Ultrahigh Resolution Mouse Optical Coherence Tomography to Aid Intraocular Injection in Retinal Gene Therapy Research

Butler, Mark C.; Sullivan, Jack

doi:10.3791/55894

Cited by 5 publications

(2 citation statements)

References 30 publications

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“…Overall retinal morphology and ONL thickness were periodically monitored and recorded noninvasively in age-matched C57BL/6J control and Dhdds flx/flx iCre + mice, using a Bioptigen 840 nm UHR-SD-OCT instrument (Bioptigen, NC, USA) essentially as described previously (Butler and Sullivan, 2018). Mice were anesthetized using ketamine-xylazine (75 mg/kg-5 mg/kg) by intraperitoneal injection.…”

Section: Spectral Doman Optical Coherence Tomography (Sd-oct) Analysismentioning

confidence: 99%

Retinal Degeneration Caused by Rod-Specific Dhdds Ablation Occurs without Concomitant Inhibition of Protein N-Glycosylation

Rao

Skelton

et al. 2020

iScience

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Dehydrodolichyl diphosphate synthase (DHDDS) catalyzes the committed step in dolichol synthesis. Recessive mutations in DHDDS cause retinitis pigmentosa (RP59), resulting in blindness. We hypothesized that rod photoreceptor-specific ablation of Dhdds would cause retinal degeneration due to diminished dolicholdependent protein N-glycosylation. Dhdds flx/flx mice were crossed with rod-specific Cre recombinase-expressing (Rho-iCre75) mice to generate rod-specific Dhdds knockout mice (Dhdds flx/flx iCre + ). In vivo morphological and electrophysiological evaluation of Dhdds flx/flx iCre + retinas revealed mild retinal dysfunction at postnatal (PN) 4 weeks, compared with age-matched controls; however, rapid photoreceptor degeneration ensued, resulting in almost complete loss of rods and cones by PN 6 weeks. Retina dolichol levels were markedly decreased by PN 4 weeks in Dhdds flx/flx iCre + mice, relative to controls; despite this, N-glycosylation of retinal proteins, including opsin (the dominant rod-specific glycoprotein), persisted in Dhdds flx/flx iCre + mice. These findings challenge the conventional mechanistic view of RP59 as a congenital disorder of glycosylation.

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Section: Spectral Doman Optical Coherence Tomography (Sd-oct) Analysismentioning

confidence: 99%

Retinal Degeneration Caused by Rod-Specific Dhdds Ablation Occurs without Concomitant Inhibition of Protein N-Glycosylation

Rao

Skelton

et al. 2020

iScience

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“…Pressure-regulated microinjectors allow for more accurate and constant injection volumes and hence a more controlled way of creating the subretinal bleb. Intraoperative optical coherence tomography (OCT) is now adopted as a diagnostic tool to help confirm the subretinal location of the needle and monitor the creation of the bleb [ 63 ].…”

Section: Subretinal Injection Techniquementioning

confidence: 99%

Subretinal Injection Techniques for Retinal Disease: A Review

Irigoyen

Alonso

Sanchez-Molina

et al. 2022

JCM

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Inherited retinal dystrophies (IRDs) affect an estimated 1 in every 2000 people, this corresponding to nearly 2 million cases worldwide. Currently, 270 genes have been associated with IRDs, most of them altering the function of photoreceptors and retinal pigment epithelium. Gene therapy has been proposed as a potential tool for improving visual function in these patients. Clinical trials in animal models and humans have been successful in various types of IRDs. Recently, voretigene neparvovec (Luxturna®) has been approved by the US Food and Drug Administration for the treatment of biallelic mutations in the RPE65 gene. The current state of the art in gene therapy involves the delivery of various types of viral vectors into the subretinal space to effectively transduce diseased photoreceptors and retinal pigment epithelium. For this, subretinal injection is becoming increasingly popular among researchers and clinicians. To date, several approaches for subretinal injection have been described in the scientific literature, all of them effective in accessing the subretinal space. The growth and development of gene therapy give rise to the need for a standardized procedure for subretinal injection that ensures the efficacy and safety of this new approach to drug delivery. The goal of this review is to offer an insight into the current subretinal injection techniques and understand the key factors in the success of this procedure.

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iOCT-guided simulated subretinal injections: a comparison between manual and robot-assisted techniques in an ex-vivo porcine model

Maierhofer,

Jablonka,

Roodaki

et al. 2023

J Robotic Surg

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The purpose of this study is to compare robot-assisted and manual subretinal injections in terms of successful subretinal blistering, reflux incidences and damage of the retinal pigment epithelium (RPE). Subretinal injection was simulated on 84 ex-vivo porcine eyes with half of the interventions being carried out manually and the other half by controlling a custom-built robot in a master–slave fashion. After pars plana vitrectomy (PPV), the retinal target spot was determined under a LUMERA 700 microscope with microscope-integrated intraoperative optical coherence tomography (iOCT) RESCAN 700 (Carl Zeiss Meditec, Germany). For injection, a 1 ml syringe filled with perfluorocarbon liquid (PFCL) was tipped with a 40-gauge metal cannula (Incyto Co., Ltd., South Korea). In one set of trials, the needle was attached to the robot’s end joint and maneuvered robotically to the retinal target site. In another set of trials, approaching the retina was performed manually. Intraretinal cannula-tip depth was monitored continuously via iOCT. At sufficient depth, PFCL was injected into the subretinal space. iOCT images and fundus video recordings were used to evaluate the surgical outcome. Robotic injections showed more often successful subretinal blistering (73.7% vs. 61.8%, p > 0.05) and a significantly lower incidence of reflux (23.7% vs. 58.8%, p < 0.01). Although larger tip depths were achieved in successful manual trials, RPE penetration occurred in 10.5% of robotic but in 26.5% of manual cases (p > 0.05). In conclusion, significantly less reflux incidences were achieved with the use of a robot. Furthermore, RPE penetrations occurred less and successful blistering more frequently when performing robotic surgery.

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Ultrahigh Resolution Mouse Optical Coherence Tomography to Aid Intraocular Injection in Retinal Gene Therapy Research

Cited by 5 publications

References 30 publications

Retinal Degeneration Caused by Rod-Specific Dhdds Ablation Occurs without Concomitant Inhibition of Protein N-Glycosylation

Retinal Degeneration Caused by Rod-Specific Dhdds Ablation Occurs without Concomitant Inhibition of Protein N-Glycosylation

Subretinal Injection Techniques for Retinal Disease: A Review

iOCT-guided simulated subretinal injections: a comparison between manual and robot-assisted techniques in an ex-vivo porcine model

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