In vivo biodistribution study of TAT-L-Sco2 fusion protein, developed as protein therapeutic for mitochondrial disorders attributed to SCO2 mutations
Autor: | Lefkothea C. Papadopoulou, Aikaterini C. Tsika, Georgios A. Spyroulias, Dionysia Papagiannopoulou, Georgios C Kaiafas, Parthenopi C. Chalkidou, Αndroulla N. Miliotou, Asterios S. Tsiftsoglou, Anastasia Tsingotjidou |
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Jazyk: | angličtina |
Rok vydání: | 2020 |
Předmět: |
ID
Injected dose Biodistribution SD Standard Deviation 99mTc Technetium-99 m p.i. Post-injection Mitochondrial disease TAT-L-Sco2 10xHis-XaSITE-TAT-L-Sco2-HA PBS Phosphate buffered saline Mitochondrion law.invention COX Cytochrome c oxidase Endocrinology Protein replacement therapy Western blot law Genetics medicine l-Arg l-Arginine RT Room Temperature Receptor lcsh:QH301-705.5 Molecular Biology lcsh:R5-920 FBS Fetal bovine serum medicine.diagnostic_test Chemistry medicine.disease Fusion protein Cell biology PTD Protein Transduction Domain i.p. Intraperitoneal PRT Protein Replacement Therapy lcsh:Biology (General) i.v. Intravenous Recombinant DNA IBs Inclusion bodies SEC Size Exclusion Chromatography lcsh:Medicine (General) Research Paper BSA Bovine Serum Albumin RA Radioactivity |
Zdroj: | Molecular Genetics and Metabolism Reports Molecular Genetics and Metabolism Reports, Vol 25, Iss, Pp 100683-(2020) |
ISSN: | 2214-4269 |
Popis: | The rapid progress achieved in the development of many biopharmaceuticals had a tremendous impact on the therapy of many metabolic/genetic disorders. This type of fruitful approach, called protein replacement therapy (PRT), aimed to either replace the deficient or malfunctional protein in human tissues that act either in plasma membrane or via a specific cell surface receptor. However, there are also many metabolic/genetic disorders attributed to either deficient or malfunctional proteins acting intracellularly. The recent developments of Protein Transduction Domain (PTD) technology offer new opportunities by allowing the intracellular delivery of recombinant proteins of a given therapeutic interest into different subcellular sites and organelles, such as mitochondria and other entities. Towards this pathway, we applied successfully PTD Technology as a protein therapeutic approach, in vitro, in SCO2 deficient primary fibroblasts, derived from patient with mutations in human SCO2 gene, responsible for fatal, infantile cardioencephalomyopathy and cytochrome c oxidase deficiency. In this work, we radiolabeled the recombinant TAT-L-Sco2 fusion protein with technetium-99 m to assess its in vivo biodistribution and fate, by increasing the sensitivity of detection of even low levels of the transduced recombinant protein. The biodistribution pattern of [99mTc]Tc-TAT-L-Sco2 in mice demonstrated fast blood clearance, significant hepatobiliary and renal clearance. In addition, western blot analysis detected the recombinant TAT-L-Sco2 protein in the isolated mitochondria of several mouse tissues, including heart, muscle and brain. These results pave the way to further consider this PTD-mediated Protein Therapy Approach as a potentially alternative treatment of genetic/metabolic disorders. Graphical abstract Unlabelled Image Highlights • Radiolabeling of human recombinant mitochondrial TAT-L-Sco2 fusion protein with 99mTc for the first time. • [99mTc]Tc-TAT-L-Sco2 can be successfully transduced into the mitochondria of peripheral tissues upon injection into animals. • Protein Replacement Therapy, through PTD technology, can be a potential therapeutic approach for mitochondrial disorders. |
Databáze: | OpenAIRE |
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