FUS-ERG induces late-onset azacitidine resistance in acute myeloid leukaemia cells
Abstract FUS-ERG is a chimeric gene with a poor prognosis, found in myelodysplastic syndromes (MDS) and acute myeloid leukaemia (AML). It remains unclear whether DNA hypomethylating agents, including azacitidine (Aza), are effective in FUS-ERG-harbouring AML and how FUS-ERG induces chemoresistance....
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-09-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-41230-1 |
| _version_ | 1797452874691641344 |
|---|---|
| author | Ai Asai-Nishishita Masahiro Kawahara Goichi Tatsumi Masaki Iwasa Aya Fujishiro Rie Nishimura Hitoshi Minamiguchi Katsuyuki Kito Makoto Murata Akira Andoh |
| author_facet | Ai Asai-Nishishita Masahiro Kawahara Goichi Tatsumi Masaki Iwasa Aya Fujishiro Rie Nishimura Hitoshi Minamiguchi Katsuyuki Kito Makoto Murata Akira Andoh |
| author_sort | Ai Asai-Nishishita |
| collection | DOAJ |
| description | Abstract FUS-ERG is a chimeric gene with a poor prognosis, found in myelodysplastic syndromes (MDS) and acute myeloid leukaemia (AML). It remains unclear whether DNA hypomethylating agents, including azacitidine (Aza), are effective in FUS-ERG-harbouring AML and how FUS-ERG induces chemoresistance. Stable Ba/F3 transfectants with FUS-ERG were repeatedly exposed to Aza for 7 days of treatment and at 21-day intervals to investigate Aza sensitivity. Stable FUS-ERG transfectants acquired resistance acquired resistance after three courses of Aza exposure. RNA sequencing (RNA-seq) was performed when Aza susceptibility began to change; genes with altered expression or transcript variants were identified. Molecular signatures of these genes were analysed using gene ontology. RNA-seq analyses identified 74 upregulated and 320 downregulated genes involved in cell motility, cytokine production, and kinase activity. Additionally, 1321 genes with altered transcript variants were identified, revealing their involvement in chromatin organisation. In a clinical case of AML with FUS-ERG, we compared whole-genome alterations between the initial MDS diagnosis and AML recurrence after Aza treatment. Genes with non-synonymous or near mutations in transcription regulatory areas (TRAs), additionally detected in AML recurrence, were collated with the gene list from RNA-seq to identify genes involved in acquiring Aza resistance in the presence of FUS-ERG. Whole-genome sequencing of clinical specimens identified 29 genes with non-synonymous mutations, including BCOR, and 48 genes located within 20 kb of 54 TRA mutations in AML recurrence. These genes were involved in chromatin organisation and included NCOR2 as an overlapping gene with RNA-seq data. Transcription regulators involved in mutated TRAs were skewed and included RCOR1 in AML recurrence. We tested the efficacy of BH3 mimetics, including venetoclax and S63845, in primary Aza-resistant AML cells treated with FUS-ERG. Primary FUS-ERG-harbouring AML cells acquiring Aza resistance affected the myeloid cell leukaemia-1 (MCL1) inhibitor S63845 but not while using venetoclax, despite no mutations in BCL2. FUS-ERG promoted Aza resistance after several treatments. The disturbance of chromatin organisation might induce this by co-repressors, including BCOR, NCOR2, and RCOR1. MCL1 inhibition could partially overcome Aza resistance in FUS-ERG-harbouring AML cells. |
| first_indexed | 2024-03-09T15:14:58Z |
| format | Article |
| id | doaj.art-c8ba6cb7c7764ed3b3d4020ab2466e9a |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-09T15:14:58Z |
| publishDate | 2023-09-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-c8ba6cb7c7764ed3b3d4020ab2466e9a2023-11-26T13:09:51ZengNature PortfolioScientific Reports2045-23222023-09-0113111110.1038/s41598-023-41230-1FUS-ERG induces late-onset azacitidine resistance in acute myeloid leukaemia cellsAi Asai-Nishishita0Masahiro Kawahara1Goichi Tatsumi2Masaki Iwasa3Aya Fujishiro4Rie Nishimura5Hitoshi Minamiguchi6Katsuyuki Kito7Makoto Murata8Akira Andoh9Division of Hematology, Department of Medicine, Shiga University of Medical ScienceDivision of Hematology, Department of Medicine, Shiga University of Medical ScienceDepartment of Hematology and Oncology, Graduate School of Medicine, Kyoto UniversityDivision of Hematology, Department of Medicine, Shiga University of Medical ScienceDivision of Hematology, Department of Medicine, Shiga University of Medical ScienceDivision of Hematology, Department of Medicine, Shiga University of Medical ScienceDivision of Hematology, Department of Medicine, Shiga University