The retinal G protein-coupled receptor (RGR) is a visual cycle photoisomerase that photopically regenerates 11-cis-retinal (11cRAL). It plays a crucial role in sustaining vision. Here, we investigated the in vivo role of RGR in the cone photoreceptor-dominant, zebrafish retina, focusing predominantly on how visual function is impacted in the absence of RGR. There are two zebrafish RGR paralogs, rgra and rgrb, both with predominant expression in retinal pigment epithelium (RPE) and Müller glia cells. Under standard light rearing conditions, bespoke rgrb-/-; rgra-/- double knockout zebrafish present with a ~21% reduction in optokinetic response (OKR) saccades per minute relative to wild-type (WT). This impaired visual behavior worsens in higher photopic conditions ranging from 20 000-81 000 lx. In contrast, no significant OKR defect is observed under dark-adapted conditions, consolidating the light-dependent role of RGR in vision. Retinoid profiling of rgrb-/-; rgra-/- zebrafish larvae demonstrated significant decreases in 11cRAL levels under standard and brighter light rearing conditions. Proteomic profiling validated the successful generation of rgrb-/-; rgra-/- zebrafish and revealed an unanticipated upregulation in ocular extracellular matrix proteins. From polarized light microscopy, increased collagen fiber abundance with dysregulated organization in Bruch's membrane at the interface between the retina and choroid was observed. These novel findings demonstrate the role of RGR in sustaining visual function under cone-mediated photopic conditions, a concomitant deficit in the photopic visual cycle and a novel role in maintaining the integrity of Bruch's membrane.

Mutations in ABCA4 cause Stargardt disease (STGD1) by disrupting retinoid handling and retinal pigment epithelium (RPE) homeostasis, yet the metabolic drivers of RPE degeneration remain unclear. Given that photoreceptor health relies heavily on the support of the RPE, loss of the RPE cells is central to STGD1 pathology. In this study, we show that dysregulation of diacylglycerol O-acyltransferase-1 (DGAT1) is associated with disrupted retinoid-lipid metabolism in STGD1 RPE cells, accompanied by excess retinyl esters and neutral lipid accumulation, impaired lipid processing, and reduced mitochondrial activity. These findings implicate DGAT1-mediated lipid remodeling as a contributing factor to RPE dysfunction in ABCA4-associated retinopathies.

Stargardt disease (STGD1), the most common inherited juvenile macular degeneration, is caused by biallelic mutations in the ABCA4 gene. Currently, there is no approved treatment. In this study, we investigated early-stage epigenomic changes in the retinal pigment epithelium (RPE) of Abca4-/- mice, a well-established model of STGD1. Reduced representation bisulfite sequencing (RRBS) revealed hypermethylation of gene regions associated with disease-related pathways, implicating methyl-CpG-binding protein 2 (MeCP2) and RE1-silencing transcription factor (REST) as potential regulators. Notably, DNA methylation of a subset of genes preceded their transcriptional change and disease phenotypes in Abca4-/- RPE. Together with the detected age-dependent increase in MeCP2 levels in Abca4-/- RPE, these findings suggest that early DNA methylation changes may contribute to RPE dysfunction and eventual cell loss in STGD1.

Stargardt disease (STGD1), the most common inherited juvenile macular degeneration, is caused by biallelic mutations in the ABCA4 gene. Currently, there is no approved treatment. In this study, we investigated early-stage epigenomic changes in the retinal pigment epithelium (RPE) of Abca4-/- mice, a well-established model of STGD1. Reduced representation bisulfite sequencing (RRBS) revealed hypermethylation of gene regions associated with disease-related pathways, implicating methyl-CpG-binding protein 2 (MeCP2) and RE1-silencing transcription factor (REST) as potential regulators. Notably, DNA methylation of a subset of genes preceded their transcriptional change and disease phenotypes in Abca4-/- RPE. Together with the detected age-dependent increase in MeCP2 levels in Abca4-/- RPE, these findings suggest that early DNA methylation changes may contribute to RPE dysfunction and eventual cell loss in STGD1.

