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Identifying novel therapeutic targets in gastric cancer using genome-wide CRISPR-Cas9 screening
Zhi Zeng1, Xu Zhang2, Cong-Qing Jiang3
1Department of Pathology, Renmin Hospital of Wuhan University, Wuhan, Hubei, China.
Abstract:
Genome-scale CRISPR-Cas9 screening technology is a powerful tool to systematically identify genes essential for cancer cell survival. Herein, TKOv3, a genome-scale CRISPR-Cas9 knock-out library, was screened in the gastric cancer (GC) cells, and relevant validation experiments were performed. We obtained 854 essential genes for the AGS cell line, and 184 were novel essential genes. After knocking down essential genes: SPC25, DHX37, ABCE1, SNRPB, TOP3A, RUVBL1, CIT, TACC3 and MTBP, cell viability and proliferation were significantly decreased. Then, we analysed the detected essential genes at different time points and proved more characteristic genes might appear with the extension of selection. After progressive selection using a series of open datasets, 41 essential genes were identified as potential drug targets. Among them, methyltransferase 1 (METTL1) was over expressed in GC tissues. High METTL1 expression was associated with poor prognosis among 3 of 6 GC cohorts. Furthermore, GC cells growth was significantly inhibited after the down-regulation of METTL1 in vitro and in vivo. Function analysis revealed that METTL1 might play a role in the cell cycle through AKT/STAT3 pathways. In conclusion, compared with existing genome-scale screenings, we obtained 184 novel essential genes. Among them, METTL1 was validated as a potential therapeutic target of GC.
Insights
This study used CRISPR-Cas9 screening to find essential genes in gastric cancer cells, identifying 184 novel genes. Methyltransferase 1 (METTL1) was validated as a potential therapeutic target for gastric cancer.
Area of Science:
- Genomics
- Cancer Biology
- Molecular Oncology
Background:
- Genome-scale CRISPR-Cas9 screening is vital for identifying genes crucial for cancer cell survival.
- Gastric cancer (GC) remains a significant health challenge, necessitating novel therapeutic targets.
Purpose of the Study:
- To systematically identify essential genes in gastric cancer cells using a genome-scale CRISPR-Cas9 knockout library.
- To validate novel essential genes and potential therapeutic targets for gastric cancer.
Main Methods:
- Utilized the TKOv3 genome-scale CRISPR-Cas9 knockout library for screening in gastric cancer cells.
- Performed validation experiments, including gene knockdown, and analyzed essential genes at different time points.
- Integrated open datasets for progressive selection of potential drug targets.
Main Results:
- Identified 854 essential genes in AGS gastric cancer cells, including 184 novel genes.
- Knockdown of specific genes (e.g., SPC25, DHX37, METTL1) significantly reduced cell viability and proliferation.
- Methyltransferase 1 (METTL1) was found to be overexpressed in GC tissues, associated with poor prognosis, and its downregulation inhibited GC cell growth in vitro and in vivo.
- Functional analysis suggested METTL1's role in the cell cycle via AKT/STAT3 pathways.
Conclusions:
- This screening identified 184 novel essential genes in gastric cancer, expanding the understanding of GC biology.
- Methyltransferase 1 (METTL1) is a promising therapeutic target for gastric cancer, with potential roles in cell cycle regulation.
