Delivery of Adeno-Associated Virus Vectors to the Fetal Retina: Impact of Viral Capsid Proteins on Retinal Neuronal Progenitor Transduction

Surace, Enrico Maria; Auricchio, Alberto; Reich, S.J.; Rex, Tonia S.; Glover, Ernest; Pineles, S; Tang, Waixing; O’Connor, Erin E.; Lyubarsky, Arkady; Savchenko, A.; Pugh, Edward N.; Maguire, Albert M.; Wilson, James M.; Bennett, Jean

doi:10.1128/jvi.77.14.7957-7963.2003

Cited by 40 publications

(35 citation statements)

References 41 publications

(44 reference statements)

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“…6). We were able to detect GFP expression in the RPE of injected retinal areas (data not shown), in agreement with the results previously reported for AAV-mediated subretinal delivery at early postnatal stages (Surace et al 2003;Colella and Auricchio 2010).…”

Section: In Vivo Misexpression Of Vax2os1 Affects Retinal Progenitor supporting

confidence: 81%

The long noncoding RNA Vax2os1 controls the cell cycle progression of photoreceptor progenitors in the mouse retina

Meola¹,

Pizzo²,

Alfano³

et al. 2011

RNA

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Long noncoding RNAs (lncRNAs) are emerging as regulators of many basic cellular pathways. Several lncRNAs are selectively expressed in the developing retina, although little is known about their functional role in this tissue. Vax2os1 is a retina-specific lncRNA whose expression is restricted to the mouse ventral retina. Here we demonstrate that spatiotemporal misexpression of Vax2os1 determines cell cycle alterations in photoreceptor progenitor cells. In particular, the overexpression of Vax2os1 in the developing early postnatal mouse retina causes an impaired cell cycle progression of photoreceptor progenitors toward their final committed fate and a consequent delay of their differentiation processes. At later developmental stages, this perturbation is accompanied by an increase of apoptotic events in the photoreceptor cell layer, in comparison with control retinas, without affecting the proper cell layering in the adult retina. Similar results are observed in mouse photoreceptor-derived 661W cells in which Vax2os1 overexpression results in an impairment of the cell cycle progression rate and cell differentiation. Based on these results, we conclude that Vax2os1 is involved in the control of cell cycle progression of photoreceptor progenitor cells in the ventral retina. Therefore, we propose Vax2os1 as the first example of lncRNA that acts as a cell cycle regulator in the mammalian retina during development.

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Section: In Vivo Misexpression Of Vax2os1 Affects Retinal Progenitor supporting

confidence: 81%

The long noncoding RNA Vax2os1 controls the cell cycle progression of photoreceptor progenitors in the mouse retina

Meola¹,

Pizzo²,

Alfano³

et al. 2011

RNA

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“…Earlier investigations that compared the transduction efficiency of rAAV serotypes in the eye used in vivo fluorescent imaging, fundus luminescence quantification, and in some studies, quantitative PCR. 3,4,[6][7][8]12 Quantitative analysis of immunostained retinal sections has been carried out after intravitreal rAAV2/2 injections, 11 but to our knowledge, this type of analysis has not been used to compare the transduction efficiency and tropism of different rAAV serotypes after injection into the vitreous. When analyzed 10 weeks after a single titre-matched intravitreal injection, we here confirm earlier tropism and transduction findings for rAAV2/2 11 and report new data on six additional rAAV vectors.…”

Section: Discussionmentioning

confidence: 99%

“…2 In the eye, rAAV has been used in experimental studies aimed at treating retinal degenerative diseases or for cell rescue following trauma. [3][4][5][6][7][8][9][10][11][12][13][14] On the basis of the success of some of these animal studies, clinical trials with AAV vectors have recently been initiated in humans with genetically linked ophthalmic diseases. 15,16 Human AAV serotype 2 was originally found as a contaminant in adenovirus preparations 17 and has not been associated with any human pathogenicity.…”

Section: Introductionmentioning

confidence: 99%

“…23 Variation in capsid structure affects viral tropism and expression kinetics in terms of the onset and expression levels of therapeutic genes, and the use of different serotypes may allow for a more cellspecific gene transfer. 3,6,8,12,24,25 Robust photoreceptor transduction has been reported after subretinal injection of many vector types; however, photoreceptors are only infrequently transduced after intravitreal injection of the serotypes tested to date. 3,4,[6][7][8]11,12,26 Intravitreal vector delivery is a less invasive technique compared with subretinal delivery.…”

