Splice-switching of the insulin receptor pre-mRNA alleviates tumorigenic hallmarks in rhabdomyosarcoma
Abstract Rhabdomyosarcoma (RMS) is an aggressive pediatric tumor with a poor prognosis for metastasis and recurrent disease. Large-scale sequencing endeavors demonstrate that Rhabdomyosarcomas have a dearth of precisely targetable driver mutations. However, IGF-2 signaling is known to be grossly alt...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2022-01-01
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| Series: | npj Precision Oncology |
| Online Access: | https://doi.org/10.1038/s41698-021-00245-5 |
| _version_ | 1797643041739112448 |
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| author | Safiya Khurshid Matias Montes Daniel F. Comiskey Brianne Shane Eleftheria Matsa Francesca Jung Chelsea Brown Hemant Kumar Bid Ruoning Wang Peter J. Houghton Ryan Roberts Frank Rigo Dawn Chandler |
| author_facet | Safiya Khurshid Matias Montes Daniel F. Comiskey Brianne Shane Eleftheria Matsa Francesca Jung Chelsea Brown Hemant Kumar Bid Ruoning Wang Peter J. Houghton Ryan Roberts Frank Rigo Dawn Chandler |
| author_sort | Safiya Khurshid |
| collection | DOAJ |
| description | Abstract Rhabdomyosarcoma (RMS) is an aggressive pediatric tumor with a poor prognosis for metastasis and recurrent disease. Large-scale sequencing endeavors demonstrate that Rhabdomyosarcomas have a dearth of precisely targetable driver mutations. However, IGF-2 signaling is known to be grossly altered in RMS. The insulin receptor (IR) exists in two alternatively spliced isoforms, IR-A and IR-B. The IGF-2 signaling molecule binds both its innate IGF-1 receptor as well as the insulin receptor variant A (IR-A) with high affinity. Mitogenic and proliferative signaling via the canonical IGF-2 pathway is, therefore, augmented by IR-A. This study shows that RMS patients express increased IR-A levels compared to control tissues that predominantly express the IR-B isoform. We also found that Hif-1α is significantly increased in RMS tumors, portraying their hypoxic phenotype. Concordantly, the alternative splicing of IR adapts to produce more IR-A in response to hypoxic stress. Upon examining the pre-mRNA structure of the gene, we identified a potential hypoxia-responsive element, which is also the binding site for the RNA-binding protein CUG-BP1 (CELF1). We designed Splice Switching Oligonucleotides (SSO) against this binding site to decrease IR-A levels in RMS cell lines and, consequently, rescue the IR-B expression levels. SSO treatment resulted in a significant reduction in cell proliferation, migration, and angiogenesis. Our data shows promising insight into how impeding the IGF-2 pathway by reducing IR-A expression mitigates tumor growth. It is evident that Rhabdomyosarcomas use IR alternative splicing as yet another survival strategy that can be exploited as a therapeutic intervention in conjunction with already established anti-IGF-1 receptor therapies. |
| first_indexed | 2024-03-11T14:09:58Z |
| format | Article |
| id | doaj.art-289d316b2b0e4cb78d5e10b41c23146a |
| institution | Directory Open Access Journal |
| issn | 2397-768X |
| language | English |
| last_indexed | 2024-03-11T14:09:58Z |
| publishDate | 2022-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | npj Precision Oncology |
