A5066681422- Retinal Development and Disorders
- Photoreceptor and optogenetics research
- Retinal Diseases and Treatments
- Neuroscience and Neural Engineering
- Connexins and lens biology
- CRISPR and Genetic Engineering
- Single-cell and spatial transcriptomics
- Photochromic and Fluorescence Chemistry
- Pluripotent Stem Cells Research
- Nicotinic Acetylcholine Receptors Study
- Corneal Surgery and Treatments
- Corneal surgery and disorders
- Neuroscience and Neuropharmacology Research
- Cell Adhesion Molecules Research
- Ocular Oncology and Treatments
- bioluminescence and chemiluminescence research
- Renal and related cancers
- Health, Environment, Cognitive Aging
- Retinal Imaging and Analysis
- Angiogenesis and VEGF in Cancer
- Ubiquitin and proteasome pathways
- Receptor Mechanisms and Signaling
- Cancer-related molecular mechanisms research
- Retinal and Macular Surgery
- Reproductive Biology and Fertility
Newcastle University
2015-2025
University of Newcastle Australia
2021
StemCells (United States)
2020
Centre for Life
2019
Carl von Ossietzky Universität Oldenburg
2012-2016
Zero to Three
2012
Single cell (sc) analyses of key embryonic, fetal and adult stages were performed to generate a comprehensive single atlas all the corneal adjacent conjunctival types from development adulthood. Four human seventeen embryonic corneas 10 21 post conception week (PCW) specimens dissociated cells subjected scRNA- and/or ATAC-Seq using 10x Genomics platform. These embedded Uniform Manifold Approximation Projection (UMAP) clustered Seurat graph-based clustering. Cluster identification was based...
The availability of in vitro models the human retina which to perform pharmacological and toxicological studies is an urgent unmet need. An essential step for developing ability generate laminated, physiologically functional, light-responsive retinal organoids from renewable patient specific sources. We investigated five different human-induced pluripotent stem cell (iPSC) lines showed a significant variability their efficiency organoids. Despite this variability, by month 5 differentiation,...
The high infection rate of SARS-CoV-2 necessitates the need for multiple studies identifying molecular mechanisms that facilitate viral entry and propagation. Currently potential extra-respiratory transmission routes remain unclear.
Abstract The rapid improvements in single cell sequencing technologies and analyses afford greater scope for dissecting organoid cultures composed of multiple types create an opportunity to interrogate these models understand tissue biology, cellular behavior interactions. To this end, retinal organoids generated from human embryonic stem cells (hESCs) were analyzed by RNA-sequencing (scRNA-Seq) at three time points differentiation. Combinatorial data all revealed the presence nine clusters,...
ABSTRACT The scarcity of embryonic/foetal material as a resource for direct study means that there is still limited understanding human retina development. Here, we present an integrated transcriptome analysis combined with immunohistochemistry in eye and retinal samples from 4 to 19 post-conception weeks. This reveals three developmental windows specific gene expression patterns informed the sequential emergence cell types enabled identification stage-specific cellular biological processes,...
The emergence of retinal progenitor cells and differentiation to various cell types represent fundamental processes during development. Herein, we provide a comprehensive single characterisation transcriptional chromatin accessibility changes that underline specification over the course human development up midgestation. Our lineage trajectory data demonstrate presence early progenitors, which transit late, further transient neurogenic give rise all neurons. Combining RNA-Seq with spatial...
Molecular information on the early stages of human retinal development remains scarce due to limitations in obtaining eye samples. Pluripotent stem cell-derived organoids (ROs) provide an unprecedented opportunity for studying retinogenesis. Using a combination single cell RNA-seq and spatial transcriptomics we present first-time spatiotemporal transcriptome RO development. Our data demonstrate that ROs recapitulate key events retinogenesis including optic vesicle/cup formation, presence...
A major goal in the stem cell field is to generate tissues that can be utilized as a universal tool for vitro models of development and disease, drug development, or resource patients suffering from disease injury. Great efforts are being made differentiate human pluripotent cells toward retinal tissue, which akin native retina its cytoarchitecture function, yet numerous existing induction protocols remain variable their efficiency do not routinely produce morphologically functionally mature...
Tissue specific extracellular matrices (ECM) provide structural support and enable access to molecular signals metabolites, which are essential for directing stem cell renewal differentiation. To mimic this phenomenon in vitro, tissue decellularisation approaches have been developed, resulting the generation of natural ECM scaffolds that comparable physical biochemical properties tissues currently gaining traction engineering regenerative therapies due ease standardised production, constant...
Retinal dystrophies often lead to blindness. Developing therapeutic interventions restore vision is therefore of paramount importance. Here we demonstrate the ability pluripotent stem cell-derived cone precursors engraft and light responses in Pde6brd1 mouse, an end-stage photoreceptor degeneration model. Our data show that up 1.5% integrate into host retina, differentiate cones, close apposition bipolar cells. Half transplanted mice exhibited visual behavior these 33% showed binocular...
Retinal drug toxicity screening is essential for the development of safe treatment strategies a large number diseases. To this end, retinal organoids derived from human pluripotent stem cells (hPSCs) provide suitable platform due to their similarity retina and ease generation in large-scale formats. In study, two hPSC cell lines were differentiated organoids, which comprised all key types multiple nuclear synaptic layers. Single-cell RNA-Seq indicated maintenance ganglion bipolar cells: both...
