miR-183/96/182 cluster is an important morphogenetic factor targeting<i>PAX6</i>expression in differentiating human retinal organoids

Pešková, Lucie; Jurčíková, Denisa; Váňová, Tereza; Křivánek, Jan; Capandová, Michaela; Šrámková, Zuzana; Kolouskova, Magdalena; Kotasová, Hana; Streit, Libor; Bárta, Tomáš

doi:10.1101/2020.04.01.019539

Cited by 4 publications

(6 citation statements)

References 31 publications

(39 reference statements)

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“…The detection and expression patterns of these markers highlight extensive laminar formation by day 35, as described in previous studies, suggesting high replicability of this protocol to generate hiPSC-derived ocular vesicles faithful to human eye development. 22,32,[38][39][40] Importantly, we detect the expression of more mature photoreceptor and retinal ganglion cell progenitor markers such as CRX, OTX2 and BRN3b by day 50, suggesting this protocol can generate retinal lineage-specific organoids. Similar expression patterns have been detected in human foetal tissue (HFT), suggesting a high fidelity between in vitro optic vesicles and HFT.…”

Section: Discussionmentioning

confidence: 75%

Efficient embryoid-based method to improve generation of optic vesicles from human induced pluripotent stem cells

et al. 2022

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Animal models have provided many insights into ocular development and disease, but they remain suboptimal for understanding human oculogenesis. Eye development requires spatiotemporal gene expression patterns and disease phenotypes can differ significantly between humans and animal models, with patient-associated mutations causing embryonic lethality reported in some animal models. The emergence of human induced pluripotent stem cell (hiPSC) technology has provided a new resource for dissecting the complex nature of early eye morphogenesis through the generation of three-dimensional (3D) cellular models. By using patient-specific hiPSCs to generate in vitro optic vesicle-like models, we can enhance the understanding of early developmental eye disorders and provide a pre-clinical platform for disease modelling and therapeutics testing. A major challenge of in vitro optic vesicle generation is the low efficiency of differentiation in 3D cultures. To address this, we adapted a previously published protocol of retinal organoid differentiation to improve embryoid body formation using a microwell plate. Established morphology, upregulated transcript levels of known early eye-field transcription factors and protein expression of standard retinal progenitor markers confirmed the optic vesicle/presumptive optic cup identity of in vitro models between day 20 and 50 of culture. This adapted protocol is relevant to researchers seeking a physiologically relevant model of early human ocular development and disease with a view to replacing animal models.

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Section: Discussionmentioning

confidence: 75%

Efficient embryoid-based method to improve generation of optic vesicles from human induced pluripotent stem cells

et al. 2022

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“…The miR-183-96-182 cluster is well known to be a typical miRNA cluster because its three components have similar seed sequences and common target genes, and even play similar biological functions [10,14]. In the present study, we demonstrated that the miR-183-96-182 cluster members may share a common promoter, which is activated by the same Increasing evidence suggests that the miR-183-96-182 cluster has a common putative promoter in humans [4,30,35,36].…”

Section: Discussionmentioning

confidence: 99%

“…Previous studies have reported that miRNA clusters are prevalent in various organisms (Drosophila melanogaster, 50%; humans, 25%) and are highly conserved among different species [6]. Owing to the sharing of common promoters, members of a cluster are usually regulated by the same factors and show similar expression profiles [7][8][9][10].…”

Section: Introductionmentioning

confidence: 99%

“…Due to similar seed sequences, all members of this cluster tend to have common targets and similar biological functions [12][13][14]. A recent study revealed that the miR-183-96-182 cluster regulates neuroectoderm specification and retinal tissue morphogenesis by targeting PAX6 [10]. In human ovaries, increased miR-183-96-182 cluster expression has been shown to enhance luteal cell survival and progesterone release by targeting FoxO1 [15].…”

Section: Introductionmentioning

confidence: 99%

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SMAD4 Inhibits Granulosa Cell Apoptosis via the miR-183-96-182 Cluster and FoxO1 Axis

Wang

et al. 2021

Reprod. Sci.

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The miR-183-96-182 cluster is a polycistronic miRNA cluster necessary for ovarian functions in mammals. However, its transcriptional regulation in the ovary is largely unclear. In this study, we characterized the promoter region of the porcine miR-183-96-182 cluster, and showed that SMAD4 may function as a transcriptional activator of the miR-183-96-182 cluster in GCs through direct binding to SBE motifs in its promoter. SMAD4 may inhibit GC apoptosis via suppression of FoxO1, an effector of GC apoptosis and a direct target of the miR-183-96-182 cluster, by inducing the miR-183-96-182 cluster, and this process may be regulated by the TGF-β/SMAD signaling pathway. Our findings uncovered the regulatory mechanism of miR-183-96-182 cluster expression in GCs and demonstrated that TGF-β1/SMAD4/miR-183-96-182 cluster/FoxO1 may be a potential pathway for regulating follicular atresia and female reproduction.

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“…This cluster is expressed in embryonic stem cells, and in the inner ear[11] and retina[12]. It was also observed that a temporal reduction in the expression of miR183 cluster is necessary for epidermal differentiation to neuroectodermal lineage[13].…”

Section: Introductionmentioning

confidence: 99%

Merkel cell polyomavirus regulates miR183 cluster and piR62011 in Merkel cell carcinoma

Arora

Dohadwala

Nanavati

et al. 2022

Preprint

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Merkel cell carcinoma (MCC) is a rare, aggressive skin cancer, a major subset of which is caused by the clonal integration of Merkel Cell Polyomavirus (MCV). Recent studies by Cheng et al. (2017) reported that virus-derived small T antigen protein-bound EP400 complex drives expression of genes essential for cellular transformation. On close analysis of their ChIP-Seq data, we uncovered that the complex binds to the promoter region of the microRNA-183 cluster. The miRNA183 cluster is a cluster of 3 miRNAs (miR183, 182 & 96) expressed and regulated together. These miRNAs are conserved across species, highly expressed in human embryonic stem cells and necessary for sensory/ mechanosensory organ development. We hypothesized that the MCV oncoproteins regulate host miRNA expression directly; an interaction novel in polyomaviruses. We tested miRNA expression via qPCR in both virus positive and negative MCC cell lines and found the former showed a much higher level. Further, fibroblasts expressing T antigens displayed an increase in miR182 expression in comparison to control. Knock-down of T antigens in MCC cells correspondingly decreased miR182 levels. To investigate its regulation we performed luciferase assays for the miRNA predicted promoter that showed increased activity in the presence of T antigens. Intriguingly, the seed sequence of miR182 completely matches to a piRNA called piR62011. Upon reanalysis of a MCC small RNA library, piR62011 emerged as the highest expressed. We found it expressed in MKL-1 , a MCV positive cell line as well. Finally, to translate our findings into therapy for MCC, we screened small molecule (CMBL) library by performing surface plasmon resonance (SPR) assay and identified small molecules that binds to pre-miRNA182 and are testing them for their activity to kill MCC.

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miR-183/96/182 cluster is an important morphogenetic factor targetingPAX6expression in differentiating human retinal organoids

Cited by 4 publications

References 31 publications

Efficient embryoid-based method to improve generation of optic vesicles from human induced pluripotent stem cells

Efficient embryoid-based method to improve generation of optic vesicles from human induced pluripotent stem cells

SMAD4 Inhibits Granulosa Cell Apoptosis via the miR-183-96-182 Cluster and FoxO1 Axis

Merkel cell polyomavirus regulates miR183 cluster and piR62011 in Merkel cell carcinoma

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