Josephine Q N Nguyen, Wojtek Drabarek, Aïsha M C H J Leeflang, Tom Brands, Thierry P P van den Bosch, Robert M Verdijk, Harmen J G van de Werken, Job van Riet, Dion Paridaens, Annelies de Klein, Erwin Brosens, Emine Kiliç
{"title":"抑制剪接体对 SF3B1 基因突变的葡萄膜黑色素瘤的影响","authors":"Josephine Q N Nguyen, Wojtek Drabarek, Aïsha M C H J Leeflang, Tom Brands, Thierry P P van den Bosch, Robert M Verdijk, Harmen J G van de Werken, Job van Riet, Dion Paridaens, Annelies de Klein, Erwin Brosens, Emine Kiliç","doi":"10.1167/iovs.65.12.11","DOIUrl":null,"url":null,"abstract":"<p><strong>Purpose: </strong>Unfortunately, treatment of patients with uveal melanoma (UM) with metastatic disease is limited. Twenty percent of patients with UM harbor a mutation in the splicing factor gene SF3B1, suggesting that aberrant spliceosome function plays a vital role in tumorigenesis. Splicing inhibitors exploit the preferential sensitivity of spliceosome-compromised leukemic cells to these compounds.</p><p><strong>Methods: </strong>We studied the effect of the splicing inhibitor E7107 using two UM cell lines and ex vivo cultured SF3B1- and BAP1-mutated primary UM tumor slices. These UM cell lines and ex vivo tumor slices were exposed for 24 hours to different concentrations of E7107. Tumor slices were stained with hematoxylin and eosin (H&E) and incubated with BAP1, MelanA, MIB-1, and caspase-3 antisera.</p><p><strong>Results: </strong>The E7107-exposed UM cell lines exhibited decreased cell viability and increased apoptosis, with the greatest effect on SF3B1-mutated UM cells. A similar effect on UM tumor slices was observed upon exposure to E7107. Additionally, RNA was isolated for differential isoform expression analysis. No significant difference in isoform usage was found genome-wide. However, specific genes were differentially expressed after E7107 treatment in the SF3B1-mutated samples. Moreover, E7107 had the greatest effect on intron retention.</p><p><strong>Conclusions: </strong>This study indicates/suggests that mutated SF3B1 UM cells are more sensitive to the splicing inhibitor E7107 than wild-type SF3B1 UM cells.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"65 12","pages":"11"},"PeriodicalIF":5.0000,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11463709/pdf/","citationCount":"0","resultStr":"{\"title\":\"The Impact of Spliceosome Inhibition in SF3B1-Mutated Uveal Melanoma.\",\"authors\":\"Josephine Q N Nguyen, Wojtek Drabarek, Aïsha M C H J Leeflang, Tom Brands, Thierry P P van den Bosch, Robert M Verdijk, Harmen J G van de Werken, Job van Riet, Dion Paridaens, Annelies de Klein, Erwin Brosens, Emine Kiliç\",\"doi\":\"10.1167/iovs.65.12.11\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Purpose: </strong>Unfortunately, treatment of patients with uveal melanoma (UM) with metastatic disease is limited. Twenty percent of patients with UM harbor a mutation in the splicing factor gene SF3B1, suggesting that aberrant spliceosome function plays a vital role in tumorigenesis. Splicing inhibitors exploit the preferential sensitivity of spliceosome-compromised leukemic cells to these compounds.</p><p><strong>Methods: </strong>We studied the effect of the splicing inhibitor E7107 using two UM cell lines and ex vivo cultured SF3B1- and BAP1-mutated primary UM tumor slices. These UM cell lines and ex vivo tumor slices were exposed for 24 hours to different concentrations of E7107. Tumor slices were stained with hematoxylin and eosin (H&E) and incubated with BAP1, MelanA, MIB-1, and caspase-3 antisera.