CRISPR/Cas9-mediated genome editing in nonhuman primates

Disease Models & Mechanisms, Journal Year: 2019, Volume and Issue: 12(10)

Published: Oct. 1, 2019

ABSTRACT Owing to their high similarity humans, non-human primates (NHPs) provide an exceedingly suitable model for the study of human disease. In this Review, we summarize history transgenic NHP models and progress CRISPR/Cas9-mediated genome editing in NHPs, from first proof-of-principle green fluorescent protein-expressing monkeys sophisticated neurodegenerative disease that accurately phenocopy several complex features. We discuss not only breakthroughs advantages, but also potential shortcomings application CRISPR/Cas9 system NHPs have emerged expanded understanding technology recent years. Although off-target mosaic mutations are main concerns modeling, techniques make it likely these technical limitations will be overcome soon, bringing excellent prospects studies.

Language: Английский

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One-stop assembly of adherent 3D retinal organoids from hiPSCs based on 3D-printed derived PDMS microwell platform

Biofabrication, Journal Year: 2023, Volume and Issue: 15(3), P. 035005 - 035005

Published: March 24, 2023

Abstract The three-dimensional (3D) retinal organoids (ROs) derived from human induced pluripotent stem cells (hiPSCs), mimicking the growth and development of retina, is a promising model for investigating inherited diseases in vitro . However, efficient generation homogenous ROs remains challenge. Here we introduce novel polydimethylsiloxane (PDMS) microwell platform containing 62 V-bottom micro-cavities differentiation hiPSCs. uniform adherent 3D could spontaneously form using neural retina (NR) induction. Our results showed that complex NR (expressing VSX2), ciliary margin (CM) RDH10), pigment epithelium (RPE) ZO-1, MITF, RPE65) developed PDMS after differentiation. It important to note platforms not only enable one-stop assembly but also maintain homogeneity mature over period more than 25 weeks without use BMP4 Matrigel. Retinal ganglion BRN3a), amacrine AP2a), horizontal PROX1 AP2 α ), photoreceptor cone S-opsin L/M-opsin) rod Rod opsin), bipolar VSX2 PKC Müller glial GS Sox9) gradually emerged. Furthermore, replaced fetal bovine serum with platelet lysate established xeno-free culture workflow facilitates clinical application. Thus, our long-term favorable disease modeling, drug screening, manufacturing translation.

Language: Английский

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Presynaptic Expression of LRIT3 Transsynaptically Organizes the Postsynaptic Glutamate Signaling Complex Containing TRPM1

Cell Reports, Journal Year: 2019, Volume and Issue: 27(11), P. 3107 - 3116.e3

Published: June 1, 2019

Throughout the CNS, interactions between pre- and postsynaptic adhesion molecules establish normal synaptic structure function. Leucine-rich repeat (LRR) domain-containing proteins are a large family that has diversity of ligands, their absence can cause disease. At first retinal synapse, LRIT3 expression leads to disassembly glutamate signaling complex (signalplex) expressed on depolarizing bipolar cell (DBC) dendrites. The prevalent view is assembly signalplex results from direct protein:protein interactions. In contrast, we demonstrate presynaptically, in rod photoreceptors (rods), when restore Lrit3-/- rods, rod-driven vision. Our that, retina, LRR-containing protein acts as transsynaptic organizer required for

Language: Английский

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Retinal Cell Transplantation, Biomaterials, and In Vitro Models for Developing Next-generation Therapies of Age-related Macular Degeneration

Stem Cells Translational Medicine, Journal Year: 2022, Volume and Issue: 11(3), P. 269 - 281

