Bio-Inspired Approach to Modelling Retinal Ganglion Cells using System Identification Techniques

The processing capabilities of biological vision systems are still vastly superior to artificial vision, even though this has been an active area of research for over half a century. Current artificial vision techniques integrate many insights from biology yet they remain far-off the capabilities of animals and humans in terms of speed, power, and performance. A key aspect to modeling the human visual system is the ability to accurately model the behavior and computation within the retina. In particular, we focus on modeling the retinal ganglion cells (RGCs) as they convey the accumulated data of real world images as action potentials onto the visual cortex via the optic nerve. Computational models that approximate the processing that occurs within RGCs can be derived by quantitatively fitting the sets of physiological data using an input–output analysis where the input is a known stimulus and the output is neuronal recordings. Currently, these input–output responses are modeled using computational combinations of linear and nonlinear models that are generally complex and lack any relevance to the underlying biophysics. In this paper, we illustrate how system identification techniques, which take inspiration from biological systems, can accurately model retinal ganglion cell behavior, and are a viable alternative to traditional linear–nonlinear approaches

Induced Pluripotent Stem Cells for Inherited Optic Neuropathies—Disease Modeling and Therapeutic Development

Background: Inherited optic neuropathies (IONs) cause progressive irreversible visual loss in children and young adults. There are limited disease-modifying treatments, and most patients progress to become severely visually impaired, fulfilling the legal criteria for blind registration. The seminal discovery of the technique for reprogramming somatic nondividing cells into induced pluripotent stem cells (iPSCs) has opened several exciting opportunities in the field of ION research and treatment. / Evidence Acquisition: A systematic review of the literature was conducted with PubMed using the following search terms: autosomal dominant optic atrophy, ADOA, dominant optic atrophy, DOA, Leber hereditary optic neuropathy, LHON, optic atrophy, induced pluripotent stem cell, iPSC, iPSC derived, iPS, stem cell, retinal ganglion cell, and RGC. Clinical trials were identified on the ClinicalTrials.gov website. / Results: This review article is focused on disease modeling and the therapeutic strategies being explored with iPSC technologies for the 2 most common IONs, namely, dominant optic atrophy and Leber hereditary optic neuropathy. The rationale and translational advances for cell-based and gene-based therapies are explored, as well as opportunities for neuroprotection and drug screening. / Conclusions: iPSCs offer an elegant, patient-focused solution to the investigation of the genetic defects and disease mechanisms underpinning IONs. Furthermore, this group of disorders is uniquely amenable to both the disease modeling capability and the therapeutic potential that iPSCs offer. This fast-moving area will remain at the forefront of both basic and translational ION research in the coming years, with the potential to accelerate the development of effective therapies for patients affected with these blinding diseases

Investigating disease mechanisms in autosomal dominant optic atrophy with retinal ganglion cells derived from induced pluripotent stem cells

Dominant Optic Atrophy (DOA) is the most common inherited optic neuropathy in the UK, characterised by the preferential loss of retinal ganglion cells (RGCs) and progressive blindness. 60-70 % of DOA patients harbour mutations in the OPA1 gene, encoding a mitochondrial protein that regulates mitochondrial morphology, bioenergetics and mitochondrial DNA (mtDNA) quality. Currently, DOA has no therapeutic options and the mechanisms driving RGC degeneration are poorly understood. In this study, a biobank of induced pluripotent stem cells iPSCs (iPSCs) encompassing the clinical and genetic DOA spectrum was created using patient-derived OPA1 mutant fibroblast cell lines, and CRISPR/Cas9 gene editing to generate isogenic cell lines. RGC differentiation was optimized and characterised in 2D and 3D in vitro methods, demonstrating expression of RGC-associated genes including BRN3B and ISL1. OPA1 mutant iPSCs showed no differentiation deficit compared to wild-type control cell lines, exhibiting comparable expression of all relevant markers. 2D-RGCs demonstrated enrichment of neuronal associated markers, including ELAVL3 and TAU, when compared to 3D retinal organoids. Phenotypic analysis demonstrated significant deficits in respiration, ATP production and increased mtDNA mutation in fibroblasts, iPSCs and 2D-RGCs compared to isogenic controls. Characterisation of mitochondrial stress through induction of stress associated gene expression demonstrated significant levels of upregulation in iPSCs, however, 3D- and 2D-RGCs exhibited fewer upregulated genes indicating that mitochondrial stress may be a cell type specific response. Importantly, correction of patient-derived iPSCs restored mitochondrial homeostasis, demonstrating that restoration of WT OPA1 expression is able to mitigate mutant associated phenotypes. Thus, an OPA1 mutant iPSC biobank has been established encompassing the clinical disease spectrum, enabling effective in vitro modelling to establish RGC specific disease mechanisms. OPA1 mutant RGCs demonstrate significant reductions in mitochondrial homeostasis, including reduced bioenergetic output and mtDNA quality. This work provides a platform for further investigation of OPA1-mediated disease mechanisms and therapeutic design

