Development of a novel PTD-mediated IVT-mRNA delivery platform for potential protein replacement therapy of metabolic/genetic disorders
| Autor: | Ioannis S. Pappas, Asterios S. Tsiftsoglou, Ioannis S. Vizirianakis, George Spyroulias, Lefkothea C. Papadopoulou, Efthimia Vlachaki, Androulla N. Miliotou |
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| Rok vydání: | 2021 |
| Předmět: |
IVT-mRNA
delivery platform RM1-950 β-globin Gene mutation protein transduction domain technology Protein replacement therapy protein therapy Drug Discovery medicine Cytochrome c oxidase Messenger RNA biology SCO2 business.industry RNA Translation (biology) Cell biology medicine.anatomical_structure biology.protein PTD technology Molecular Medicine Original Article Therapeutics. Pharmacology Bone marrow business Intracellular |
| Zdroj: | Molecular Therapy. Nucleic Acids Molecular Therapy: Nucleic Acids, Vol 26, Iss, Pp 694-710 (2021) |
| ISSN: | 2162-2531 |
| Popis: | 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. Graphical abstract The corresponding author and colleagues successfully developed a universal delivery platform of therapeutic in vitro-transcribed (IVT)-mRNAs conjugated to a selected PTD through an innovative patent-pending methodology. PTD-IVT-mRNAs exhibited significant stability, intracellular transduction, and protein expression efficiency and were applied in patients’ cells of two different monogenic/metabolic disorders, revealing powerful future perspectives. |
| Databáze: | OpenAIRE |
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