A Compensatory U1snRNA Partially Rescues FAH Splicing and Protein Expression in a Splicing-Defective Mouse Model of Tyrosinemia Type I
The elucidation of aberrant splicing mechanisms, frequently associated with disease has led to the development of RNA therapeutics based on the U1snRNA, which is involved in 5′ splice site (5′ss) recognition. Studies in cellular models have demonstrated that engineered U1snRNAs c...
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MDPI AG
2020-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/6/2136 |
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doaj-7d2b03228a504040b8b832911c05fb852020-11-25T03:10:06ZengMDPI AGInternational Journal of Molecular Sciences1422-00672020-03-01216213610.3390/ijms21062136ijms21062136A Compensatory U1snRNA Partially Rescues FAH Splicing and Protein Expression in a Splicing-Defective Mouse Model of Tyrosinemia Type IDario Balestra0Daniela Scalet1Mattia Ferrarese2Silvia Lombardi3Nicole Ziliotto4Chrystal C. Croes5Naomi Petersen6Piter Bosma7Federico Riccardi8Franco Pagani9Mirko Pinotti10Stan F. J. van de Graaf11Department of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, ItalyDepartment of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, ItalyDepartment of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, ItalyDepartment of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, ItalyDepartment of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, ItalyTytgat Institute for Liver and Intestinal Research, Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The NetherlandsTytgat Institute for Liver and Intestinal Research, Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The NetherlandsTytgat Institute for Liver and Intestinal Research, Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The NetherlandsHuman Molecular Genetics, International Centre for Genetic Engineering and Biotechnology, 34149 Trieste, ItalyHuman Molecular Genetics, International Centre for Genetic Engineering and Biotechnology, 34149 Trieste, ItalyDepartment of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, ItalyTytgat Institute for Liver and Intestinal Research, Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The NetherlandsThe elucidation of aberrant splicing mechanisms, frequently associated with disease has led to the development of RNA therapeutics based on the U1snRNA, which is involved in 5′ splice site (5′ss) recognition. Studies in cellular models have demonstrated that engineered U1snRNAs can rescue different splicing mutation types. However, the assessment of their correction potential in vivo is limited by the scarcity of animal models with the targetable splicing defects. Here, we challenged the U1snRNA in the <i>FAH5961SB</i> mouse model of hepatic fumarylacetoacetate hydrolase (FAH) deficiency (Hereditary Tyrosinemia type I, HT1) due to the <i>FAH</i> c.706G>A splicing mutation. Through minigene expression studies we selected a compensatory U1snRNA (U1<sup>F</sup>) that was able to rescue this mutation. Intriguingly, adeno-associated virus-mediated delivery of U1<sup>F</sup> (AAV8-U1<sup>F</sup>), but not of U1<sup>wt</sup>, partially rescued FAH splicing in mouse hepatocytes. Consistently, FAH protein was detectable only in the liver of AAV8-U1<sup>F</sup> treated mice, which displayed a slightly prolonged survival. Moreover, RNA sequencing revealed the negligible impact of the U1<sup>F</sup> on the splicing profile and overall gene expression, thus pointing toward gene specificity. These data provide early in vivo proof-of-principle of the correction potential of compensatory U1snRNAs in HTI and encourage further optimization on a therapeutic perspective, and translation to other splicing-defective forms of metabolic diseases.https://www.mdpi.com/1422-0067/21/6/2136fahfumarylacetoacetate hydrolase deficiencytyrosinemia type iaberrant splicingmouse modelsu1snrnarna therapeutics |
| collection |
DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Dario Balestra Daniela Scalet Mattia Ferrarese Silvia Lombardi Nicole Ziliotto Chrystal C. Croes Naomi Petersen Piter Bosma Federico Riccardi Franco Pagani Mirko Pinotti Stan F. J. van de Graaf |
| spellingShingle |
Dario Balestra Daniela Scalet Mattia Ferrarese Silvia Lombardi Nicole Ziliotto Chrystal C. Croes Naomi Petersen Piter Bosma Federico Riccardi Franco Pagani Mirko Pinotti Stan F. J. van de Graaf A Compensatory U1snRNA Partially Rescues FAH Splicing and Protein Expression in a Splicing-Defective Mouse Model of Tyrosinemia Type I International Journal of Molecular Sciences fah fumarylacetoacetate hydrolase deficiency tyrosinemia type i aberrant splicing mouse models u1snrna rna therapeutics |
| author_facet |
Dario Balestra Daniela Scalet Mattia Ferrarese Silvia Lombardi Nicole Ziliotto Chrystal C. Croes Naomi Petersen Piter Bosma Federico Riccardi Franco Pagani Mirko Pinotti Stan F. J. van de Graaf |
| author_sort |
Dario Balestra |
| title |
A Compensatory U1snRNA Partially Rescues FAH Splicing and Protein Expression in a Splicing-Defective Mouse Model of Tyrosinemia Type I |
| title_short |
A Compensatory U1snRNA Partially Rescues FAH Splicing and Protein Expression in a Splicing-Defective Mouse Model of Tyrosinemia Type I |
| title_full |
A Compensatory U1snRNA Partially Rescues FAH Splicing and Protein Expression in a Splicing-Defective Mouse Model of Tyrosinemia Type I |
| title_fullStr |
A Compensatory U1snRNA Partially Rescues FAH Splicing and Protein Expression in a Splicing-Defective Mouse Model of Tyrosinemia Type I |
| title_full_unstemmed |
A Compensatory U1snRNA Partially Rescues FAH Splicing and Protein Expression in a Splicing-Defective Mouse Model of Tyrosinemia Type I |
| title_sort |
compensatory u1snrna partially rescues fah splicing and protein expression in a splicing-defective mouse model of tyrosinemia type i |
| publisher |
MDPI AG |
| series |
International Journal of Molecular Sciences |
| issn |
1422-0067 |
| publishDate |
2020-03-01 |
| description |
The elucidation of aberrant splicing mechanisms, frequently associated with disease has led to the development of RNA therapeutics based on the U1snRNA, which is involved in 5′ splice site (5′ss) recognition. Studies in cellular models have demonstrated that engineered U1snRNAs can rescue different splicing mutation types. However, the assessment of their correction potential in vivo is limited by the scarcity of animal models with the targetable splicing defects. Here, we challenged the U1snRNA in the <i>FAH5961SB</i> mouse model of hepatic fumarylacetoacetate hydrolase (FAH) deficiency (Hereditary Tyrosinemia type I, HT1) due to the <i>FAH</i> c.706G>A splicing mutation. Through minigene expression studies we selected a compensatory U1snRNA (U1<sup>F</sup>) that was able to rescue this mutation. Intriguingly, adeno-associated virus-mediated delivery of U1<sup>F</sup> (AAV8-U1<sup>F</sup>), but not of U1<sup>wt</sup>, partially rescued FAH splicing in mouse hepatocytes. Consistently, FAH protein was detectable only in the liver of AAV8-U1<sup>F</sup> treated mice, which displayed a slightly prolonged survival. Moreover, RNA sequencing revealed the negligible impact of the U1<sup>F</sup> on the splicing profile and overall gene expression, thus pointing toward gene specificity. These data provide early in vivo proof-of-principle of the correction potential of compensatory U1snRNAs in HTI and encourage further optimization on a therapeutic perspective, and translation to other splicing-defective forms of metabolic diseases. |
| topic |
fah fumarylacetoacetate hydrolase deficiency tyrosinemia type i aberrant splicing mouse models u1snrna rna therapeutics |
| url |
https://www.mdpi.com/1422-0067/21/6/2136 |
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