Light‐activated gene transduction enhances adeno‐associated virus vector–mediated gene expression in human articular chondrocytes

Ulrich-Vinther, Michael; Maloney, Michael D.; Goater, J. Jeffrey; Søballe, Kjeld; Goldring, Mary B.; O’Keefe, Regis J.; Schwarz, Edward M.

doi:10.1002/art.10433

Cited by 35 publications

(27 citation statements)

References 52 publications

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“…However, because of the dense cartilage ECM that surrounds these cells, they have been typically unavailable for genetic modification by direct intraarticular injection of most recombinant vectors. 33,[40][41][42] There have been reports of AAV-mediated transduction of chondrocytes in cartilage explants in culture, [43][44][45] which has been attributed to the smaller size of the AAV particle relative to other viral systems. However, since AAV does not specifically target chondrocytes following direct injection into the joint, it will similarly transduce receptive cells in the synovium.…”

Section: Chondrocytes As Targets For Gene Deliverymentioning

confidence: 99%

“…44,48,51 Although chondrocytes have been somewhat resistant to transfection with plasmid DNA, formulation with certain commercially available lipid-based reagents such as FuGENE6t 52,53 and Lipofectint has been found to enhance the efficiency of DNA uptake, as has pretreatment of the cells with hyaluronidase 54 and mild detergent. 55 Similar to gene delivery in most cell types, viral-based vectors are capable of generating far higher levels of transgene expression with greater persistence.…”

Section: Chondrocytes As Targets For Gene Deliverymentioning

confidence: 99%

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Gene-based approaches for the repair of articular cartilage

2004

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Section: Chondrocytes As Targets For Gene Deliverymentioning

confidence: 99%

Section: Chondrocytes As Targets For Gene Deliverymentioning

confidence: 99%

Gene-based approaches for the repair of articular cartilage

2004

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“…Recombinant AAV is a promising delivery vehicle for articular cartilage with the advantages of sustained transgene expression, reduced potential for host immune response, and the capacity to transduce both dividing and nondividing cells in vivo and in vitro (Ulrich-Vinther et al, 2002;Lee et al, 2011). The viral vector is derived from an endemic, nonpathogenic parvovirus, and intra-articular injection of AAV has been used in clinical trials for delivery of bioactive substances (Mease et al, 2010;High and Aubourg, 2011).…”

Section: Introductionmentioning

confidence: 99%

Persistence, Localization, and External Control of Transgene Expression After Single Injection of Adeno-Associated Virus into Injured Joints

et al. 2013

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A single intra-articular injection of adeno-associated virus (AAV) results in stable and controllable transgene expression in normal rat knees. Because undamaged joints are unlikely to require treatment, the study of AAV delivery in joint injury models is crucial to potential therapeutic applications. This study tests the hypotheses that persistent and controllable AAV-transgene expression are (1) highly localized to the cartilage when AAV is injected postinjury and (2) localized to the intra-articular soft tissues when AAV is injected preinjury. Two AAV injection time points, postinjury and preinjury, were investigated in osteochondral defect and anterior cruciate ligament transection models of joint injury. Rats injected with AAV tetracycline response element (TRE)-luciferase received oral doxycycline for 7 days. Luciferase expression was evaluated longitudinally for 6 months. Transgene expression was persistent and controllable with oral doxycycline for 6 months in all groups. However, the location of transgene expression was different: postinjury AAV-injected knees had luciferase expression highly localized to the cartilage, while preinjury AAV-injected knees had more widespread signal from intraarticular soft tissues. The differential transgene localization between preinjury and postinjury injection can be used to optimize treatment strategies. Highly localized postinjury injection appears advantageous for treatments targeting repair cells. The more generalized and controllable reservoir of transgene expression following AAV injection before anterior cruciate ligament transection (ACLT) suggests an intriguing concept for prophylactic delivery of joint protective factors to individuals at high risk for early osteoarthritis (OA). Successful external control of intra-articular transgene expression provides an added margin of safety for these potential clinical applications.

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“…36 In contrast to gamma irradiation or other cell-damaging agents, we found that as little as 30 J/m 2 of UV irradiation causes a significant increase in gene transduction.…”

Section: Introductionmentioning

confidence: 60%

“…These results are consistent with data obtained in experiments with primary human articular chondrocytes. 36 They also indicated that the level of UV exposure predicted to induce LAGT is well below the cytotoxic threshold.…”

Section: Sensitivity Of Humscs To Uv Irradiationmentioning

confidence: 99%

Light-activated gene transduction of recombinant adeno-associated virus in human mesenchymal stem cells

et al. 2003

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Deficiencies in skeletal tissue repair and regeneration lead to conditions like osteoarthritis, osteoporosis and degenerative disc disease. While no cure for these conditions is available, the use of human bone marrow derived-mesenchymal stem cells (HuMSCs) has been shown to have potential for cellbased therapy. Furthermore, recombinant adeno-associated viruses (rAAV) could be used together with HuMSCs for in vivo or ex vivo gene therapy. Unfortunately, the poor transduction efficiency of these cells remains a significant obstacle. Here, we describe the properties of ultraviolet (UV) light-activated gene transduction (LAGT) with rAAV in HuMSCs, an advance toward overcoming this limitation. Using direct fluorescent image analysis and real-time quantitative PCR to evaluate enhanced green fluorescent protein (eGFP) gene expression, we found that the optimal effects of LAGT with limited cytotoxicity occurred at a UV dose of 200 J/m 2 . Furthermore, this UV irradiation had no effect on either the chondrogenic or osteogenic potential of HuMSCs. Significant effects of LAGT in HuMSCs could be detected as early as 12 h after exposure and persisted over 21 days, in a time and energy-dependent manner. This LAGT effect was maintained for more than 8 h after irradiation and required only a 10-min exposure to rAAV after UV irradiation. Finally, we show that the production of secreted TGFb1 protein from rAAV-TGFb1-IRES-eGFP infected to HuMSCs is highly inducible by UV irradiation. These results demonstrate that LAGT combined with rAAV is a promising procedure to facilitate gene induction in HuMSCs for human gene therapy.

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Light‐activated gene transduction enhances adeno‐associated virus vector–mediated gene expression in human articular chondrocytes

Cited by 35 publications

References 52 publications

Gene-based approaches for the repair of articular cartilage

Gene-based approaches for the repair of articular cartilage

Persistence, Localization, and External Control of Transgene Expression After Single Injection of Adeno-Associated Virus into Injured Joints

Light-activated gene transduction of recombinant adeno-associated virus in human mesenchymal stem cells

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