The Argus® II Retinal Prosthesis System

Progress in Retinal and Eye Research, Journal Year: 2015, Volume and Issue: 50, P. 89 - 107

Published: Sept. 25, 2015

Language: Английский

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Optogenetics for light control of biological systems

Nature Reviews Methods Primers, Journal Year: 2022, Volume and Issue: 2(1)

Published: July 21, 2022

Language: Английский

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Early and late stage gene therapy interventions for inherited retinal degenerations

Progress in Retinal and Eye Research, Journal Year: 2021, Volume and Issue: 86, P. 100975 - 100975

Published: May 29, 2021

Inherited and age-related retinal degeneration is the hallmark of a large group heterogeneous diseases main cause untreatable blindness today. Genetic factors play major pathogenic role in degenerations for both monogenic (such as retinitis pigmentosa) complex with established genetic risk macular degeneration). Progress genotyping techniques back eye imaging are completing our understanding these their manifestations patient populations suffering from degenerations. It clear that whatever cause, majority vision loss results photoreceptor function. The timing circumstances surrounding function determine adequate therapeutic approach to use each patient. Among such approaches, gene therapy rapidly becoming reality applicable clinic. This massive move laboratory work towards clinical application has been propelled by advances disease genetics mechanisms, delivery vectors, editing systems, compensatory strategies Here, we provide an overview existing modalities relevance based on needs inherited

Language: Английский

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Red‐shifted channelrhodopsin stimulation restores light responses in blind mice, macaque retina, and human retina

EMBO Molecular Medicine, Journal Year: 2016, Volume and Issue: 8(11), P. 1248 - 1264

