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Hepatic DR5 induces apoptosis and limits adenovirus gene therapy product expression in the liver
Huang-Ge Zhang1, Jinfu Xie, Liang Xu
1Division of Clinical Immunology and Rheumatology, Department of Medicine, The University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
Abstract:
A major limitation of adenovirus (Ad) gene therapy product expression in the liver is subsequent elimination of the hepatocytes expressing the gene therapy product. This elimination is caused by both necrosis and apoptosis related to the innate and cell-mediated immune response to the Ad. Apoptosis of hepatocytes can be induced by the innate immune response by signaling through death domain receptors on hepatocytes including the tumor necrosis factor alpha (TNF-alpha) receptor (TNFR), Fas, and death domain receptors DR4 and DR5. We have previously shown that blocking signaling through TNFR enhances and prolongs gene therapy product expression in the liver. In the present study, we constructed an Ad that produces a soluble DR5-Fc (AdsDR5), which is capable of neutralizing TNF-related apoptosis-inducing ligand (TRAIL). AdsDR5 prevents TRAIL-mediated apoptosis of CD3-activated T cells and decreases hepatocyte apoptosis after AdCMVLacZ administration and enhances the level and duration of lacZ transgene expression in the liver. In addition to blocking TRAIL and directly inhibiting apoptosis, AdsDR5 decreases production of gamma interferon (IFN-gamma) and TNF-alpha and decreases NK cell activation, all of which limit Ad-mediated transgene expression in the liver. These results indicate that (i) AdsDR5 produces a DR5-Fc capable of neutralizing TRAIL, (ii) AdsDR5 can reduce activation of NK cells and reduce induction of IFN-gamma and TNF-alpha after Ad administration, and (iii) administration of AdsDR5 can enhance Ad gene therapy in the liver.
Insights
Adenovirus gene therapy faces limitations due to immune responses causing cell death. A novel adenovirus expressing soluble DR5-Fc (AdsDR5) neutralizes TRAIL, reducing cell death and enhancing gene therapy effectiveness in the liver.
Area of Science:
- Hepatocyte Biology
- Immunology
- Gene Therapy Vector Development
Background:
- Adenovirus (Ad) gene therapy in the liver is limited by immune-mediated elimination of transduced hepatocytes.
- Hepatocyte apoptosis, triggered by innate immune responses via death domain receptors like TNF-alpha receptor (TNFR) and DR5, contributes to this limitation.
- Previous work demonstrated that blocking TNFR signaling improves gene therapy outcomes.
Purpose of the Study:
- To construct and evaluate an adenovirus vector expressing a soluble DR5-Fc fusion protein (AdsDR5) designed to neutralize TNF-related apoptosis-inducing ligand (TRAIL).
- To assess the efficacy of AdsDR5 in preventing TRAIL-mediated apoptosis and enhancing transgene expression in the liver.
- To investigate the impact of AdsDR5 on immune responses, including NK cell activation and cytokine production (IFN-gamma, TNF-alpha).
Main Methods:
- Construction of an adenovirus vector (AdsDR5) encoding a soluble DR5-Fc fusion protein.
- Administration of AdsDR5 to assess its ability to neutralize TRAIL and prevent apoptosis.
- Evaluation of AdsDR5's effect on hepatocyte apoptosis following AdCMVLacZ administration.
- Measurement of transgene expression levels and duration in the liver.
- Assessment of immune cell activation (NK cells) and cytokine production (IFN-gamma, TNF-alpha).
Main Results:
- AdsDR5 successfully produced a soluble DR5-Fc protein capable of neutralizing TRAIL.
- AdsDR5 administration reduced hepatocyte apoptosis and prevented TRAIL-mediated apoptosis of T cells.
- AdsDR5 enhanced both the level and duration of lacZ transgene expression in the liver.
- AdsDR5 decreased the production of gamma interferon (IFN-gamma) and TNF-alpha and reduced NK cell activation.
Conclusions:
- AdsDR5 effectively neutralizes TRAIL, thereby inhibiting apoptosis.
- AdsDR5 mitigates key immune responses (NK cell activation, IFN-gamma, TNF-alpha production) that limit adenovirus-mediated gene expression.
- Administration of AdsDR5 represents a promising strategy to enhance adenovirus gene therapy efficacy in the liver.
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