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Genetic switch selectively kills hepatocellular carcinoma cell based on microRNA and tissue-specific promoter
Yuan-Yuan Lu1, Yi Li2, Zhi-Li Chen3
1Institutes of Physical Science and Information Technology, Anhui University, Hefei, 230000, China; Academy of Military Medical Sciences, Beijing, 100850, China.
Abstract:
The clinical treatment of hepatocellular carcinoma (HCC) is still a heavy burden worldwide. Intracellular microRNAs (miRNAs) commonly express abnormally in cancers, thus they are potential therapeutic targets for cancer treatment. miR-21 is upregulated in HCC whereas miR-122 is enriched in normal hepatocyte but downregulated in HCC. In our study, we first generated a reporter genetic switch compromising of miR-21 and miR-122 sponges as sensor, green fluorescent protein (GFP) as reporter gene and L7Ae:K-turn as regulatory element. The reporter expression was turned up in miR-21 enriched environment while turned down in miR-122 enriched environment, indicating that the reporter switch is able to respond distinctly to different miRNA environment. Furthermore, an AAT promoter, which is hepatocyte-specific, is applied to increase the specificity to hepatocyte. A killing switch with AAT promoter and an apoptosis-inducing element, Bax, in addition to miR-21 and miR-122 significantly inhibited cell viability in Huh-7 by 70 % and in HepG2 by 60 %. By contrast, cell viability was not affected in five non-HCC cells. Thus, we provide a novel feasible strategy to improve the safety of miRNA-based therapeutic agent to cancer.
Insights
This study developed a novel genetic switch for hepatocellular carcinoma (HCC) therapy. The system precisely targets cancer cells, significantly reducing viability while sparing healthy cells, improving miRNA-based treatment safety.
Area of Science:
- Biotechnology
- Molecular Biology
- Oncology
Background:
- Hepatocellular carcinoma (HCC) presents a significant global health challenge.
- Intracellular microRNAs (miRNAs) show altered expression in cancers, offering therapeutic targets.
- miR-21 is elevated in HCC, while miR-122 is decreased, presenting a therapeutic opportunity.
Purpose of the Study:
- To engineer a novel miRNA-responsive genetic switch for targeted HCC therapy.
- To enhance the specificity and safety of miRNA-based cancer treatments.
- To develop a system that can distinguish between HCC and normal liver cells based on miRNA profiles.
Main Methods:
- Constructed a reporter genetic switch using miRNA sponges, a reporter gene (GFP), and a regulatory element (L7Ae:K-turn).
- Incorporated a hepatocyte-specific promoter (AAT) for targeted delivery.
- Integrated an apoptosis-inducing element (Bax) to create a therapeutic killing switch.
Main Results:
- The reporter switch demonstrated distinct responses to varying miRNA environments (miR-21 vs. miR-122).
- The AAT promoter ensured hepatocyte specificity.
- The therapeutic system significantly inhibited HCC cell viability (Huh-7 by 70%, HepG2 by 60%) with no effect on non-HCC cells.
Conclusions:
- A novel, feasible strategy for enhancing the safety of miRNA-based cancer therapeutics was developed.
- The engineered system shows high specificity for HCC cells, minimizing off-target effects.
- This approach holds promise for improving the clinical treatment of hepatocellular carcinoma.
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