Adeno-associated virus (AAV) vector antisense gene transfer in vivo decreases GABAAα1 containing receptors and increases inferior collicular seizure sensitivity

Background: Reduced activity and expression of sarcoplasmic reticulum Ca 2+ ATPase (SERCA2a) is known to occur in HF. Method: Our 4-month study examined the effects of SERCA2a gene transfer in a swine volume-overload HF (VO-HF) model. Using Yorkshire-Landrace swine, HF was created by severing mitral apparatus chordae to induce mitral regurgitation. Results: At 2 months (M), a compensated state of VO-HF was found: prolongation of the rate of isovolumic relaxation (Tau), increased left ventricular internal diameter diastolic and systolic diameters (LVIDd, LVIDs). At 2M, intracoronary injection of adeno-associated virus serotype 1 vector carrying SERCA2a under a cytomegalovirus promoter (AAV1.SERCA2a) (n = 10) vs. saline (n = 6) was performed. At 4M, gene transfer resulted in (A) positive LV inotropic effects: (dP/dt)/P, 15.5 ± 3.0 sec − 1 SERCA2a-group vs. 21.2 ± 3.2 sec − 1 controls; p < 0.01; (B) a favorable trend in LV lusitropic effects: Tau, 0.037 ± 0.019 vs. 0.051 ± 0.01 msec, p = 0.09; (C) improvement in LV geometry: % change in LVIDs, +15 ± 11% controls vs. −3.0 ± 10% SERCA2a-group, p < 0.01. At 4M, BNP levels remained stable in post- SERCA2a gene transfer, in contrast to the progressive rising levels among controls. Further, cardiac SERCA2a expression was significantly decreased in controls whereas it was restored to normal levels in the SERCA2a group (Figure ). Lastly, there was no histopathological evidence of myocardial inflammatory reaction or necrosis. Conclusion: Overexpression of SERCA2a by in vivo AAV1-mediated intracoronary gene transfer preserved systolic function, potentially prevented diastolic dysfunction and improved ventricular remodeling.

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Nonrandom Transduction of Recombinant Adeno-Associated Virus Vectors in Mouse Hepatocytes In Vivo: Cell Cycling Does Not Influence Hepatocyte Transduction

Journal of Virology ◽

10.1128/jvi.74.8.3793-3803.2000 ◽

2000 ◽

Vol 74 (8) ◽

pp. 3793-3803 ◽

Cited By ~ 101

Author(s):

Carol H. Miao ◽

Hiroyuki Nakai ◽

Arthur R. Thompson ◽

Theresa A. Storm ◽

Winnie Chiu ◽

...

Keyword(s):

In Situ Hybridization ◽

Gene Transfer ◽

Genetic Diseases ◽

Critical Event ◽

Dimer Formation ◽

Adeno Associated Virus ◽

Virus Vectors ◽

Preclinical Trials

ABSTRACT Recombinant adeno-associated virus vectors (rAAV) show promise in preclinical trials for the treatment of genetic diseases including hemophilia. Liver-directed gene transfer results in a slow rise in transgene expression, reaching steady-state levels over a period of 5 weeks concomitant with the conversion of the single-stranded rAAV molecules into high-molecular-weight concatemers in about 5% of hepatocytes. Immunohistochemistry and RNA in situ hybridization show that the transgene product is made in about ∼5% of hepatocytes, suggesting that most rAAV-mediated gene expression occurs in hepatocytes containing the double-stranded concatemers. In this study, the mechanism(s) involved in stable transduction in vivo was evaluated. While only ∼5% of hepatocytes are stably transduced, in situ hybridization experiments demonstrated that the vast majority of the hepatocytes take up AAV-DNA genomes after portal vein infusion of the vector. Two different vectors were infused together or staggered by 1, 3, or 5 weeks, and two-color fluorescent in situ hybridization and molecular analyses were performed 5 weeks after the infusion of the second vector. These experiments revealed that a small but changing subpopulation of hepatocytes were permissive to stable transduction. Furthermore, in animals that received a single infusion of two vectors, about one-third of the transduced cells contained heteroconcatemers, suggesting that dimer formation was a critical event in the process of concatemer formation. To determine if the progression through the cell cycle was important for rAAV transduction, animals were continuously infused with 5′-bromo-2′-deoxyuridine (BrdU), starting at the time of administration of a rAAV vector that expressed cytoplasmic β-galactosidase. Colabeling for β-galactosidase and BrdU revealed that there was no preference for transduction of cycling cells. This was further confirmed by demonstrating no increase in rAAV transduction efficiencies in animals whose livers were induced to cycle at the time of or after vector administration. Taken together, our studies suggest that while virtually all hepatocytes take up vector, unknown cellular factors are required for stable transduction, and that dimer formation is a critical event in the transduction pathway. These studies have important implications for understanding the mechanism of integration and may be useful for improving liver gene transfer in vivo.

