Versatile Genome Engineering Techniques Advance Human Ocular Disease Researches in Zebrafish

Zheng, Simin; Han, Ru-Yi; Xiang, Lan; Zhuang, Youyuan; Jin, Zi‐Bing

doi:10.3389/fcell.2018.00075

Cited by 11 publications

(21 citation statements)

References 90 publications

(95 reference statements)

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“…To validate the functional relevance of the prioritised genes to AMD pathogenesis, we sought an animal model that is amenable to manipulating gene expression and that can reliably evaluate ocular phenotypes. We chose zebrafish also because it has been extensively used to model ocular and other disorders 25–27 . Accounting for sequence homology between human and zebrafish, we obtained 4 of the 9 prioritised genes ( CNN2 , SARM1 , BLOC1S1 replicated at P SMR < 5.6e−3 and MMP9 at P SMR < 0.05 using the retina eQTL data; Table 1 ) with orthologue similarity above 60%, reported by either Ensembl or GeneCards, for functional follow-up ( URLs, Supplementary Table 2 ).…”

Section: Resultsmentioning

confidence: 99%

Towards the identification of causal genes for age-related macular degeneration

Cheng

Zhuang

Wen

et al. 2019

Preprint

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18Age-related macular degeneration (AMD) is a leading cause of visual impairment in ageing 19 populations and has no radical treatment or prevention. Although genome-wide association studies 20 (GWAS) have identified many susceptibility loci for AMD, the underlying causal genes remain 21 elusive. Here, we prioritized nine putative causal genes by integrating expression quantitative trait 22 locus (eQTL) data from blood ( = 2,765) with AMD GWAS data (16,144 cases vs. 17,832 controls) 23 and replicated six of them using retina eQTL data ( = 523). Of the six genes, altering expression of 24 cnn2, sarm1 and bloc1s1 led to ocular phenotype, impaired vision and retinal pigment epithelium 25 (RPE) loss in zebrafish. Essential photoreceptor and RPE genes were downregulated in cnn2-and 26 sarm1-knockdown zebrafishes. Through integration of GWAS and eQTL data followed by functional 27 validation, our study reveals potential roles of CNN2, SARM1 and BLOC1S1 in AMD pathogenesis 28 and demonstrates an efficient platform to prioritise causal genes for human complex diseases. 29 2 Introduction 30 Age-related macular degeneration (AMD) is an incurable blinding disorder caused by dysfunction of the 31 retinal pigment epithelium (RPE) and progressive loss of photoreceptors in the macula 1 . It results in visual 32 impairment of central vision and disability of daily life activities, such as reading, walking and face 33recognition. The prevalence of AMD is 8.69% in the age range of 45-85 years globally, and it is projected 34 to affect 196 million people worldwide in 2020 2 . As such, AMD is highly endorsed as a major health and 35 social problem for both individuals and communities, especially in elderly populations 3 . 36 37 AMD is one of the most genetically well-defined complex diseases. Genome-wide association studies 38 (GWAS) with increasing sample sizes have identified 52 susceptibility loci which together explain more 39 than 50% of the heritability of liability 4-6 . These findings provide important clues for understanding the 40 genetic architecture of the disease, but the causal genes at those susceptibility loci and underlying 41 mechanisms remain largely unclear. For example, a nonsynonymous variant, CFH p. Arg1210Cys (allele 42 frequency = 0.00017 in ExAC), increases AMD risk by >20-fold 7 , but there has been no evidence showing 43 its functional impact on the regulation of gene expression, structural and functional integrity of the protein-44 coding region, or interplay with the genes nearby 8 . This is partly because of linkage disequilibrium (LD) 45 between single-nucleotide polymorphisms (SNPs) and causative variants that GWAS mapping resolution 9 . 46This could also be the reason that the trait-associated variants, especially those residing in non-coding 47

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Section: Resultsmentioning

confidence: 99%

Towards the identification of causal genes for age-related macular degeneration

Cheng

Zhuang

Wen

et al. 2019

Preprint

Self Cite

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“…Although gene-based therapy could improve vision of RPE65-LCA patients, it failed to prevent degeneration process of this disease. Patient-derived retinal organoids or RPE cells can be utilized for regenerative medicine in combination with genome-editing technology (Deng et al, 2018; Zheng et al, 2018). Genome editing tools, the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system, zinc finger nucleases (ZFNs), and transcription activator-like effector nucleases (TALENs), are capable to correct mutations that lead to genetic diseases (Kime et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Generation and Characterization of Induced Pluripotent Stem Cells and Retinal Organoids From a Leber’s Congenital Amaurosis Patient With Novel RPE65 Mutations

Gao

Wang

et al. 2019

Front. Mol. Neurosci.

