Ocular localization and transduction by adenoviral vectors are serotype-dependent and can be modified by inclusion of rgd fiber modifications

Kazuhiro Ueyama; Keisuke Mori; Takuhei Shoji; Hidekazu Omata; Peter L. Gehlbach; Douglas E. Brough; Lisa L. Wei; Shin Yoneya

doi:10.1371/journal.pone.0108071

Ocular localization and transduction by adenoviral vectors are serotype-dependent and can be modified by inclusion of rgd fiber modifications

Kazuhiro Ueyama, Keisuke Mori, Takuhei Shoji, Hidekazu Omata, Peter L. Gehlbach, Douglas E. Brough, Lisa L. Wei, Shin Yoneya

School of Medicine

Research output: Contribution to journal › Article › peer-review

12 Scopus citations

Abstract

Purpose: To evaluate localization and transgene expression from adenoviral vector of serotypes 5, 35, and 28, 6 an RGD motif in the fiber following intravitreal or subretinal administration.

Methods: Ocular transduction by adenoviral vector serotypes 6 RGD was studied in the eyes of mice receiving an intravitreous or subretinal injection. Each serotype expressed a CMV-GFP expression cassette and histological sections of eyes were examined. Transgene expression levels were examined using luciferase (Luc) regulated by the CMV promoter.

Results: GFP localization studies revealed that serotypes 5 and 28 given intravitreously transduced corneal endothelial, trabecular, and iris cells. Intravitreous delivery of the unmodified Ad35 serotype transduced only trabecular meshwork cells, but, the modification of the RGD motif into the fiber of the Ad35 viral vector base expanded transduction to corneal endothelial and iris cells. Incorporation of the RGD motif into the fiber knob with deletion of RGD from the penton base did not affect the transduction ability of the Ad5 vector base. Subretinal studies showed that RGD in the Ad5 knob shifted transduction from RPE cells to photoreceptor cells. Using a CMV-Luc expression cassette, intravitreous delivery of all the tested vectors, such as Ad5-, Ad35- and Ad28- resulted in an initial rapid induction of luciferase activity that thereafter declined. Subretinal administration of vectors showed a marked difference in transgene activity. Ad35-Luc gene expression peaked at 7 days and remained elevated for 6 months. Ad28-Luc expression was high after 1 day and remained sustained for one month.

Conclusions: Different adenoviral vector serotypes 6 modifications transduce different cells within the eye. Transgene expression can be brief or extended and is serotype and delivery route dependent. Thus, adenoviral vectors provide a versatile platform for the delivery of therapeutic agents for ocular diseases. Copyright:

Original language	English (US)
Article number	A1358
Journal	PloS one
Volume	9
Issue number	9
DOIs	https://doi.org/10.1371/journal.pone.0108071
State	Published - Sep 18 2014

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology
General Agricultural and Biological Sciences
General

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@article{5ac9944fd8d64ebd9c5f27f1ddd8a2e0,

title = "Ocular localization and transduction by adenoviral vectors are serotype-dependent and can be modified by inclusion of rgd fiber modifications",

abstract = "Purpose: To evaluate localization and transgene expression from adenoviral vector of serotypes 5, 35, and 28, 6 an RGD motif in the fiber following intravitreal or subretinal administration.Methods: Ocular transduction by adenoviral vector serotypes 6 RGD was studied in the eyes of mice receiving an intravitreous or subretinal injection. Each serotype expressed a CMV-GFP expression cassette and histological sections of eyes were examined. Transgene expression levels were examined using luciferase (Luc) regulated by the CMV promoter.Results: GFP localization studies revealed that serotypes 5 and 28 given intravitreously transduced corneal endothelial, trabecular, and iris cells. Intravitreous delivery of the unmodified Ad35 serotype transduced only trabecular meshwork cells, but, the modification of the RGD motif into the fiber of the Ad35 viral vector base expanded transduction to corneal endothelial and iris cells. Incorporation of the RGD motif into the fiber knob with deletion of RGD from the penton base did not affect the transduction ability of the Ad5 vector base. Subretinal studies showed that RGD in the Ad5 knob shifted transduction from RPE cells to photoreceptor cells. Using a CMV-Luc expression cassette, intravitreous delivery of all the tested vectors, such as Ad5-, Ad35- and Ad28- resulted in an initial rapid induction of luciferase activity that thereafter declined. Subretinal administration of vectors showed a marked difference in transgene activity. Ad35-Luc gene expression peaked at 7 days and remained elevated for 6 months. Ad28-Luc expression was high after 1 day and remained sustained for one month.Conclusions: Different adenoviral vector serotypes 6 modifications transduce different cells within the eye. Transgene expression can be brief or extended and is serotype and delivery route dependent. Thus, adenoviral vectors provide a versatile platform for the delivery of therapeutic agents for ocular diseases. Copyright:",

author = "Kazuhiro Ueyama and Keisuke Mori and Takuhei Shoji and Hidekazu Omata and Gehlbach, {Peter L.} and Brough, {Douglas E.} and Wei, {Lisa L.} and Shin Yoneya",

year = "2014",

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day = "18",

doi = "10.1371/journal.pone.0108071",

language = "English (US)",

volume = "9",

journal = "PloS one",

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T1 - Ocular localization and transduction by adenoviral vectors are serotype-dependent and can be modified by inclusion of rgd fiber modifications

AU - Ueyama, Kazuhiro

AU - Mori, Keisuke

AU - Shoji, Takuhei

AU - Omata, Hidekazu

AU - Gehlbach, Peter L.