Abstract Age-related macular degeneration (AMD) is the most prevalent cause of blindness in developed world. Vision loss advanced stages disease caused by atrophy retinal photoreceptors, overlying pigment epithelium (RPE) and choroidal endothelial cells. The molecular events that underline development these cell types from utero to adult as well progression intermediate AMD are not yet fully understood. We performed single-cell RNA-sequencing (RNA-Seq) human fetal RPE–choroidal tissues,...
Abstract Microglia are the primary resident immune cells in retina. They regulate neuronal survival and synaptic pruning making them essential for normal development. Following injury, they mediate adaptive responses under pathological conditions can trigger neurodegeneration exacerbating effect of a disease. Retinal organoids derived from human induced pluripotent stem (hiPSCs) increasingly being used range applications, including disease modelling, development new therapies study...
Cell replacement therapy is a promising treatment for irreversible retinal cell death in diverse diseases such as Stargardt's disease, age-related macular degeneration, and retinitis pigmentosa. The final impact of all dystrophies the loss photoreceptors; hence, there pressing need research into replacement. Seminal work has shown that simple three-dimensional culture system enables differentiation human pluripotent stem cells to organoids containing large numbers photoreceptors developing...
Retinoblastoma (Rb) is a childhood cancer of the developing retina, accounting for up to 17% all tumors in infancy. To gain insights into transcriptional events cell state transitions during Rb development, we established 2 disease models via retinal organoid differentiation pRB (retinoblastoma protein)-depleted human embryonic stem line (RB1-null hESCs) and patient-specific induced pluripotent (iPSC) harboring RB1 biallelic mutation (c.2082delC). Both were characterized by depletion...
Abstract The carboxy-terminus of the spliceosomal protein PRPF8 , which regulates RNA helicase Brr2, is a hotspot for mutations causing retinitis pigmentosa-type 13, with unclear role in human splicing and tissue-specificity mechanism. We used patient induced pluripotent stem cells-derived cells, carrying heterozygous c.6926 A > C (p.H2309P) mutation to demonstrate retinal-specific endophenotypes comprising photoreceptor loss, apical-basal polarity ciliary defects. Comprehensive...
Spatial transcriptomics (ST) has the potential to provide unprecedented insights into gene expression across tissue architecture, but existing analytical methods often overlook full complexity of spatial dimension. We present STExplorer, an R package that adapts well-established computational geography (CG) explore micro-geography omics data. By incorporating techniques like Geographically Weighted Principal Component Analysis (GWPCA), Fuzzy Clustering (FGWC), Regression (GWR), and analyses...
Abstract Stargardt disease is an inherited retinopathy affecting approximately 1:8000 individuals. It characterised by biallelic variants in ABCA4 which encodes a vital protein for the recycling of retinaldehydes retina. Despite its prevalence and impact, there are currently no treatments available this condition. Furthermore, 35% STGD1 cases remain genetically unsolved. To investigate cellular molecular characteristics associated with STGD1, we generated iPSCs from two monoallelic...
In the brain, including retina, interneurons show an enormous structural and functional diversity. Retinal horizontal cells represent a class of that form triad synapses with photoreceptors ON bipolar cells. At this first retinal synapse, modulate signal transmission from to by feedback feedforward inhibition. To test how fully developed retina reacts specific loss cells, these were specifically ablated adult mice using diphtheria toxin (DT)/DT-receptor system <i>connexin57</i> promoter....
The extracellular matrix (ECM) plays an important role in numerous processes including cellular proliferation, differentiation, migration, maturation, adhesion guidance and axonal growth. To date, there has been no detailed analysis of the ECM distribution during retinal ontogenesis humans functional importance many components is poorly understood. In this study, expression key adult mouse monkey retina, developing human retina organoids derived from pluripotent stem cells was studied. Our...
Pannexin1 (Panx1) belongs to a class of vertebrate proteins that exhibits sequence homology innexins, the invertebrate gap junction proteins, and which also shares topological similarities with connexins. Unlike junctional channels, Panx1 forms single-membrane whose functional role in neuronal circuits is still unsettled. We therefore investigated subcellular distribution mouse retina wildtype Panx1-null mice by reverse-transcription polymerase chain reaction (RT-PCR), immunohistochemistry,...
Electrical synapses (gap junctions) rapidly transmit signals between neurons and are composed of connexins. In neurons, connexin36 (Cx36) is the most abundant isoform; however, mechanisms underlying formation Cx36-containing electrical unknown. We focus on homocellular heterocellular gap junctions formed by an AII amacrine cell, a key interneuron found in all mammalian retinas. mice lacking native Cx36 but expressing variant tagged with enhanced green fluorescent protein at C-terminus...
The purpose of this study was to generate human embryonic stem cell (hESC) lines harboring the green fluorescent protein (GFP) reporter at endogenous loci Cone-Rod Homeobox (CRX) gene, a key transcription factor in retinal development. Zinc finger nucleases (ZFNs) designed cleave 3' UTR CRX were transfected into hESCs along with donor construct containing homology target region, eGFP reporter, and puromycin selection cassette. Following selection, polymerase chain reaction (PCR) sequencing...
Abstract This unit describes a protocol for generating retinal organoids that contain all major cell types and are responsive to light from human pluripotent stem cells (hPSCs). hPSCs differentiated in 96‐well plates allow large‐scale production of could be used multiple applications, including study development, disease modeling, compound screening. The differentiation approach is based on the knowledge insulin‐like growth factor 1 signaling together with retinoic acid triiodothyronine...