nanoparticles are prepared through the self-assembly of complementary DNA strands. Because the DNA strands are easily programmable, the size of the nanoparticles and the spatial orientation and density of cancer-targeting ligands (such as peptides and folate) on the nanoparticle surface can be controlled precisely. We show that at least three folate molecules per nanoparticle are required for optimal delivery of the siRNAs into cells and, gene silencing occurs only when the ligands are in the appropriate spatial orientation. In vivo, these nanoparticles showed a longer blood circulation time (t(1/2) ≈ 24.2 min) than the parent siRNA (t(1/2) ≈ 6 min).

Article	Year
Nature\| PubMed ID: 15538359	2004
Therapeutic RNAi targeting PCSK9 acutely lowers plasma cholesterol in rodents and LDL cholesterol in nonhuman primates. Proceedings of the National Academy of Sciences of the United States of America\| PubMed ID: 18695239	2008
Molecular neurodegeneration\| PubMed ID: 18976489	2008
Oligonucleotides\| PubMed ID: 19093781	2009
Analytical biochemistry\| PubMed ID: 20156415	2010
Nucleic acids research\| PubMed ID: 20403811	2010
Nucleic acids research\| PubMed ID: 20610434	2010
The Journal of organic chemistry\| PubMed ID: 21299239	2011
Angewandte Chemie (International ed. in English)\| PubMed ID: 21351337	2011
Experimental neurology\| PubMed ID: 22119622	2012
RNA (New York, N.Y.)\| PubMed ID: 22294662	2012
Molecularly self-assembled nucleic acid nanoparticles for targeted in vivo siRNA delivery. Nature nanotechnology\| PubMed ID: 22659608	2012

Klaus Charisse

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