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Lentiviral Vector-mediated Gene Therapy of Hepatocytes Ex Vivo for Autologous Transplantation in Swine
Published on: November 4, 2018
Hepatoma-specific gene therapy through retrovirus-mediated and targeted gene transfer using an adenovirus carrying
1Department of Internal Medicine II, Miyazaki Medical College, 5200 Kihara, Kiyotake, Miyazaki, 889-1692, Japan.
Abstract:
Tissue-specific transcriptional regulatory sequences have been inserted in retrovirus vectors for therapeutic gene expression in cancer gene therapy. However, the transcriptional activity of these sequences is generally low, and the proviral DNA integration appears to increase the possibility of genomic DNA alteration in nontarget cells. Therefore, retrovirus-mediated targeted gene transduction into human hepatoma cells was evaluated using transient expression of an ecotropic receptor gene, mouse cationic amino acid transporter-1 (MCAT-1). Two recombinant adenoviruses, AxCAMCAT and AxAFMCAT, carrying the MCAT-1 gene under the control of the CAG and human alpha-fetoprotein (AFP) promoter, respectively, were generated. The preinfection with AxCAMCAT allowed highly efficient ecotropic retroviral infection of human cells. In addition, after AxAFMCAT infection, retroviral infection occurred only in AFP-producing hepatoma cells, resulting in selective cytotoxicity induced by the herpes simplex virus thymidine kinase (HSV-tk)/ganciclovir (GCV) system. Transient expression of the MCAT-1 gene under the control of the AFP promoter permits ecotropic retrovirus-mediated gene transduction specifically in AFP-producing human hepatoma cells, resulting in selective induction of the suicide killing effect while the therapeutic gene is driven from ubiquitously expressed promoters.
Insights
This study introduces a novel method for targeted gene therapy in liver cancer. By using adenoviruses to deliver a receptor gene, researchers achieved specific retroviral gene delivery to hepatoma cells, enabling selective cancer cell killing.
Area of Science:
- Molecular Biology
- Gene Therapy
- Oncology
Background:
- Retrovirus vectors are used for cancer gene therapy but often have low transcriptional activity and risk off-target genomic alterations.
- Targeted gene delivery to cancer cells remains a challenge in developing effective gene therapies.
Purpose of the Study:
- To evaluate retrovirus-mediated targeted gene transduction into human hepatoma cells.
- To enhance the efficiency and specificity of gene delivery for cancer gene therapy.
Main Methods:
- Generated two recombinant adenoviruses, AxCAMCAT and AxAFMCAT, to express the mouse cationic amino acid transporter-1 (MCAT-1) receptor gene.
- Utilized the CAG and human alpha-fetoprotein (AFP) promoters to control MCAT-1 expression.
- Employed the herpes simplex virus thymidine kinase (HSV-tk)/ganciclovir (GCV) suicide system for selective cancer cell killing.
Main Results:
- Preinfection with AxCAMCAT enabled highly efficient ecotropic retroviral infection of human cells.
- AxAFMCAT infection led to retroviral infection exclusively in AFP-producing hepatoma cells.
- This targeted delivery resulted in selective cytotoxicity in hepatoma cells via the HSV-tk/GCV system.
Conclusions:
- Transient expression of MCAT-1 under the AFP promoter allows specific retroviral gene transduction in AFP-producing hepatoma cells.
- This approach enables targeted induction of a suicide killing effect in cancer cells.
- The strategy offers a promising method for selective cancer gene therapy, overcoming limitations of previous retroviral vectors.
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