Repeat Administration of Proteins to the Eye With a Single Intraocular Injection of an Adenovirus Vector

McVey, Duncan; Hamilton, Melissa; Hsu, Cheng-Ying; King, C. Richter; Brough, Douglas E.; Wei, Lisa L.

doi:10.1038/mt.2008.124

Cited by 5 publications

(3 citation statements)

References 39 publications

(43 reference statements)

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“…After vector delivery to macular organ culture, both DNA and RNA were extracted at 1, 4, 7, and 14 days postvector delivery. The amount of vector particles and level of transgene expression was determined by TaqMan assay, as previously described . (SplicePrimer: TGC CAA GAG TGA C∧GT GTC CA; gfp,EgfpaPrimer: TCC TCG CCC TTG CTC A; SpliceProbe: 6FAM CCC AGG TCC AAC TGC AGC CGG MGBNFQ).…”

Section: Methodsmentioning

confidence: 99%

“…The amount of vector particles and level of transgene expression was determined by TaqMan assay, as previously described. 15 (SplicePrimer: TGC CAA GAG TGA CÙGT GTC CA; gfp,EgfpaPrimer: TCC TCG CCC TTG CTC A; SpliceProbe: 6FAM CCC AGG TCC AAC TGC AGC CGG MGBNFQ). For LCM specimens, detection of gfp DNA sequences was carried out to determine the presence of vector-delivered transgenes.…”

Section: Quantitative Pcr and Rt-pcrmentioning

confidence: 99%

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Optimizing atoh1‐induced vestibular hair cell regeneration

et al. 2014

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Objectives/Hypothesis Determine the optimal design characteristics of an adenoviral vector to deliver atoh1 and induce regeneration of vestibular hair cells. Study Design Evaluation of a mouse model of intra-labyrinthine gene delivery. Tissue culture of mouse and human macular organs. Methods Macular organs from adult C57Bl/6 mice were treated with binding modified and alternate adenovectors expressing green fluorescent protein (gfp) or luciferase (L). Expression of marker genes was determined over time to determine vector transfection efficiency. The inner ear of adult mice was then injected with modified vectors. Expression of gfp and distribution of vector DNA was followed. Hearing and balance function was evaluated in normal animals to ensure safety of the novel vector designs. An optimized vector was identified and tested for its ability to induce hair cell regeneration in a mouse vestibulopathy model. Finally this vector was tested for its ability to induce hair cell regeneration in human tissue. Results Ad5 serotype based vectors were identified as having a variety of different binding capacities for inner ear tissue. This makes it difficult to limit the dose of vector due to entry into non-targeted cells. Screening of rare adenovector serotypes demonstrated that Ad28 based vectors were ideally suited for delivery to supporting cells and therefore useful for hair cell regeneration studies. Utilization of an Ad28 based vector to deliver atoh1 to a mouse model of vestibular loss resulted significant functional recovery of balance. This vector was also capable of transfecting human macular organs and inducing regeneration of human vestibular hair cells in vitro. Conclusions Improvement in vector design can lead to more specific cell based delivery and reduction of non specific delivery of the trans gene leading to the development of optimized molecular therapeutics to induce hair cell regeneration. Level of Evidence N/A Controlled basic science study.

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Section: Methodsmentioning

confidence: 99%

Section: Quantitative Pcr and Rt-pcrmentioning

confidence: 99%

Optimizing atoh1‐induced vestibular hair cell regeneration

et al. 2014

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“…Various therapeutic proteins have been proposed in the literature with a wide range of roles and functions in the body (4-7): formation of receptor domains on the cell surface, improvement of the intracellular and/or extracellular molecular transport, enzymatic catalysis of biochemical reactions, enzymatic or regulatory activity, targeting, vaccines (8,9), and diagnostics (10)(11)(12). Protein drugs are able to act selectively on biological pathways but often require repeated administration, making their clinical use even more challenging than that of conventional drugs (13)(14)(15)(16). The controlled and sustained release of proteins may enhance their therapeutic efficacy and reduce the pain and inconvenience of frequent injections.…”

Section: Introductionmentioning

confidence: 99%