Enhancement of rAAV2-Mediated Transgene Expression in Retina Cells In Vitro and In Vivo by Coadministration of Low-Dose Chemotherapeutic Drugs

Zhang, Shenghai; Wu, Jihong; Wu, Xiaobing; Xu, Ping; Tian, Ye; Yi, Miaoying; Liu, Xinjian; Dong, Xiaoyan; Wolf, Frank de; Li, Chuan-Yuan; Huang, Qian

doi:10.1167/iovs.11-8856

Cited by 13 publications

(10 citation statements)

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“…Several technical methods of enhancing the efficiency and specificity of AAV transduction in the eye have been investigated, such as use of cell-specific transgene promoters (61), engineered serotypes of AAV (24, 25), ultrasound micro bubbles (62), and co-administration of either chemo-therapeutic drugs (63) or adenovirus (64). It is unknown how these adjunctive techniques could affect the immune response in humans, but it could be speculated that additional antigens and disruption of cellular barriers could influence antigen presentation and immune infiltration.…”

Section: Methods To Enhance Efficiencymentioning

confidence: 99%

Immunology of AAV-Mediated Gene Transfer in the Eye

Willett¹,

Bennett²

2013

Front. Immunol.

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The eye has been at the forefront of translational gene therapy largely owing to suitable disease targets, anatomic accessibility, and well-studied immunologic privilege. These advantages have fostered research culminating in several clinical trials and adeno-associated virus (AAV) has emerged as the vector of choice for many ocular therapies. Pre-clinical and clinical investigations have assessed the humoral and cellular immune responses to a variety of naturally occurring and engineered AAV serotypes as well as their delivered transgenes and these data have been correlated to potential clinical sequelae. Encouragingly, AAV appears safe and effective with clinical follow-up surpassing 5 years in some studies. As disease targets continue to expand for AAV in the eye, thorough and deliberate assessment of immunologic safety is critical. With careful study, the development of these technologies should concurrently inform the biology of the ocular immune response.

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Section: Methods To Enhance Efficiencymentioning

confidence: 99%

Immunology of AAV-Mediated Gene Transfer in the Eye

Willett¹,

Bennett²

2013

Front. Immunol.

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“…Subretinal AAV2 localization was first tested with well-established reporter genes: AcGFP, which has a fluorescent product (Cereso et al, 2014; Cronin et al, 2012; Li et al, 2008; Zhang et al, 2012, 2015), and LacZ, which has a chromogenic product (Hojo et al, 2004), to validate successful inoculation of the targeted tissue.…”

Section: Resultsmentioning

confidence: 99%

“…1 month duration) and intense immunogenicity (Campochiaro, 2011;Wang et al, 2004; Zhang et al, 2012). The intraocular humoral and cellular response to the nonpathogenic AAV, conversely, is minimal and benign (Amado et al, 2010; Cheng et al, 2013; Daya and Berns, 2008; Dismuke et al, 2013; Roy et al, 2010; Wang et al, 2004; Zhang et al, 2012). In human studies, the AAV2 serotype was shown to be safe for human retinal disorders, and effective for up to 3.5 years (Roy et al, 2010; Simonelli et al, 2010; Zhang et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

“…With specificity for the outer retina and the RPE/choroid complex (Cronin et al, 2012; Muhlfriedel et al, 2013; Zhang et al, 2012, 2015), a subretinal route for AAV2 offers stronger and more diffuse transfection of these layers, along with a milder immune reaction, compared to an intravitreal route (Campochiaro, 2011; Li et al, 2008). Targeting this space may also mitigate the danger of medication-induced global retinal toxicity linked with the intravitreal approach (Campochiaro, 2011; Grunwald et al, 2014; Rofagha et al, 2013; Zhang et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Subretinal AAV2.COMP-Ang1 suppresses choroidal neovascularization and vascular endothelial growth factor in a murine model of age-related macular degeneration