</p><p><strong>Results: </strong>The E7107-exposed UM cell lines exhibited decreased cell viability and increased apoptosis, with the greatest effect on SF3B1-mutated UM cells. A similar effect on UM tumor slices was observed upon exposure to E7107. Additionally, RNA was isolated for differential isoform expression analysis. No significant difference in isoform usage was found genome-wide. However, specific genes were differentially expressed after E7107 treatment in the SF3B1-mutated samples. Moreover, E7107 had the greatest effect on intron retention.</p><p><strong>Conclusions: </strong>This study indicates/suggests that mutated SF3B1 UM cells are more sensitive to the splicing inhibitor E7107 than wild-type SF3B1 UM cells.</p>\",\"PeriodicalId\":14620,\"journal\":{\"name\":\"Investigative ophthalmology & visual science\",\"volume\":\"65 12\",\"pages\":\"11\"},\"PeriodicalIF\":5.0000,\"publicationDate\":\"2024-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11463709/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Investigative ophthalmology & visual science\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1167/iovs.65.12.11\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"OPHTHALMOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Investigative ophthalmology & visual science","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1167/iovs.65.12.11","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"OPHTHALMOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
目的:遗憾的是,葡萄膜黑色素瘤(UM)转移性疾病患者的治疗受到限制。20%的葡萄膜黑色素瘤患者携带剪接因子基因 SF3B1 突变,这表明剪接体功能异常在肿瘤发生中起着至关重要的作用。剪接抑制剂利用了剪接体受损的白血病细胞对这些化合物的优先敏感性:我们使用两种 UM 细胞系和体外培养的 SF3B1 和 BAP1 突变的原发性 UM 肿瘤切片研究了剪接抑制剂 E7107 的作用。将这些 UM 细胞系和体外肿瘤切片暴露于不同浓度的 E7107 中 24 小时。肿瘤切片用苏木精和伊红(H&E)染色,并与 BAP1、MelanA、MIB-1 和 caspase-3 抗血清孵育:结果:暴露于 E7107 的 UM 细胞系的细胞存活率降低,细胞凋亡增加,其中对 SF3B1 突变的 UM 细胞影响最大。接触 E7107 对 UM 肿瘤切片也有类似影响。此外,还分离了 RNA 进行差异同工酶表达分析。在全基因组范围内没有发现同工酶表达量的明显差异。然而,在SF3B1突变样本中,E7107处理后特定基因的表达出现差异。此外,E7107对内含子保留的影响最大:本研究表明/暗示突变的SF3B1 UM细胞比野生型SF3B1 UM细胞对剪接抑制剂E7107更敏感。
The Impact of Spliceosome Inhibition in SF3B1-Mutated Uveal Melanoma.
Purpose: Unfortunately, treatment of patients with uveal melanoma (UM) with metastatic disease is limited. Twenty percent of patients with UM harbor a mutation in the splicing factor gene SF3B1, suggesting that aberrant spliceosome function plays a vital role in tumorigenesis. Splicing inhibitors exploit the preferential sensitivity of spliceosome-compromised leukemic cells to these compounds.
Methods: We studied the effect of the splicing inhibitor E7107 using two UM cell lines and ex vivo cultured SF3B1- and BAP1-mutated primary UM tumor slices. These UM cell lines and ex vivo tumor slices were exposed for 24 hours to different concentrations of E7107. Tumor slices were stained with hematoxylin and eosin (H&E) and incubated with BAP1, MelanA, MIB-1, and caspase-3 antisera.
Results: The E7107-exposed UM cell lines exhibited decreased cell viability and increased apoptosis, with the greatest effect on SF3B1-mutated UM cells. A similar effect on UM tumor slices was observed upon exposure to E7107. Additionally, RNA was isolated for differential isoform expression analysis. No significant difference in isoform usage was found genome-wide. However, specific genes were differentially expressed after E7107 treatment in the SF3B1-mutated samples. Moreover, E7107 had the greatest effect on intron retention.
Conclusions: This study indicates/suggests that mutated SF3B1 UM cells are more sensitive to the splicing inhibitor E7107 than wild-type SF3B1 UM cells.
期刊介绍:
Investigative Ophthalmology & Visual Science (IOVS), published as ready online, is a peer-reviewed academic journal of the Association for Research in Vision and Ophthalmology (ARVO). IOVS features original research, mostly pertaining to clinical and laboratory ophthalmology and vision research in general.