Cargando…
Pathological mechanism and antisense oligonucleotide-mediated rescue of a non-coding variant suppressing factor 9 RNA biogenesis leading to hemophilia B
Loss-of-function mutations in the human coagulation factor 9 (F9) gene lead to hemophilia B. Here, we dissected the consequences and the pathomechanism of a non-coding mutation (c.2545A>G) in the F9 3’ untranslated region. Using wild type and mutant factor IX (FIX) minigenes we revealed that the...
Autores principales: | , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7141619/ https://www.ncbi.nlm.nih.gov/pubmed/32267853 http://dx.doi.org/10.1371/journal.pgen.1008690 |
_version_ | 1783519227923136512 |
---|---|
author | Krooss, Simon Werwitzke, Sonja Kopp, Johannes Rovai, Alice Varnholt, Dirk Wachs, Amelie S. Goyenvalle, Aurelie Aarstma-Rus, Annemieke Ott, Michael Tiede, Andreas Langemeier, Jörg Bohne, Jens |
author_facet | Krooss, Simon Werwitzke, Sonja Kopp, Johannes Rovai, Alice Varnholt, Dirk Wachs, Amelie S. Goyenvalle, Aurelie Aarstma-Rus, Annemieke Ott, Michael Tiede, Andreas Langemeier, Jörg Bohne, Jens |
author_sort | Krooss, Simon |
collection | PubMed |
description | Loss-of-function mutations in the human coagulation factor 9 (F9) gene lead to hemophilia B. Here, we dissected the consequences and the pathomechanism of a non-coding mutation (c.2545A>G) in the F9 3’ untranslated region. Using wild type and mutant factor IX (FIX) minigenes we revealed that the mutation leads to reduced F9 mRNA and FIX protein levels and to lower coagulation activity of cell culture supernatants. The phenotype could not be compensated by increased transcription. The pathomechanism comprises the de novo creation of a binding site for the spliceosomal component U1snRNP, which is able to suppress the nearby F9 poly(A) site. This second, splicing-independent function of U1snRNP was discovered previously and blockade of U1snRNP restored mutant F9 mRNA expression. In addition, we explored the vice versa approach and masked the mutation by antisense oligonucleotides resulting in significantly increased F9 mRNA expression and coagulation activity. This treatment may transform the moderate/severe hemophilia B into a mild or subclinical form in the patients. This antisense based strategy is applicable to other mutations in untranslated regions creating deleterious binding sites for cellular proteins. |
format | Online Article Text |
id | pubmed-7141619 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-71416192020-04-09 Pathological mechanism and antisense oligonucleotide-mediated rescue of a non-coding variant suppressing factor 9 RNA biogenesis leading to hemophilia B Krooss, Simon Werwitzke, Sonja Kopp, Johannes Rovai, Alice Varnholt, Dirk Wachs, Amelie S. Goyenvalle, Aurelie Aarstma-Rus, Annemieke Ott, Michael Tiede, Andreas Langemeier, Jörg Bohne, Jens PLoS Genet Research Article Loss-of-function mutations in the human coagulation factor 9 (F9) gene lead to hemophilia B. Here, we dissected the consequences and the pathomechanism of a non-coding mutation (c.2545A>G) in the F9 3’ untranslated region. Using wild type and mutant factor IX (FIX) minigenes we revealed that the mutation leads to reduced F9 mRNA and FIX protein levels and to lower coagulation activity of cell culture supernatants. The phenotype could not be compensated by increased transcription. The pathomechanism comprises the de novo creation of a binding site for the spliceosomal component U1snRNP, which is able to suppress the nearby F9 poly(A) site. This second, splicing-independent function of U1snRNP was discovered previously and blockade of U1snRNP restored mutant F9 mRNA expression. In addition, we explored the vice versa approach and masked the mutation by antisense oligonucleotides resulting in significantly increased F9 mRNA