title: Lipid nanoparticle delivery of a microRNA-145 inhibitor improves experimental pulmonary hypertension.
abstract: Therapies that exploit RNA interference (RNAi) hold great potential for improving disease outcomes. However, there are several challenges that limit the application of RNAi therapeutics. One of the most important challenges is effective delivery of oligonucleotides to target cells and reduced delivery to non-target cells. We have previously developed a functionalized cationic lipopolyamine (Star:Star-mPEG-550) for in vivo delivery of siRNA to pulmonary vascular cells. This optimized lipid formulation enhances the retention of siRNA in mouse lungs and achieves significant knockdown of target gene expression for at least 10days following a single intravenous injection. Although this suggests great potential for developing lung-directed RNAi-based therapies, the application of Star:Star-mPEG mediated delivery of RNAi based therapies for pulmonary vascular diseases such as pulmonary arterial hypertension (PAH) remains unknown. We identified differential expression of several microRNAs known to regulate cell proliferation, cell survival and cell fate that are associated with development of PAH, including increased expression of microRNA-145 (miR-145). Here we test the hypothesis that Star:Star-mPEG mediated delivery of an antisense oligonucleotide against miR-145 (antimiR-145) will improve established PAH in rats. We performed a series of experiments testing the in vivo distribution, toxicity, and efficacy of Star:Star-mPEG mediated delivery of antimiR-145 in rats with Sugen-5416/hypoxia induced PAH. We showed that after subchronic therapy of three intravenous injections over 5weeks at 2mg/kg, antimiR-145 accumulated in rat lung tissue and reduced expression of endogenous miR-145. Using a novel in situ hybridization approach, we demonstrated substantial distribution of antimiR-145 in the lungs as well as the liver, kidney, and spleen. We assessed toxic effects of Star:Star-mPEG/antimiR-145 with serial complete blood counts of leukocytes and serum metabolic panels, gross pathology, and histopathology and did not detect significant off-target effects. AntimiR-145 reduced the degree of pulmonary arteriopathy, reduced the severity of pulmonary hypertension, and reduced the degree of cardiac dysfunction. The results establish effective and low toxicity of lung delivery of a miRNA-145 inhibitor using functionalized cationic lipopolyamine nanoparticles to repair pulmonary arteriopathy and improve cardiac function in rats with severe PAH.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of South Alabama College of Medicine, Mobile, AL 36688, USA; Center for Lung Biology, University of South Alabama College of Medicine, Mobile, AL 36688, USA. Electronic address: jmclendon@jagmail.southalabama.edu.
authors.forename: Jared M
authors.initials: JM
authors.lastname: McLendon
authors.affiliation: Department of Biochemistry and Molecular Biology, University of South Alabama College of Medicine, Mobile, AL 36688, USA; Center for Lung Biology, University of South Alabama College of Medicine, Mobile, AL 36688, USA.
authors.forename: Sachindra R
authors.initials: SR
authors.lastname: Joshi
authors.affiliation: Celsion-EGEN, 601 Genome Way, Huntsville, AL 35806, USA.
authors.forename: Jeff
authors.initials: J
authors.lastname: Sparks
authors.affiliation: Celsion-EGEN, 601 Genome Way, Huntsville, AL 35806, USA.
authors.forename: Majed
authors.initials: M
authors.lastname: Matar
authors.affiliation: Celsion-EGEN, 601 Genome Way, Huntsville, AL 35806, USA.
authors.forename: Jason G
authors.initials: JG
authors.lastname: Fewell
authors.affiliation: Center for Lung Biology, University of South Alabama College of Medicine, Mobile, AL 36688, USA.
authors.forename: Kohtaro
authors.initials: K
authors.lastname: Abe
authors.affiliation: Department of Internal Medicine, University of South Alabama College of Medicine, Mobile, AL 36688, USA; Center for Lung Biology, University of South Alabama College of Medicine, Mobile, AL 36688, USA.
authors.forename: Masahiko
authors.initials: M
authors.lastname: Oka
authors.affiliation: Department of Pharmacology, University of South Alabama College of Medicine, Mobile, AL 36688, USA; Department of Internal Medicine, University of South Alabama College of Medicine, Mobile, AL 36688, USA; Center for Lung Biology, University of South Alabama College of Medicine, Mobile, AL 36688, USA.
authors.forename: Ivan F
authors.initials: IF
authors.lastname: McMurtry
authors.affiliation: Department of Biochemistry and Molecular Biology, University of South Alabama College of Medicine, Mobile, AL 36688, USA; Center for Lung Biology, University of South Alabama College of Medicine, Mobile, AL 36688, USA.
authors.forename: William T
authors.initials: WT
authors.lastname: Gerthoffer
mesh_terms: D000818:Animals; D006976:Hypertension, Pulmonary / Q000188:drug therapy* / Q000378:metabolism; D000860:Hypoxia / Q000150:complications; D007211:Indoles; D008055:Lipids / Q000737:chemistry; D008081:Liposomes; D008168:Lung / Q000378:metabolism; D008297:Male; D035683:MicroRNAs / Q000037:antagonists & inhibitors* / Q000378:metabolism; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry; D009841:Oligonucleotides / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics; D011758:Pyrroles; D017207:Rats, Sprague-Dawley
chemical_list: D007211:Indoles; D008055:Lipids; D008081:Liposomes; D035683:MicroRNAs; D009841:Oligonucleotides; D011758:Pyrroles; C477371:locked nucleic acid; C116890:Semaxinib
keywords: Antisense oligonucleotide; Lipid nanoparticle; Lung delivery; MicroRNA-145; Pulmonary hypertension; Sugen5416/hypoxia