Safety of Adeno-associated virus-based vector-mediated gene therapy—impact of vector dose

Background:: Lung cancer remains the most common cause of cancer-related deaths in China and worldwide. Traditional surgery and chemotherapy do not offer an effective cure although gene therapy may be a promising future alter-native. Kallistatin (Kal) is an endogenous inhibitor of angiogenesis and tumorigenesis. Recombinant adeno-associated virus (rAAV) is considered the most promising vector for gene therapy of many diseases due to persistent and long-term transgen-ic expression. Objective:: The aim of this study was to investigate whether rAAV9-Kal inhibited NCI-H446 subcutaneous xenograft tumor growth in mice. Method:: The subcutaneous xenograft mode were induced by subcutaneous injection of 2×106 H446 cells into the dorsal skin of BALB/c nude mice. The mice were administered with ssrAAV9-Kal (single-stranded rAAV9) or dsrAAV9-Kal (double-stranded rAAV9）by intraperitoneal injection (I.P.). Tumor microvessel density (MVD) was examined by anti-CD34 stain-ing to evaluate tumor angiogenesis. Results:: Compared with the PBS (blank control) group, tumor growth in the high-dose ssrAAV9-Kal group was inhibited by 40% by day 49, and the MVD of tumor tissues was significantly decreased. Conclusion:: The results indicate that this therapeutic strategy is a promising approach for clinical cancer therapy and impli-cate rAAV9-Kal as a candidate for gene therapy of lung cancer.

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Adeno-Associated Virus (AAV)-Mediated Gene Therapy for Disorders of Inherited and Non-Inherited Origin

In Vivo and Ex Vivo Gene Therapy for Inherited and Non-Inherited Disorders ◽

10.5772/intechopen.80317 ◽

2019 ◽

Author(s):

Indu Rajapaksha ◽

Peter Angus ◽

Chandana Herath

Keyword(s):

Gene Therapy ◽

Adeno Associated Virus

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Quantitation of Trace Levels of DNA Released from Disrupted Adeno-Associated Virus Gene Therapy Vectors

Journal of Pharmaceutical Sciences ◽

10.1016/j.xphs.2021.06.010 ◽

2021 ◽

Author(s):

Jared S. Bee ◽

Kristin O'Berry ◽

Yu (Zoe) Zhang ◽

Megan Kuhn Phillippi ◽

Akanksha Kaushal ◽

...

Keyword(s):

Gene Therapy ◽

Adeno Associated Virus ◽

Virus Gene ◽

Gene Therapy Vectors

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Bombyx mori Pupae Efficiently Produce Recombinant AAV2/HBoV1 Vectors with a Bombyx mori Nuclear Polyhedrosis Virus Expression System

Viruses ◽

10.3390/v13040704 ◽

2021 ◽

Vol 13 (4) ◽

pp. 704

Author(s):

Qian Yu ◽

Pengfei Chang ◽

Xiaoxuan Liu ◽

Peng Lü ◽

Qi Tang ◽

...

Keyword(s):

Gene Therapy ◽

Bombyx Mori ◽

Nuclear Polyhedrosis Virus ◽

Expression System ◽

Efficient Production ◽

Human Bocavirus ◽

Adeno Associated Virus ◽

Silkworm Pupae ◽

Polyhedrosis Virus ◽

Nuclear Polyhedrosis

Recombinant adeno-associated virus (AAV) vectors have broad application prospects in the field of gene therapy. The establishment of low-cost and large-scale manufacturing is now the general agenda for industry. The baculovirus-insect cell/larva expression system has great potential for these applications due to its scalability and predictable biosafety. To establish a more efficient production system, Bombyx mori pupae were used as a new platform and infected with recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV). The production of a chimeric recombinant adeno-associated virus (rAAV) serotype 2/human bocavirus type-1 (HBoV1) vector was used to evaluate the efficiency of this new baculovirus expression vector (BEV)–insect expression system. For this purpose, we constructed two recombinant BmNPVs, which were named rBmNPV/AAV2Rep-HBoV1Cap and rBmNPV/AAV2ITR-eGFP. The yields of rAAV2/HBoV1 derived from the rBmNPV/AAV2Rep-HBoV1Cap and rBmNPV/AAV2ITR-eGFP co-infected BmN cells exceeded 2 × 104 vector genomes (VG) per cell. The rBmNPV/AAV2Rep-HBoV1Cap and rBmNPV/AAV2ITR-eGFP can express stably for at least five passages. Significantly, rAAV2/HBoV1 could be efficiently generated from BmNPV-infected silkworm larvae and pupae at average yields of 2.52 × 1012 VG/larva and 4.6 × 1012 VG/pupa, respectively. However, the vectors produced from the larvae and pupae had a high percentage of empty particles, which suggests that further optimization is required for this platform in the future. Our work shows that silkworm pupae, as an efficient bioreactor, have great potential for application in the production of gene therapy vectors.

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Next Step in Gene Delivery: Modern Approaches and Further Perspectives of AAV Tropism Modification

Pharmaceutics ◽

10.3390/pharmaceutics13050750 ◽

2021 ◽

Vol 13 (5) ◽

pp. 750

Author(s):

Maxim A. Korneyenkov ◽

Andrey A. Zamyatnin

Keyword(s):

Gene Therapy ◽

Genome Organization ◽

Rational Design ◽

Structural Data ◽

Rational Approach ◽

Tissue Tropism ◽

Adeno Associated Virus ◽

Chemical Conjugation ◽

The Usa ◽

Therapy Delivery

Today, adeno-associated virus (AAV) is an extremely popular choice for gene therapy delivery. The safety profile and simplicity of the genome organization are the decisive advantages which allow us to claim that AAV is currently among the most promising vectors. Several drugs based on AAV have been approved in the USA and Europe, but AAV serotypes’ unspecific tissue tropism is still a serious limitation. In recent decades, several techniques have been developed to overcome this barrier, such as the rational design, directed evolution and chemical conjugation of targeting molecules with a capsid. Today, all of the abovementioned approaches confer the possibility to produce AAV capsids with tailored tropism, but recent data indicate that a better understanding of AAV biology and the growth of structural data may theoretically constitute a rational approach to most effectively produce highly selective and targeted AAV capsids. However, while we are still far from this goal, other approaches are still in play, despite their drawbacks and limitations.

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