Human organotypic retinal cultures (HORCs) as a chronic experimental model for investigation of retinal ganglion cell degeneration

There is a growing need for models of human diseases that utilise native, donated human tissue in order to model disease processes and develop novel therapeutic strategies. In this paper we assessed the suitability of adult human retinal explants as a potential model of chronic retinal ganglion cell (RGC) degeneration. Our results confirmed that RGC markers commonly used in rodent studies (NeuN, bIII Tubulin and Thy-1) were appropriate for labelling human RGCs and followed the expected differential expression patterns across, as well as throughout, the macular and para-macular regions of the retina. Furthermore, we showed that neither donor age nor post-mortem time (within 24 h) significantly affected the initial expression levels of RGC markers. In addition, the feasibility of using human post mortem donor tissue as a long-term model of RGC degeneration was determined with RGC protein being detectable up to 4 weeks in culture with an associated decline in RGC mRNA and significant, progressive, apoptotic labelling of NeuNþ cells. Differences in RGC apoptosis might have been influenced by medium compositions indicating that media constituents could play a role in supporting axotomised RGCs. We propose that using ex vivo human explants may prove to be a useful model for testing the effectiveness of neuroprotective strategies

Müller glia activation in response to inherited retinal degeneration is highly varied and disease-specific

Despite different aetiologies, most inherited retinal disorders culminate in photoreceptor loss, which induces concomitant changes in the neural retina, one of the most striking being reactive gliosis by Müller cells. It is typically assumed that photoreceptor loss leads to an upregulation of glial fibrilliary acidic protein (Gfap) and other intermediate filament proteins, together with other gliosis-related changes, including loss of integrity of the outer limiting membrane (OLM) and deposition of proteoglycans. However, this is based on a mix of both injury-induced and genetic causes of photoreceptor loss. There are very few longitudinal studies of gliosis in the retina and none comparing these changes across models over time. Here, we present a comprehensive spatiotemporal assessment of features of gliosis in the degenerating murine retina that involves Müller glia. Specifically, we assessed Gfap, vimentin and chondroitin sulphate proteoglycan (CSPG) levels and outer limiting membrane (OLM) integrity over time in four murine models of inherited photoreceptor degeneration that encompass a range of disease severities (Crb1rd8/rd8, Prph2+/Δ307, Rho-/-, Pde6brd1/rd1). These features underwent very different changes, depending upon the disease-causing mutation, and that these changes are not correlated with disease severity. Intermediate filament expression did indeed increase with disease progression in Crb1rd8/rd8 and Prph2+/Δ307, but decreased in the Prph2+/Δ307 and Pde6brd1/rd1 models. CSPG deposition usually, but not always, followed the trends in intermediate filament expression. The OLM adherens junctions underwent significant remodelling in all models, but with differences in the composition of the resulting junctions; in Rho-/- mice, the adherens junctions maintained the typical rod-Müller glia interactions, while in the Pde6brd1/rd1 model they formed predominantly between Müller cells in late stage of degeneration. Together, these results show that gliosis and its associated processes are variable and disease-dependent