A more efficient method for generating glioblastoma-multiforme model in mice using genome editing technology

Area of Science:

• Neuroscience
• Oncology
• Genetics

Background:

• Developing accurate glioblastoma multiforme mouse models is crucial for understanding malignant glioma properties and therapeutic development.
• Previous methods utilized p53 knockout mice to induce glioma-initiating cells (GICs) via HRasL61 overexpression in neural stem/progenitor cells (NSCs).
• Orthotopic transplantation of these induced GICs resulted in glioblastoma multiforme-like tumors in immunodeficient mice.

Purpose of the Study:

• To establish an efficient method for inducing glioma-initiating cells (GICs) from wild-type fetal neural stem cells (NSCs).
• To create a more effective mouse model for studying glioblastoma multiforme.
• To compare the efficiency of CRISPR/Cas9-based induction versus knockout mice-based induction of GICs.

Main Methods:

• Utilized CRISPR/Cas9 technology to generate p53 knockout (KO) neural stem cells (NSCs).
• Overexpressed active HRas (HRasL61) in the p53 KO NSCs.
• Cloned the resulting p53-/HRasL61+ cells and orthotopically transplanted them into immunodeficient mice.

Main Results:

• Successfully induced glioma-initiating cells (GICs) from wild-type fetal NSCs using the CRISPR/Cas9 method.
• Transplanted p53-/HRasL61+ cells formed glioblastoma multiforme-like tumors in mice.
• Strong expression of GIC markers was observed in the induced p53-/HRasL61+ cells, confirming their GIC nature.

Conclusions:

• The CRISPR/Cas9-based method for inducing GICs is significantly more efficient than methods relying on knockout mice.
• The p53-/HRasL61+ cell line represents a validated induced GIC.
• This study provides a valuable and efficient framework for generating glioblastoma mouse models for research.