Section: Introductionmentioning

confidence: 99%

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Cellular tropism and transduction properties of seven adeno-associated viral vector serotypes in adult retina after intravitreal injection

et al. 2008

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Recombinant adeno-associated virus (rAAV) vectors are increasingly being used as tools for gene therapy, and clinical trials have begun in patients with genetically linked retinal disorders. Intravitreal injection is optimal for the transduction of retinal ganglion cells (RGCs), although complete selectivity has not been achieved. There may also be advantages in using intravitreal approaches for the transduction of photoreceptors. Here we compared the cellular tropism and transduction efficiency of rAAV2/1, -2/2, -2/3, -2/4, -2/5, -2/6 and -2/8 in adult rat retina after intravitreal injection. Each vector encoded green fluorescent protein (GFP), and the number, laminar distribution and morphology of transduced GFP + cells were determined using fluorescent microscopy. Assessment of transduced cell phenotype was based on cell morphology and immunohistochemistry. rAAV2/2 and rAAV2/6 transduced the greatest number of cells, whereas rAAV2/5 and rAAV2/8 were least efficient. Most vectors primarily transduced RGCs; however, rAAV2/6 had a more diverse tropism profile, with 46% identified as amacrine or bipolar cells, 23% as RGCs and 22% as Mü ller cells. Mü ller cells were also frequently transduced by rAAV2/4. The highest photoreceptor transduction was seen after intravitreal rAAV2/3 injection. These data facilitate the design and selection of rAAV vectors to target specific retinal cells, potentially leading to an improved gene therapy for various human retinal pathologies.

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“…Detection must depend upon a brief window of time after surgery needed to introduce the vector, perhaps related to a concurrent but transient inflammation. This strategy may be useful in managing other types of rejection in the retina.Numerous investigators have used viral vectors to transduce retinal cells with enhanced green fluorescent protein (EGFP), with no evidence of rejection of this foreign protein (2,3,15,20,21). Similar results have been reported with other reporter genes that express foreign proteins in the retina, such as LacZ (6,8,14).…”

mentioning

confidence: 99%

Transient Immunosuppression Stops Rejection of Virus-Transduced Enhanced Green Fluorescent Protein in Rabbit Retina

Doi¹,

Kong²,

Hargitai³

et al. 2004

J Virol

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The expression of lentivirus-transduced enhanced green fluorescent protein (EGFP) was detectable in rabbit retinal pigment epithelium (RPE) within 3 to 5 days after subretinal injection of the vector. Within 2 to 3 weeks, EGFP-expressing cells were eliminated by rejection. In the current experiments, we monitor serum antibody titers for EGFP before and after transduction and determine whether systemic immunosuppression prevents recognition of EGFP by the immune system. While all control rabbits developed antibodies against EFGP and showed signs of rejection, no such evidence was observed with animals which received immunosuppression. One month of systemic immunosuppression permanently prevented rejection of RPE with EGFP expression. Fluorescence has been maintained for more than a year. If a control eye was injected with the same virus after terminating immunosuppression, both eyes showed signs of rejection. The lack of rejection is not due to tolerance but to a failure of the animals to detect the foreign protein. Detection must depend upon a brief window of time after surgery needed to introduce the vector, perhaps related to a concurrent but transient inflammation. This strategy may be useful in managing other types of rejection in the retina.Numerous investigators have used viral vectors to transduce retinal cells with enhanced green fluorescent protein (EGFP), with no evidence of rejection of this foreign protein (2,3,15,20,21). Similar results have been reported with other reporter genes that express foreign proteins in the retina, such as LacZ (6,8,14). We have found that with relatively high retinal expression of EGFP, transduced by lentivirus, rejection will occur (7). GFP has been shown to be immunogenic (9) and will be rejected in mice (19). In order to examine this phenomenon more completely, we have determined whether antibodies to EGFP can be detected in rabbits during the course of rejection in the retina. We have also determined whether this rejection can be prevented by immunosuppression. Our results indicate that serum antibodies to EGFP are detectable concurrent with retinal rejection of this fluorescent protein. If the rabbits are immunosuppressed, rejection does not occur. Surprisingly, if immunosuppression is stopped after 1 month, rejection never occurs and antibodies to EGFP are not detectable. These results imply that foreign proteins transduced in the retina are only detectable immunologically during a brief period of time after the surgical procedure needed to introduce the viral vector into the retina. MATERIALS AND METHODSPreparation of viral stocks. The viral stocks were produced as described previously (16) by cotransfection of human kidney-derived 293T cells with three plasmids by the calcium phosphate method (4). The packaging construct designated pCMV⌬R8.2 contained the cytomegalovirus (CMV) promoter and the insulin polyadenylation signal to express all of the viral proteins in trans form, except the envelope and Vpu. The second plasmid, pHRЈ-CMV-GFP, provided a vector with all ...

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Delivery of Adeno-Associated Virus Vectors to the Fetal Retina: Impact of Viral Capsid Proteins on Retinal Neuronal Progenitor Transduction

Cited by 40 publications

References 41 publications

The long noncoding RNA Vax2os1 controls the cell cycle progression of photoreceptor progenitors in the mouse retina

The long noncoding RNA Vax2os1 controls the cell cycle progression of photoreceptor progenitors in the mouse retina

Cellular tropism and transduction properties of seven adeno-associated viral vector serotypes in adult retina after intravitreal injection

Transient Immunosuppression Stops Rejection of Virus-Transduced Enhanced Green Fluorescent Protein in Rabbit Retina

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