AAV-Mediated Expression Targeting of Rod and Cone Photoreceptors with a Human Rhodopsin Kinase Promoter

Khani, Shahrokh C.; Pawlyk, Basil; Bulgakov, Oleg V.; Kasperek, Eileen M.; Young, Justin G.; Adamian, Michael; Sun, Xun; Smith, Alexander J.; Ali, Robin R.; Li, Tiansen

doi:10.1167/iovs.07-0257

Cited by 113 publications

(130 citation statements)

References 39 publications

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“…To prevent expression in the retinal pigment epithelium, we used a photoreceptor-specific promoter, human GRK1 in AAV2/5 GRK1-CRE-GFP for expression of CRE-GFP specifically in photoreceptors (23). Subretinal administration of the AAV2/5 GRK1-driven constructs restricted the expression of GFP and CRE-GFP to the photoreceptor cells, no GFP-positive retinal pigment epithelial or inner retinal cells were detected ( Fig.…”

Section: Human Molecularmentioning

confidence: 99%

Targeted ablation of Crb2 in photoreceptor cells induces retinitis pigmentosa

Alves¹,

Pellissier²,

Vos³

et al. 2014

Human Molecular Genetics

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In humans, the Crumbs homolog-1 (CRB1) gene is mutated in autosomal recessive Leber congenital amaurosis and early-onset retinitis pigmentosa. In mammals, the Crumbs family is composed of: CRB1, CRB2, CRB3A and CRB3B. Recently, we showed that removal of mouse Crb2 from retinal progenitor cells, and consequent removal from Mü ller glial and photoreceptor cells, results in severe and progressive retinal degeneration with concomitant loss of retinal function that mimics retinitis pigmentosa due to mutations in the CRB1 gene. Here, we studied the effects of cell-type-specific loss of CRB2 from the developing mouse retina using targeted conditional deletion of Crb2 in photoreceptors or Mü ller cells. We analyzed the consequences of targeted loss of CRB2 in the adult mouse retina using adeno-associated viral vectors encoding Cre recombinase and short hairpin RNA against Crb2. In vivo retinal imaging by means of optical coherence tomography on retinas lacking CRB2 in photoreceptors showed progressive thinning of the photoreceptor layer and cellular mislocalization. Electroretinogram recordings under scotopic conditions showed severe attenuation of the a-wave, confirming the degeneration of photoreceptors. Retinas lacking CRB2 in developing photoreceptors showed early onset of abnormal lamination, whereas retinas lacking CRB2 in developing Mü ller cells showed late onset retinal disorganization. Our data suggest that in the developing retina, CRB2 has redundant functions in Mü ller glial cells, while CRB2 has essential functions in photoreceptors. Our data suggest that short-term loss of CRB2 in adult mouse photoreceptors, but not in Mü ller glial cells, causes sporadic loss of adhesion between photoreceptors and Mü ller cells.

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Section: Human Molecularmentioning

confidence: 99%

Targeted ablation of Crb2 in photoreceptor cells induces retinitis pigmentosa

Alves¹,

Pellissier²,

Vos³

et al. 2014

Human Molecular Genetics

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“…In contrast, foveal, parafoveal, perifoveal, and peripheral cones of nonhuman primates were all effectively transduced following subretinal delivery of AAV5-hGRK1-GFP (10 10 vg delivered) . As in rodent retinas (Khani et al 2007;Tan et al 2009;Boye et al 2010Boye et al , 2011Pawlyk et al 2010;Sun et al 2010), hGRK1 has exclusive activity in cones and rods of nonhuman primates . Taken together, these results support the use of a subretinally delivered AAV5 vector containing the hGRK1 promoter for delivering GUCY2D in a clinical setting.…”

Section: Discussionmentioning

confidence: 99%

“…Indeed, subretinal delivery of murine Gucy2e with AAV5 containing either the smCBA or photoreceptor-specific, human rhodopsin kinase (hGRK1) promoter (Khani et al 2007) led to robust rescue of the GC1KO phenotype (AAV5-smCBAGucy2e, 4 Â 10 10 vg delivered; AAV5-hGRK1-Gucy2e, 4 Â 10 10 vg delivered; [Boye et al 2010]). AAV-mediated GC1 expression was found exclusively in rods and cones of treated mice, and cone photoreceptors were preserved during the course of the 3-mo study.…”

