Cell Types of the Human Retina and Its Organoids at Single-Cell Resolution

Cell, Год журнала: 2020, Номер 182(6), С. 1623 - 1640.e34

Опубликована: Сен. 1, 2020

Human organoids recapitulating the cell-type diversity and function of their target organ are valuable for basic translational research. We developed light-sensitive human retinal with multiple nuclear synaptic layers functional synapses. sequenced RNA 285,441 single cells from these at seven developmental time points periphery, fovea, pigment epithelium choroid light-responsive adult retinas, performed histochemistry. Cell types in matured vitro to a stable "developed" state rate similar retina development vivo. Transcriptomes organoid cell converged toward transcriptomes peripheral types. Expression disease-associated genes was cell-type-specific retina, specificity retained organoids. implicate unexpected diseases such as macular degeneration. This resource identifies cellular targets studying disease mechanisms targeted repair retinas.

Язык: Английский

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Retinal organoids: a window into human retinal development

Development, Год журнала: 2020, Номер 147(24)

Опубликована: Дек. 15, 2020

ABSTRACT Retinal development and maturation are orchestrated by a series of interacting signalling networks that drive the morphogenetic transformation anterior developing brain. Studies in model organisms continue to elucidate these complex events. However, human retina shows many differences from other investigation eye now benefits stem cell-derived organoids. differentiation methods have progressed simple 2D adherent cultures self-organising micro-physiological systems. As models development, collectively offered new insights into previously unexplored early informed our knowledge key cell fate decisions govern specification light-sensitive photoreceptors. Although developmental trajectories retinal types remain more elusive, collation omics datasets, combined with advanced culture methodology, will enable modelling intricate process retinogenesis disease vitro.

Язык: Английский

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Pluripotent stem cell-derived retinal organoids for disease modeling and development of therapies

Stem Cells, Год журнала: 2020, Номер 38(10), С. 1206 - 1215

Опубликована: Июнь 7, 2020

Abstract Retinal diseases constitute a genetically and phenotypically diverse group of clinical conditions leading to vision impairment or blindness with limited treatment options. Advances in reprogramming somatic cells induced pluripotent stem generation three-dimensional organoids resembling the native retina offer promising tools interrogate disease mechanisms evaluate potential therapies for currently incurable retinal neurodegeneration. Next-generation sequencing, single-cell analysis, advanced electrophysiology, high-throughput screening approaches are expected greatly expand utility cell-derived developing personalized treatments. In this review, we discuss current status future combining as human models recent technologies advance development gene, cell, drug retinopathies.

Язык: Английский

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Primary cilia biogenesis and associated retinal ciliopathies

Seminars in Cell and Developmental Biology, Год журнала: 2020, Номер 110, С. 70 - 88

Опубликована: Июль 31, 2020

The primary cilium is a ubiquitous microtubule-based organelle that senses external environment and modulates diverse signaling pathways in different cell types tissues. originates from the mother centriole through complex set of cellular events requiring hundreds distinct components. Aberrant ciliogenesis or ciliary transport leads to broad spectrum clinical entities with overlapping yet highly variable phenotypes, collectively called ciliopathies, which include sensory defects syndromic disorders multi-organ pathologies. For efficient light detection, photoreceptors retina elaborate modified known as outer segment, packed membranous discs enriched for components phototransduction machinery. Retinopathy phenotype involves dysfunction and/or degeneration sensing penetrant ciliopathies. This review will discuss cilia biogenesis focus on retina, role CP110-CEP290-CC2D2A network. We also explore how recent technologies can advance our understanding biology new paradigms developing potential therapies retinal

Язык: Английский

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Gene Therapy of Dominant CRX-Leber Congenital Amaurosis using Patient Stem Cell-Derived Retinal Organoids

Stem Cell Reports, Год журнала: 2021, Номер 16(2), С. 252 - 263

Опубликована: Янв. 28, 2021

Mutations in the photoreceptor transcription factor gene cone-rod homeobox (CRX) lead to distinct retinopathy phenotypes, including early-onset vision impairment dominant Leber congenital amaurosis (LCA). Using induced pluripotent stem cells (iPSCs) from a patient with CRX-I138fs48 mutation, we established an vitro model of CRX-LCA retinal organoids that showed defective maturation by histology and profiling, diminished expression visual opsins. Adeno-associated virus (AAV)-mediated CRX augmentation therapy partially restored phenotype phototransduction-related genes as determined single-cell RNA-sequencing. Retinal derived iPSCs second carrying K88N mutation revealed loss opsin common phenotype, which was alleviated AAV-mediated CRX. Our studies provide proof-of-concept for developing other retinopathies.

