A Dual Approach with Organoid and CRISPR Screening Reveals ERCC6 as a Determinant of Cisplatin Resistance in Osteosarcoma
Abstract Osteosarcoma (OS), the most prevalent primary bone malignancy in adolescents, is typically treated with cisplatin‐based chemotherapy. However, the development of cisplatin resistance often leads to relapse or metastasis, significantly impairing therapeutic efficacy. To tackle this issue, pa...
Saved in:
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2025-07-01
|
| Series: | Advanced Science |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/advs.202500632 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849417822221893632 |
|---|---|
| author | Ruiling Xu Sai Zhu Wenchao Zhang Haodong Xu Chao Tu Honghui Wang Lu Wang Na He Tang Liu Xiaoning Guo Xiaolei Ren Zhihong Li |
| author_facet | Ruiling Xu Sai Zhu Wenchao Zhang Haodong Xu Chao Tu Honghui Wang Lu Wang Na He Tang Liu Xiaoning Guo Xiaolei Ren Zhihong Li |
| author_sort | Ruiling Xu |
| collection | DOAJ |
| description | Abstract Osteosarcoma (OS), the most prevalent primary bone malignancy in adolescents, is typically treated with cisplatin‐based chemotherapy. However, the development of cisplatin resistance often leads to relapse or metastasis, significantly impairing therapeutic efficacy. To tackle this issue, patient‐derived osteosarcoma organoids (OSOs) is established that accurately reflect the cellular composition and heterogeneity of the original tumors, as validated by single‐cell RNA sequencing, bulk RNA sequencing, and histopathology analysis. Cisplatin resistance is successfully induced in these OSOs, creating a clinically relevant model for investigating chemoresistance. Utilizing RNA sequencing in cisplatin‐resistance OSOs and CRISPR screening in OS cell line, ERCC6 is identified as a pivotal regulator of cisplatin resistance. Knockdown of ERCC6 markedly enhanced cisplatin sensitivity in vitro and in vivo. Mechanistically, ERCC6 interacts with HNRNPM, influencing the PI3K/AKT signaling pathway and alternative splicing of pre‐mRNA for BAX. Notably, the knockdown of ERCC6 and HNRNPM increased expression of full‐length BAX and reduced skipping of exon 2, thus promoting apoptosis. This exon skipping in BAX results in a frameshift and introduces a premature stop codon (TGA) within the BH3 domain. These findings underscore the utility of OSOs in elucidating resistance mechanisms and highlight ERCC6 and HNRNPM as potential therapeutic targets. |
| format | Article |
| id | doaj-art-898a8a25b28b4a6a9de6142b52b35829 |
| institution | Kabale University |
| issn | 2198-3844 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Wiley |
| record_format | Article |
| series | Advanced Science |
| spelling | doaj-art-898a8a25b28b4a6a9de6142b52b358292025-08-20T03:32:37ZengWileyAdvanced Science2198-38442025-07-011228n/an/a10.1002/advs.202500632A Dual Approach with Organoid and CRISPR Screening Reveals ERCC6 as a Determinant of Cisplatin Resistance in OsteosarcomaRuiling Xu0Sai Zhu1Wenchao Zhang2Haodong Xu3Chao Tu4Honghui Wang5Lu Wang6Na He7Tang Liu8Xiaoning Guo9Xiaolei Ren10Zhihong Li11Department of Orthopedics The Second Xiangya Hospital Central South University Changsha Hunan 410011 P. R. ChinaDepartment of Orthopedics The Second Xiangya Hospital Central South University Changsha Hunan 410011 P. R. ChinaDepartment of Orthopedics The Second Xiangya Hospital Central South University Changsha Hunan 410011 P. R. ChinaDepartment of Orthopedics The Second Xiangya Hospital Central South University Changsha Hunan 410011 P. R. ChinaDepartment of Orthopedics The Second Xiangya Hospital Central South University Changsha Hunan 410011 P. R. ChinaCollege of Biology Hunan University Changsha 410082 P. R. ChinaDepartment of Orthopedics The Second Xiangya Hospital Central South University Changsha Hunan 410011 P. R. ChinaDepartment of Orthopedics The Second Xiangya Hospital Central South University Changsha Hunan 410011 P. R. ChinaDepartment of Orthopedics The Second Xiangya Hospital Central South University Changsha Hunan 410011 P. R. ChinaDepartment of Orthopedics The Second Xiangya Hospital Central South University Changsha Hunan 410011 P. R. ChinaDepartment of Orthopedics The Second Xiangya Hospital Central South University Changsha Hunan 410011 P. R. ChinaDepartment of Orthopedics The Second Xiangya Hospital Central South University Changsha Hunan 410011 P. R. ChinaAbstract Osteosarcoma (OS), the most prevalent primary bone malignancy in adolescents, is