Development of a novel PTD-mediated IVT-mRNA delivery platform for potential protein replacement therapy of metabolic/genetic disorders

Molecular Therapy: Nucleic Acids

Lefkothea C. Papadopoulou¹  Efthimia Vlachaki²  George Spyroulias³  Ioannis S. Pappas⁴  Asterios S. Tsiftsoglou⁵  Androulla N. Miliotou⁵  Ioannis S. Vizirianakis⁵ 

[1] Department of Life and Health Sciences, University of Nicosia, 1700 Nicosia, Cyprus;Adult Thalassemia Unit, Hippokrateion General Hospital, Thessaloniki, 546 42 Macedonia, Greece;Department of Pharmacy, University of Patras, 265 04 Patras, Greece;Laboratory of Pharmacology and Toxicology, Faculty of Veterinary Science, University of Thessaly, Karditsa, 431 00 Thessaly, Greece;Laboratory of Pharmacology, School of Pharmacy, Faculty of Health Sciences, Aristotle University of Thessaloniki, Thessaloniki, 546 42 Macedonia, Greece;

关键词: IVT-mRNA;    protein transduction domain technology;    PTD technology;    delivery platform;    protein therapy;    SCO2;   

DOI  :  

来源: DOAJ

【 摘 要 】

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 cytochrome c oxidase (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, whereas the PTD-IVT-mRNA of β-globin was transduced and translated in bone marrow cells, derived from three β-thalassemic patients. The transducibility and the structural stability of the PDT-IVT-mRNAs, in both cases, were 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.

【 授权许可】

Unknown