| spelling | doaj.art-289d316b2b0e4cb78d5e10b41c23146a2023-11-02T00:33:54ZengNature Portfolionpj Precision Oncology2397-768X2022-01-016111110.1038/s41698-021-00245-5Splice-switching of the insulin receptor pre-mRNA alleviates tumorigenic hallmarks in rhabdomyosarcomaSafiya Khurshid0Matias Montes1Daniel F. Comiskey2Brianne Shane3Eleftheria Matsa4Francesca Jung5Chelsea Brown6Hemant Kumar Bid7Ruoning Wang8Peter J. Houghton9Ryan Roberts10Frank Rigo11Dawn Chandler12Department of Pediatrics and the Center for RNA Biology, The Ohio State UniversityDepartment of Pediatrics and the Center for RNA Biology, The Ohio State UniversityDepartment of Pediatrics and the Center for RNA Biology, The Ohio State UniversityDepartment of Pediatrics and the Center for RNA Biology, The Ohio State UniversityDepartment of Pediatrics and the Center for RNA Biology, The Ohio State UniversityDepartment of Pediatrics and the Center for RNA Biology, The Ohio State UniversityDepartment of Pediatrics and the Center for RNA Biology, The Ohio State UniversityResonant Therapeutics, Inc.Department of Pediatrics and the Center for RNA Biology, The Ohio State UniversityGreenhey Children’s Cancer Research Institute, UT HealthDepartment of Pediatrics and the Center for RNA Biology, The Ohio State UniversityIonis PharmaceuticalsDepartment of Pediatrics and the Center for RNA Biology, The Ohio State UniversityAbstract Rhabdomyosarcoma (RMS) is an aggressive pediatric tumor with a poor prognosis for metastasis and recurrent disease. Large-scale sequencing endeavors demonstrate that Rhabdomyosarcomas have a dearth of precisely targetable driver mutations. However, IGF-2 signaling is known to be grossly altered in RMS. The insulin receptor (IR) exists in two alternatively spliced isoforms, IR-A and IR-B. The IGF-2 signaling molecule binds both its innate IGF-1 receptor as well as the insulin receptor variant A (IR-A) with high affinity. Mitogenic and proliferative signaling via the canonical IGF-2 pathway is, therefore, augmented by IR-A. This study shows that RMS patients express increased IR-A levels compared to control tissues that predominantly express the IR-B isoform. We also found that Hif-1α is significantly increased in RMS tumors, portraying their hypoxic phenotype. Concordantly, the alternative splicing of IR adapts to produce more IR-A in response to hypoxic stress. Upon examining the pre-mRNA structure of the gene, we identified a potential hypoxia-responsive element, which is also the binding site for the RNA-binding protein CUG-BP1 (CELF1). We designed Splice Switching Oligonucleotides (SSO) against this binding site to decrease IR-A levels in RMS cell lines and, consequently, rescue the IR-B expression levels. SSO treatment resulted in a significant reduction in cell proliferation, migration, and angiogenesis. Our data shows promising insight into how impeding the IGF-2 pathway by reducing IR-A expression mitigates tumor growth. It is evident that Rhabdomyosarcomas use IR alternative splicing as yet another survival strategy that can be exploited as a therapeutic intervention in conjunction with already established anti-IGF-1 receptor therapies.https://doi.org/10.1038/s41698-021-00245-5 |
| spellingShingle | Safiya Khurshid Matias Montes Daniel F. Comiskey Brianne Shane Eleftheria Matsa Francesca Jung Chelsea Brown Hemant Kumar Bid Ruoning Wang Peter J. Houghton Ryan Roberts Frank Rigo Dawn Chandler Splice-switching of the insulin receptor pre-mRNA alleviates tumorigenic hallmarks in rhabdomyosarcoma npj Precision Oncology |
| title | Splice-switching of the insulin receptor pre-mRNA alleviates tumorigenic hallmarks in rhabdomyosarcoma |
| title_full | Splice-switching of the insulin receptor pre-mRNA alleviates tumorigenic hallmarks in rhabdomyosarcoma |
| title_fullStr | Splice-switching of the insulin receptor pre-mRNA alleviates tumorigenic hallmarks in rhabdomyosarcoma |
| title_full_unstemmed | Splice-switching of the insulin receptor pre-mRNA alleviates tumorigenic hallmarks in rhabdomyosarcoma |
| title_short | Splice-switching of the insulin receptor pre-mRNA alleviates tumorigenic hallmarks in rhabdomyosarcoma |
| title_sort | splice switching of the insulin receptor pre mrna alleviates tumorigenic hallmarks in rhabdomyosarcoma |
| url | https://doi.org/10.1038/s41698-021-00245-5 |
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