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Dose response results of self complementary adeno-associated virus (AAV) vector-mediated factor IX gene transfer in non-human primates

Haemophilia ◽

10.1111/j.1365-2516.2009.01996_6.x ◽

2009 ◽

Vol 15 (2) ◽

pp. 635-635

Author(s):

ULRIKE REISS ◽

ANDREW DAVIDOFF ◽

JOHN GRAY ◽

ARTHUR NIENHUIS ◽

AMIT NATHWANI

Keyword(s):

Gene Transfer ◽

Dose Response ◽

Factor Ix ◽

Aav Vector ◽

Adeno Associated Virus

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Cloning and Characterization of a Bovine Adeno-Associated Virus

Journal of Virology ◽

10.1128/jvi.78.12.6509-6516.2004 ◽

2004 ◽

Vol 78 (12) ◽

pp. 6509-6516 ◽

Cited By ~ 55

Author(s):

Michael Schmidt ◽

Hisako Katano ◽

Ioannis Bossis ◽

John A. Chiorini

Keyword(s):

Gene Transfer ◽

Human Cancer ◽

Adeno Associated Virus ◽

Reading Frame ◽

Human Cancer Cell Lines ◽

Left Hand ◽

Serotype 5 ◽

Unique Cell ◽

The Right

ABSTRACT To better understand the relationship between primate adeno-associated viruses (AAVs) and those of other mammals, we have cloned and sequenced the genome of an AAV found as a contaminant in two isolates of bovine adenovirus that was reported to be serologically distinct from primate AAVs. The bovine AAV (BAAV) genome has 4,693 bp, and its organization is similar to that of other AAV isolates. The left-hand open reading frame (ORF) and both inverted terminal repeats (ITRs) have the highest homology with the rep ORF and ITRs of AAV serotype 5 (AAV-5) (89 and 96%, respectively). However, the right-hand ORF was only 55% identical to the AAV-5 capsid ORF; it had the highest homology with the capsid ORF of AAV-4 (76%). By comparing the BAAV cap sequence with a model of an AAV-4 capsid, we mapped the regions of BAAV VP1 that are divergent from AAV-4. These regions are located on the outside of the capsid and are partially located in exposed loops. BAAV was not neutralized by antisera raised against recombinant AAV-2, AAV-4, or AAV-5, and it demonstrated a unique cell tropism profile in four human cancer cell lines, suggesting that BAAV might have transduction activity distinct from that of other isolates. A murine model of salivary gland gene transfer was used to evaluate the in vivo performance of recombinant BAAV. Recombinant BAAV-mediated gene transfer was 11 times more efficient than that with AAV-2. Overall, these data suggest that vectors based on BAAV could be useful for gene transfer applications.

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Incorporation of Adeno-Associated Virus in a Calcium Phosphate Coprecipitate Improves Gene Transfer to Airway Epithelia In Vitro and In Vivo

Journal of Virology ◽

10.1128/jvi.74.1.535-540.2000 ◽

2000 ◽

Vol 74 (1) ◽

pp. 535-540 ◽

Cited By ~ 27

Author(s):

Robert W. Walters ◽

Dongsheng Duan ◽

John F. Engelhardt ◽

Michael J. Welsh

Keyword(s):

Calcium Phosphate ◽

Gene Transfer ◽

Mouse Lung ◽

Apical Surface ◽

Airway Epithelia ◽

Adeno Associated Virus ◽

Human Airway

ABSTRACT Adeno-associated virus (AAV) is inefficient at infecting differentiated airway epithelia because of a lack of receptors at the apical surface. We hypothesized that incorporation of AAV in a calcium phosphate coprecipitate would circumvent this barrier. Interestingly, coprecipitation of AAV type 2 improved gene transfer to differentiated human airway epithelia in vitro and to the mouse lung in vivo. These results suggest that delivery of AAV as a CaPicoprecipitate may significantly enhance its utility for gene transfer to the airway epithelia in vivo.

