Herpes Simplex Virus Mediated Gene Transfer to Primate Ocular Tissues

doi:10.1006/exer.1999.0711

Experimental Eye Research

Volume 69, Issue 4, October 1999, Pages 385-395

https://doi.org/10.1006/exer.1999.0711 Get rights and content

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 (2×10⁷pfu) and/or the vitreous (5×10⁷pfu), 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.

References (50)

A. Bill
Aqueous humor dynamics in monkeys (Macaca irus and Cercopithecus ethiops)
Exp. Eye Res.
(1971)
L.Z. Bito
Species differences in the response of the eye to irritation and trauma: a hypothesis of divergence in ocular defense mechanisms, and the choice of the experimental animals for eye research
Exp. Eye Res.
(1984)
B.T. Gabelt et al.
Prostaglandin F_2αincreases uveoscleral outflow in the cynomolgus monkey
Exp. Eye Res.
(1989)
D.J. Goldstein et al.
Factor (s) present in the herpes simplex virus type-1-infected cells can compensate for the loss of the large subunit of the virus ribonucleotide reductase: characterization of an ICP6 deletion mutant
Virology
(1988)
J. Huard et al.
Gene transfer to muscle using herpes simplex virus-based vectors
Neuromuscular Disorders
(1997)
A.D. Idowu et al.
Deletion of the herpes simplex virus type-1 ribonucleotide reductase gene alters virulence and latency in vivo
Antiviral Res.
(1992)
J.G. Jacobson et al.
A herpes simplex virus ribonucleotide reductase deletion mutant is defective for productive acute and reactivatable latent infections of mice and for replication in mouse cells
Virology
(1989)
C. Jomary et al.
Adenovirus-mediated gene transfer to murine retinal cells in vitro and in vivo
FEBS Lett.
(1994)
J.R. Polansky et al.
Trabecular meshwork cell culture in glaucoma research: Evaluation of biological activity and structural properties of human trabecular cells in vitro
Ophthalmology
(1984)
J.W. Rohen
The histologic structure of the chamber angle in primates
Am. J. Ophthalmol.
(1961)

R.J. Visalli et al.

The HSV-1 UL45 18 kDa gene product is a true late protein and a component of the virion

Virus Res.

(1993)

N.G. Abraham et al.

Adenovirus-mediated heme oxygenase-1 gene transfer into rabbit ocular tissue

Invest. Ophthalmol. Vis. Sci.

(1995)

R.R. Ali et al.

Gene transfer into the mouse retina mediated by an adeno-associated viral vector

Hum. Mol. Genetics

(1996)

R.R. Ali et al.

Adeno-associated virus gene transfer to mouse retina

Hum. Gene Ther.

(1998)

E.H. Bárány

Simultaneous measurement of changing intraocular pressure and outflow facility in the vervet monkey by constant pressure infusion

Invest. Ophthalmol.

(1964)

E.H. Bárány

Relative importance of autonomic nervous tone and structure as determinants of outflow resistance in normal monkey eyes (Cercopithecus ethiops and Macaca irus)

J. Bennett et al.

Adenovirus vector-mediated in vivo gene transfer into adult murine retina

Invest. Ophthalmol. Vis. Sci.

(1994)

T. Borrás et al.

Adenovirus gene transfer varies in efficiency and inflammatory response

Invest. Ophthalmol. Vis. Sci.

(1996)

T. Borrás et al.

Gene transfer into human trabecular meshwork by anterior segment perfusion

Invest. Ophthalmol. Vis. Sci.

(1998)

C.R. Brandt et al.

The herpes simplex virus ribonucleotide reductase is require for ocular virulence

J. Gen. Virol.

(1991)

C.R. Brandt et al.

The herpes simplex virus type 1 ribonucleotide reductase is required for acute retinal disease

Arch. Virol.

(1997)

X.O. Breakefield et al.

Herpes simplex virus for gene delivery to neurons

New Biologist

(1991)

D.L. Budenz et al.

In vivo gene transfer into murine corneal endothelial and trabecular meshwork cells

Invest. Ophthalmol. Vis. Sci.

(1995)

J.M. Cameron et al.

Ribonucleotide reductase encoded by herpes simplex virus is a determinant of the pathogenicity of the virus in mice and a valid antiviral target

J. Gen. Virol.

(1988)

M. Cayouette et al.

Adenovirus-mediated gene transfer to retinal ganglion cells

Invest. Ophthalmol. Vis. Sci.

