CLN7 neuronal ceroid lipofuscinosis is an inherited lysosomal storage neurodegenerative disease highly prevalent in children. CLN7/MFSD8 gene encodes a lysosomal membrane glycoprotein, but the biochemical processes affected by CLN7-loss of function are unexplored thus preventing development of potential treatments. Here, we found, in the Cln7∆ex2 mouse model of CLN7 disease, that failure in autophagy causes accumulation of structurally and bioenergetically impaired neuronal mitochondria. In vivo genetic approach reveals elevated mitochondrial reactive oxygen species (mROS) in Cln7∆ex2 neurons that mediates glycolytic enzyme PFKFB3 activation and contributes to CLN7 pathogenesis. Mechanistically, mROS sustains a signaling cascade leading to protein stabilization of PFKFB3, normally unstable in healthy neurons. Administration of the highly selective PFKFB3 inhibitor AZ67 in Cln7∆ex2 mouse brain in vivo and in CLN7 patients-derived cells rectifies key disease hallmarks. Thus, aberrant upregulation of the glycolytic enzyme PFKFB3 in neurons may contribute to CLN7 pathogenesis and targeting PFKFB3 could alleviate this and other lysosomal storage diseases.

The application of induced pluripotent stem cells (iPSCs) in advanced therapies is increasing at pace, but concerns remain over their clinical safety profile. We report the first-ever application of doggybone DNA (dbDNA) vectors to generate human iPSCs. dbDNA vectors are closed-capped linear double-stranded DNA gene expression cassettes that contain no bacterial DNA and are amplified by a chemically defined, current good manufacturing practice (cGMP)-compliant methodology. We achieved comparable iPSC reprogramming efficiencies using transiently expressing dbDNA vectors with the same iPSC reprogramming coding sequences as the state-of-the-art OriP/EBNA1 episomal vectors but, crucially, in the absence of p53 shRNA repression. Moreover, persistent expression of EBNA1 from bacterially derived episomes resulted in stimulation of the interferon response, elevated DNA damage, and increased spontaneous differentiation. These cellular activities were diminished or absent in dbDNA-iPSCs, resulting in lines with a greater stability and safety potential for cell therapy.

Inhibition of E-cad in mouse embryonic stem cells (mESCs) leads to a switch from LIF-BMP to Activin/Nodal-dependent pluripotency, consistent with transition from a naïve to primed pluripotent phenotype. We have used both genetic ablation and steric inhibition of E-cad function in mESCs to assess alterations to phenotype using quantitative mass spectrometry analysis, network models, and functional assays. Proteomic analyses revealed that one third of detected proteins were altered in E-cad null mESCs (Ecad −/− mESCs) compared to wild type (624 proteins were downregulated and 705 were proteins upregulated). Network pathway analysis and subsequent cellular flux assays confirmed a metabolic shift from oxidative phosphorylation (OXPHOS) to aerobic glycolysis, specifically through mitochondrial complex III downregulation and hypoxia inducible factor 1a target upregulation. Central to this was the transcriptional coactivator EP300. E-cad is a well-known tumor suppressor, its downregulation during cancer initiation and metastasis can be linked to the metabolic switch known as Warburg effect. This study highlights a phenomena found in both primed pluripotent state and cancer stemness and links it to loss of E-cad. Data are available via ProteomeXchange with identifier PXD012679.

Aims or ObjectivesThe Syrian crisis has resulted in a significant impact on refugee oral health status, but the scientific literature lacks information characterizing the pattern of tooth extraction in refugees.Materials and MethodsThe current study analyzed the pattern of tooth extraction and the associated sociodemographic factors among 626 (53.4% females, 46.6% males) Syrian refugees who were treated in dental clinics in the Zaatari camp. All adult Syrian refugees attending the dental unit at Zaatari camp were eligible to participate. Clinical and sociodemographic data were collected from the study sample using a validated semi‐structured survey over a period of 4 months. Descriptive statistics were generated and Chi‐square test, independent sample t‐test, ANOVA test and Bonferroni Post Hoc test were used to examine associations between the different variables.ResultsMore than one third of the attendees (33.9%) were young adults (18–29 years), and nearly two thirds (69.8%) had no or very basic education. The mean number of teeth that needed extraction was (1.46 ± 0.84) and was significantly higher in males, smokers, older patients, and those with no or basic education. The most common cause for tooth extraction was dental caries (54.8%), followed by tooth fracture/un‐restorability (38.5%), and periodontitis (3.4%); while the least common reason was tooth extraction for orthodontic treatment (0.20%).ConclusionsDental caries and its sequelae were the leading cause of tooth extraction in this refugee population. Molar teeth were found to be the most frequently extracted and lower anteriors were the least. Older age, smoking, patients with lower educational level, poor oral hygiene and male gender all predispose to a higher incidence of tooth extractions.