471. In Vivo Potency Assay for AAV-Based Gene Therapy Vectors

Adeno-Associated Virus is the leading vector for gene therapy. Although it is the vector for all in vivo gene therapies approved for clinical use by the US Food and Drug Administration, its biology is still not yet fully understood. It has been shown that different serotypes of AAV bind to their cellular receptor, AAVR, in different ways. Previously we have reported a 2.4Å structure of AAV2 bound to AAVR that shows ordered structure for only one of the two AAVR domains with which AAV2 interacts. In this study we present a 2.5Å resolution structure of AAV5 bound to AAVR. AAV5 binds to the first polycystic kidney disease (PKD) domain of AAVR that was not ordered in the AAV2 structure. Interactions of AAV5 with AAVR are analyzed in detail, and the implications for AAV2 binding are explored through molecular modeling. Moreover, we find that binding sites for the antibodies ADK5a, ADK5b, and 3C5 on AAV5 overlap with the binding site of AAVR. These insights provide a structural foundation for development of gene therapy agents to better evade immune neutralization without disrupting cellular entry.

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181. Optimization of In Vivo Delivery of Baculovirus Gene Therapy Vectors for the Treatment of Alpha-1 Antitrypsin Deficiency

Molecular Therapy ◽

10.1016/s1525-0016(16)32990-2 ◽

2016 ◽

Vol 24 ◽

pp. S71

Author(s):

Laura vanLieshout ◽

Jondavid de Jong ◽

Jacob vanVloten ◽

Sarah K. Wootton

Keyword(s):

Gene Therapy ◽

Baculovirus Gene ◽

Gene Therapy Vectors ◽

Antitrypsin Deficiency ◽

Alpha 1 Antitrypsin Deficiency ◽

Alpha 1 Antitrypsin ◽

In Vivo Delivery

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542. New Amplifier Gene Therapy Vectors: Higher Levels of Gene Expression In Vitro and Improved Therapeutic Outcomes In Vivo

Molecular Therapy ◽

10.1016/s1525-0016(16)40984-6 ◽

2003 ◽

Vol 7 (5) ◽

pp. S211

Keyword(s):

Gene Expression ◽

Gene Therapy ◽

Therapeutic Outcomes ◽

Gene Therapy Vectors

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1040. A Direct Comparison of Two Non-Viral Gene Therapy Vectors for Somatic Integration: In Vivo Evaluation of the Bacteriophage Integrase PhiC31 and the Sleeping Beauty Transposase

Molecular Therapy ◽

10.1016/j.ymthe.2004.06.985 ◽

2004 ◽

Vol 9 ◽

pp. S398-S399

Keyword(s):

Gene Therapy ◽

Sleeping Beauty ◽

Viral Gene ◽

In Vivo Evaluation ◽

Viral Gene Therapy ◽

Gene Therapy Vectors

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In Vivo Evaluation of Gene Therapy Vectors in Ex Vivo-Derived Marrow Stromal Cells for Bone Regeneration in a Rat Critical-Size Calvarial Defect Model

Human Gene Therapy ◽

10.1089/104303403322611719 ◽

2003 ◽

Vol 14 (18) ◽

pp. 1689-1701 ◽

Cited By ~ 66

Author(s):

Jeremy S. Blum ◽

Michael A. Barry ◽

Antonios G. Mikos ◽

John A. Jansen

Keyword(s):

Gene Therapy ◽

Stromal Cells ◽

Critical Size ◽

Ex Vivo ◽

Marrow Stromal Cells ◽

Calvarial Defect ◽

Defect Model ◽

In Vivo Evaluation ◽

Gene Therapy Vectors

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153. Adenoviral Gene Therapy Vectors Expressing the Adenovirus Death Protein Demonstrate Increased Transgene Expression In Vivo as Demonstrated by Dose-Volume Histogram Analysis of Reporter Gene Activity

Molecular Therapy ◽

10.1016/j.ymthe.2005.06.157 ◽

2005 ◽

Vol 11 ◽

pp. S61-S62

Keyword(s):

Gene Therapy ◽

Reporter Gene ◽

Transgene Expression ◽

Histogram Analysis ◽

Dose Volume Histogram ◽

Reporter Gene Activity ◽

Dose Volume ◽

Gene Therapy Vectors ◽

Adenoviral Gene Therapy

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Impact of Medium-Sized Extracellular Vesicles on the Transduction Efficiency of Adeno-Associated Viruses in Neuronal and Primary Astrocyte Cell Cultures

International Journal of Molecular Sciences ◽

10.3390/ijms22084221 ◽

2021 ◽

Vol 22 (8) ◽

pp. 4221

Author(s):

Orsolya Tünde Kovács ◽

Eszter Soltész-Katona ◽

Nikolett Marton ◽

Eszter Baricza ◽

László Hunyady ◽

...

Keyword(s):

Gene Therapy ◽

Extracellular Vesicles ◽

Neutralizing Antibodies ◽

Phospholipid Bilayer ◽

Transduction Efficiency ◽

Target Cells ◽

Primary Astrocyte ◽

Efficient Gene ◽

Gene Therapy Vectors

(1) Adeno-associated viruses (AAV) are safe and efficient gene therapy vectors with promising results in the treatment of several diseases. Extracellular vesicles (EV) are phospholipid bilayer-surrounded structures carrying several types of lipids, proteins, and nucleic acids with the ability to cross biological barriers. EV-associated AAVs might serve as new and efficient gene therapy vectors considering that they carry the benefits of both AAVs and EVs. (2) We tested vesicle-associated AAVs and vesicles mixed with AAVs on two major cell types of the central nervous system: a neural cell line (N2A) and primary astrocyte cells. (3) In contrast to previously published in vivo observations, the extracellular vesicle packaging did not improve but, in the case of primary astrocyte cells, even inhibited the infection capacity of the AAV particles. The observed effect was not due to the inhibitory effects of the vesicles themselves, since mixing the AAVs with extracellular vesicles did not change the effectiveness. (4) Our results suggest that improvement of the in vivo efficacy of the EV-associated AAV particles is not due to the enhanced interaction between the AAV and the target cells, but most likely to the improved delivery of the AAVs through tissue barriers and to the shielding of AAVs from neutralizing antibodies.

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