The efficacy of simultaneous action of several antisense oligonucleotide derivatives by the example of lipid metabolism in mice

   Motivation and Aim: Demonstration of the possibility of lipid metabolism regulation by joint action of several antisense oligonucleotide derivatives (ASOs) targeted to mRNAs of closely associated proteins involved in lipid metabolism.

   Material and Methods: Female mice of the C57BL/6J strain; 13- and 20-mer oligonucleotide derivatives (ASOs) protected from nucleases by the presence of inter-nucleotide thiophosphate bonds and locked nucleic acid (LNA) fragments at their 5’- and 3’- ends; conventional methods of assaying blood serum ALT, total cholesterol, TG, apolipoprotein apoB and lipoproteins: LDL-C, HDL-C.

   Results: Solutions of ASOs in isotonic solution were prepared in combinations: (1) anti-apoB; (2) anti-apoB + anti-PCSK9; (3) anti-apoB + anti-PCSK9 + anti-apoCIII. Four injections to the tail veins of C57BL/6J mice were done. After three injections of combination (3), done at weekly intervals, the LDL-C level decreased by 25 % with reference to the control, and the HDL-C/LDL-C ratio improved by 42 %. After subsequent injection of the ASOs accompanied by change to high-fat diet, mice exposed to combinations (2) and (3) showed lower LDL-C levels, by factors of 1.5 and 3.5 with reference to the control, and their HDL-C/LDL-C ratios were better two- and fivefold, respectively. In cases of combinations (2) and (3), a significant decrease in TG
concentration was recorded, twofold in case of (3). These effects were observed at threefold fat content in the diet. The ALT level decreased significantly with transition from combination (1) to (3), i. e., when the liver experienced heavier load of oligonucleotide matter, which indicates to the absence of hepatotoxicity.

   Conclusions: The possibility of efficient regulatory joint action of several ASOs targeted to mRNAs of associated proteins is demonstrated by the example of lipid metabolism.

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