711. Intravenous Neonatal Gene Therapy Corrects GM2 Gangliosidosis in Sandhoff Mice for Long-Term, By Using AAV Viral Vector Expressing a New Hexosaminidase Variant

Intravitreal delivery of brain-derived neurotrophic factor (BDNF) by injection of recombinant protein or by gene therapy can alleviate retinal ganglion cell (RGC) loss after optic nerve injury (ONI) or laser-induced ocular hypertension (OHT). In models of glaucoma, BDNF therapy can delay or halt RGCs loss, but this protection is time-limited. The decreased efficacy of BDNF supplementation has been in part attributed to BDNF TrkB receptor downregulation. However, whether BDNF overexpression causes TrkB downregulation, impairing long-term BDNF signaling in the retina, has not been conclusively proven. After ONI or OHT, when increased retinal BDNF was detected, a concomitant increase, no change or a decrease in TrkB was reported. We examined quantitatively the retinal concentrations of the TrkB protein in relation to BDNF, in a course of adeno-associated viral vector gene therapy (AAV2-BDNF), using a microbead trabecular occlusion model of glaucoma. We show that unilateral glaucoma, with intraocular pressure ( IOP) increased for five weeks, leads to a bilateral decrease of BDNF in the retina at six weeks, accompanied by up to four-fold TrkB upregulation, while a moderate BDNF overexpression in a glaucomatous eye triggers changes that restore normal TrkB concentrations, driving signaling towards long-term RGCs neuroprotection. We conclude that for glaucoma therapy, the careful selection of the appropriate BDNF concentration is the main factor securing the long-term responsiveness of RGCs and the maintenance of normal TrkB levels.

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Long-term restoration of visual function in end-stage retinal degeneration using subretinal human melanopsin gene therapy

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1701589114 ◽

2017 ◽

Vol 114 (42) ◽

pp. 11211-11216 ◽

Cited By ~ 26

Author(s):

Samantha R. De Silva ◽

Alun R. Barnard ◽

Steven Hughes ◽

Shu K. E. Tam ◽

Chris Martin ◽

...

Keyword(s):

Gene Therapy ◽

Retinal Degeneration ◽

Viral Vector ◽

Therapeutic Option ◽

Ectopic Expression ◽

Action Potential Firing ◽

Pupil Light Reflex ◽

Potential Firing ◽

End Stage

Optogenetic strategies to restore vision in patients who are blind from end-stage retinal degenerations aim to render remaining retinal cells light sensitive once photoreceptors are lost. Here, we assessed long-term functional outcomes following subretinal delivery of the human melanopsin gene (OPN4) in the rd1 mouse model of retinal degeneration using an adeno-associated viral vector. Ectopic expression of OPN4 using a ubiquitous promoter resulted in cellular depolarization and ganglion cell action potential firing. Restoration of the pupil light reflex, behavioral light avoidance, and the ability to perform a task requiring basic image recognition were restored up to 13 mo following injection. These data suggest that melanopsin gene therapy via a subretinal route may be a viable and stable therapeutic option for the treatment of end-stage retinal degeneration in humans.

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459. Long-Term Correction of Canine Leukocyte Adhesion Deficiency With Foamy Viral Vector-Mediated Gene Therapy Using Reduced Intensity Busulfan Conditioning

Molecular Therapy ◽

10.1016/s1525-0016(16)35472-7 ◽

2014 ◽

Vol 22 ◽

pp. S176

Keyword(s):

Gene Therapy ◽

Viral Vector ◽

Leukocyte Adhesion ◽

Leukocyte Adhesion Deficiency ◽

Reduced Intensity

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Gene Therapy Applications of Non-Human Lentiviral Vectors

Viruses ◽

10.3390/v12101106 ◽

2020 ◽

Vol 12 (10) ◽

pp. 1106

Author(s):

Altar M. Munis

Keyword(s):

Gene Therapy ◽

Gene Transfer ◽

Transgene Expression ◽

Viral Vector ◽

Lentiviral Vector ◽

Cell Therapies ◽

Dividing Cells ◽

Cell Genome ◽

Hiv 1

Recent commercialization of lentiviral vector (LV)-based cell therapies and successful reports of clinical studies have demonstrated the untapped potential of LVs to treat diseases and benefit patients. LVs hold notable and inherent advantages over other gene transfer agents based on their ability to transduce non-dividing cells, permanently transform target cell genome, and allow stable, long-term transgene expression. LV systems based on non-human lentiviruses are attractive alternatives to conventional HIV-1-based LVs due to their lack of pathogenicity in humans. This article reviews non-human lentiviruses and highlights their unique characteristics regarding virology and molecular biology. The LV systems developed based on these lentiviruses, as well as their successes and shortcomings, are also discussed. As the field of gene therapy is advancing rapidly, the use of LVs uncovers further challenges and possibilities. Advances in virology and an improved understanding of lentiviral biology will aid in the creation of recombinant viral vector variants suitable for translational applications from a variety of lentiviruses.

