Development of an AAV-Based MicroRNA Gene Therapy to Treat Machado-Joseph Disease
| Autor: | Jeannette Hübener-Schmid, Jiri Klima, Sonay Keskin, Huu Phuc Nguyen, Pavlina Konstantinova, Janice Stricker-Shaver, Eva Haas, Marina Sogorb-Gonzalez, Stefan Juhas, Lodewijk J.A. Toonen, Melvin M. Evers, Jana Juhasova, Sander J. H. van Deventer, Zdenka Ellederova, Jan Motlik, Raygene Martier |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Small RNA congenital hereditary and neonatal diseases and abnormalities lcsh:QH426-470 MicroRNA Gene Biology Article 03 medical and health sciences 0302 clinical medicine microRNA Genetics medicine Gene silencing lcsh:QH573-671 Induced pluripotent stem cell Molecular Biology Gene lcsh:Cytology medicine.disease 3. Good health lcsh:Genetics 030104 developmental biology 030220 oncology & carcinogenesis Cancer research Spinocerebellar ataxia Molecular Medicine Machado–Joseph disease |
| Zdroj: | Molecular Therapy: Methods & Clinical Development, Vol 15, Iss, Pp 343-358 (2019) Molecular Therapy. Methods & Clinical Development |
| ISSN: | 2329-0501 |
| Popis: | Spinocerebellar ataxia type 3 (SCA3) or Machado-Joseph disease (MJD) is a progressiveneurodegenerative disorder caused by a CAG expansion in the ATXN3 gene. The expanded CAGrepeat is translated into a prolonged polyglutamine repeat in the ataxin-3 protein and accumulateswithin inclusions, acquiring toxic properties, which results in degeneration of the cerebellum and brainstem.In the current study, a non-allele specific ATXN3 silencing approach was investigated using artificialmicroRNAs engineered to target various regions of the ATXN3 gene (miATXN3). The miATXN3candidates were screened in vitro based on their silencing efficacy on a luciferase reporter co-expressing ATXN3. The three best miATXN3 candidates were further tested for target engagement andpotential off-target activity in induced-pluripotent stem cells (iPSC) differentiated into frontal brain-like neurons and in a SCA3 knock-in mouse model. Besides a strong reduction of ATXN3 mRNA andprotein, small RNA sequencing revealed efficient guide strand processing without passenger strandsbeing produced. We used different methods to predict alteration of off-target genes upon AAV5-miATXN3 treatment and found no evidence for unwanted effects. Furthermore, we demonstrated in alarge animal model, the minipig, that intrathecal delivery of AAV5 can transduce the main areasaffected in SCA3 patients. These results proved a strong basis to move forward to investigatedistribution, efficacy and safety of AAV5-miATXN3 in large animals. |
| Databáze: | OpenAIRE |
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