of Medical ScienceDivision of Hematology, Department of Medicine, Shiga University of Medical ScienceDivision of Hematology, Department of Medicine, Shiga University of Medical ScienceDivision of Gastroenterology, Department of Medicine, Shiga University of Medical ScienceAbstract FUS-ERG is a chimeric gene with a poor prognosis, found in myelodysplastic syndromes (MDS) and acute myeloid leukaemia (AML). It remains unclear whether DNA hypomethylating agents, including azacitidine (Aza), are effective in FUS-ERG-harbouring AML and how FUS-ERG induces chemoresistance. Stable Ba/F3 transfectants with FUS-ERG were repeatedly exposed to Aza for 7 days of treatment and at 21-day intervals to investigate Aza sensitivity. Stable FUS-ERG transfectants acquired resistance acquired resistance after three courses of Aza exposure. RNA sequencing (RNA-seq) was performed when Aza susceptibility began to change; genes with altered expression or transcript variants were identified. Molecular signatures of these genes were analysed using gene ontology. RNA-seq analyses identified 74 upregulated and 320 downregulated genes involved in cell motility, cytokine production, and kinase activity. Additionally, 1321 genes with altered transcript variants were identified, revealing their involvement in chromatin organisation. In a clinical case of AML with FUS-ERG, we compared whole-genome alterations between the initial MDS diagnosis and AML recurrence after Aza treatment. Genes with non-synonymous or near mutations in transcription regulatory areas (TRAs), additionally detected in AML recurrence, were collated with the gene list from RNA-seq to identify genes involved in acquiring Aza resistance in the presence of FUS-ERG. Whole-genome sequencing of clinical specimens identified 29 genes with non-synonymous mutations, including BCOR, and 48 genes located within 20 kb of 54 TRA mutations in AML recurrence. These genes were involved in chromatin organisation and included NCOR2 as an overlapping gene with RNA-seq data. Transcription regulators involved in mutated TRAs were skewed and included RCOR1 in AML recurrence. We tested the efficacy of BH3 mimetics, including venetoclax and S63845, in primary Aza-resistant AML cells treated with FUS-ERG. Primary FUS-ERG-harbouring AML cells acquiring Aza resistance affected the myeloid cell leukaemia-1 (MCL1) inhibitor S63845 but not while using venetoclax, despite no mutations in BCL2. FUS-ERG promoted Aza resistance after several treatments. The disturbance of chromatin organisation might induce this by co-repressors, including BCOR, NCOR2, and RCOR1. MCL1 inhibition could partially overcome Aza resistance in FUS-ERG-harbouring AML cells.https://doi.org/10.1038/s41598-023-41230-1 |
| spellingShingle | Ai Asai-Nishishita Masahiro Kawahara Goichi Tatsumi Masaki Iwasa Aya Fujishiro Rie Nishimura Hitoshi Minamiguchi Katsuyuki Kito Makoto Murata Akira Andoh FUS-ERG induces late-onset azacitidine resistance in acute myeloid leukaemia cells Scientific Reports |
| title | FUS-ERG induces late-onset azacitidine resistance in acute myeloid leukaemia cells |
| title_full | FUS-ERG induces late-onset azacitidine resistance in acute myeloid leukaemia cells |
| title_fullStr | FUS-ERG induces late-onset azacitidine resistance in acute myeloid leukaemia cells |
| title_full_unstemmed | FUS-ERG induces late-onset azacitidine resistance in acute myeloid leukaemia cells |
| title_short | FUS-ERG induces late-onset azacitidine resistance in acute myeloid leukaemia cells |
| title_sort | fus erg induces late onset azacitidine resistance in acute myeloid leukaemia cells |
| url | https://doi.org/10.1038/s41598-023-41230-1 |
| work_keys_str_mv | AT aiasainishishita fuserginduceslateonsetazacitidineresistanceinacutemyeloidleukaemiacells AT masahirokawahara fuserginduceslateonsetazacitidineresistanceinacutemyeloidleukaemiacells AT goichitatsumi fuserginduceslateonsetazacitidineresistanceinacutemyeloidleukaemiacells AT masakiiwasa fuserginduceslateonsetazacitidineresistanceinacutemyeloidleukaemiacells AT ayafujishiro fuserginduceslateonsetazacitidineresistanceinacutemyeloidleukaemiacells AT rienishimura fuserginduceslateonsetazacitidineresistanceinacutemyeloidleukaemiacells AT hitoshiminamiguchi fuserginduceslateonsetazacitidineresistanceinacutemyeloidleukaemiacells AT katsuyukikito fuserginduceslateonsetazacitidineresistanceinacutemyeloidleukaemiacells AT makotomurata fuserginduceslateonsetazacitidineresistanceinacutemyeloidleukaemiacells AT akiraandoh fuserginduceslateonsetazacitidineresistanceinacutemyeloidleukaemiacells |