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Abstract 5423: rAAV-Mediated Gene Therapy with Inducible Nitric Oxide Synthase (iNOS) Affords Permanent Cardioprotection (1 Year) without Adverse Functional Consequences

Circulation ◽

10.1161/circ.118.suppl_18.s_547-d ◽

2008 ◽

Vol 118 (suppl_18) ◽

Author(s):

Qianhong Li ◽

Yiru Guo ◽

Wen-Jian Wu ◽

Qinghui Ou ◽

Santosh K Sanganalmath ◽

...

Keyword(s):

Gene Therapy ◽

Gene Transfer ◽

Viral Vector ◽

Long Term Effects ◽

Inos Gene ◽

Inos Protein Expression ◽

Functional Consequences ◽

And Function ◽

Oxide Synthase

The ultimate goal of prophylactic gene therapy is to confer permanent protection against ischemia. Although gene therapy with iNOS is known to protect against myocardial infarction at 3 days and up to 2 months, the long-term effects of iNOS gene therapy on myocardial ischemic injury and function are unknown. To address this issue, we created a recombinant adeno-associated viral vector carrying the iNOS gene (rAAV/iNOS) which enables long-lasting transgene expression. Mice received injections in anterior LV wall of rAAV/LacZ or rAAV/iNOS; 1 year later, they underwent a 30-min coronary occlusion (O) and 4 h of reperfusion (R). iNOS gene transfer resulted in elevated iNOS protein expression (+ 2.9-fold vs. LacZ group, n=6, P<0.05; Fig ) and iNOS activity (+ 3.3-fold vs. LacZ group, n=4, P<0.05) 1 year later. Infarct size (% of risk region) was dramatically reduced at 1 year after iNOS gene transfer (13.5+/−2.2%, n=12, vs. 42.9+/−2.6%, n=12, in LacZ group; Fig ). The infarct-sparing effects of iNOS gene therapy at 1 year were as powerful as those observed 24 h after ischemic PC (six 4-min O/4-min R cycles) (16.3+/−2.3%, n=8; Fig ). Importantly, compared with the LacZ group (n=11), iNOS gene transfer (n=10) had no effect on LV dimensions or function for up to 1 year (at 1 year: LVEDD 4.4+/−0.1 vs. 4.2+/−0.2 mm; LVESD 2.9+/−0.1 vs. 2.9+/−0.2 mm; FS 34+/−1.8 vs. 32+/−2.6%; EF 56+/−2.3 vs. 60+/−2.9%) (echocardiography). These data demonstrate, for the first time, that rAAV-mediated iNOS gene transfer affords long-term, probably permanent (1 year) cardioprotection without adverse functional consequences, providing a strong rationale for further preclinical testing of prophylactic gene therapy.

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SMALL ALPHAHERPESVIRUS LATENCY-ASSOCIATED PROMOTERS DRIVE EFFICIENT AND LONG-TERM TRANSGENE EXPRESSION IN THE CENTRAL NERVOUS SYSTEM

10.1101/2019.12.31.891903 ◽

2019 ◽

Author(s):

Carola J. Maturana ◽

Jessica L. Verpeut ◽

Thomas J. Pisano ◽

Zahra M. Dhanerawala ◽

Andrew Esteves ◽

...

Keyword(s):

Gene Therapy ◽

Central Nervous System ◽

Nervous System ◽

Transgene Expression ◽

Pseudorabies Virus ◽

Viral Vector ◽

Gene Promoters ◽

Systemic Delivery ◽

Light Sheet Microscopy

AbstractRecombinant adeno-associated viral vectors (rAAV) are used as gene therapy vectors to treat central nervous system (CNS) diseases. Despite their safety and broad tropism, important issues need to be corrected such as the limited payload capacity and the lack of small gene promoters providing long-term, pan-neuronal transgene expression in the CNS. Commonly used gene promoters are relatively large and can be repressed a few months after CNS transduction, risking the long-term performance of single-dose gene therapy applications. We used a whole-CNS screening approach based on systemic delivery of AAV-PHP.eB, iDisco+ tissue-clearing and light-sheet microscopy, to identify three small latency-associated promoters (LAP) from the herpesvirus pseudorabies virus (PRV). These promoters are LAP1 (404bp), LAP2 (498bp) and LAP1_2 (880bp). They drive chronic transcription of the virus encoded latency-associated transcript (LAT) during productive and latent phases of PRV infection. We observed stable, pan-neuronal transgene transcription and translation from AAV-LAP in the CNS for six months post AAV transduction. In several CNS areas, the number of cells expressing the transgene was higher for LAP2 than the large conventional EF1α promoter (1264bp). Our data suggests that the LAP are suitable candidates for viral vector-based CNS gene therapies requiring chronic transgene expression after one-time viral-vector administration.

