Development of a novel PTD-mediated IVT-mRNA Delivery Platform for potential Clinical Application as a Protein Therapy approach for Metabolic/Genetic disorders

Abstract The potential clinical applications of the powerful in vitro transcribed (IVT)-mRNAs, to restore defective protein functions, strongly depend on their successful intracellular delivery and transient translation through the development of safe and efficient delivery platforms. In this study, an innovative (international patent-pending) methodology was developed, combining the IVT-mRNAs with the Protein Transduction Domain (PTD) technology, as an efficient delivery platform. Based on the PTD technology, which enables the intracellular delivery of various cargoes intracellularly, successful conjugation of a PTD to the IVT-mRNAs was achieved and evaluated by band shift assay and NMR spectroscopy. In addition, the PTD-IVT-mRNAs were applied and evaluated in two protein-disease-models, including the mitochondrial disorder fatal infantile cardioencephalomyopathy and COX deficiency (attributed to SCO2 gene mutations) and β-thalassemia. The PTD-IVT-mRNA of SCO2 was successfully transduced and translated to the corresponding Sco2 protein inside the primary fibroblasts of a SCO2/COX-deficient patient, while the PTD-IVT-mRNA of β-globin was transduced and translated in the bone-marrow cells, derived from three β-thalassemic patients. The transducibility and the structural stability of the PDT-IVT-mRNAs, in both cases, was confirmed at the RNA and protein levels. We propose that our novel delivery platform could be clinically applicable as a Protein Therapy for Metabolic/Genetic Disorders.