CRISPR-Cas9 knockout screen identifies novel treatment targets in childhood high-grade glioma
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Published on: June 14, 2018
Anna Wenger1, Ida Karlsson1, Teresia Kling1
1Sahlgrenska Center for Cancer Research, Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Medicinaregatan 1F, 405 30, Gothenburg, Sweden.
Background:
Brain tumours are the leading cause of cancer-related death in children, and there is no effective treatment. A growing body of evidence points to deregulated epigenetics as a tumour driver, particularly in paediatric cancers as they have relatively few genomic alterations, and key driver mutations have been identified in histone 3 (H3). Cancer stem cells (CSC) are implicated in tumour development, relapse and therapy resistance and thus particularly important to target. We therefore aimed to identify novel epigenetic treatment targets in CSC derived from H3-mutated high-grade glioma (HGG) through a CRISPR-Cas9 knockout screen.
Results:
The knockout screen identified more than 100 novel genes essential for the growth of CSC derived from paediatric HGG with H3K27M mutation. We successfully validated 12 of the 13 selected hits by individual knockout in the same two CSC lines, and for the top six hits we included two additional CSC lines derived from H3 wild-type paediatric HGG. Knockout of these genes led to a significant decrease in CSC growth, and altered stem cell and differentiation markers.
Conclusions:
The screen robustly identified essential genes known in the literature, but also many novel genes essential for CSC growth in paediatric HGG. Six of the novel genes (UBE2N, CHD4, LSM11, KANSL1, KANSL3 and EED) were validated individually thus demonstrating their importance for CSC growth in H3-mutated and wild-type HGG. These genes should be further studied and evaluated as novel treatment targets in paediatric HGG.
Insights
Researchers identified novel epigenetic targets in pediatric brain tumors by screening cancer stem cells. Six key genes were validated, offering potential new treatments for high-grade glioma.
Area of Science:
- Oncology
- Epigenetics
- Pediatric Cancer Research
Background:
- Brain tumors are the leading cause of cancer death in children, with limited treatment options.
- Epigenetic dysregulation is a key driver in pediatric cancers, particularly those with histone 3 (H3) mutations.
- Cancer stem cells (CSCs) are crucial in tumor development, relapse, and therapy resistance, necessitating targeted approaches.
Purpose of the Study:
- To identify novel epigenetic treatment targets in pediatric high-grade glioma (HGG) cancer stem cells (CSCs).
- To utilize a CRISPR-Cas9 knockout screen to discover genes essential for CSC growth.
Main Methods:
- Conducted a CRISPR-Cas9 knockout screen in CSCs derived from H3-mutated pediatric HGG.
- Validated essential genes through individual knockouts in multiple CSC lines (H3-mutated and wild-type).
- Assessed the impact of gene knockouts on CSC growth and differentiation markers.
Main Results:
- The screen identified over 100 novel genes critical for CSC growth in pediatric HGG.
- Twelve of thirteen selected genes were validated, with six novel genes (UBE2N, CHD4, LSM11, KANSL1, KANSL3, EED) showing significant impact.
- Knockout of these validated genes markedly reduced CSC proliferation and altered stem cell markers.
Conclusions:
- The study successfully identified novel genes essential for pediatric HGG CSC growth.
- Six validated genes represent promising therapeutic targets for both H3-mutated and wild-type pediatric HGG.
- Further research into these genes is warranted for developing new treatment strategies.
