Epigenetic-focused CRISPR/Cas9 screen identifies (absent, small, or homeotic)2-like protein (ASH2L) as a regulator of glioblastoma cell survival
<p><strong>Background:</strong> Glioblastoma is the most common and aggressive primary brain tumor with extremely poor prognosis, highlighting an urgent need for developing novel treatment options. Identifying epigenetic vulnerabilities of cancer cells can provide excellen...
| Main Authors: | , , , , , , , , , , , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
BioMed Central
2023
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| _version_ | 1826311616501645312 |
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| author | Ozyerli-Goknar, E Kala, EY Aksu, AC Bulut, I Cingöz, A Nizamuddin, S Biniossek, M Seker-Polat, F Morova, T Aztekin, C Kung, SHY Syed, H Tuncbag, N Gönen, M Philpott, M Cribbs, AP Acilan, C Lack, NA Onder, TT Timmers, HTM Bagci-Onder, T |
| author_facet | Ozyerli-Goknar, E Kala, EY Aksu, AC Bulut, I Cingöz, A Nizamuddin, S Biniossek, M Seker-Polat, F Morova, T Aztekin, C Kung, SHY Syed, H Tuncbag, N Gönen, M Philpott, M Cribbs, AP Acilan, C Lack, NA Onder, TT Timmers, HTM Bagci-Onder, T |
| author_sort | Ozyerli-Goknar, E |
| collection | OXFORD |
| description | <p><strong>Background:</strong> Glioblastoma is the most common and aggressive primary brain tumor with extremely poor prognosis, highlighting an urgent need for developing novel treatment options. Identifying epigenetic vulnerabilities of cancer cells can provide excellent therapeutic intervention points for various types of cancers.</p>
<p><strong>Method:</strong> In this study, we investigated epigenetic regulators of glioblastoma cell survival through CRISPR/Cas9 based genetic ablation screens using a customized sgRNA library EpiDoKOL, which targets critical functional domains of chromatin modifiers.</p>
<p><strong>Results:</strong> Screens conducted in multiple cell lines revealed <em>ASH2L</em>, a histone lysine methyltransferase complex subunit, as a major regulator of glioblastoma cell viability. <em>ASH2L</em> depletion led to cell cycle arrest and apoptosis. RNA sequencing and greenCUT&RUN together identified a set of cell cycle regulatory genes, such as <em>TRA2B, BARD1, KIF20B, ARID4A and SMARCC1</em> that were downregulated upon <em>ASH2L</em> depletion. Mass spectrometry analysis revealed the interaction partners of ASH2L in glioblastoma cell lines as SET1/MLL family members including SETD1A, SETD1B, MLL1 and MLL2. We further showed that glioblastoma cells had a differential dependency on expression of <em>SET1/MLL</em> family members for survival. The growth of <em>ASH2L</em>-depleted glioblastoma cells was markedly slower than controls in orthotopic in vivo models. TCGA analysis showed high ASH2L expression in glioblastoma compared to low grade gliomas and immunohistochemical analysis revealed significant ASH2L expression in glioblastoma tissues, attesting to its clinical relevance. Therefore, high throughput, robust and affordable screens with focused libraries, such as EpiDoKOL, holds great promise to enable rapid discovery of novel epigenetic regulators of cancer cell survival, such as <em>ASH2L</em>.</p>
<p><strong>Conclusion:</strong> Together, we suggest that targeting <em>ASH2L</em> could serve as a new therapeutic opportunity for glioblastoma.</p> |
| first_indexed | 2024-03-07T08:13:54Z |
| format | Journal article |
| id | oxford-uuid:4620e400-db55-4c20-9ef7-0fd8477feb75 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T08:13:54Z |
| publishDate | 2023 |
| publisher | BioMed Central |
| record_format | dspace |
