Comprehensive characterization of fetal and mature retinal cell identity to assess the fidelity of retinal organoids

Kim, Hani Jieun; O’Hara-Wright, Michelle; Kim, Daniel; Loi, To Ha; Lim, Benjamin Y; Jamieson, Robyn V; Gonzalez‐Cordero, Anai; Yang, Pengyi

doi:10.1016/j.stemcr.2022.12.002

Cited by 12 publications

(9 citation statements)

References 50 publications

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“…For either Cas13bt3-valid or -invalid cells, we further used Seurat v4.3 for data pre-processing, data normalization, feature selection, data scaling, dimensional reduction and clustering. To refine the characterization of cell types within each cluster, ScType ( 62 ) was employed, leveraging genesets specific to eight retinal cell types— amacrine, bipolar, cones, horizontal, macroglial, microglial, rods, and RPE—validated by Kim et al ( 27 ). The magnitude (log2-transformed fold change) and significance (P-value) of differential expression between cell types were then calculated.…”

Section: Methodsmentioning

confidence: 99%

Characterisation of RNA editing and gene therapy with a compact CRISPR-Cas13 in the retina

Kumar,

Hsiao,

Wong

et al. 2024

Preprint

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CRISPR-Cas gene editing technology has revolutionised the development of novel genetic therapies. CRISPR-Cas13 nucleases are programmable RNA-targeting effectors that can silence gene expression in a reversible manner. Recent iterations of Cas13 nucleases are compact and adequate for adeno-associated virus (AAV) delivery to achieve strong and persistent transduction of various organs in a safe manner. Here, we report significant transcriptomic signatures of Cas13bt3 expression and demonstrate that all-in-one AAV gene therapy with Cas13bt3 can effectively silenceVEGFAin human retinal organoids and humanised mouse models. Specifically, human embryonic stem cells (hESC)-derived retinal pigment epithelium cells show high expression of Cas13bt3 after virus delivery corresponding to significant reduction ofVEGFAmRNA. We further show that intravitreal delivery of Cas13bt3 can transduce mouse retinal cells efficiently, reaching the photoreceptor cells for specific knockdown of humanVEGFA, without impacting mouseVEGFAin the trVEGF029 (Kimba) mouse. Our results reveal important considerations for assessing Cas13 activity and establish Cas13bt3 as a potential anti-VEGF agent that can achieve long-term control of VEGF for treatment of diabetic retinopathy and wet age-related macular degeneration.

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Section: Methodsmentioning

confidence: 99%

Characterisation of RNA editing and gene therapy with a compact CRISPR-Cas13 in the retina

Kumar,

Hsiao,

Wong

et al. 2024

Preprint

Self Cite

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“…Retinal organoids are complex organoids containing a rich population of rod and cone cells, and they are capable of recapitulating the human retina's developmental process from the optic vesicle to the optic cup 165 , 166 . The development of retinal organoids is strongly correlated with retinal differentiation programs and genes related to development, both in terms of their chemical composition and cellular makeup 167 . The integration of ST with single-cell transcriptomics and temporal transcriptomics has enabled the precise reconstruction of the process of retinal neuron differentiation and localization during retinal organoid development 168 .…”

Section: St For Drug Researchmentioning

confidence: 99%

Spatial Transcriptomics: A Powerful Tool in Disease Understanding and Drug Discovery

Cao,

Li,

Cui

et al. 2024

Theranostics

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Recent advancements in modern science have provided robust tools for drug discovery. The rapid development of transcriptome sequencing technologies has given rise to single-cell transcriptomics and single-nucleus transcriptomics, increasing the accuracy of sequencing and accelerating the drug discovery process. With the evolution of single-cell transcriptomics, spatial transcriptomics (ST) technology has emerged as a derivative approach. Spatial transcriptomics has emerged as a hot topic in the field of omics research in recent years; it not only provides information on gene expression levels but also offers spatial information on gene expression. This technology has shown tremendous potential in research on disease understanding and drug discovery. In this article, we introduce the analytical strategies of spatial transcriptomics and review its applications in novel target discovery and drug mechanism unravelling. Moreover, we discuss the current challenges and issues in this research field that need to be addressed. In conclusion, spatial transcriptomics offers a new perspective for drug discovery.

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“… 8 , 10 While the development rate of organoids in vitro and fetal retina in vivo is similar, the molecular and cellular composition of the organoids is highly dependent on the differentiation protocol. 11 …”

Section: Introductionmentioning

confidence: 99%

Deciphering the spatiotemporal transcriptional and chromatin accessibility of human retinal organoid development at the single-cell level

Dorgau,

Collin,

Rozanska

et al. 2024

iScience

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Comprehensive characterization of fetal and mature retinal cell identity to assess the fidelity of retinal organoids

Cited by 12 publications

References 50 publications

Characterisation of RNA editing and gene therapy with a compact CRISPR-Cas13 in the retina

Characterisation of RNA editing and gene therapy with a compact CRISPR-Cas13 in the retina

Spatial Transcriptomics: A Powerful Tool in Disease Understanding and Drug Discovery

Deciphering the spatiotemporal transcriptional and chromatin accessibility of human retinal organoid development at the single-cell level

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