Oncolytic virus-mediated base editing for targeted killing of cervical cancer cells.

Huanhuan XU; Siwei LI; Xi LUO; Zuping ZHOU; Changhao BI

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Oncolytic virus-mediated base editing for targeted killing of cervical cancer cells.

Author: Huanhuan XU ¹ ; Siwei LI ² ; Xi LUO ¹ ; Zuping ZHOU ¹ ; Changhao BI ²
Author Information

1. Key Laboratory of Stem Cells and Pharmaceutical Biotechnology of Guangxi Universities, College of Life Sciences, Guangxi Normal University, Guilin 541004, Guangxi, China.
2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300307, China.
Publication Type:Journal Article
Keywords: base editor; cervical cancer; oncogene; oncolytic virus
MeSH: Humans; Uterine Cervical Neoplasms/pathology*; Oncolytic Viruses/genetics*; Female; HEK293 Cells; Oncolytic Virotherapy/methods*; Adenoviridae/genetics*; Gene Editing/methods*; Telomerase/genetics*; Adenovirus E1A Proteins/genetics*; Genetic Vectors/genetics*; HeLa Cells
From: Chinese Journal of Biotechnology 2025;41(4):1382-1394
CountryChina
Language:Chinese
Abstract: Conventional cancer therapies, such as radiotherapy and chemotherapy, often damage normal cells and may induce new tumors. Oncolytic viruses (OVs) selectively target tumor cells while sparing normal cells. Most OVs used in clinical trials have been genetically engineered to enhance their ability to target tumor cells and activate immune responses. To develop a specific OV-based approach for treating cervical cancer, this study constructed an oncolytic adenovirus that delivered a base editor targeting oncogenes to achieve efficient killing of tumor cells through inhibiting tumor growth and directly lysing tumor cells. We utilized the human telomerase reverse transcriptase (TERT) promoter to drive the expression of adenovirus early region 1A (E1A) and successfully constructed the P-hTERT-E1A-GFP vector, which was validated for its activity in cervical cancer cells. Given the critical role of the MYC oncogene in the research of oncology, identifying efficient editing sites for the MYC oncogene is a key step in this study.Three MYC-targeting gRNAs were engineered and co-delivered with ABE8e base editor plasmids into HEK293T cells. Following puromycin selection, Sanger sequencing demonstrated differential editing efficiencies: MYC-1 (43%), MYC-2 (25%), and MYC-3 (35%), identifying MYC-1 as the most efficient editing locus. By constructing the P-ABEs-hTERT-E1A-GFP and P-MYC gRNA-hTERT-E1A-GFP vectors, we successfully packaged the virus and confirmed its specificity and efficacy. The experimental results demonstrate that this novel oncolytic adenovirus effectively inhibits the growth of HeLa cells in vitro, providing new experimental evidence and potential strategies for treating cervical cancer based on the HeLa cell model.