(1996)

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Ninety percent of humans already have antibodies to HSV-1 or HSV-2 (Wald and Corey, 2011). In nonhuman primates, nearly 100% of the TM and non-pigmented ciliary epithelium cells were transduced (Liu et al., 1999), but achieved only 10%–20% transduction efficiency in rats and none in mice following an intracameral bolus (Spencer et al., 2000). While early generation vectors had short term expression measured in days (Liu et al., 1999), long-term expression has been achieved in the CNS with later generation vectors (Zhang et al., 2012).
Elevated intraocular pressure is the primary cause of open angle glaucoma. Outflow resistance exists within the trabecular meshwork but also at the level of Schlemm's canal and further downstream within the outflow system. Viral vectors allow to take advantage of naturally evolved, highly efficient mechanisms of gene transfer, a process that is termed transduction. They can be produced at biosafety level 2 in the lab using protocols that have evolved considerably over the last 15–20 years. Applied by an intracameral bolus, vectors follow conventional as well as uveoscleral outflow pathways. They may affect other structures in the anterior chamber depending on their transduction kinetics which can vary among species when using the same vector. Not all vectors can express long-term, a desirable feature to address the chronicity of glaucoma. Vectors that integrate into the genome of the target cell can achieve transgene function for the life of the transduced cell but are mutagenic by definition. The most prominent long-term expressing vector systems are based on lentiviruses that are derived from HIV, FIV, or EIAV. Safety considerations make non-primate lentiviral vector systems easier to work with as they are not derived from human pathogens. Non-integrating vectors are subject to degradation and attritional dilution during cell division. Lentiviral vectors have to integrate in order to express while adeno-associated viral vectors (AAV) often persist as intracellular concatemers but may also integrate. Adeno- and herpes viral vectors do not integrate and earlier generation systems might be relatively immunogenic. Nonviral methods of gene transfer are termed transfection with few restrictions of transgene size and type but often a much less efficient gene transfer that is also short-lived. Traditional gene transfer delivers exons while some vectors (lentiviral, herpes and adenoviral) allow transfer of entire genes that include introns. Recent insights have highlighted the role of non-coding RNA, most prominently, siRNA, miRNA and lncRNA. SiRNA is highly specific, miRNA is less specific, while lncRNA uses highly complex mechanisms that involve secondary structures and intergenic, intronic, overlapping, antisense, and bidirectional location. Several promising preclinical studies have targeted the RhoA or the prostaglandin pathway or modified the extracellular matrix. TGF-β and glaucoma myocilin mutants have been transduced to elevate the intraocular pressure in glaucoma models. Cell based therapies have started to show first promise. Past approaches have focused on the trabecular meshwork and the inner wall of Schlemm's canal while new strategies are concerned with modification of outflow tract elements that are downstream of the trabecular meshwork.
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^f1: Address correspondence to: Paul L. Kaufman, MD, Department of Ophthalmology and Visual Sciences, 600 Highland Avenue, Madison, WI 53792-3220, U.S.A.

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Regular articleHerpes Simplex Virus Mediated Gene Transfer to Primate Ocular Tissues

Abstract

Exp. Eye Res.

Exp. Eye Res.

Exp. Eye Res.

Virology

Neuromuscular Disorders

Antiviral Res.

Virology

FEBS Lett.

Ophthalmology

Am. J. Ophthalmol.

Virus Res.

Adenovirus-mediated heme oxygenase-1 gene transfer into rabbit ocular tissue

Invest. Ophthalmol. Vis. Sci.

Gene transfer into the mouse retina mediated by an adeno-associated viral vector

Hum. Mol. Genetics

Adeno-associated virus gene transfer to mouse retina

Hum. Gene Ther.

Simultaneous measurement of changing intraocular pressure and outflow facility in the vervet monkey by constant pressure infusion

Invest. Ophthalmol.

Relative importance of autonomic nervous tone and structure as determinants of outflow resistance in normal monkey eyes (Cercopithecus ethiops and Macaca irus)

Adenovirus vector-mediated in vivo gene transfer into adult murine retina

Invest. Ophthalmol. Vis. Sci.

Adenovirus gene transfer varies in efficiency and inflammatory response

Invest. Ophthalmol. Vis. Sci.

Gene transfer into human trabecular meshwork by anterior segment perfusion

Invest. Ophthalmol. Vis. Sci.

The herpes simplex virus ribonucleotide reductase is require for ocular virulence

J. Gen. Virol.

The herpes simplex virus type 1 ribonucleotide reductase is required for acute retinal disease

Arch. Virol.

Herpes simplex virus for gene delivery to neurons

New Biologist

In vivo gene transfer into murine corneal endothelial and trabecular meshwork cells

Invest. Ophthalmol. Vis. Sci.

Ribonucleotide reductase encoded by herpes simplex virus is a determinant of the pathogenicity of the virus in mice and a valid antiviral target

J. Gen. Virol.

Adenovirus-mediated gene transfer to retinal ganglion cells

Invest. Ophthalmol. Vis. Sci.

Regular article
Herpes Simplex Virus Mediated Gene Transfer to Primate Ocular Tissues