Inhibition of miR-92a normalizes vascular gene expression and prevents diastolic dysfunction in heart failure with preserved ejection fraction
Inhibition of miR-92a normalizes vascular gene expression and prevents diastolic dysfunction in heart failure with preserved ejection fraction
To analyze the cardiac transcriptome on single nuclear level, myocardium was collected from explanted hearts of healthy controls (n = 3) or mice with HFpEF either untreated (n = 3) or treated with LNA-92a (n = 4) or LNA-control (n = 4) after 10 weeks of HFD/L-NAME exposure or chow diet. All procedures involving the use and care of animals were performed according to the German animal protection code. Approval was granted by the local state authorities (state of Hessen). To induce HFpEF, male C57Bl/6J wild-type mice from Janvier (Le Genest Saint Isle, 239 France) and from Charles River (Sulzfeld, Germany) were subjected to a combination of high-fat diet (HFD) (60% kilocalories from fat (lard)) and N[w]-nitro-l-arginine methyl ester (L-NAME; 0.5 g/l in drinking water, Sigma-Aldrich) for 10 weeks Following two weeks of HFpEF induction, locked nucleic acid (LNA)-modified antisense oligonucleotides-92a (1.5 mg/kg BW weekly) or LNA-control were injected intraperitoneal, which are directed to inhibit miR-92a-3p (miRagen Therapeutics) Tissue was lysed in Qiazol and RNA isolated using Qiagen miRNeasy Mini Kits following the manufactures protocol. Expression analysis of miR-92a-3p was performed using Taqman Assays primers and relative miR-92a-3p expression was calculated using 2−ΔCT Library quantification and quality assessment was performed using Bioanalyzer Agilent 2100 using a High Sensitivity DNA chip (5067-4627 Agilent Genomics).