typically treated with cisplatin‐based chemotherapy. However, the development of cisplatin resistance often leads to relapse or metastasis, significantly impairing therapeutic efficacy. To tackle this issue, patient‐derived osteosarcoma organoids (OSOs) is established that accurately reflect the cellular composition and heterogeneity of the original tumors, as validated by single‐cell RNA sequencing, bulk RNA sequencing, and histopathology analysis. Cisplatin resistance is successfully induced in these OSOs, creating a clinically relevant model for investigating chemoresistance. Utilizing RNA sequencing in cisplatin‐resistance OSOs and CRISPR screening in OS cell line, ERCC6 is identified as a pivotal regulator of cisplatin resistance. Knockdown of ERCC6 markedly enhanced cisplatin sensitivity in vitro and in vivo. Mechanistically, ERCC6 interacts with HNRNPM, influencing the PI3K/AKT signaling pathway and alternative splicing of pre‐mRNA for BAX. Notably, the knockdown of ERCC6 and HNRNPM increased expression of full‐length BAX and reduced skipping of exon 2, thus promoting apoptosis. This exon skipping in BAX results in a frameshift and introduces a premature stop codon (TGA) within the BH3 domain. These findings underscore the utility of OSOs in elucidating resistance mechanisms and highlight ERCC6 and HNRNPM as potential therapeutic targets.https://doi.org/10.1002/advs.202500632alternative splicingchemoresistanceERCC6organoidosteosarcoma |
| spellingShingle | Ruiling Xu Sai Zhu Wenchao Zhang Haodong Xu Chao Tu Honghui Wang Lu Wang Na He Tang Liu Xiaoning Guo Xiaolei Ren Zhihong Li A Dual Approach with Organoid and CRISPR Screening Reveals ERCC6 as a Determinant of Cisplatin Resistance in Osteosarcoma Advanced Science alternative splicing chemoresistance ERCC6 organoid osteosarcoma |
| title | A Dual Approach with Organoid and CRISPR Screening Reveals ERCC6 as a Determinant of Cisplatin Resistance in Osteosarcoma |
| title_full | A Dual Approach with Organoid and CRISPR Screening Reveals ERCC6 as a Determinant of Cisplatin Resistance in Osteosarcoma |
| title_fullStr | A Dual Approach with Organoid and CRISPR Screening Reveals ERCC6 as a Determinant of Cisplatin Resistance in Osteosarcoma |
| title_full_unstemmed | A Dual Approach with Organoid and CRISPR Screening Reveals ERCC6 as a Determinant of Cisplatin Resistance in Osteosarcoma |
| title_short | A Dual Approach with Organoid and CRISPR Screening Reveals ERCC6 as a Determinant of Cisplatin Resistance in Osteosarcoma |
| title_sort | dual approach with organoid and crispr screening reveals ercc6 as a determinant of cisplatin resistance in osteosarcoma |
| topic | alternative splicing chemoresistance ERCC6 organoid osteosarcoma |
| url | https://doi.org/10.1002/advs.202500632 |
| work_keys_str_mv | AT ruilingxu adualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT saizhu adualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT wenchaozhang adualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT haodongxu adualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT chaotu adualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT honghuiwang adualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT luwang adualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT nahe adualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT tangliu adualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT xiaoningguo adualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT xiaoleiren adualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT zhihongli adualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT ruilingxu dualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT saizhu dualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT wenchaozhang dualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT haodongxu dualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT chaotu dualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT honghuiwang dualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT luwang dualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT nahe dualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT tangliu dualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT xiaoningguo dualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT xiaoleiren dualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma AT zhihongli dualapproachwithorganoidandcrisprscreeningrevealsercc6asadeterminantofcisplatinresistanceinosteosarcoma |