Herpes simplex virus mediated gene transfer to primate ocular tissues

Xuyang Liu; Curtis R. Brandt; B'Ann T. Gabelt; Paul J. Bryar; Morton E. Smith; Paul L. Kaufman

doi:10.1006/exer.1999.0711

Herpes simplex virus mediated gene transfer to primate ocular tissues

Xuyang Liu, Curtis R. Brandt, B'Ann T. Gabelt, Paul J. Bryar, Morton E. Smith, Paul L. Kaufman^*

^*Corresponding author for this work

Ophthalmology

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

60 Scopus citations

Abstract

We evaluated the feasibility of delivering a gene into monkey eyes using a replication-competent herpes simplex virus (HSV) type 1 ribonucleotide reductase mutant (hrR3) expressing the Escherichia coli lacZ gene. To determine the efficiency of in vitro HSV-mediated gene transfer, cultured human trabecular meshwork (HTM) and human ciliary muscle (HCM) cells were infected with hrR3 and β-galactosidase activity was measured histochemically. Six cynomolgus monkey eyes received viral injections into the anterior chamber (2 x 10⁷ pfu) and/or the vitreous (5 x 10⁷ pfu), and the distribution of cells expressing lacZ was evaluated. In vitro, both cultured HTM and HCM cells displayed multiplicity-dependent β-galactosidase activity. In vivo, intracameral and/or intravitreal injection resulted in transgene expression in TM cells and in non-pigmented ciliary epithelial cells (NPE), but not in CM cells. Transgene expression was also detected in retinal pigmented epithelial (RPE) cells and sporadic retinal ganglion cells (RGC) in eyes receiving virus intracamerally and intravitreally respectively. We observed significant inflammation in the anterior chamber, TM and CM in virus-injected eyes, along with mild vitritis and retinitis. This study demonstrates successful gene transfer using hrR3 as a vector in human ocular cells and in ocular tissues in living monkeys. Further investigation of the etiology of the inflammatory response, possible cytotoxicity, and limited duration of transgene expression is necessary in order to make this technique clinically applicable.

Original language	English (US)
Pages (from-to)	385-395
Number of pages	11
Journal	Experimental eye research
Volume	69
Issue number	4
DOIs	https://doi.org/10.1006/exer.1999.0711
State	Published - Oct 1999

Funding

The authors thank Janice Lokken, Vicky L. Rogness, Gretchen Poulsen, William Hubbard, Bernadette E. Bull and Dr Sina J. Sabet for technical assistance. This work was supported by the National Institutes of Health (EY02698, PLK); the Glaucoma Research Foundation, San Francisco, CA, U.S.A. (PLK); the Retina Research Foundation, Houston, TX, U.S.A. (CRB), and Research to Prevent Blindness, New York, NY, U.S.A. (PLK, CRB). The HCM and HTM cells were provided respectively by Dr Robert Weinreb (EY05990) and Dr Jon R. Polansky (EY02477).

Keywords

Ciliary epithelium
Ciliary muscle
Cynomolgus monkey
Gene therapy
Gene transfer
Herpes simplex virus type 1
LacZ
Retinal ganglion cells
Ribonucleotide reductase mutant
Trabecular meshwork
β- galactosidase

ASJC Scopus subject areas

Ophthalmology
Sensory Systems
Cellular and Molecular Neuroscience

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10.1006/exer.1999.0711

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@article{7ebd40794046456e9cab773806009360,

title = "Herpes simplex virus mediated gene transfer to primate ocular tissues",

abstract = "We evaluated the feasibility of delivering a gene into monkey eyes using a replication-competent herpes simplex virus (HSV) type 1 ribonucleotide reductase mutant (hrR3) expressing the Escherichia coli lacZ gene. To determine the efficiency of in vitro HSV-mediated gene transfer, cultured human trabecular meshwork (HTM) and human ciliary muscle (HCM) cells were infected with hrR3 and β-galactosidase activity was measured histochemically. Six cynomolgus monkey eyes received viral injections into the anterior chamber (2 x 107 pfu) and/or the vitreous (5 x 107 pfu), and the distribution of cells expressing lacZ was evaluated. In vitro, both cultured HTM and HCM cells displayed multiplicity-dependent β-galactosidase activity. In vivo, intracameral and/or intravitreal injection resulted in transgene expression in TM cells and in non-pigmented ciliary epithelial cells (NPE), but not in CM cells. Transgene expression was also detected in retinal pigmented epithelial (RPE) cells and sporadic retinal ganglion cells (RGC) in eyes receiving virus intracamerally and intravitreally respectively. We observed significant inflammation in the anterior chamber, TM and CM in virus-injected eyes, along with mild vitritis and retinitis. This study demonstrates successful gene transfer using hrR3 as a vector in human ocular cells and in ocular tissues in living monkeys. Further investigation of the etiology of the inflammatory response, possible cytotoxicity, and limited duration of transgene expression is necessary in order to make this technique clinically applicable.",

keywords = "Ciliary epithelium, Ciliary muscle, Cynomolgus monkey, Gene therapy, Gene transfer, Herpes simplex virus type 1, LacZ, Retinal ganglion cells, Ribonucleotide reductase mutant, Trabecular meshwork, β- galactosidase",