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Long-Term Follow-up of Foamy Viral Vector-Mediated Gene Therapy for Canine Leukocyte Adhesion Deficiency

Molecular Therapy ◽

10.1038/mt.2013.34 ◽

2013 ◽

Vol 21 (5) ◽

pp. 964-972 ◽

Cited By ~ 24

Author(s):

Thomas R Bauer ◽

Laura M Tuschong ◽

Katherine R Calvo ◽

Heather R Shive ◽

Tanya H Burkholder ◽

...

Keyword(s):

Gene Therapy ◽

Viral Vector ◽

Leukocyte Adhesion ◽

Leukocyte Adhesion Deficiency ◽

Long Term Follow Up

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Development of gene therapy for blood disorders: an update

Blood ◽

10.1182/blood-2013-04-453209 ◽

2013 ◽

Vol 122 (9) ◽

pp. 1556-1564 ◽

Cited By ~ 28

Author(s):

Arthur W. Nienhuis

Keyword(s):

Gene Therapy ◽

Gene Transfer ◽

Viral Infections ◽

Viral Vector ◽

Suicide Gene ◽

Retroviral Vectors ◽

Specific Protein ◽

Current Status ◽

Blood Disorders

Abstract This review addresses the current status of gene therapy for immunodeficiencies, chronic granulomatous disease, suicide gene therapy for graft-versus-host disease, viral infections, malignant hematologic disorders, hemophilia, and the hemoglobin disorders. New developments in vector design have fostered improved expression as well as enhanced safety, particularly of integrating retroviral vectors. Several immunodeficiencies have been treated successfully by stem cell–targeted, retroviral-mediated gene transfer with reconstitution of the immune system following infusion of the transduced cells. In a trial for hemophilia B, long-term expression of human FIX has been observed following adeno-associated viral vector–mediated gene transfer into the liver. This approach should be successful in treating any disorder in which liver production of a specific protein is therapeutic.

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Long-term, antidiabetogenic effects of GLP-1 gene therapy using a double-stranded, adeno-associated viral vector

Gene Therapy ◽

10.1038/gt.2010.119 ◽

2010 ◽

Vol 18 (2) ◽

pp. 155-163 ◽

Cited By ~ 8

Author(s):

S H Choi ◽

H C Lee

Keyword(s):

Gene Therapy ◽

Viral Vector

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Enhanced Stability and Controlled Delivery of MOF Encapsulated Vaccines and Their Immunogenic Response In Vivo

10.26434/chemrxiv.7335122.v1 ◽

2018 ◽

Author(s):

Michael Luzuriaga ◽

Raymond P. Welch ◽

Madushani Dharmawardana ◽

Candace Benjamin ◽

Shaobo Li ◽

...

Keyword(s):

Viral Vector ◽

Controlled Delivery ◽

Conformational Structure ◽

Spectroscopy Analysis ◽

Zeolitic Imidazolate Framework ◽

Structural Conformation ◽

Depth Study ◽

Viral Nanoparticle

<div><div><div><p>Vaccines have an innate tendency to lose their structural conformation upon environmental and chemical stressors. A loss in conformation reduces the therapeutic ability to prevent the spread of a pathogen. Herein, we report an in-depth study of zeolitic imidazolate framework-8 (ZIF-8) and its ability to provide protection for a model viral vector against dena- turing conditions. The immunoassay and spectroscopy analysis together demonstrate enhanced thermal and chemical stability to the conformational structure of the encapsulated viral nanoparticle. The long-term biological activity of this virus-ZIF composite was investigated in animal models to further elucidate the integrity of the encapsulated virus, the bio-safety, and immunogenicity of the overall composite. Additionally, histological analysis found no observable tissue damage in the skin or vital organs in mice, following multiple subcutaneous administrations. This study shows that ZIF-based protein composites are strong candidates for improved preservation of proteinaceous drugs, are biocompatible, and capable of controlling the release and adsorption of drugs in vivo.</p></div></div></div>

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