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RPE65-associated Leber congenital amaurosis (LCA) is one of highly heterogeneous, early onset, severe retinal dystrophies with at least 130 gene mutation sites identified. Their pathogenicity has not been directly clarified due to lack of diseased cells. Here, we generated human-induced pluripotent stem cells (hiPSCs) from one putative LCA patient carrying two novel RPE65 mutations with c.200T>G (p.L67R) and c.430T>C (p.Y144H), named RPE65-hiPSCs, which were confirmed to contain the same mutations. The RPE65-hiPSCs presented typical morphological features with normal karyotype, expressed pluripotency markers, and developed teratoma in NOD-SCID mice. Moreover, the patient hiPSCs were able to differentiate toward retinal lineage fate and self-form retinal organoids with layered neural retina. All major retinal cell types including photoreceptor and retinal pigment epithelium (RPE) cells were also acquired overtime. Compared to healthy control, RPE cells from patient iPSCs had lower expression of RPE65, but similar phagocytic activity and VEGF secretion level. This study provided the valuable patient specific, disease targeted retinal organoids containing photoreceptor and RPE cells, which would facilitate the study of personalized pathogenic mechanisms of disease, drug screening, and cell replacement therapy.

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“…The characteristics mentioned above, and the great genetic homology enable a widespread investigation of human ocular diseases in this vertebrate model (75). Several researchers search for genetic modifications in Zebrafish.…”

Section: Zebrafish Model In Ocular Diseasesmentioning

confidence: 99%

Alterações Genotóxicas, Citotóxicas E Mutagênicas: Um Conteúdo a Ser Ilustrado E Trabalhado No Ensino Médio

Brito¹,

Chagas²,

Messias³

et al. 2021

A Genética E a Construção De Novos Paradigmas Nas Ciências Da Vida

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Direitos para esta edição cedidos à Atena Editora pelos autores. Todo o conteúdo deste livro está licenciado sob uma Licença de Atribuição Creative Commons. Atribuição-Não-Comercial-NãoDerivativos 4.0 Internacional (CC BY-NC-ND 4.0). O conteúdo dos artigos e seus dados em sua forma, correção e confiabilidade são de responsabilidade exclusiva dos autores, inclusive não representam necessariamente a posição oficial da Atena Editora. Permitido o download da obra e o compartilhamento desde que sejam atribuídos créditos aos autores, mas sem a possibilidade de alterá-la de nenhuma forma ou utilizá-la para fins comerciais. Todos os manuscritos foram previamente submetidos à avaliação cega pelos pares, membros do Conselho Editorial desta Editora, tendo sido aprovados para a publicação com base em critérios de neutralidade e imparcialidade acadêmica. A Atena Editora é comprometida em garantir a integridade editorial em todas as etapas do processo de publicação, evitando plágio, dados ou resultados fraudulentos e impedindo que interesses financeiros comprometam os padrões éticos da publicação. Situações suspeitas de má conduta científica serão investigadas sob o mais alto padrão de rigor acadêmico e ético.

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Versatile Genome Engineering Techniques Advance Human Ocular Disease Researches in Zebrafish

Cited by 11 publications

References 90 publications

Towards the identification of causal genes for age-related macular degeneration

Towards the identification of causal genes for age-related macular degeneration

Generation and Characterization of Induced Pluripotent Stem Cells and Retinal Organoids From a Leber’s Congenital Amaurosis Patient With Novel RPE65 Mutations

Alterações Genotóxicas, Citotóxicas E Mutagênicas: Um Conteúdo a Ser Ilustrado E Trabalhado No Ensino Médio

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