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The Potential of eCD4-Ig, Delivered by Adeno-Associated Virus (AAV) Vector as a Novel Vaccine for HIV/AIDS Infection

SCRIPTA SCORE Scientific Medical Journal ◽

10.32734/scripta.v2i2.3922 ◽

2021 ◽

Vol 2 (2) ◽

pp. 133-9

Author(s):

Ghea Mangkuliguna

Keyword(s):

Hiv Infection ◽

Immunoglobulin G ◽

Rhesus Macaques ◽

Therapeutic Vaccine ◽

Hiv Vaccine ◽

Aav Vector ◽

Adeno Associated Virus ◽

Dependent Cell ◽

Hiv Aids

Background: HIV/AIDS has already become one of the world's major health issues taking its toll on millions of lives each year. Developing an HIV vaccine with excellent efficacy has become a global urgency that must be addressed immediately. Recently, researchers have successfully developed a more self-like molecule which is a fusion protein between human CD4 domains and immunoglobulin G (IgG) Fc with a CCR5-mimetic sulfopeptide in the carboxy terminus. This molecule, eCD4-Ig, targets only the conserved regions of HIV Env and thus demonstrated the most remarkable potency and breadth so far. By using adeno-associated virus (AAV) vector, eCD4-Ig’s long-term expression in vivo can be achieved. Objectives: Evaluate the efficacy of AAV-eCD4-Ig as both preventive and therapeutic vaccine for HIV/AIDS infection. Methods: A systematic literature study was conducted with the database in PubMed, ScienceDirect, and Proquest. No time and language restriction were applied. Discussion: This review shows that eCD4-Ig eliminates HIV-infected cells through neutralization and antibody-dependent cell-mediated cytotoxicity (ADCC). Moreover, eCD4-Ig is also capable of preventing HIV infection in vivo. Delivered with AAV, eCD4-Ig is maintained stably at both protective and therapeutic levels, as well as gives robust protection for rhesus macaques for almost a year long through a single injection. Conclusion: This study offers evidences that AAV-eCD4-Ig appears to have the potential to be an effective vaccine to prevent HIV infection. Keywords: AAV, AIDS, eCD4-Ig, HIV, vaccine Latar Belakang: Pengembangan vaksin HIV yang efektif menjadi sangat penting mengingat tingginya angka kematian yang ditimbulkan oleh HIV/AIDS. Beberapa tahun terakhir, peneliti berhasil menemukan sebuah molekul yang tersusun atas domain CD4 manusia, immunoglobulin G (IgG) Fc, dan sulfopeptida yang menyerupai CCR5. Molekul yang dinamakan eCD4-Ig ini menargetkan area konservatif dari HIV Env sehingga berpotensi untuk menjadi vaksin HIV yang efektif. Ekspresi eCD4-Ig akan dipertahankan menggunakan Adeno-associated Virus Vector (AAV). Tujuan: Evaluasi efektivitas AAV-eCD4-Ig sebagai vaksin untuk HIV/AIDS. Metode: Penelitian dilakukan dengan melakukan tinjauan pustaka dari beberapa database jurnal, yakni PubMed, ScienceDirect, dan Proquest tanpa ada batasan waktu dan bahasa. Pembahasan: eCD4-Ig membunuh sel-sel yang terinfeksi HIV melalui proses netralisasi dan antibody-dependent cell-mediated cytotoxicity (ADCC). eCD4-Ig juga memberikan perlindungan terhadap infeksi HIV. Ekspresi AAV-eCD4-Ig sangat stabil untuk dosis protektif dan terapeutik, sekaligus melindungi rhesus macaques dari infeksi HIV selama hampir 1 tahun lamanya hanya dengan sekali injeksi. Kesimpulan: AAV-eCD4-Ig memiliki potensi yang menjanjikan untuk menjadi vaksin HIV yang efektif bagi seluruh penderita HIV/AIDS di seluruh dunia. Kata Kunci: AAV, AIDS, eCD4-Ig, HIV, vaksin

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