author = "Xuyang Liu and Brandt, {Curtis R.} and Gabelt, {B'Ann T.} and Bryar, {Paul J.} and Smith, {Morton E.} and Kaufman, {Paul L.}",

note = "Funding Information: The authors thank Janice Lokken, Vicky L. Rogness, Gretchen Poulsen, William Hubbard, Bernadette E. Bull and Dr Sina J. Sabet for technical assistance. This work was supported by the National Institutes of Health (EY02698, PLK); the Glaucoma Research Foundation, San Francisco, CA, U.S.A. (PLK); the Retina Research Foundation, Houston, TX, U.S.A. (CRB), and Research to Prevent Blindness, New York, NY, U.S.A. (PLK, CRB). The HCM and HTM cells were provided respectively by Dr Robert Weinreb (EY05990) and Dr Jon R. Polansky (EY02477).",

year = "1999",

month = oct,

doi = "10.1006/exer.1999.0711",

language = "English (US)",

volume = "69",

pages = "385--395",

journal = "Experimental eye research",

issn = "0014-4835",

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AU - Brandt, Curtis R.

AU - Gabelt, B'Ann T.

AU - Bryar, Paul J.

AU - Smith, Morton E.

AU - Kaufman, Paul L.

N1 - Funding Information: The authors thank Janice Lokken, Vicky L. Rogness, Gretchen Poulsen, William Hubbard, Bernadette E. Bull and Dr Sina J. Sabet for technical assistance. This work was supported by the National Institutes of Health (EY02698, PLK); the Glaucoma Research Foundation, San Francisco, CA, U.S.A. (PLK); the Retina Research Foundation, Houston, TX, U.S.A. (CRB), and Research to Prevent Blindness, New York, NY, U.S.A. (PLK, CRB). The HCM and HTM cells were provided respectively by Dr Robert Weinreb (EY05990) and Dr Jon R. Polansky (EY02477).

PY - 1999/10

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N2 - We evaluated the feasibility of delivering a gene into monkey eyes using a replication-competent herpes simplex virus (HSV) type 1 ribonucleotide reductase mutant (hrR3) expressing the Escherichia coli lacZ gene. To determine the efficiency of in vitro HSV-mediated gene transfer, cultured human trabecular meshwork (HTM) and human ciliary muscle (HCM) cells were infected with hrR3 and β-galactosidase activity was measured histochemically. Six cynomolgus monkey eyes received viral injections into the anterior chamber (2 x 107 pfu) and/or the vitreous (5 x 107 pfu), and the distribution of cells expressing lacZ was evaluated. In vitro, both cultured HTM and HCM cells displayed multiplicity-dependent β-galactosidase activity. In vivo, intracameral and/or intravitreal injection resulted in transgene expression in TM cells and in non-pigmented ciliary epithelial cells (NPE), but not in CM cells. Transgene expression was also detected in retinal pigmented epithelial (RPE) cells and sporadic retinal ganglion cells (RGC) in eyes receiving virus intracamerally and intravitreally respectively. We observed significant inflammation in the anterior chamber, TM and CM in virus-injected eyes, along with mild vitritis and retinitis. This study demonstrates successful gene transfer using hrR3 as a vector in human ocular cells and in ocular tissues in living monkeys. Further investigation of the etiology of the inflammatory response, possible cytotoxicity, and limited duration of transgene expression is necessary in order to make this technique clinically applicable.

AB - We evaluated the feasibility of delivering a gene into monkey eyes using a replication-competent herpes simplex virus (HSV) type 1 ribonucleotide reductase mutant (hrR3) expressing the Escherichia coli lacZ gene. To determine the efficiency of in vitro HSV-mediated gene transfer, cultured human trabecular meshwork (HTM) and human ciliary muscle (HCM) cells were infected with hrR3 and β-galactosidase activity was measured histochemically. Six cynomolgus monkey eyes received viral injections into the anterior chamber (2 x 107 pfu) and/or the vitreous (5 x 107 pfu), and the distribution of cells expressing lacZ was evaluated. In vitro, both cultured HTM and HCM cells displayed multiplicity-dependent β-galactosidase activity. In vivo, intracameral and/or intravitreal injection resulted in transgene expression in TM cells and in non-pigmented ciliary epithelial cells (NPE), but not in CM cells. Transgene expression was also detected in retinal pigmented epithelial (RPE) cells and sporadic retinal ganglion cells (RGC) in eyes receiving virus intracamerally and intravitreally respectively. We observed significant inflammation in the anterior chamber, TM and CM in virus-injected eyes, along with mild vitritis and retinitis. This study demonstrates successful gene transfer using hrR3 as a vector in human ocular cells and in ocular tissues in living monkeys. Further investigation of the etiology of the inflammatory response, possible cytotoxicity, and limited duration of transgene expression is necessary in order to make this technique clinically applicable.

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KW - Retinal ganglion cells

KW - Ribonucleotide reductase mutant

KW - Trabecular meshwork

KW - β- galactosidase

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DO - 10.1006/exer.1999.0711

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JO - Experimental eye research

JF - Experimental eye research

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ER -

Herpes simplex virus mediated gene transfer to primate ocular tissues

Abstract

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Keywords

ASJC Scopus subject areas

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