Published: Sept. 27, 2016

Research Article27 September 2016Open Access Transparent process Red-shifted channelrhodopsin stimulation restores light responses in blind mice, macaque retina, and human retina Abhishek Sengupta INSERM, U968, Paris, France Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, CNRS, UMR_7210, Search for more papers by this author Antoine Chaffiol Emilie Macé Romain Caplette Mélissa Desrosiers Maruša Lampič Valérie Forster Olivier Marre John Y Lin School of Medicine, University Tasmania, Hobart, Australia José-Alain Sahel Hôpital des Quinze-Vingts, Serge Picaud Deniz Dalkara Corresponding Author [email protected] Jens Duebel orcid.org/0000-0002-9906-5597 Information Sengupta1,2,3,6,‡, Chaffiol1,2,3,‡, Macé1,2,3, Caplette1,2,3, Desrosiers1,2,3, Lampič1,2,3, Forster1,2,3, Marre1,2,3, Lin4, Sahel1,2,3,5, Picaud1,2,3, *,1,2,3 1INSERM, 2Sorbonne 3CNRS, 4School 5Hôpital 6Present address: Unit on Retinal Neurophysiology, National Eye Institute Graduate Partnerships Program, Institutes Health, Bethesda, MD, USA ‡These authors contributed equally to work *Corresponding author. Tel: +33 6 52 56 24 00; E-mail: 1 53 46 25 32; EMBO Mol Med (2016)8:1248-1264https://doi.org/10.15252/emmm.201505699 PDFDownload PDF article text main figures. Peer ReviewDownload a summary the editorial decision including letters, reviewer comments feedback. ToolsAdd favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract Targeting photosensitive ion channel channelrhodopsin-2 (ChR2) retinal circuitry downstream photoreceptors holds promise treating vision loss caused degeneration. However, high intensity blue necessary activate exceeds safety threshold illumination because its strong potential induce photochemical damage. In contrast, damage red-shifted is vastly lower than that light. Here, we show (ReaChR), delivered AAV injections rd1 enables restoration at retinal, cortical, behavioral levels, using orange intensities below retina. We further postmortem retinae infected with AAV-ReaChR can respond spike trains safe intensities. Finally, directly address question translatability subjects, demonstrate first time, AAV- lentivirus-mediated optogenetic ganglion cells Synopsis A (ReaChR) was targeted three models parallel: mouse, macaque, human. Safe able trigger all systems. The ReaChR restored levels limit Optogenetic were demonstrated explanted ex vivo viral vectors encoding ReaChR. study presents electrophysiological recordings obtained from fovea as well far peripheral Introduction Pioneering studies have targeting microbial opsins, such (Nagel et al, 2003), surviving neurons sensitivity degenerated retinae, light-driven information transmitted higher visual centers visually guided behavior be elicited (Bi 2006; Lagali 2008; Thyagarajan 2010; Tomita Doroudchi 2011). major drawback ChR2 requires intensities, having risk inducing pigment epithelium (Ham 1978; Chen, 1993; Rozanowska 1995; Sparrow 2000, 2002; Lamb 2001; Wu Hunter 2012). Indeed, sufficient would exceed artificial radiation permitted applied (European Commission, ICNIRP, 2013). solution problem use variants. key point eye has an exponentially decaying correlation wavelength, implying even small changes wavelength impact threshold. For example, shifting 470 nm (blue light) 590 (orange light), are increase orders magnitude 2013), thus making variants ideal candidates restoration. derived Volvox carteri (VChR1) exhibits peak response ∼530 but shows poor membrane trafficking when expressed mammalian (Zhang 2008). Molecular engineering efforts led novel mutants enhanced expression action spectra (Lin 2013; Klapoetke 2014; 2014). recently developed variant, called red-activatable ChR which displays improved robust spectral up 600 By vector, (RGCs) mouse model retinitis pigmentosa (rd1 mouse). Upon wavelengths, allowed eye, observed ReaChR-induced relayed cortex mediated behavior. also functional, Results mice (4–5 weeks old) AAV2 ReaChR-mCitrine under pan-neuronal hSyn promoter via intravitreal injections. Four post-injection, mCitrine revealed fundus imaging (Fig 1A). Live 2-photon ReaChR-treated 1B) confocal (Figs 1C, EV1A) exhibited endogenous fluorescence localized RGC dense labeling dendritic arbors inner plexiform layer. Next, tested required stimulate ReaChR-expressing RGCs (ReaChR-RGCs) (> 4 months old). ReaChR-evoked photocurrents different spanning five logarithmic units measured patch-clamp electrodes 2A). wild-type cones (Nikonov 2006) 2B). (n = 7 cells) regulatory dashed line). comparison, nm, normally used ChR-2, ~1,000× line) This highlights importance strategy (see Table 1). stimulating wavelengths constant (~1016 photons cm−2 s−1, see 4), spectrum seen representative 2C). normalized 2D) calculated both (solid line; n 8 multi-electrode array (dashed 168 ReaChR-RGCs activated ranging 500 nm. To assess temporal properties RGCs, recorded pulses 2 22 Hz. Despite rather slow offset (τOff) EV1C) consistently persisting periodicity current Hz 2E). Then, quantified flicker stimulations 2F). RGC-spike discharges could follow increasing frequencies 30 standard cinema movie frame refresh rate Hz, rd1-RGCs achieve resolution vision. Figure 1. efficiently arbor image AAV-injected expressing driven (AAV2-hSyn:ReaChR-mCitrine); transduced visible green puncta. showing restricted membranes their processes. Scale bar, 10 μm. Confocal vertical section ReaChR-mCitrine. Endogenous (mCit) processes Fig EV1A); blue: DAPI (to visualize nuclei); nuclear layer (INL); (IPL); cell (GCL). Download figure PowerPoint Click here expand figure. EV1. Membrane/dendrite-targeted treated Vertical through AAV2-hSyn:ReaChR-mCitrine-injected IPL. stain label layers, (INL), (IPL), mCit not amplified. Example mCitrine-positive being approached patch pipette flat-mounted scale On off time constants (τ) photocurrent response; 1.2 × 1016 s−1 550 (left) results (right). τOn 26 ± ms, τOff 153 15 ms (mean SEM). Marks indicate needed reach 63% onset value, decay 37% maxima (τ 2. triggers injected AAV2-hSyn:ReaChR-mCitrine Light sensitivity: Photocurrents stimulated varying (˜1013 − 1018 s−1). function (590 cells, mean amplitude 4.2 1017 −349 192 pA). At bottom, range shown comparison RGCs. lines maximum respectively Photocurrent (1.2 Normalized spectrums ReaChR-RGCs. Solid line represents data cells; −324 97 pA. Dashed ˜1016 Temporal properties. Modulation (patch-clamp recording RGC; magnified trace shown; 1.3 Spiking modulation ratio demonstrates reliable triggering (297 Data information: Graphs (B), (D), (F) presented SEM. Safety thresholds specific (ICNIRP, 2013) Wavelength: λ (nm) Spectral weighting factor: Bλ retina: (photons s−1) 440 1.0 ≤ 4.43 1014 0.62 7.62 0.1 5.03 1015 0.01 5.53 0.001 5.94 order therapeutic approaches useful patients, triggered must brain. this, performed age-matched control (WT) 3A–C). 200 (595 nm) pulses. maximal local field (LFP) −281.2 68.4 μV; 3A) flat-lined untreated (−10.3 2.9 μV, 3; vs. P < 0.03, two-tailed t-test) 3A). cortical profile WT similarly 595 provided ON OFF responses, while displayed pure responses. Additionally, stimuli produced 3B) failed generate 3B). assessed onsets 3D) 2.5 1013 modulations LFP beginning suggesting detect 3. ReaChR-triggered AAV2-hSyn:ReaChR-mCitrine, detected Examples traces flashes (light stimulus bar; 1.25 Spike raster plots repetitions same stimulus. Averaged peristimulus histograms (PSTH, bin size: 5 ms) based plots. Top row: middle bottom mouse. (spikes LFP) demonstrating 4) Plots (A–C) been constructed single WT, respectively. (B C) single-unit neural Could ReaChR, modulated light? investigate placed circular open-field like arena fitted 590-nm LED sources. showed 4A rd1, Movie EV1) characterized sharp reduction velocity movement respect (2.18 0.33 cm 11 3.55 0.37 12 rd1; 0.006, one-tailed during min after 4B). remained unaffected upon EV2). Since only our approach, asked whether these discriminate dark fields. Therefore, light/dark box, illuminated 2.1 s−1. avoidance starting 2.11 spent 14.43 4.4% total duration test compartment 4C, EV3). significantly less performance increased elevated. did any light-induced Independent intensity, average ~59% 4C). 4D animals 59.3 2.8% 9), 37.7 2.44% 14.4 7) compartment. Both (P 0.0001, one-way ANOVA followed Tukey's multiple comparisons compare ReaChR-treated). 4. modulate Locomotory (irradiance level mice's eyes ˜1015 s−1; cumulative activity trajectories red illumination). See Movies EV1 EV2. Mean velocities analyzed over onset. reduced compared (*P t-test, ReaChR) illumination. SEM each group. 11), 12). Light/dark box test. Percentage (WT, 9; ReaChR-treated, 7). 9) ordinary ANOVA). ***P ****P 0.0001 significance determined EV3 evaluate clinical primate prepared explants mid-peripheral parts them tissue culture (Fradot After days incubation AAV2-hSyn:ReaChR-mCit, membrane-bound dendrites stratifying 5A). repeated trials, attempted record construct. stronger vector conditions, utilized ubiquitous CAG instead packaged it AAV8 Y447 733. 3 733-CAG:ReaChR-GFP, GFP 5B) 5C). characteristics 2-s 580 array. stimulation, sustained 5C EV2A), evoked reconstruct larger population, averaged AAV-infected (28 ~550 5D). evaluated cell-attached recording, 5E). All experiments bath application L-AP4 block possible remaining input photoreceptors. Control multielectrode uninfected (three two animals) presence complete abolition spontaneous spiking EV2C voltage traces). 5. Evaluation translational explant transfected AAV2-hSyn:ReaChR-mCit exhibiting densely neuronal GCL. 733F-CAG:ReaChR-GFP incubation. primarily bodies RGCs; green: fluorescence; DAPI; outer (ONL). PSTH 4, 8, respectively, macaques (580 nm; 1.24 PSTHs lines, responding cells). Raster individual (10 repetitions). Action recordings. firing rate, error bars cells), EV2B Responses (Hz) mode) (2–22 Hz; represented Peristimulus (PSTH) B built full-field 1.59 (performed AMES supplemented L-AP4) lines). individually A, B, C