Section: Gc1ko Mousementioning

confidence: 99%

Leber Congenital Amaurosis Caused by Mutations in GUCY2D

Boye

2014

Cold Spring Harbor Perspectives in Medicine

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“…As a result, our therapeutic viral construct led to clear improvement in the rod system but little cone improvement. However, given a wider therapeutic time window and an appropriate promoter (27)(28)(29), there is no obvious reason why cones could not also be rescued. Unlike the mouse used in this study, the Bbs1 M390R knockin has relatively normal cone ERG function at 11 wk of age (16) and may therefore be an appropriate model for future evaluation of cone rescue.…”

Section: Discussionmentioning

confidence: 99%

Gene therapy prevents photoreceptor death and preserves retinal function in a Bardet-Biedl syndrome mouse model

Simons

Boye

Hauswirth

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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Patients with Bardet-Biedl syndrome (BBS) experience severe retinal degeneration as a result of impaired photoreceptor transport processes that are not yet fully understood. To date, there is no effective treatment for BBS-associated retinal degeneration, and blindness is imminent by the second decade of life. Here we report the development of an adeno-associated viral (AAV) vector that rescues rhodopsin mislocalization, maintains nearly normalappearing rod outer segments, and prevents photoreceptor death in the Bbs4-null mouse model. Analysis of the electroretinogram awave indicates that rescued rod cells are functionally indistinguishable from wild-type rods. These results demonstrate that gene therapy can prevent retinal degeneration in a mammalian BBS model. ciliopathies | intra-flagellar transport | electroretinography B ardet-Biedl syndrome (BBS) is clinically diagnosed by the presence of at least four of the following signs: retinal dystrophy, polydactyly, obesity, learning disabilities, male hypogonadism, and renal anomalies (1). Of these, the visual phenotype is particularly devastating: the average BBS patient will progress to legal blindness before his or her 16th birthday. There are currently 15 genetic loci (Online Mendelian Inheritance in Man #209900) that are known to be associated with BBS, and although inheritance of this disease was once thought to follow a classic autosomal recessive pattern, recent evidence suggests that a more complex pattern, termed "triallelic inheritance," is involved at certain BBS loci (2). Only within the last decade has evidence emerged that BBS proteins play a role in ciliary function (3). More specifically, Bbs4 and the other BBSome components seem to be involved in both recruitment of cargo toward the ciliary basal body (4, 5) and in intraflagellar transport along the cilium (5-7). Several reports have elucidated the mechanisms by which disruption of BBS proteins give rise to the individual phenotypes seen in this highly pleiotropic syndrome (8-11).Retinal degeneration is a central feature of all BBS mouse models generated to date (8,(12)(13)(14)(15)(16). In rod and cone photoreceptors, the connecting cilium is a highly specialized ciliary structure that serves as the sole conduit from the inner segment to the outer segment. Because protein synthesis occurs proximal to the outer segment, rhodopsin and other visual proteins must be trafficked through the connecting cilium to reach their site of action in the outer segment. Abd-El-Barr et al. (11) showed that when Bbs4 was deleted in mice, rhodopsin and cone opsins became grossly mislocalized in rod and cone photoreceptors, respectively. This was followed by apoptotic photoreceptor death and deterioration of the electroretinogram (ERG) a-and b-waves (14). Ultrastructural analysis of rods from young animals revealed normal-appearing connecting cilia and basal body structures. This latter finding has important implications for gene therapy: because the structural transport apparatus seems to develop normally in Bbs4-nu...

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AAV-Mediated Expression Targeting of Rod and Cone Photoreceptors with a Human Rhodopsin Kinase Promoter

Abstract: The hRK promoter is active and specific for rod and cone photoreceptors. Because of its small size and proven activity in cones, it is a promoter of choice for somatic gene transfer and gene therapy targeting rods and cones.

Cited by 113 publications

References 39 publications

Targeted ablation of Crb2 in photoreceptor cells induces retinitis pigmentosa

Targeted ablation of Crb2 in photoreceptor cells induces retinitis pigmentosa

Leber Congenital Amaurosis Caused by Mutations in GUCY2D

Gene therapy prevents photoreceptor death and preserves retinal function in a Bardet-Biedl syndrome mouse model

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