Recessive Stargardt disease (STGD1) is an inherited juvenile maculopathy caused by mutations in the ABCA4 gene, for which there is no suitable treatment. Loss of functional ABCA4 in the retinal pigment epithelium (RPE) alone, without contribution from photoreceptor cells, was shown to induce STGD1 pathology. Here, we identified cathepsin D (CatD), the primary RPE lysosomal protease, as a key molecular player contributing to endo-lysosomal dysfunction in STGD1 using a newly developed "disease-in-a-dish" RPE model from confirmed STGD1 patients. Induced pluripotent stem cell (iPSC)-derived RPE originating from three STGD1 patients exhibited elevated lysosomal pH, as previously reported in Abca4-/- mice. CatD protein maturation and activity were impaired in RPE from STGD1 patients and Abca4-/- mice. Consequently, STGD1 RPE cells have reduced photoreceptor outer segment degradation and abnormal accumulation of α-synuclein, the natural substrate of CatD. Furthermore, dysfunctional ABCA4 in STGD1 RPE cells results in intracellular accumulation of autofluorescent material and phosphatidylethanolamine (PE). The altered distribution of PE associated with the internal membranes of STGD1 RPE cells presumably compromises LC3-associated phagocytosis, contributing to delayed endo-lysosomal degradation activity. Drug-mediated re-acidification of lysosomes in the RPE of STGD1 restores CatD functional activity and reduces the accumulation of immature CatD protein loads. This preclinical study validates the contribution of CatD deficiencies to STGD1 pathology and provides evidence for an efficacious therapeutic approach targeting RPE cells. Our findings support a cell-autonomous RPE-driven pathology, informing future research aimed at targeting RPE cells to treat ABCA4-mediated retinopathies.

A major limitation to developing chimeric antigen receptor (CAR)-T cell therapies for solid tumors is identifying surface proteins highly expressed in tumors but not in normal tissues. Here, we identify Tyrosinase Related Protein 1 (TYRP1) as a CAR-T cell therapy target to treat patients with cutaneous and rare melanoma subtypes unresponsive to immune checkpoint blockade. TYRP1 is primarily located intracellularly in the melanosomes, with a small fraction being trafficked to the cell surface via vesicular transport. We develop a highly sensitive CAR-T cell therapy that detects surface TYRP1 in tumor cells with high TYRP1 overexpression and presents antitumor activity in vitro and in vivo in murine and patient-derived cutaneous, acral and uveal melanoma models. Furthermore, no systemic or off-tumor severe toxicities are observed in an immunocompetent murine model. The efficacy and safety profile of the TYRP1 CAR-T cell therapy supports the ongoing preparation of a phase I clinical trial.

MicroRNAs (miRs) are short, evolutionarily conserved non-coding RNAs that canonically downregulate expression of target genes. The miR family composed of miR-204 and miR-211 is among the most highly expressed in the retinal pigment epithelium (RPE) in both mouse and human, and also retains high sequence identity. To assess the role of this miR family in the developed mouse eye, we generated two floxed conditional knockout mouse lines crossed to the RPE65-ERT2-Cre driver mouse line to perform an RPE-specific conditional knockout of this miR family in adult mice. After Cre-mediated deletion, we observed retinal structural changes by optical coherence tomography; dysfunction and loss of photoreceptors by retinal imaging; and retinal inflammation marked by subretinal infiltration of immune cells by imaging and immunostaining. Single-cell RNA sequencing of diseased RPE and retinas showed potential miR-regulated target genes, as well as changes in non-coding RNAs in the RPE, rod photoreceptors, and Müller glia. This work thus highlights the role of miR-204 and miR-211 in maintaining RPE function and how the loss of miRs in the RPE exerts effects on the neural retina, leading to inflammation and retinal degeneration.

Digital transformation has received widespread academic, policy and media attention in the last few years, amid rapid technological developments, geopolitical tensions and an aggressive media strategy launched by a number of Silicon Valley CEOs. Post-pandemic, the task of tracing the relevant scenarios for technological change and evaluating their social consequences has become more difficult, partly due to the rise of artificial intelligence. This special issue explores novel lines of inquiry in the study of Internet governance at a critical juncture in its evolution, zooming in on three important trends: (1) the public attempts to refocus on infrastructure (control); (2) the stricter regulation of social media, partly in response to the push from civil society to address violent behaviour online; and (3) the renewed contestation over representation in Internet governance, where youth and other historically underrepresented groups are struggling to achieve better visibility for their interests. The articles in this special issue underline that new forms of contention by a variety of actors are emerging from “within” the existing governance mechanisms. However, the most successful initiatives are mainly from governments.

Vacuolar protein sorting 35 (VPS35), the core component of the retromer complex which regulates endosomal trafficking, is genetically linked with Parkinson’s disease (PD). Impaired vision is a common non-motor manifestation of PD. Here, we show mouse retinas with VPS35-deficient rods exhibit synapse loss and visual deficit, followed by progressive degeneration concomitant with the emergence of Lewy body-like inclusions and phospho-α-synuclein (P-αSyn) aggregation. Ultrastructural analyses reveal VPS35-deficient rods accumulate aggregates in late endosomes, deposited as lipofuscins bound to P-αSyn. Mechanistically, we uncover a protein network of VPS35 and its interaction with HSC70. VPS35 deficiency promotes sequestration of HSC70 and P-αSyn aggregation in late endosomes. Microglia which engulf lipofuscins and P-αSyn aggregates are activated, displaying autofluorescence, observed as bright dots in fundus imaging of live animals, coinciding with pathology onset and progression. The Rod∆Vps35 mouse line is a valuable tool for further mechanistic investigation of αSyn lesions and retinal degenerative diseases.

