Oktriani, Risky and Pirona, Anna Chiara and Kalmár, Lili and Rahadian, Ariani S. and Miao, Beiping and Bauer, Andrea S. and Hoheisel, Jörg D. and Boettcher, Michael and Du, Haoqi (2024) Genome-Wide CRISPR Screen Identifies Genes Involved in Metastasis of Pancreatic Ductal Adenocarcinoma. Cancers, 16 (21). p. 3684. ISSN 20726694
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Genome-Wide CRISPR Screen Identifies Genes Involved in Metastasis of pancreatic Ductal Adenocarcinoma.pdf - Published Version
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Abstract
Background/Objectives: Early and aggressive metastasis is a major feature of pancreatic ductal adenocarcinoma. Understanding the processes underlying metastasis is crucial for making a difference to disease outcome. Towards these ends, we looked in a comprehensive manner for genes that are metastasis-specific. Methods: A genome-wide CRISPR-Cas9 gene knockout screen with 259,900 single guide RNA constructs was performed on pancreatic cancer cell lines with very high or very low metastatic capacity, respectively. Functional aspects of some of the identified genes were analysed in vitro. The injection of tumour cells with or without a gene knockout into mice was used to confirm the effect on metastasis. Results: The knockout of 590 genes—and, with higher analysis stringency, 67 genes—affected the viability of metastatic cells substantially, while these genes were not vital to non-metastasizing cells. Further evaluations identified different molecular processes related to this observation. One of the genes was MYBL2, encoding for a well-known transcription factor involved in the regulation of cell survival, proliferation, and differentiation in cancer tissues. In our metastasis-focussed study, no novel functional activity was detected for MYBL2, however. Instead, a metastasis-specific transformation of its genetic interaction with FOXM1 was observed. The interaction was synergistic in cells of low metastatic capacity, while there was a strong switch to a buffering mode in metastatic cells. In vivo analyses confirmed the strong effect of MYBL2 on metastasis. Conclusions: The genes found to be critical for the viability of metastatic cells form a basis for further investigations of the processes responsible for triggering and driving metastasis. As shown for MYBL2, unexpected processes of regulating metastasis might also be involved. © 2024 by the authors.
Item Type: | Article |
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Additional Information: | Cited by: 1; All Open Access, Gold Open Access |
Uncontrolled Keywords: | protein Myb; puromycin; transcription factor; Article; cell invasion; cell viability; cell viability assay; clustered regularly interspaced short palindromic repeat; colony formation; controlled study; CRISPR-CAS9 system; flow cytometry; gene interaction; gene knockout; gene sequence; genetic transduction; HEK293T cell line; human; human cell; immunoassay; immunohistochemistry; in vitro study; KEGG; metastasis; pancreas cancer; pancreatic cancer cell line; pancreatic ductal carcinoma; quality control; s2-007 cell line; spliceosome; Western blotting |
Subjects: | R Medicine > RN Non Surgical Divisions R Medicine > RC Internal medicine > RC0254 Neoplasms. Tumors. Oncology (including Cancer) |
Divisions: | Faculty of Medicine, Public Health and Nursing > Non Surgical Divisions |
Depositing User: | Ani PURWANDARI |
Date Deposited: | 16 Apr 2025 01:57 |
Last Modified: | 16 Apr 2025 01:57 |
URI: | https://ir.lib.ugm.ac.id/id/eprint/16052 |