Language: Английский

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Persistent remodeling and neurodegeneration in late-stage retinal degeneration

Progress in Retinal and Eye Research, Journal Year: 2019, Volume and Issue: 74, P. 100771 - 100771

Published: July 26, 2019

Language: Английский

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A New Promoter Allows Optogenetic Vision Restoration with Enhanced Sensitivity in Macaque Retina

Molecular Therapy, Journal Year: 2017, Volume and Issue: 25(11), P. 2546 - 2560

Published: July 20, 2017

The majority of inherited retinal degenerations converge on the phenotype photoreceptor cell death. Second- and third-order neurons are spared in these diseases, making it possible to restore light responses using optogenetics. Viral expression channelrhodopsin under ubiquitous promoters was previously shown visual function, albeit at intensities above illumination safety thresholds. Here, we report (to our knowledge, for first time) activation macaque retinas, up 6 months post-injection, channelrhodopsin-Ca2+-permeable (CatCh) safe intensities. High-level CatCh achieved due a new promoter based regulatory region gamma-synuclein gene (SNCG) allowing strong ganglion cells across species. Our promoter, combination with clinically proven adeno-associated virus 2 (AAV2), provides peri-foveolar responding robustly thresholds human eye. On contrary, threshold proportion unresponsive were much higher when (cytomegalovirus [CMV]) used express CatCh. results study suggest that inclusion optimized is key path clinical translation