| spelling | oxford-uuid:4620e400-db55-4c20-9ef7-0fd8477feb752023-12-13T06:10:11ZEpigenetic-focused CRISPR/Cas9 screen identifies (absent, small, or homeotic)2-like protein (ASH2L) as a regulator of glioblastoma cell survivalJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:4620e400-db55-4c20-9ef7-0fd8477feb75EnglishSymplectic ElementsBioMed Central2023Ozyerli-Goknar, EKala, EYAksu, ACBulut, ICingöz, ANizamuddin, SBiniossek, MSeker-Polat, FMorova, TAztekin, CKung, SHYSyed, HTuncbag, NGönen, MPhilpott, MCribbs, APAcilan, CLack, NAOnder, TTTimmers, HTMBagci-Onder, T<p><strong>Background:</strong> Glioblastoma is the most common and aggressive primary brain tumor with extremely poor prognosis, highlighting an urgent need for developing novel treatment options. Identifying epigenetic vulnerabilities of cancer cells can provide excellent therapeutic intervention points for various types of cancers.</p> <p><strong>Method:</strong> In this study, we investigated epigenetic regulators of glioblastoma cell survival through CRISPR/Cas9 based genetic ablation screens using a customized sgRNA library EpiDoKOL, which targets critical functional domains of chromatin modifiers.</p> <p><strong>Results:</strong> Screens conducted in multiple cell lines revealed <em>ASH2L</em>, a histone lysine methyltransferase complex subunit, as a major regulator of glioblastoma cell viability. <em>ASH2L</em> depletion led to cell cycle arrest and apoptosis. RNA sequencing and greenCUT&RUN together identified a set of cell cycle regulatory genes, such as <em>TRA2B, BARD1, KIF20B, ARID4A and SMARCC1</em> that were downregulated upon <em>ASH2L</em> depletion. Mass spectrometry analysis revealed the interaction partners of ASH2L in glioblastoma cell lines as SET1/MLL family members including SETD1A, SETD1B, MLL1 and MLL2. We further showed that glioblastoma cells had a differential dependency on expression of <em>SET1/MLL</em> family members for survival. The growth of <em>ASH2L</em>-depleted glioblastoma cells was markedly slower than controls in orthotopic in vivo models. TCGA analysis showed high ASH2L expression in glioblastoma compared to low grade gliomas and immunohistochemical analysis revealed significant ASH2L expression in glioblastoma tissues, attesting to its clinical relevance. Therefore, high throughput, robust and affordable screens with focused libraries, such as EpiDoKOL, holds great promise to enable rapid discovery of novel epigenetic regulators of cancer cell survival, such as <em>ASH2L</em>.</p> <p><strong>Conclusion:</strong> Together, we suggest that targeting <em>ASH2L</em> could serve as a new therapeutic opportunity for glioblastoma.</p> |
| spellingShingle | Ozyerli-Goknar, E Kala, EY Aksu, AC Bulut, I Cingöz, A Nizamuddin, S Biniossek, M Seker-Polat, F Morova, T Aztekin, C Kung, SHY Syed, H Tuncbag, N Gönen, M Philpott, M Cribbs, AP Acilan, C Lack, NA Onder, TT Timmers, HTM Bagci-Onder, T Epigenetic-focused CRISPR/Cas9 screen identifies (absent, small, or homeotic)2-like protein (ASH2L) as a regulator of glioblastoma cell survival |
| title | Epigenetic-focused CRISPR/Cas9 screen identifies (absent, small, or homeotic)2-like protein (ASH2L) as a regulator of glioblastoma cell survival |
| title_full | Epigenetic-focused CRISPR/Cas9 screen identifies (absent, small, or homeotic)2-like protein (ASH2L) as a regulator of glioblastoma cell survival |
| title_fullStr | Epigenetic-focused CRISPR/Cas9 screen identifies (absent, small, or homeotic)2-like protein (ASH2L) as a regulator of glioblastoma cell survival |
| title_full_unstemmed | Epigenetic-focused CRISPR/Cas9 screen identifies (absent, small, or homeotic)2-like protein (ASH2L) as a regulator of glioblastoma cell survival |
| title_short | Epigenetic-focused CRISPR/Cas9 screen identifies (absent, small, or homeotic)2-like protein (ASH2L) as a regulator of glioblastoma cell survival |
| title_sort | epigenetic focused crispr cas9 screen identifies absent small or homeotic 2 like protein ash2l as a regulator of glioblastoma cell survival |
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