Язык: Английский

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Therapeutic Options in Hereditary Optic Neuropathies

Drugs, Год журнала: 2020, Номер 81(1), С. 57 - 86

Опубликована: Ноя. 7, 2020

Options for the effective treatment of hereditary optic neuropathies have been a long time coming. The successful launch antioxidant idebenone Leber's Hereditary Optic Neuropathy (LHON), followed by its introduction into clinical practice across Europe, was an important step forward. Nevertheless, other options, especially variety mitochondrial such as dominant atrophy (DOA), are needed, and number pharmaceutical agents, acting on different molecular pathways, currently under development. These include gene therapy, which has reached Phase III development LHON, but is expected to be developed also DOA, whilst most agents (other antioxidants, anti-apoptotic drugs, activators mitobiogenesis, etc.) almost all at II or preclinical stage research. Here, we review proposed target mechanisms, evidence, available trials with primary endpoints results, wide range tested molecules, give overview field, providing landscape future scenarios, including editing, reproductive options prevent transmission DNA mutations.

Язык: Английский

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A high-risk retinoblastoma subtype with stemness features, dedifferentiated cone states and neuronal/ganglion cell gene expression

Nature Communications, Год журнала: 2021, Номер 12(1)

Опубликована: Сен. 22, 2021

Abstract Retinoblastoma is the most frequent intraocular malignancy in children, originating from a maturing cone precursor developing retina. Little known on molecular basis underlying biological and clinical behavior of this cancer. Here, using multi-omics data, we demonstrate existence two retinoblastoma subtypes. Subtype 1, earlier onset, includes heritable forms. It harbors few genetic alterations other than initiating RB1 inactivation corresponds to differentiated tumors expressing mature markers. By contrast, subtype 2 harbor recurrent including MYCN -amplification. They express markers less together with neuronal/ganglion cell marked inter- intra-tumor heterogeneity. The dedifferentiation associated stemness features low immune interferon response, E2F MYC/MYCN activation higher propensity for metastasis. recognition these subtypes, one maintaining cone-differentiated state, other, more aggressive, expression neuronal markers, opens up important perspectives retinoblastomas.

Язык: Английский

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A look into retinal organoids: methods, analytical techniques, and applications

Cellular and Molecular Life Sciences, Год журнала: 2021, Номер 78(19-20), С. 6505 - 6532

Опубликована: Авг. 22, 2021

Inherited retinal diseases (IRDs) cause progressive loss of light-sensitive photoreceptors in the eye and can lead to blindness. Gene-based therapies for IRDs have shown remarkable progress past decade, but vast majority forms remain untreatable. In era personalised medicine, induced pluripotent stem cells (iPSCs) emerge as a valuable system cell replacement model IRD because they retain specific patient genome differentiate into any adult type. Three-dimensional (3D) iPSCs-derived retina-like tissue called organoid contains all major retina-specific types: amacrine, bipolar, horizontal, ganglion cells, Müller glia, well rod cone photoreceptors. Here, we describe main applications organoids provide comprehensive overview state-of-art analysis methods that apply this system. Finally, will discuss outlook improvements would bring cellular step closer become an established research treatment development IRDs.

Язык: Английский

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Cone photoreceptors in human stem cell-derived retinal organoids demonstrate intrinsic light responses that mimic those of primate fovea

Cell stem cell, Год журнала: 2022, Номер 29(3), С. 460 - 471.e3

Опубликована: Янв. 31, 2022

Язык: Английский

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Allele-specific gene editing to rescue dominant CRX-associated LCA7 phenotypes in a retinal organoid model

Stem Cell Reports, Год журнала: 2021, Номер 16(11), С. 2690 - 2702

Опубликована: Окт. 16, 2021

Cases of Leber congenital amaurosis caused by mutations in CRX (LCA7) exhibit an early form the disease and show signs significant photoreceptor dysfunction eventual loss. To establish a translational vitro model system to study gene-editing-based therapies, we generated LCA7 retinal organoids harboring dominant disease-causing mutation CRX. Our develop immature dysfunctional cells, providing us with reliable recapitulate LCA7. Furthermore, performed proof-of-concept which utilize allele-specific CRISPR/Cas9-based gene editing knock out mutant saw moderate rescue phenotypes our organoids. This work provides evidence for effective approach treat LCA7, can be applied more broadly other genetic diseases.

Язык: Английский

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