Lambert¹,

Zhang

Ruju

et al. 2016

Experimental Eye Research

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To assess whether Tie2-mediated vascular stabilization ameliorates neovascular age-related macular degeneration (AMD), we investigated the impact of adeno-associated virus-mediated gene therapy with cartilage oligomeric matrix protein angiopoietin-1 (AAV2.COMP-Ang1) on choroidal neovascularization (CNV), vascular endothelial growth factor (VEGF), and hypoxia-inducible factor (HIF) in a mouse model of the disease. We treated mice with subretinal injections of AAV2.COMP-Ang1 or control (AAV2.AcGFP, AAV2.LacZ, and phosphate-buffered saline). Subretinal AAV2 localization and plasmid protein expression was verified in the retinal pigment epithelium (RPE)/choroid of mice treated with all AAV2 constructs. Laser-assisted simulation of neovascular AMD was performed and followed by quantification of HIF, VEGF, and CNV in each experimental group. We found that AAV2.COMP-Ang1 was associated with a significant reduction in VEGF levels (29–33%, p < 0.01) and CNV volume (60–70%, p < 0.01), without a concomitant decrease in HIF1-α, compared to all controls. We concluded that a) AAV2 is a viable vector for delivering COMP-Ang1 to subretinal tissues, b) subretinal COMP-Ang1 holds promise as a prospective treatment for neovascular AMD, and c) although VEGF suppression in the RPE/choroid may be one mechanism by which AAV2.COMP-Ang1 reduces CNV, this therapeutic effect may be hypoxia-independent. Taken together, these findings suggest that AAV2.COMP-Ang1 has potential to serve as an alternative or complementary option to anti-VEGF agents for the long-term amelioration of neovascular AMD.

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“…Finally, enhancing transduction efficiency through the use of pharmacological agents has been explored. Previous work has shown that topoisomerase inhibitors and anthracyclines can enhance transduction both in vitro and in vivo (35)(36)(37)(38)(39)(40)(41)(42). Proteasome inhibitors, particularly the FDA-approved bortezomib (Velcade), have been shown to enhance transduction in vitro as well as in both small-and large-animal models (39,40,43).…”

mentioning

confidence: 99%

Identification and Validation of Small Molecules That Enhance Recombinant Adeno-associated Virus Transduction following High-Throughput Screens

Nicolson

Hirsch

et al. 2016

J Virol

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While the recent success of adeno-associated virus (AAV)-mediated gene therapy in clinical trials is promising, challenges still face the widespread applicability of recombinant AAV(rAAV). A major goal is to enhance the transduction efficiency of vectors in order to achieve therapeutic levels of gene expression at a vector dose that is below the immunological response threshold. In an attempt to identify novel compounds that enhance rAAV transduction, we performed two high-throughput screens comprising 2,396 compounds. We identified 13 compounds that were capable of enhancing transduction, of which 12 demonstrated vector-specific effects and 1 could also enhance vector-independent transgene expression. Many of these compounds had similar properties and could be categorized into five groups: epipodophyllotoxins (group 1), inducers of DNA damage (group 2), effectors of epigenetic modification (group 3), anthracyclines (group 4), and proteasome inhibitors (group 5). We optimized dosing for the identified compounds in several immortalized human cell lines as well as normal diploid cells. We found that the group 1 epipodophyllotoxins (teniposide and etoposide) consistently produced the greatest transduction enhancement. We also explored transduction enhancement among single-stranded, self-complementary, and fragment vectors and found that the compounds could impact fragmented rAAV2 transduction to an even greater extent than single-stranded vectors. In vivo analysis of rAAV2 and all of the clinically relevant compounds revealed that, consistent with our in vitro results, teniposide exhibited the greatest level of transduction enhancement. Finally, we explored the capability of teniposide to enhance transduction of fragment vectors in vivo using an AAV8 capsid that is known to exhibit robust liver tropism. Consistent with our in vitro results, teniposide coadministration greatly enhanced fragmented rAAV8 transduction at 48 h and 8 days. This study provides a foundation based on the rAAV small-molecule screen methodology, which is ideally used for more-diverse libraries of compounds that can be tested for potentiating rAAV transduction. IMPORTANCEThis study seeks to enhance the capability of adeno-associated viral vectors for therapeutic gene delivery applicable to the treatment of diverse diseases. To do this, a comprehensive panel of FDA-approved drugs were tested in human cells and in animal models to determine if they increased adeno-associated virus gene delivery. The results demonstrate that particular groups of drugs enhance adeno-associated virus gene delivery by unknown mechanisms. In particular, the enhancement of gene delivery was approximately 50 to 100 times better with than without teniposide, a compound that is also used as chemotherapy for cancer. Collectively, these results highlight the potential for FDA-approved drug enhancement of adeno-associated virus gene therapy, which could result in safe and effective treatments for diverse acquired or genetic diseases.A deno-associated viral (AAV) vec...

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Enhancement of rAAV2-Mediated Transgene Expression in Retina Cells In Vitro and In Vivo by Coadministration of Low-Dose Chemotherapeutic Drugs

Cited by 13 publications

References 31 publications

Immunology of AAV-Mediated Gene Transfer in the Eye

Immunology of AAV-Mediated Gene Transfer in the Eye

Subretinal AAV2.COMP-Ang1 suppresses choroidal neovascularization and vascular endothelial growth factor in a murine model of age-related macular degeneration

Identification and Validation of Small Molecules That Enhance Recombinant Adeno-associated Virus Transduction following High-Throughput Screens

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