Published: Jan. 7, 2022

Retinal pigment epithelium (RPE) cells grown on a scaffold, an RPE patch, have potential to ameliorate visual impairment in limited number of retinal degenerative conditions. This tissue-replacement therapy is suited for age-related macular degeneration (AMD), and related diseases. must be transplanted before the disease reaches point no return, represented by loss photoreceptors. Photoreceptors are specialized, terminally differentiated neurosensory that interact with RPE's apical processes functional. Human photoreceptors not known regenerate. On basal side, transplant induce reformation choriocapillaris, thereby re-establishing outer blood-retinal barrier. Because scaffold positioned between it should ideally degrade replaced natural extracellular matrix separates these tissues. Besides biodegradable, scaffolds need nontoxic, thin enough affect focal length eye, strong survive procedure, yet flexible conform curvature retina. The challenge patients progressing AMD treasure their remaining vision fear risky surgical procedure will further vision. Accordingly, clinical trials only treat eyes severe few patch. Although safety has been demonstrated, cell-replacement mechanism efficacy remain difficult validate. review covers structure retina, pathology AMD, limitations cell approaches, recent progress developing therapies using biomaterials.

Language: Английский

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Consensus guidelines for nomenclature of companion animal inherited retinal disorders

Veterinary Ophthalmology, Journal Year: 2024, Volume and Issue: unknown

Published: Feb. 9, 2024

Companion animals, namely dogs, cats, and horses, can be affected with many forms of hereditary retinal disease. The number such diseases characterized in the last decade has increased substantially, nomenclature is nonstandardized, heterogenous, confusing. We provide this viewpoint article consensus guidelines for naming companion animal diseases, either prospectively or retrospectively. These have been developed purpose standardizing nomenclature. examples iterative process a comprehensive File S1 on proposed renaming previously described diseases.

Language: Английский

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Outer Retinal Cell Replacement: Putting the Pieces Together

Translational Vision Science & Technology, Journal Year: 2021, Volume and Issue: 10(10), P. 15 - 15

Published: Nov. 1, 2021

Retinal degenerative diseases (RDDs) affecting photoreceptors (PRs) are one of the most prevalent sources incurable blindness worldwide. Due to a lack endogenous repair mechanisms, functional cell replacement PRs and/or retinal pigmented epithelium (RPE) cells among anticipated approaches for restoring vision in advanced RDD. Human pluripotent stem (hPSC) technologies have accelerated development outer therapies as they provide theoretically unlimited source donor cells. PSC-RPE progressed rapidly, with several completed and ongoing clinical trials. Although potentially more promising, hPSC-PR still their infancy. A first-in-human trial hPSC-derived neuroretinal transplantation has recently begun, but number questions regarding survival, reproducibility, integration, mechanism action remain. The discovery biomaterial transfer between PR highlighted need rigorous safety efficacy studies replacement. In this review, we briefly discuss history identify remaining challenges outline stepwise approach address specific pieces puzzle.

Language: Английский

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Biology, Pathobiology and Gene Therapy of CNG Channel-Related Retinopathies

Biomedicines, Journal Year: 2023, Volume and Issue: 11(2), P. 269 - 269

Published: Jan. 19, 2023

The visual process begins with the absorption of photons by photopigments cone and rod photoreceptors in retina. In this process, signal is first amplified a cyclic guanosine monophosphate (cGMP)-based signaling cascade then converted into an electrical nucleotide-gated (CNG) channels. CNG channels are purely ligand-gated whose activity can be controlled cGMP, which induces depolarizing Na+/Ca2+ current upon binding to channel. Structurally, belong superfamily pore-loop cation share structural similarities hyperpolarization-activated nucleotide (HCN) voltage-gated potassium (KCN) Cone express distinct encoded homologous genes. Mutations genes encoding channel (CNGA1 CNGB1) result retinitis-pigmentosa-type blindness. (CNGA3 CNGB3) lead achromatopsia. Here, we review molecular properties describe their physiological pathophysiological roles Moreover, summarize recent activities field gene therapy aimed at developing therapies for channelopathies.

