Multispanning membrane protein SIDT2 increases knockdown activity of gapmer antisense oligonucleotides
Abstract Recent advances in the clinical development of oligonucleotide therapeutics, such as antisense oligonucleotides (ASOs) and small interfering RNAs, have attracted attention as promising therapeutic modalities for genetic and intractable diseases. These oligonucleotide therapeutics exert thei...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-01-01
|
| Series: | Scientific Reports |
| Subjects: | |
| Online Access: | https://doi.org/10.1038/s41598-024-84310-6 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1841559686161104896 |
|---|---|
| author | Kohshi Kusumoto Kiyomi Sasaki Yasunori Uchida Ayaka Utsumi Tokuyuki Yoshida Satoshi Obika Takao Inoue Keiichiro Okuhira |
| author_facet | Kohshi Kusumoto Kiyomi Sasaki Yasunori Uchida Ayaka Utsumi Tokuyuki Yoshida Satoshi Obika Takao Inoue Keiichiro Okuhira |
| author_sort | Kohshi Kusumoto |
| collection | DOAJ |
| description | Abstract Recent advances in the clinical development of oligonucleotide therapeutics, such as antisense oligonucleotides (ASOs) and small interfering RNAs, have attracted attention as promising therapeutic modalities for genetic and intractable diseases. These oligonucleotide therapeutics exert their efficacy by binding to target RNAs present within cells; however, the mechanisms underlying their cellular uptake, especially their passage through membranes, remain largely unclear. In the nematode, Caenorhabditis elegans, the multi-pass transmembrane protein, SID-1, is involved in the cellular uptake of double-stranded RNAs. In mammals, SIDT1 and SIDT2 (SID-1 transmembrane family, members 1 and 2, respectively) are homologs of SID-1, yet their functional differences are not fully understood. In this study, we conducted a comparative analysis of the amino acid sequences of mammalian SIDT1 and SIDT2 to identify regions characteristic to each. By inducing SIDT1 or SIDT2 expression in human cell lines, we demonstrated that SIDT2 enhanced the knockdown activity of gapmer ASOs and potentially promoted their endosomal escape into the cytosol. Furthermore, by analyzing chimeric proteins of SIDT2 and SIDT1, we identified a region in SIDT2 that might be crucial for the enhancement of gapmer ASO activity. These findings elucidate the novel role of SIDT2 in the transport mechanism of gapmer ASOs and are expected to contribute to further development of oligonucleotide therapeutics. |
| format | Article |
| id | doaj-art-bc572341c23147aa888ae31dce15121f |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-bc572341c23147aa888ae31dce15121f2025-01-05T12:18:42ZengNature PortfolioScientific Reports2045-23222025-01-0115111210.1038/s41598-024-84310-6Multispanning membrane protein SIDT2 increases knockdown activity of gapmer antisense oligonucleotidesKohshi Kusumoto0Kiyomi Sasaki1Yasunori Uchida2Ayaka Utsumi3Tokuyuki Yoshida4Satoshi Obika5Takao Inoue6Keiichiro Okuhira7Osaka Medical and Pharmaceutical UniversityDivision of Molecular Target and Gene Therapy Products, National Institute of Health SciencesDivision of Molecular Target and Gene Therapy Products, National Institute of Health SciencesInstitute of Biomedical Sciences, Tokushima UniversityDivision of Molecular Target and Gene Therapy Products, National Institute of Health SciencesGraduate School of Pharmaceutical Sciences, Osaka UniversityDivision of Molecular Target and Gene Therapy Products, National Institute of Health SciencesOsaka Medical and Pharmaceutical UniversityAbstract Recent advances in the clinical development of oligonucleotide therapeutics, such as antisense oligonucleotides (ASOs) and small interfering RNAs, have attracted attention as promising therapeutic modalities for genetic and intractable diseases. These oligonucleotide therapeutics exert their efficacy by binding to target RNAs present within cells; however, the mechanisms underlying their cellular uptake, especially their passage through membranes, remain largely unclear. In the nematode, Caenorhabditis elegans, the multi-pass transmembrane protein, SID-1, is involved in the cellular uptake of double-stranded RNAs. In mammals, SIDT1 and SIDT2 (SID-1 transmembrane family, members 1 and 2, respectively) are homologs of SID-1, yet their functional differences are not fully understood. In this study, we conducted a comparative analysis of the amino acid sequences of mammalian SIDT1 and SIDT2 to identify regions characteristic to each. By inducing SIDT1 or SIDT2 expression in human cell lines, we demonstrated that SIDT2 enhanced the knockdown activity of gapmer ASOs and potentially promoted their endosomal escape into the cytosol. Furthermore, by analyzing chimeric proteins of SIDT2 and SIDT1, we identified a region in SIDT2 that might be crucial for the enhancement of gapmer ASO activity. These findings elucidate the novel role of SIDT2 in the transport mechanism of gapmer ASOs and are expected to contribute to further development of oligonucleotide therapeutics.https://doi.org/10.1038/s41598-024-84310-6Antisense oligonucleotideGapmerEndosomal escapeOligonucleotide therapeuticsSIDT1SIDT2 |
| spellingShingle | Kohshi Kusumoto Kiyomi Sasaki Yasunori Uchida Ayaka Utsumi Tokuyuki Yoshida Satoshi Obika Takao Inoue Keiichiro Okuhira Multispanning membrane protein SIDT2 increases knockdown activity of gapmer antisense oligonucleotides Scientific Reports Antisense oligonucleotide Gapmer Endosomal escape Oligonucleotide therapeutics SIDT1 SIDT2 |
| title | Multispanning membrane protein SIDT2 increases knockdown activity of gapmer antisense oligonucleotides |
| title_full | Multispanning membrane protein SIDT2 increases knockdown activity of gapmer antisense oligonucleotides |
| title_fullStr | Multispanning membrane protein SIDT2 increases knockdown activity of gapmer antisense oligonucleotides |
| title_full_unstemmed | Multispanning membrane protein SIDT2 increases knockdown activity of gapmer antisense oligonucleotides |
| title_short | Multispanning membrane protein SIDT2 increases knockdown activity of gapmer antisense oligonucleotides |
| title_sort | multispanning membrane protein sidt2 increases knockdown activity of gapmer antisense oligonucleotides |
| topic | Antisense oligonucleotide Gapmer Endosomal escape Oligonucleotide therapeutics SIDT1 SIDT2 |
| url | https://doi.org/10.1038/s41598-024-84310-6 |
| work_keys_str_mv | AT kohshikusumoto multispanningmembraneproteinsidt2increasesknockdownactivityofgapmerantisenseoligonucleotides AT kiyomisasaki multispanningmembraneproteinsidt2increasesknockdownactivityofgapmerantisenseoligonucleotides AT yasunoriuchida multispanningmembraneproteinsidt2increasesknockdownactivityofgapmerantisenseoligonucleotides AT ayakautsumi multispanningmembraneproteinsidt2increasesknockdownactivityofgapmerantisenseoligonucleotides AT tokuyukiyoshida multispanningmembraneproteinsidt2increasesknockdownactivityofgapmerantisenseoligonucleotides AT satoshiobika multispanningmembraneproteinsidt2increasesknockdownactivityofgapmerantisenseoligonucleotides AT takaoinoue multispanningmembraneproteinsidt2increasesknockdownactivityofgapmerantisenseoligonucleotides AT keiichirookuhira multispanningmembraneproteinsidt2increasesknockdownactivityofgapmerantisenseoligonucleotides |