Mutations in KEOPS-Complex Genes Cause Nephrotic Syndrome with Primary Microcephaly

Galloway-Mowat syndrome (GAMOS) is an autosomal-recessive disease characterized by the combination of early-onset nephrotic syndrome (SRNS) and microcephaly with brain anomalies. Here we identified recessive mutations in OSGEP, TP53RK, TPRKB, and LAGE3, genes encoding the four subunits of the KEOPS complex, in 37 individuals from 32 families with GAMOS. CRISPR-Cas9 knockout in zebrafish and mice recapitulated the human phenotype of primary microcephaly and resulted in early lethality. Knockdown of OSGEP, TP53RK, or TPRKB inhibited cell proliferation, which human mutations did not rescue. Furthermore, knockdown of these genes impaired protein translation, caused endoplasmic reticulum stress, activated DNA-damage-response signaling, and ultimately induced apoptosis. Knockdown of OSGEP or TP53RK induced defects in the actin cytoskeleton and decreased the migration rate of human podocytes, an established intermediate phenotype of SRNS. We thus identified four new monogenic causes of GAMOS, describe a link between KEOPS function and human disease, and delineate potential pathogenic mechanisms

The genomic landscape of balanced cytogenetic abnormalities associated with human congenital anomalies

Despite the clinical significance of balanced chromosomal abnormalities (BCAs), their characterization has largely been restricted to cytogenetic resolution. We explored the landscape of BCAs at nucleotide resolution in 273 subjects with a spectrum of congenital anomalies. Whole-genome sequencing revised 93% of karyotypes and demonstrated complexity that was cryptic to karyotyping in 21% of BCAs, highlighting the limitations of conventional cytogenetic approaches. At least 33.9% of BCAs resulted in gene disruption that likely contributed to the developmental phenotype, 5.2% were associated with pathogenic genomic imbalances, and 7.3% disrupted topologically associated domains (TADs) encompassing known syndromic loci. Remarkably, BCA breakpoints in eight subjects altered a single TAD encompassing MEF2C, a known driver of 5q14.3 microdeletion syndrome, resulting in decreased MEF2C expression. We propose that sequence-level resolution dramatically improves prediction of clinical outcomes for balanced rearrangements and provides insight into new pathogenic mechanisms, such as altered regulation due to changes in chromosome topology

Text-fading based training leads to transfer effects on children\u27s sentence reading fluency

Previous studies used a text-fading procedure as a training tool with the goal to increase silent reading fluency (i.e., proficient reading rate and comprehension). In recently published studies, this procedure resulted in lasting reading enhancements for adult and adolescent research samples. However, studies working with children reported mixed results. While reading rate improvements were observable for Dutch reading children in a text-fading training study, reading fluency improvements in standardized reading tests post-training attributable to the fading manipulation were not detectable. These results raise the question of whether text-fading training is not effective for children or whether research design issues have concealed possible transfer effects. Hence, the present study sought to investigate possible transfer effects resulting from a text-fading based reading training program, using a modified research design. Over a period of 3 weeks, two groups of German third-graders read sentences either with an adaptive text-fading procedure or at their self-paced reading rate. A standardized test measuring reading fluency at the word, sentence, and text level was conducted pre- and post-training. Text level reading fluency improved for both groups equally. Post-training gains at the word level were found for the text-fading group, however, no significant interaction between groups was revealed for word reading fluency. Sentence level reading fluency gains were found for the text-fading group, which significantly differed from the group of children reading at their self-paced reading routine. These findings provide evidence for the efficacy of text-fading as a training method for sentence reading fluency improvement also for children. (DIPF/Orig.

Text-fading based training leads to transfer effects on children's sentence reading fluency

Previous studies used a text-fading procedure as a training tool with the goal to increase silent reading fluency (i.e., proficient reading rate and comprehension). In recently published studies, this procedure resulted in lasting reading enhancements for adult and adolescent research samples. However, studies working with children reported mixed results. While reading rate improvements were observable for Dutch reading children in a text-fading training study, reading fluency improvements in standardized reading tests post-training attributable to the fading manipulation were not detectable. These results raise the question of whether text-fading training is not effective for children or whether research design issues have concealed possible transfer effects. Hence, the present study sought to investigate possible transfer effects resulting from a text-fading based reading training program, using a modified research design. Over a period of 3 weeks, two groups of German third-graders read sentences either with an adaptive text-fading procedure or at their self-paced reading rate. A standardized test measuring reading fluency at the word, sentence, and text level was conducted pre- and post-training. Text level reading fluency improved for both groups equally. Post-training gains at the word level were found for the text-fading group, however, no significant interaction between groups was revealed for word reading fluency. Sentence level reading fluency gains were found for the text-fading group, which significantly differed from the group of children reading at their self-paced reading routine. These findings provide evidence for the efficacy of text-fading as a training method for sentence reading fluency improvement also for children

LPS-Induced Endotoxemia Evokes Epigenetic Alterations in Mitochondrial DNA That Impacts Inflammatory Response

Mitochondrial DNA (mtDNA) plays a vital role as a damage-associated molecular pattern in sepsis being able to shape the immune response. Since pathogen recognition receptors of innate immune cells are activated by demethylated DNA only, we set out to investigate the amount of DNA methyltransferase 1 (DNMT1) in mitochondria and the extent of mtDNA methylation in a human endotoxin model. Peripheral blood mononuclear cells of 20 healthy individuals were isolated from whole blood and stimulated with lipopolysaccharide (LPS) for 48 h. Subsequently, DNMT1 protein abundance was assessed in whole cells and a mitochondrial fraction. At the same time, methylation levels of mtDNA were quantified, and cytokine expression in the supernatant was measured. Despite increased cellular expression of DNMT1 after LPS stimulation, the degree of mtDNA methylation slightly decreased. Strikingly the mitochondrial protein abundance of DNMT1 was reduced by 50% in line with the lower degree of mtDNA methylation. Although only modest alterations were seen in the degree of mtDNA methylation, these strongly correlated with IL-6 and IL-10 expression. Our data may hint at a protein import problem for DNMT1 into the mitochondria under LPS stimulation and suggest a role of demethylated mtDNA in the regulation of the inflammatory immune response

A novel understanding of postoperative complications: In vitro study of the impact of propofol on epigenetic modifications in cholinergic genes.

BackgroundPropofol is a widely used anaesthetic drug with advantageous operating conditions and recovery profile. However, propofol could have long term effects on neuronal cells and is associated with post-operative delirium (POD). In this context, one of the contributing factors to the pathogenesis of POD is a reduction of cholinesterase activity. Accordingly, we investigated the effects of propofol on the methylation, expression and activity of cholinergic genes and proteins in an in-vitro model.ResultsWe found that propofol indeed reduced the activity of AChE / BChE in our in-vitro model, without affecting the protein levels. Furthermore, we could show that propofol reduced the methylation of a repressor region of the CHRNA7 gene without changing the secretion of pro-or anti-inflammatory cytokines. Lastly, propofol changed the expression patterns of genes responsible for maintaining the epigenetic status of the cell and accordingly reduced the tri-methylation of H3 K27.ConclusionIn conclusion we found a possible functional link between propofol treatment and POD, due to a reduced cholinergic activity. In addition to this, propofol changed the expression of different maintenance genes of the epigenome that also affected histone methylation. Thus, propofol treatment may also induce strong, long lasting changes in the brain by potentially altering the epigenetic landscape