Language: Английский

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Restoration of high-sensitivity and adapting vision with a cone opsin

Nature Communications, Journal Year: 2019, Volume and Issue: 10(1)

Published: March 15, 2019

Abstract Inherited and age-related retinal degenerative diseases cause progressive loss of rod cone photoreceptors, leading to blindness, but spare downstream neurons, which can be targeted for optogenetic therapy. However, approaches have been limited by either low light sensitivity or slow kinetics, lack adaptation changes in ambient light, not shown restore object vision. We find that the vertebrate medium wavelength opsin (MW-opsin) overcomes these limitations supports vision dim light. MW-opsin enables an otherwise blind retinitis pigmenotosa mouse discriminate temporal spatial patterns displayed on a standard LCD computer tablet, displays adaption restores open-field novel exploration under incidental room By contrast, rhodopsin, is similar slower response has greater rundown, fails tests. Thus, provides speed, needed patterned

Language: Английский

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Noninvasive gene delivery to foveal cones for vision restoration

JCI Insight, Journal Year: 2018, Volume and Issue: 3(2)

Published: Jan. 24, 2018

Intraocular injection of adeno-associated viral (AAV) vectors has been an evident route for delivering gene drugs into the retina. However, gaps in our understanding AAV transduction patterns within anatomically unique environments subretinal and intravitreal space primate eye impeded establishment noninvasive efficient delivery to foveal cones clinic. Here, we establish new vector-promoter combinations overcome limitations associated with AAV-mediated cone fovea supporting studies mouse models, human induced pluripotent stem cell–derived organoids, postmortem retinal explants, living macaques. We show that AAV9 variant provides when injected several millimeters away from fovea, without detaching this delicate region. An engineered AAV2 a well-tolerated dose administered intravitreally. Both modalities rely on cone-specific promoter result high-level transgene expression compatible optogenetic vision restoration. The model systems described here provide insight behavior across species obtain safety efficacy needed therapy neurodegenerative disorders.

Language: Английский

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Restoration of visual function by transplantation of optogenetically engineered photoreceptors

Nature Communications, Journal Year: 2019, Volume and Issue: 10(1)

Published: Oct. 4, 2019

A major challenge in the treatment of retinal degenerative diseases, with transplantation replacement photoreceptors, is difficulty inducing grafted cells to grow and maintain light sensitive outer segments host retina, which depends on proper interaction underlying pigment epithelium (RPE). Here, for an RPE-independent approach, we introduce a hyperpolarizing microbial opsin into photoreceptor precursors from newborn mice, transplant them blind mice lacking layer. These optogenetically-transformed photoreceptors are responsive their leads recovery visual function, as shown by ganglion cell recordings behavioral tests. Subsequently, generate cone human induced pluripotent stem cells, expressing chloride pump Jaws. After observe light-driven responses at levels. results demonstrate that structural functional repair possible combining therapy optogenetics.

Language: Английский

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Optogenetic therapy: high spatiotemporal resolution and pattern discrimination compatible with vision restoration in non-human primates

Communications Biology, Journal Year: 2021, Volume and Issue: 4(1)

Published: Jan. 27, 2021

Abstract Vision restoration is an ideal medical application for optogenetics, because the eye provides direct optical access to retina stimulation. Optogenetic therapy could be used diseases involving photoreceptor degeneration, such as retinitis pigmentosa or age-related macular degeneration. We describe here selection, in non-human primates, of a specific optogenetic construct currently tested clinical trial. microbial opsin ChrimsonR, and showed that AAV2.7m8 vector had higher transfection efficiency than AAV2 retinal ganglion cells (RGCs) ChrimsonR fused tdTomato (ChR-tdT) was expressed more efficiently ChrimsonR. Light at 600 nm activated RGCs transfected with ChR-tdT, from irradiance 10 15 photons.cm −2 .s −1 . Vector doses 5 × 11 vg/eye up 7000 RGCs/mm 2 perifovea, no significant immune reaction. recorded RGC responses stimulus duration 1 ms upwards. When using activity decode information, we obtained estimated visual acuity 20/249, above level legal blindness (20/400). These results lay groundwork ongoing trial - ChR-tdT vision patients pigmentosa.

Language: Английский

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