Though rod and cone photoreceptors use similar phototransduction mechanisms, previous model calculations have indicated that the most important differences in their light responses are likely to be differences in amplification of the G-protein cascade, different decay rates of phosphodiesterase (PDE) and pigment phosphorylation, and different rates of turnover of cGMP in darkness. To test this hypothesis, we constructed TrUx;GapOx rods by crossing mice with decreased transduction gain from decreased transducin expression, with mice displaying an increased rate of PDE decay from increased expression of GTPase-activating proteins (GAPs). These two manipulations brought the sensitivity of TrUx;GapOx rods to within a factor of 2 of WT cone sensitivity, after correcting for outer-segment dimensions. These alterations did not, however, change photoreceptor adaptation: rods continued to show increment saturation though at a higher background intensity. These experiments confirm model calculations that rod responses can mimic some (though not all) of the features of cone responses after only a few changes in the properties of transduction proteins.

The Domain Name System (DNS) is vital to the internet, enabling everyday uses such as browsing, emailing and chatting. One function in particular – the DNS resolution, performed by resolver operators – allows us to reach what we are looking for online. The market of recursive resolution is highly dynamic and is currently shifting towards open or public resolvers, which tend to belong to large tech companies. In 2022, the European Commission launched the DNS4EU initiative, which established a European resolver as part of the new EU cybersecurity strategy, in order to respond to the resilience, security and privacy needs of the union. This article provides a comprehensive analysis of the DNS4EU project, which provided seed funding to a European competitor in a market increasingly dominated by non-EU players. As a critical infrastructure service, the DNS4EU represents one of the first concrete steps towards enhancing the strategic autonomy of the union. But it also constitutes an unprecedented public intervention in a largely private market relying on voluntary adoption. This article contextualises the DNS4EU initiative, outlining both advantages and limitations of the European strategy and related tender process and implementation plan, concluding with a discussion on the future of DNS resolution.

This article builds on existing literature on digital inequality and the digitised welfare state to elucidate one underexplored way in which the rise of e-government platforms further disadvantages already-marginalised people: by requiring that they possess a verifiable digital footprint distributed across multiple public and commercial platforms. We illustrate the pertinence and nuances of this particular risk through lived experience research in a UK public library where limited users receive help with digital skills. Although there is a growing recognition of both the inevitability of digital welfare and the risks to marginalised communities, little work has been done to connect these abstract policy discussions to lived experience-to pinpoint how digitisation creates these exclusions, beyond simply having internet access or not. This article argues that the prerequisite of a digital footprint engenders a double disadvantage: (1) lacking a digital footprint is the result of barriers that are largely invisible to data-driven, digital-by-default systems, and (2) when marginalised users establish a sufficient footprint, this entails a disproportionately onerous responsibility for managing a distributed personal data trail in the long term. This combination of mundane barriers and the burden of responsibility for a digital identity points to policy implications for governments aiming to advance inclusive digital transformation agendas.

This special issue offers a new perspective on decision-making in internet governance, bringing together theoretical contributions and empirical knowledge on authority and influence in underexplored policy settings, from the Congress of the United States to RightsCon, a leading summit on human rights in the digital era. The articles included here shed light on specific groups underrepresented in existing research (such as civil society groups, expert activists, technical community) and on the interactions between these groups and traditional, powerful actors such as governments and industry. These contributions raise new questions, such as: is agency possible without representation in the digital space? Who can represent? Are deficiencies in technology governance affecting the representation model of our political systems more broadly? How do representation and agency manifest themselves across time and space?

Motivated by the goal of developing ultralow power, smart and multifunctional nano (micro) electronic devices, research has shown unwavering interest in synthesis methods, architectures and interphase connectivity of composites containing magnetostrictive and piezoelectric phases, known as magnetoelectric (ME) materials. Herein we report on CoFe 2 O 4 / 0.92 Bi 0.5 Na 0.5 TiO 3 – 0.08 BaTiO 3 biphasic ME composites, obtained by sol-gel chemistry, by mixing the precursor sols of the two phases into one precursor sol and further transforming it into gel, with the goal of obtaining homogeneous nanocomposites with magnetoelectric properties. The structural properties, the temperature dependance of dielectric properties, the magnetic and magnetoelectric properties of these biphasic mixtures, with various molar ratios CoFe 2 O 4 /BNT–BT 0.08 = 0.5:1, 1:1 and 1.5:1, are investigated. It is observed that the amount of CoFe 2 O 4 and the synthesis in situ of these composites influences their macroscopic properties showing a high difficulty of carrying out an efficient poling resulting in small piezoelectric and magnetoelectric response. It was concluded that the synthesis procedure, the type of architecture and the interphase connectivity are of outmost importance for magnetoelectric properties based on lead-free materials.