High Concentration or Combined Treatment of Antisense Oligonucleotides for Spinal Muscular Atrophy Perturbed <i>SMN2</i> Splicing in Patient Fibroblasts
Spinal muscular atrophy (SMA) is caused by <i>survival motor neuron 1 SMN1</i> deletion. The <i>survival motor neuron 2 (SMN2)</i> encodes the same protein as <i>SMN1</i> does, but it has a splicing defect of exon 7. Some antisense oligonucleotides (ASOs) have bee...
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MDPI AG
2022-04-01
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| author | Yogik Onky Silvana Wijaya Emma Tabe Eko Niba Hisahide Nishio Kentaro Okamoto Hiroyuki Awano Toshio Saito Yasuhiro Takeshima Masakazu Shinohara |
| author_facet | Yogik Onky Silvana Wijaya Emma Tabe Eko Niba Hisahide Nishio Kentaro Okamoto Hiroyuki Awano Toshio Saito Yasuhiro Takeshima Masakazu Shinohara |
| author_sort | Yogik Onky Silvana Wijaya |
| collection | DOAJ |
| description | Spinal muscular atrophy (SMA) is caused by <i>survival motor neuron 1 SMN1</i> deletion. The <i>survival motor neuron 2 (SMN2)</i> encodes the same protein as <i>SMN1</i> does, but it has a splicing defect of exon 7. Some antisense oligonucleotides (ASOs) have been proven to correct this defect. One of these, nusinersen, is effective in SMA-affected infants, but not as much so in advanced-stage patients. Furthermore, the current regimen may exhibit a ceiling effect. To overcome these problems, high-dose ASOs or combined ASOs have been explored. Here, using SMA fibroblasts, we examined the effects of high-concentration ASOs and of combining two ASOs. Three ASOs were examined: one targeting intronic splicing suppressor site N1 (ISS-N1) in intron 7, and two others targeting the 3′ splice site and 5′ region of exon 8. In our experiments on all ASO types, a low or intermediate concentration (50 or 100 nM) showed better splicing efficiency than a high concentration (200 nM). In addition, a high concentration of each ASO created a cryptic exon in exon 6. When a mixture of two different ASOs (100 nM each) was added to the cells, the cryptic exon was included in the mRNA. In conclusion, ASOs at a high concentration or used in combination may show less splicing correction and cryptic exon creation. |
| first_indexed | 2024-03-09T13:37:06Z |
| format | Article |
| id | doaj.art-759887d58bf54b5d87d24bc296228ff6 |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-09T13:37:06Z |
| publishDate | 2022-04-01 |
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| series | Genes |
| spelling | doaj.art-759887d58bf54b5d87d24bc296228ff62023-11-30T21:10:26ZengMDPI AGGenes2073-44252022-04-0113468510.3390/genes13040685High Concentration or Combined Treatment of Antisense Oligonucleotides for Spinal Muscular Atrophy Perturbed <i>SMN2</i> Splicing in Patient FibroblastsYogik Onky Silvana Wijaya0Emma Tabe Eko Niba1Hisahide Nishio2Kentaro Okamoto3Hiroyuki Awano4Toshio Saito5Yasuhiro Takeshima6Masakazu Shinohara7Department of Community Medicine and Social Healthcare Science, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Hyogo, JapanDepartment of Community Medicine and Social Healthcare Science, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Hyogo, JapanDepartment of Community Medicine and Social Healthcare Science, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Hyogo, JapanDepartment of Pediatrics, Ehime Prefectural Imabari Hospital, 4-5-5 Ishii-cho, Imabari, 794-0006, Ehime, JapanDepartment of Pediatrics, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Hyogo, JapanDepartment of Neurology, National Hospital Organization Osaka Toneyama Medical Center, 5-1-1 Toneyama, Toyonaka 560-8552, Osaka, JapanDepartment of Pediatrics, Hyogo College of Medicine, 1-1 Mukogawa-cho, Nishinomiya 663-8501, Hyogo, JapanDepartment of Community Medicine and Social Healthcare Science, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Hyogo, JapanSpinal muscular atrophy (SMA) is caused by <i>survival motor neuron 1 SMN1</i> deletion. The <i>survival motor neuron 2 (SMN2)</i> encodes the same protein as <i>SMN1</i> does, but it has a splicing defect of exon 7. Some antisense oligonucleotides (ASOs) have been proven to correct this defect. One of these, nusinersen, is effective in SMA-affected infants, but not as much so in advanced-stage patients. Furthermore, the current regimen may exhibit a ceiling effect. To overcome these problems, high-dose ASOs or combined ASOs have been explored. Here, using SMA fibroblasts, we examined the effects of high-concentration ASOs and of combining two ASOs. Three ASOs were examined: one targeting intronic splicing suppressor site N1 (ISS-N1) in intron 7, and two others targeting the 3′ splice site and 5′ region of exon 8. In our experiments on all ASO types, a low or intermediate concentration (50 or 100 nM) showed better splicing efficiency than a high concentration (200 nM). In addition, a high concentration of each ASO created a cryptic exon in exon 6. When a mixture of two different ASOs (100 nM each) was added to the cells, the cryptic exon was included in the mRNA. In conclusion, ASOs at a high concentration or used in combination may show less splicing correction and cryptic exon creation.https://www.mdpi.com/2073-4425/13/4/685spinal muscular atrophy<i>SMN1</i><i>SMN2</i>splicingantisense oligonucleotidecryptic exon |
| spellingShingle | Yogik Onky Silvana Wijaya Emma Tabe Eko Niba Hisahide Nishio Kentaro Okamoto Hiroyuki Awano Toshio Saito Yasuhiro Takeshima Masakazu Shinohara High Concentration or Combined Treatment of Antisense Oligonucleotides for Spinal Muscular Atrophy Perturbed <i>SMN2</i> Splicing in Patient Fibroblasts Genes spinal muscular atrophy <i>SMN1</i> <i>SMN2</i> splicing antisense oligonucleotide cryptic exon |
| title | High Concentration or Combined Treatment of Antisense Oligonucleotides for Spinal Muscular Atrophy Perturbed <i>SMN2</i> Splicing in Patient Fibroblasts |
| title_full | High Concentration or Combined Treatment of Antisense Oligonucleotides for Spinal Muscular Atrophy Perturbed <i>SMN2</i> Splicing in Patient Fibroblasts |
| title_fullStr | High Concentration or Combined Treatment of Antisense Oligonucleotides for Spinal Muscular Atrophy Perturbed <i>SMN2</i> Splicing in Patient Fibroblasts |
| title_full_unstemmed | High Concentration or Combined Treatment of Antisense Oligonucleotides for Spinal Muscular Atrophy Perturbed <i>SMN2</i> Splicing in Patient Fibroblasts |
| title_short | High Concentration or Combined Treatment of Antisense Oligonucleotides for Spinal Muscular Atrophy Perturbed <i>SMN2</i> Splicing in Patient Fibroblasts |
| title_sort | high concentration or combined treatment of antisense oligonucleotides for spinal muscular atrophy perturbed i smn2 i splicing in patient fibroblasts |
| topic | spinal muscular atrophy <i>SMN1</i> <i>SMN2</i> splicing antisense oligonucleotide cryptic exon |
| url | https://www.mdpi.com/2073-4425/13/4/685 |
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