expression and coagulation activity. This treatment may transform the moderate/severe hemophilia B into a mild or subclinical form in the patients. This antisense based strategy is applicable to other mutations in untranslated regions creating deleterious binding sites for cellular proteins. Public Library of Science 2020-04-08 /pmc/articles/PMC7141619/ /pubmed/32267853 http://dx.doi.org/10.1371/journal.pgen.1008690 Text en © 2020 Krooss et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Krooss, Simon Werwitzke, Sonja Kopp, Johannes Rovai, Alice Varnholt, Dirk Wachs, Amelie S. Goyenvalle, Aurelie Aarstma-Rus, Annemieke Ott, Michael Tiede, Andreas Langemeier, Jörg Bohne, Jens Pathological mechanism and antisense oligonucleotide-mediated rescue of a non-coding variant suppressing factor 9 RNA biogenesis leading to hemophilia B |
title | Pathological mechanism and antisense oligonucleotide-mediated rescue of a non-coding variant suppressing factor 9 RNA biogenesis leading to hemophilia B |
title_full | Pathological mechanism and antisense oligonucleotide-mediated rescue of a non-coding variant suppressing factor 9 RNA biogenesis leading to hemophilia B |
title_fullStr | Pathological mechanism and antisense oligonucleotide-mediated rescue of a non-coding variant suppressing factor 9 RNA biogenesis leading to hemophilia B |
title_full_unstemmed | Pathological mechanism and antisense oligonucleotide-mediated rescue of a non-coding variant suppressing factor 9 RNA biogenesis leading to hemophilia B |
title_short | Pathological mechanism and antisense oligonucleotide-mediated rescue of a non-coding variant suppressing factor 9 RNA biogenesis leading to hemophilia B |
title_sort | pathological mechanism and antisense oligonucleotide-mediated rescue of a non-coding variant suppressing factor 9 rna biogenesis leading to hemophilia b |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7141619/ https://www.ncbi.nlm.nih.gov/pubmed/32267853 http://dx.doi.org/10.1371/journal.pgen.1008690 |
work_keys_str_mv | AT kroosssimon pathologicalmechanismandantisenseoligonucleotidemediatedrescueofanoncodingvariantsuppressingfactor9rnabiogenesisleadingtohemophiliab AT werwitzkesonja pathologicalmechanismandantisenseoligonucleotidemediatedrescueofanoncodingvariantsuppressingfactor9rnabiogenesisleadingtohemophiliab AT koppjohannes pathologicalmechanismandantisenseoligonucleotidemediatedrescueofanoncodingvariantsuppressingfactor9rnabiogenesisleadingtohemophiliab AT rovaialice pathologicalmechanismandantisenseoligonucleotidemediatedrescueofanoncodingvariantsuppressingfactor9rnabiogenesisleadingtohemophiliab AT varnholtdirk pathologicalmechanismandantisenseoligonucleotidemediatedrescueofanoncodingvariantsuppressingfactor9rnabiogenesisleadingtohemophiliab AT wachsamelies pathologicalmechanismandantisenseoligonucleotidemediatedrescueofanoncodingvariantsuppressingfactor9rnabiogenesisleadingtohemophiliab AT goyenvalleaurelie pathologicalmechanismandantisenseoligonucleotidemediatedrescueofanoncodingvariantsuppressingfactor9rnabiogenesisleadingtohemophiliab AT aarstmarusannemieke pathologicalmechanismandantisenseoligonucleotidemediatedrescueofanoncodingvariantsuppressingfactor9rnabiogenesisleadingtohemophiliab AT ottmichael pathologicalmechanismandantisenseoligonucleotidemediatedrescueofanoncodingvariantsuppressingfactor9rnabiogenesisleadingtohemophiliab AT tiedeandreas pathologicalmechanismandantisenseoligonucleotidemediatedrescueofanoncodingvariantsuppressingfactor9rnabiogenesisleadingtohemophiliab AT langemeierjorg pathologicalmechanismandantisenseoligonucleotidemediatedrescueofanoncodingvariantsuppressingfactor9rnabiogenesisleadingtohemophiliab AT bohnejens pathologicalmechanismandantisenseoligonucleotidemediatedrescueofanoncodingvariantsuppressingfactor9rnabiogenesisleadingtohemophiliab |