Gene delivery into ischemic myocardium by double-targeted lipoplexes with anti-myosin antibody and TAT peptide

Abstract

The treatment of myocardial ischemia using gene therapy is a rather novel but promising approach. Gene delivery to target cells may be enhanced by using double-targeted delivery systems simultaneously capable of extracellular accumulation and intracellular penetration. With this in mind, we have used low cationic liposomes-plasmid DNA complexes (lipoplexes) modified with cell-penetrating transactivating transcriptional activator (TAT) peptide (TATp) and/or with monoclonal anti-myosin monoclonal antibody 2G4 (mAb 2G4) specific toward cardiac myosin, for targeted gene delivery to ischemic myocardium. In vitro transfection of both normoxic and hypoxic cardiomyocytes was enhanced by the presence of TATp as determined by fluorescence microscopy and ELISA. The in vitro transfection was further enhanced by the additional modification with mAb 2G4 antibody in the case of hypoxic, but not normoxic cardiomyocytes. However, we did not observe a synergism between TATp and mAb 2G4 ligands under our experimental condition. In in vivo experiments, we have clearly demonstrated an increased accumulation of mAb 2G4-modified TATp lipoplexes in the ischemic rat myocardium and significantly enhanced transfection of cardiomyocytes in the ischemic zone. Thus, the genetic transformation of normoxic and hypoxic cardiomyocytes can be enhanced by using lipoplexes modified with TATp and/or mAb 2G4. Such complexes also demonstrate an increased accumulation in the ischemic myocardium and effective transfection of

Myocardial Gene Transfer: Routes and Devices for Regulation of Transgene Expression by Modulation of Cellular Permeability

“…Animals that received treatment with CPP demonstrated increased left ventricular (LV) fractional shortening and reduced fibrosis of the infarcted myocardium (Bian et al, 2007). The enhanced transfection of the gene delivery using Tat-lipoplexes was demonstrated in a rat infarct model in vivo and in hypoxic cardiomyocytes in vitro (Ko et al, 2009).…”

Engineered Exosomes With Ischemic Myocardium‐Targeting Peptide for Targeted Therapy in Myocardial Infarction

“…Compared with conventional carriers, such as liposomes, nanospheres, micelles, microemulsion, and different kinds of conjugates, exosomes afford all the desirable advantages, such as low toxicity, low immunogenicity, high stability in circulation, biocompatibility, and biological barrier permeability,20 which makes them promising carriers for efficient drug or therapeutic gene delivery. Nevertheless, both the conventional carriers and exosomes are apt to be trapped in nonspecific organs, especially in the lung and the liver, leading to insufficiency in targeting myocardial ischemia area 21. Therefore, attempts to modify exosomes as effective carriers directly targeting ischemic myocardium have been considered.…”

Anti remodeling therapy: new strategies and future perspective in post-ischemic heart failure: Part I

“…Insieme a questa criticità, va sottolineato anche il basso tasso di internalizzazione nelle cellule bersaglio dovuto alla difficoltà per il plasmide di superare il sistema immunitario e le nucleasi extracellulari, la membrana cellulare, gli enzimi lisosomiali e le nucleasi citoplasmatiche e infine le membrane nucleari [34]. Una tecnica attuale riesce a proteggere in qualche modo il plasmide inserendolo in liposomi cationici che legano gli acidi nucleici carichi negativamente [35], ma tale metodica necessita di una perfusione tissutale ottimale o, in alternativa, della somministrazione per via iniettiva nell'area infartuata, procedura gravata da un alto rischio di perforazione.…”