Mutation-adapted U1 snRNA corrects a splicing error of the dopa decarboxylase gene

Ni-Chung Lee; Yu-May Lee; Pin-Wen Chen; Barry J Byrne; Wuh-Liang Hwu

doi:10.1093/hmg/ddw323

Mutation-adapted U1 snRNA corrects a splicing error of the dopa decarboxylase gene

Hum Mol Genet. 2016 Dec 1;25(23):5142-5147. doi: 10.1093/hmg/ddw323.

Authors

Ni-Chung Lee^{1

2}, Yu-May Lee¹, Pin-Wen Chen¹, Barry J Byrne³, Wuh-Liang Hwu^{1

2}

Affiliations

¹ Department of Medical Genetics, National Taiwan University Hospital, Taipei, Taiwan.
² Department of Pediatrics, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan.
³ Powell Gene Therapy Center, University of Florida, Gainesville, FL, USA.

PMID: 27658936
DOI: 10.1093/hmg/ddw323

Abstract

Aromatic l-amino acid decarboxylase (AADC) deficiency is an inborn error of monoamine neurotransmitter synthesis, which results in dopamine, serotonin, epinephrine and norepinephrine deficiencies. The DDC gene founder mutation IVS6 + 4A > T is highly prevalent in Chinese patients with AADC deficiency. In this study, we designed several U1 snRNA vectors to adapt U1 snRNA binding sequences of the mutated DDC gene. We found that only the modified U1 snRNA (IVS-AAA) that completely matched both the intronic and exonic U1 binding sequences of the mutated DDC gene could correct splicing errors of either the mutated human DDC minigene or the mouse artificial splicing construct in vitro. We further injected an adeno-associated viral (AAV) vector to express IVS-AAA in the brain of a knock-in mouse model. This treatment was well tolerated and improved both the survival and brain dopamine and serotonin levels of mice with AADC deficiency. Therefore, mutation-adapted U1 snRNA gene therapy can be a promising method to treat genetic diseases caused by splicing errors, but the efficiency of such a treatment still needs improvements.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Metabolism, Inborn Errors / genetics*
Amino Acid Metabolism, Inborn Errors / therapy
Animals
Aromatic-L-Amino-Acid Decarboxylases / deficiency*
Aromatic-L-Amino-Acid Decarboxylases / genetics
Dependovirus / genetics
Disease Models, Animal
Dopa Decarboxylase / genetics*
Exons / genetics
Gene Knock-In Techniques
Genetic Therapy*
Humans
Introns / genetics
Mice
Mutation
Neurotransmitter Agents / genetics
RNA Splicing / genetics
RNA, Small Nuclear / administration & dosage
RNA, Small Nuclear / genetics*

Substances

Neurotransmitter Agents
RNA, Small Nuclear
U1 small nuclear RNA
Dopa Decarboxylase
Aromatic-L-Amino-Acid Decarboxylases

Supplementary concepts

Aromatic amino acid decarboxylase deficiency