Hepatocyte Heparan Sulfate Is Required for Adeno-Associated Virus 2 but Dispensable for Adenovirus 5 Liver Transduction In Vivo
| Autor: | Erin M. Foley, Harvey R. Herschman, Arthur Catapang, Roger Lawrence, Jeffrey D. Esko, Ramon Alemany, Hamidreza Hoveida, Patrick Secrest, Anne K. Zaiss, Dmitry M. Shayakhmetov, Yu Yamaguchi, Lina S. Schneider |
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| Přispěvatelé: | Banks, L |
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Male viruses medicine.disease_cause Medical and Health Sciences chemistry.chemical_compound Transduction (genetics) Mice Transduction Genetic Receptors Adeno-associated virus Cells Cultured Heparan Sulfate Biosynthesis Cultured Liver Disease Gene Therapy Heparan sulfate Biological Sciences Dependovirus Virus Liver Receptors Virus Female Development of treatments and therapeutic interventions Biotechnology Cells Immunology Genetic Vectors Biology Microbiology Adenoviridae Transduction 03 medical and health sciences Gene Delivery Genetic Viral entry In vivo Cell surface receptor Virology Genetics medicine Animals 5.2 Cellular and gene therapies Agricultural and Veterinary Sciences biochemical phenomena metabolism and nutrition Molecular biology carbohydrates (lipids) 030104 developmental biology chemistry Insect Science Hepatocytes Heparitin Sulfate Digestive Diseases |
| Zdroj: | Journal of virology, vol 90, iss 1 |
| Popis: | Adeno-associated virus 2 (AAV2) and adenovirus 5 (Ad5) are promising gene therapy vectors. Both display liver tropism and are currently thought to enter hepatocytes in vivo through cell surface heparan sulfate proteoglycans (HSPGs). To test directly this hypothesis, we created mice that lack Ext1 , an enzyme required for heparan sulfate biosynthesis, in hepatocytes. Ext1 HEP mutant mice exhibit an 8-fold reduction of heparan sulfate in primary hepatocytes and a 5-fold reduction of heparan sulfate in whole liver tissue. Conditional hepatocyte Ext1 gene deletion greatly reduced AAV2 liver transduction following intravenous injection. Ad5 transduction requires blood coagulation factor X (FX); FX binds to the Ad5 capsid hexon protein and bridges the virus to HSPGs on the cell surface. Ad5.FX transduction was abrogated in primary hepatocytes from Ext1 HEP mice. However, in contrast to the case with AAV2, Ad5 transduction was not significantly reduced in the livers of Ext1 HEP mice. FX remained essential for Ad5 transduction in vivo in Ext1 HEP mice. We conclude that while AAV2 requires HSPGs for entry into mouse hepatocytes, HSPGs are dispensable for Ad5 hepatocyte transduction in vivo . This study reopens the question of how adenovirus enters cells in vivo . IMPORTANCE Our understanding of how viruses enter cells, and how they can be used as therapeutic vectors to manage disease, begins with identification of the cell surface receptors to which viruses bind and which mediate viral entry. Both adeno-associated virus 2 and adenovirus 5 are currently thought to enter hepatocytes in vivo through heparan sulfate proteoglycans (HSPGs). However, direct evidence for these conclusions is lacking. Experiments presented herein, in which hepatic heparan sulfate synthesis was genetically abolished, demonstrated that HSPGs are not likely to function as hepatocyte Ad5 receptors in vivo . The data also demonstrate that HSPGs are required for hepatocyte transduction by AAV2. These results reopen the question of the identity of the Ad5 receptor in vivo and emphasize the necessity of demonstrating the nature of the receptor by genetic means, both for understanding Ad5 entry into cells in vivo and for optimization of Ad5 vectors as therapeutic agents. |
| Databáze: | OpenAIRE |
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