AU - Brough, Douglas E.

AU - Wei, Lisa L.

AU - Yoneya, Shin

PY - 2014/9/18

Y1 - 2014/9/18

N2 - Purpose: To evaluate localization and transgene expression from adenoviral vector of serotypes 5, 35, and 28, 6 an RGD motif in the fiber following intravitreal or subretinal administration.Methods: Ocular transduction by adenoviral vector serotypes 6 RGD was studied in the eyes of mice receiving an intravitreous or subretinal injection. Each serotype expressed a CMV-GFP expression cassette and histological sections of eyes were examined. Transgene expression levels were examined using luciferase (Luc) regulated by the CMV promoter.Results: GFP localization studies revealed that serotypes 5 and 28 given intravitreously transduced corneal endothelial, trabecular, and iris cells. Intravitreous delivery of the unmodified Ad35 serotype transduced only trabecular meshwork cells, but, the modification of the RGD motif into the fiber of the Ad35 viral vector base expanded transduction to corneal endothelial and iris cells. Incorporation of the RGD motif into the fiber knob with deletion of RGD from the penton base did not affect the transduction ability of the Ad5 vector base. Subretinal studies showed that RGD in the Ad5 knob shifted transduction from RPE cells to photoreceptor cells. Using a CMV-Luc expression cassette, intravitreous delivery of all the tested vectors, such as Ad5-, Ad35- and Ad28- resulted in an initial rapid induction of luciferase activity that thereafter declined. Subretinal administration of vectors showed a marked difference in transgene activity. Ad35-Luc gene expression peaked at 7 days and remained elevated for 6 months. Ad28-Luc expression was high after 1 day and remained sustained for one month.Conclusions: Different adenoviral vector serotypes 6 modifications transduce different cells within the eye. Transgene expression can be brief or extended and is serotype and delivery route dependent. Thus, adenoviral vectors provide a versatile platform for the delivery of therapeutic agents for ocular diseases. Copyright:

AB - Purpose: To evaluate localization and transgene expression from adenoviral vector of serotypes 5, 35, and 28, 6 an RGD motif in the fiber following intravitreal or subretinal administration.Methods: Ocular transduction by adenoviral vector serotypes 6 RGD was studied in the eyes of mice receiving an intravitreous or subretinal injection. Each serotype expressed a CMV-GFP expression cassette and histological sections of eyes were examined. Transgene expression levels were examined using luciferase (Luc) regulated by the CMV promoter.Results: GFP localization studies revealed that serotypes 5 and 28 given intravitreously transduced corneal endothelial, trabecular, and iris cells. Intravitreous delivery of the unmodified Ad35 serotype transduced only trabecular meshwork cells, but, the modification of the RGD motif into the fiber of the Ad35 viral vector base expanded transduction to corneal endothelial and iris cells. Incorporation of the RGD motif into the fiber knob with deletion of RGD from the penton base did not affect the transduction ability of the Ad5 vector base. Subretinal studies showed that RGD in the Ad5 knob shifted transduction from RPE cells to photoreceptor cells. Using a CMV-Luc expression cassette, intravitreous delivery of all the tested vectors, such as Ad5-, Ad35- and Ad28- resulted in an initial rapid induction of luciferase activity that thereafter declined. Subretinal administration of vectors showed a marked difference in transgene activity. Ad35-Luc gene expression peaked at 7 days and remained elevated for 6 months. Ad28-Luc expression was high after 1 day and remained sustained for one month.Conclusions: Different adenoviral vector serotypes 6 modifications transduce different cells within the eye. Transgene expression can be brief or extended and is serotype and delivery route dependent. Thus, adenoviral vectors provide a versatile platform for the delivery of therapeutic agents for ocular diseases. Copyright:

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U2 - 10.1371/journal.pone.0108071

DO - 10.1371/journal.pone.0108071

M3 - Article

C2 - 25232844

AN - SCOPUS:84907210200

SN - 1932-6203

VL - 9

JO - PloS one

JF - PloS one

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M1 - A1358

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Ocular localization and transduction by adenoviral vectors are serotype-dependent and can be modified by inclusion of rgd fiber modifications

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