Target-Cell-Directed Bioengineering Approaches for Gene Therapy of Hemophilia A
| العنوان: | Target-Cell-Directed Bioengineering Approaches for Gene Therapy of Hemophilia A |
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| المؤلفون: | Philip M. Zakas, Harrison C. Brown, H. Trent Spencer, Stephan N. George, Ernest T. Parker, Christopher B. Doering |
| المصدر: | Molecular Therapy. Methods & Clinical Development Molecular Therapy: Methods & Clinical Development, Vol 9, Iss, Pp 57-69 (2018) |
| سنة النشر: | 2017 |
| مصطلحات موضوعية: | 0301 basic medicine, congenital, hereditary, and neonatal diseases and abnormalities, vector optimization, lcsh:QH426-470, Genetic enhancement, Transgene, Cell, Computational biology, Biology, Genome, Article, promoter design, codon optimization, 03 medical and health sciences, Transcription (biology), hemic and lymphatic diseases, hemophilia, Genetics, medicine, lcsh:QH573-671, Molecular Biology, Gene, Messenger RNA, lcsh:Cytology, AAV, lcsh:Genetics, Vector optimization, 030104 developmental biology, medicine.anatomical_structure, factor VIII, Molecular Medicine |
| الوصف: | Potency is a key optimization parameter for hemophilia A gene therapy product candidates. Optimization strategies include promoter engineering to increase transcription, codon optimization of mRNA to improve translation, and amino-acid substitution to promote secretion. Herein, we describe both rational and empirical design approaches to the development of a minimally sized, highly potent AAV-fVIII vector that incorporates three unique elements: a liver-directed 146-nt transcription regulatory module, a target-cell-specific codon optimization algorithm, and a high-expression bioengineered fVIII variant. The minimal synthetic promoter allows for the smallest AAV-fVIII vector genome known at 4,832 nt, while the tissue-directed codon optimization strategy facilitates increased fVIII transgene product expression in target cell types, e.g., hepatocytes, over traditional genome-level codon optimization strategies. As a tertiary approach, we incorporated ancient and orthologous fVIII sequence elements previously shown to facilitate improved biosynthesis through post-translational mechanisms. Together, these technologies contribute to an AAV-fVIII vector that confers sustained, curative levels of fVIII at a minimal dose in hemophilia A mice. Moreover, the first two technologies should be generalizable to all liver-directed gene therapy vector designs. Keywords: vector optimization, AAV, hemophilia, factor VIII, codon optimization, promoter design |
| تدمد: | 2329-0501 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::8057961f9965bff41367530909c46bfb https://pubmed.ncbi.nlm.nih.gov/29552578 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....8057961f9965bff41367530909c46bfb |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 23290501 |
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