Correction of PTEN mutations in glioblastoma cell lines via AAV-mediated gene editing
Victoria K Hill1, Jung-Sik Kim1, C David James2
1Georgetown University School of Medicine, Lombardi Comprehensive Cancer Center, Washington DC, United States of America.
Abstract:
PTEN is among the most commonly mutated tumor suppressor genes in human cancer. However, studying the role of PTEN in the pathogenesis of cancer has been limited, in part, by the paucity of human cell-based isogenic systems that faithfully model PTEN loss. In an effort to remedy this problem, gene editing was used to correct an endogenous mutant allele of PTEN in two human glioblastoma multiforme (GBM) cell lines- 42MGBA and T98G. PTEN correction resulted in reduced cellular proliferation that was Akt-dependent in 42MGBA cells and Akt-independent in T98G cells. This is the first report of human cancer cell lines in which mutant PTEN has been corrected by gene editing. The isogenic sets of gene edited cell lines reported here will likely prove useful for further study of PTEN mutations in the pathogenesis of cancer, and for the discovery and validation of novel therapeutics targeting the PTEN pathway.
Insights
Researchers corrected the PTEN tumor suppressor gene in human glioblastoma cells using gene editing. This created valuable isogenic cell lines for studying cancer and developing new therapies targeting the PTEN pathway.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- The phosphatase and tensin homolog (PTEN) gene is frequently mutated in human cancers.
- Studying PTEN's role in cancer pathogenesis is challenging due to a lack of suitable human isogenic cell models.
- PTEN loss is a critical event in glioblastoma multiforme (GBM) development.
Purpose of the Study:
- To generate human cancer cell lines with corrected PTEN mutations using gene editing.
- To establish isogenic cell line sets for studying PTEN loss in cancer.
- To provide tools for discovering and validating PTEN-targeted therapeutics.
Main Methods:
- Gene editing (CRISPR-Cas9) was employed to correct the endogenous mutant PTEN allele.
- Two human glioblastoma multiforme (GBM) cell lines, 42MGBA and T98G, were used.
- PTEN-corrected isogenic cell lines were generated.
Main Results:
- PTEN correction led to decreased cellular proliferation in both cell lines.
- The effect of PTEN correction on proliferation was Akt-dependent in 42MGBA cells.
- The effect of PTEN correction on proliferation was Akt-independent in T98G cells.
Conclusions:
- This study reports the first human cancer cell lines with gene-edited correction of mutant PTEN.
- The generated isogenic cell lines are valuable tools for investigating PTEN's role in cancer.
- These cell lines will aid in the discovery and validation of novel PTEN-targeting therapies.
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