Splice isoform-specific suppression of the CaV2.1 variant underlying spinocerebellar ataxia type 6
Spinocerebellar ataxia type 6 (SCA6) is an inherited neurodegenerative disease caused by a polyglutamine (polyQ) expansion in the CaV2.1 voltage-gated calcium channel subunit (CACNA1A). There is currently no treatment for this debilitating disorder and thus a pressing need to develop preventative th...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2011-09-01
|
| Series: | Neurobiology of Disease |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0969996111001331 |
| _version_ | 1818724919074095104 |
|---|---|
| author | Wei-Ling Tsou Bing-Wen Soong Henry L. Paulson Edgardo Rodríguez-Lebrón |
| author_facet | Wei-Ling Tsou Bing-Wen Soong Henry L. Paulson Edgardo Rodríguez-Lebrón |
| author_sort | Wei-Ling Tsou |
| collection | DOAJ |
| description | Spinocerebellar ataxia type 6 (SCA6) is an inherited neurodegenerative disease caused by a polyglutamine (polyQ) expansion in the CaV2.1 voltage-gated calcium channel subunit (CACNA1A). There is currently no treatment for this debilitating disorder and thus a pressing need to develop preventative therapies. RNA interference (RNAi) has proven effective at halting disease progression in several models of spinocerebellar ataxia (SCA), including SCA types 1 and 3. However, in SCA6 and other dominantly inherited neurodegenerative disorders, RNAi-based strategies that selectively suppress expression of mutant alleles may be required. Using a CaV2.1 mini-gene reporter system, we found that pathogenic CAG expansions in CaV2.1 enhance splicing activity at the 3′end of the transcript, leading to a CAG repeat length-dependent increase in the levels of a polyQ-encoding CaV2.1 mRNA splice isoform and the resultant disease protein. Taking advantage of this molecular phenomenon, we developed a novel splice isoform-specific (SIS)-RNAi strategy that selectively targets the polyQ-encoding CaV2.1 splice variant. Selective suppression of transiently expressed and endogenous polyQ-encoding CaV2.1 splice variants was achieved in a variety of cell-based models including a human neuronal cell line, using a new artificial miRNA-like delivery system. Moreover, the efficacy of gene silencing correlated with effective intracellular recognition and processing of SIS-RNAi miRNA mimics. These results lend support to the preclinical development of SIS-RNAi as a potential therapy for SCA6 and other dominantly inherited diseases. |
| first_indexed | 2024-12-17T21:34:03Z |
| format | Article |
| id | doaj.art-143c5d1060e44fc58034cbbfbed0225f |
| institution | Directory Open Access Journal |
| issn | 1095-953X |
| language | English |
| last_indexed | 2024-12-17T21:34:03Z |
| publishDate | 2011-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Neurobiology of Disease |
| spelling | doaj.art-143c5d1060e44fc58034cbbfbed0225f2022-12-21T21:31:47ZengElsevierNeurobiology of Disease1095-953X2011-09-01433533542Splice isoform-specific suppression of the CaV2.1 variant underlying spinocerebellar ataxia type 6Wei-Ling Tsou0Bing-Wen Soong1Henry L. Paulson2Edgardo Rodríguez-Lebrón3Institute of Neuroscience, School of Life Sciences, National Yang-Ming University, Taipei, Taiwan; Department of Neurology, University of Michigan, Ann Arbor, Michigan, USADepartment of Neurology, National Yang-Ming University and Taipei Veterans General Hospital, Taipei, TaiwanDepartment of Neurology, University of Michigan, Ann Arbor, Michigan, USA; Correspondence to: H. Paulson, 4001 BSRB, University of Michigan, Ann Arbor MI 48109. Fax: +1 734 647 9777.Department of Neurology, University of Michigan, Ann Arbor, Michigan, USA; Correspondence to: E. Rodríguez, Department of Internal Medicine, 200B EMRB, University of Iowa, Iowa City, Iowa 52242. Fax: +1 319 353 5572.Spinocerebellar ataxia type 6 (SCA6) is an inherited neurodegenerative disease caused by a polyglutamine (polyQ) expansion in the CaV2.1 voltage-gated calcium channel subunit (CACNA1A). There is currently no treatment for this debilitating disorder and thus a pressing need to develop preventative therapies. RNA interference (RNAi) has proven effective at halting disease progression in several models of spinocerebellar ataxia (SCA), including SCA types 1 and 3. However, in SCA6 and other dominantly inherited neurodegenerative disorders, RNAi-based strategies that selectively suppress expression of mutant alleles may be required. Using a CaV2.1 mini-gene reporter system, we found that pathogenic CAG expansions in CaV2.1 enhance splicing activity at the 3′end of the transcript, leading to a CAG repeat length-dependent increase in the levels of a polyQ-encoding CaV2.1 mRNA splice isoform and the resultant disease protein. Taking advantage of this molecular phenomenon, we developed a novel splice isoform-specific (SIS)-RNAi strategy that selectively targets the polyQ-encoding CaV2.1 splice variant. Selective suppression of transiently expressed and endogenous polyQ-encoding CaV2.1 splice variants was achieved in a variety of cell-based models including a human neuronal cell line, using a new artificial miRNA-like delivery system. Moreover, the efficacy of gene silencing correlated with effective intracellular recognition and processing of SIS-RNAi miRNA mimics. These results lend support to the preclinical development of SIS-RNAi as a potential therapy for SCA6 and other dominantly inherited diseases.http://www.sciencedirect.com/science/article/pii/S0969996111001331NeurodegenerationAutosomal dominant disorderSpinocerebellar ataxiaPolyglutamineCav2.1CACNA1A |
| spellingShingle | Wei-Ling Tsou Bing-Wen Soong Henry L. Paulson Edgardo Rodríguez-Lebrón Splice isoform-specific suppression of the CaV2.1 variant underlying spinocerebellar ataxia type 6 Neurobiology of Disease Neurodegeneration Autosomal dominant disorder Spinocerebellar ataxia Polyglutamine Cav2.1 CACNA1A |
| title | Splice isoform-specific suppression of the CaV2.1 variant underlying spinocerebellar ataxia type 6 |
| title_full | Splice isoform-specific suppression of the CaV2.1 variant underlying spinocerebellar ataxia type 6 |
| title_fullStr | Splice isoform-specific suppression of the CaV2.1 variant underlying spinocerebellar ataxia type 6 |
| title_full_unstemmed | Splice isoform-specific suppression of the CaV2.1 variant underlying spinocerebellar ataxia type 6 |
| title_short | Splice isoform-specific suppression of the CaV2.1 variant underlying spinocerebellar ataxia type 6 |
| title_sort | splice isoform specific suppression of the cav2 1 variant underlying spinocerebellar ataxia type 6 |
| topic | Neurodegeneration Autosomal dominant disorder Spinocerebellar ataxia Polyglutamine Cav2.1 CACNA1A |
| url | http://www.sciencedirect.com/science/article/pii/S0969996111001331 |
| work_keys_str_mv | AT weilingtsou spliceisoformspecificsuppressionofthecav21variantunderlyingspinocerebellarataxiatype6 AT bingwensoong spliceisoformspecificsuppressionofthecav21variantunderlyingspinocerebellarataxiatype6 AT henrylpaulson spliceisoformspecificsuppressionofthecav21variantunderlyingspinocerebellarataxiatype6 AT edgardorodriguezlebron spliceisoformspecificsuppressionofthecav21variantunderlyingspinocerebellarataxiatype6 |