Language: Английский

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Manifestations of systemic disease in the retina and fundus of cats and dogs

Frontiers in Veterinary Science, Journal Year: 2024, Volume and Issue: 11

Published: Feb. 20, 2024

The fundus is unique in that it the only part of body allows for a noninvasive and uninterrupted view vasculature nervous tissue. Utilization this can be powerful tool uncovering salient incidental findings which point to underlying systemic diseases, monitoring response therapy. Retinal venules arterioles allow clinician assess changes vascular color, diameter, outline, tortuosity. retina optic nerve may exhibit associated with increased or decreased thickness, inflammatory infiltrates, hemorrhages, detachments. While some retinal manifestations disease nonspecific, others are pathognomonic, presenting sign illness. examination an essential comprehensive physical examination. Systemic diseases present abnormalities include variety classifications, as represented by DAMNIT-V acronym, Degenerative/Developmental, Anomalous, Metabolic, Neoplastic, Nutritional, Inflammatory (Infectious/Immune-mediated/ischemic), Toxic, Traumatic Vascular. This review details illnesses syndromes have been reported manifest companion animals discusses key aspects differentiating their cause. Normal variations anatomy morphology also considered.

Language: Английский

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GTPBP2 in-frame deletion in canine model with non-syndromic progressive retinal atrophy

Scientific Reports, Journal Year: 2025, Volume and Issue: 15(1)

Published: Feb. 19, 2025

Progressive retinal atrophy (PRA), caused by aberrant functioning of rod/cone photoreceptors, leads to blindness affecting mammals, including dogs. We identified a litter three Labrador retrievers affected non-syndromic PRA; the parents and other siblings were unaffected. Homozygosity mapping whole-genome sequencing detected homozygous 3-bp deletion in coding region GTPBP2, located CFA12 (NC_049233.1:12,264,348_12,264,350del, c.1606_1608del, p.Ala536del). The variant was absent from online European Variation Archive (EVA) database, Dog Biomedical Variants Database Consortium, Dog10k database. tested 91 non-affected dogs same kennel found 75 wild-type (WT) 16 carriers, all clinically normal, 569 Labradors general population (USA), WT. GTPBP2 is associated with Jaberi-Elahi syndrome (JES) Homo sapiens, splice variants Mus musculus are neurodegeneration; both cases photoreceptor degeneration may be included its manifestation. Heterologous cellular systems transfected cDNA encoding WT or A536del mutant protein immunoblot analysis total cell lysate anti-GTPBP2 antibodies showed that expression level slightly but not significantly reduced compared Immunofluorescent methods confocal cells revealed form diffuse throughout cytosol, while resulted formation cytoplasmic aggregates ~70–80% cells. deleted amino acid falls within conserved interval outside GTP domain suggesting potentially novel role sequence on localization protein.

Language: Английский

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Choroid and Retinal Pigmented Epithelium in Pigs: A Comprehensive Light and Ultrastructural Study

Anatomia Histologia Embryologia, Journal Year: 2025, Volume and Issue: 54(2)

Published: March 1, 2025

This study was conducted on 12 adult pigs of a local mixed breed to examine the histology, histochemistry and ultrastructure choroid retinal pigmented epithelium (RPE). The tissues were fixed in Davidson's Fluid for light microscopy Karnovsky's fluid electron microscopy. Due physiological, anatomical metabolic similarities between humans, are suitable animal model various ophthalmic studies. Bruch's membrane exhibited five distinct layers: basement RPE, inner collagenous layer, elastic lamina, outer layer choriocapillaris. Age-related changes, such as vesicular formation, also observed. Patches collagen fibre lamellae noted stroma, indicating presence vestigial tapetum. apical part intercellular space RPE cells completely sealed off by zonula occludens adherens. Hemidesmosomes not present at basal surface cells. Numerous melanosomes, lipofuscin, mitochondria, phagolysosomes, melanolysosomal bodies, both smooth rough endoplasmic reticulum observed granular cytoplasm RPE. Double-membrane-bound round vesicles cytoplasm, characterised uniform intermembranous central core space, potentially representing liposome-like structures, although this type vesicle has been previously described Overall, demonstrated that have choroid, morphology similar providing valuable reference studies using model. Thus, can be utilised an experimental research.

Language: Английский

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