Search

Clear cell sarcoma of the kidney (CCSK) is a rare pediatric renal tumor with a worse prognosis than Wilms’ tumor. Although recently, BCOR internal tandem duplication (ITD) has been found as a driver mutation in more than 80% of cases, a deep molecular characterization of this tumor is still lacking, as well as its correlation with the clinical course. The aim of this study was to investigate the differential molecular signature between metastatic and localized BCOR-ITD-positive CCSK at diagnosis. Whole-exome sequencing (WES) and whole-transcriptome sequencing (WTS) were performed on six localized and three metastatic BCOR-ITD-positive CCSKs, confirming that this tumor carries a low mutational burden. No significant recurrences of somatic or germline mutations other than BCOR-ITD were identified among the evaluated samples. Supervised analysis of gene expression data showed enrichment of hundreds of genes, with a significant overrepresentation of the MAPK signaling pathway in metastatic cases (p < 0.0001). Within the molecular signature of metastatic CCSK, five genes were highly and significantly over-expressed: FGF3, VEGFA, SPP1, ADM, and JUND. The role of FGF3 in the acquisition of a more aggressive phenotype was investigated in a cell model system obtained by introducing the ITD into the last exon of BCOR by Crispr/Cas9 gene editing of the HEK-293 cell line. Treatment with FGF3 of BCOR-ITD HEK-293 cell line induced a significant increase in cell migration versus both untreated and scramble cell clone. The identification of over-expressed genes in metastatic CCSKs, with a particular focus on FGF3, could offer new prognostic and therapeutic targets in more aggressive cases.

Gaucher disease is a lysosomal storage disorder characterized by β-glucosidase enzyme deficiency and substrate accumulation, especially in cells of the reticuloendothelial system. Typical features of the disease are the unrestrained activation of inflammatory mechanisms, whose molecular pathways are still unclear. To investigate biological mechanisms underlying the macrophage activation in GD, we derived iPSCs from a healthy donor and a GD patient line and differentiated them into hematopoietic progenitors. While GD iPSCs are able to efficiently give rise to CD33+/CD45+ myeloid progenitors, the maturation towards the CD14+/CD163+ monocyte/macrophages fate resulted enhanced in the GD lines, that in addition displayed a decreased growth potential compared to control cells either in semisolid or in liquid culture. The GD lines growth impairment was associated with a significant upregulation of RIPK3 and MLKL, two key effectors of necroptosis, the inflammation related cell death pathway. The activation of necroptosis, which has already been linked to neuronopathic GD, may play a role in the disease proinflammatory condition and in the identified cell growth defects. Understanding the GD macrophage role in the alteration of mechanisms linked to cellular metabolism imbalance, cell death and inflammation are crucial in identifying new ways to approach the disease.

Summary In an attempt to explore the role of the gut microbiome during recent canine evolutionary history, we sequenced the metagenome of 13 canine coprolites dated ca. 3,600–3,450 years ago from the Bronze Age archaeological site of Solarolo (Italy), which housed a complex farming community. The microbiome structure of Solarolo dogs revealed continuity with that of modern dogs, but it also shared some features with the wild wolf microbiome, as a kind of transitional state between them. The dietary niche, as also inferred from the microbiome composition, was omnivorous, with evidence of consumption of starchy agricultural foods. Of interest, the Solarolo dog microbiome was particularly enriched in sequences encoding alpha-amylases and complemented a low copy number of the host amylase gene. These findings suggest that Neolithic dogs could have responded to the transition to a starch-rich diet by expanding microbial functionalities devoted to starch catabolism, thus compensating for delayed host response., Graphical abstract, Highlights • Ancient DNA of Bronze Age canine coprolites from Solarolo was sequenced • Solarolo dogs share gut microbiome features with modern wolves and dogs • The gut microbiome of Solarolo dogs shows high number of reads for alpha-amylase • Neolithic canine gut microbiome complemented delay in host genome adaptation, Biological sciences; Microbiome; Evolutionary biology; Evolutionary processes; Phylogenetics; Phylogeny; Evolutionary history; Genomic analysis; Sequence analysis; Omics; Genomics

Dog evolution has long been debated, particularly with regards to the origin of domestication and its effects on the genome, but little is known about the role of the gut microbiome during the recent canine evolutionary history. We sequenced the metagenome of 13 canine coprolites dated ca. 3,600-3,450 y ago from the Bronze Age archaeological site of Solarolo (Italy), which housed a complex farming community. The microbiome structure of Solarolo dogs revealed continuity with that of modern dogs, but it also shared some features with the wild wolf microbiome, as a kind of transitional state between them. The dietary niche, as also inferred from the microbiome composition, was omnivorous, with evidence of consumption of starchy agricultural foods. Interestingly, the microbiome from Solarolo dogs was particularly enriched in sequences encoding alpha-amylases and complemented a low copy number of the host amylase (AMY2B) gene. These findings suggest that Neolithic dogs could have responded to the transition to a starch-rich diet by expanding microbial functionalities devoted to starch catabolism. Such a microbiome adaptive mechanism would have compensated for delayed host response to the new primarily abundant plant food resources, available from companionship with Bronze Age agriculturalists.

Gaucher disease (GD) is a monogenic disorder characterized byβ-glucocerebrosidase enzyme deficiency. Typical features of the diseaseare the unrestrained activation of inflammatory mechanisms andneuronal cell death in neuronopathic conditions, whose molecularpathways are still not fully understood. Recent evidences show thatupregulation of the Hippo pathway is associated with neuroinflamma-tion and neuronal cell death. We analyzed the Hippo pathway activitytaking advantage of a Drosophila Gaucher-like model characterized bydGBA1b knock-out (GBA1bKO).WefoundderegulationofCycE,dIAPandMYC, direct targets of the pathway both in Drosophila and mammals,which were severely reduced at transcript and protein level,suggestingacell growth impairment. Analyzing upstream components we foundthat Fat, an atypical cadherin upstream of the kinase complex, wasupregulated. Moreover the glypicans Dally and Dally-like, known to benegatively regulated by Fat and involved in glial and synapsedevelopment, were found downregulated in the GBA1bKObackground,supporting a role in neurological damage. On the other hand, theaberrant inflammatory condition of the hematological compartmentwas investigated in the human model of induced Pluripotent Stem Cells(iPSC), differentiated into hematopoietic progenitors. While GD iPSCswere able to differentiate into CD34+/CD43+/CD45+ progenitors, theyshowed a decreased proliferative potential compared to healthy donoriPSC either in semisolid and liquid culture, therefore exhibiting a growthimpairment. This effect was coupled with the activation of necroptosispathway with a significant upregulation of RIPK3 and MLKL both in thepluripotent and differentiated state. Correction of the mutation to rescuethe phenotype is ongoing, taking advantage of AAVS1 editing approachin order to develop a fully comparable cell model useful in a cell therapyperspective. These data obtained with suitable GD models support therole of Hippo and necroptosis effectors in GD, amenable of furtherconsideration as potential therapeutic targets.

Gaucher disease (GD) is an autosomic recessive genetic disorder, known as the most common lysosomal storage disease. The causative genetic alteration involves mutation of the GBA gene, encoding for the β-glucocerebrosidase enzyme, whose loss-of-function causes the accumulation of glucosylceramide and glucosylsphingosine especially in cells primarily of the reticulo-endothelial system. More than 90% of patients show a systemic disorder and only fewer has an impairment of the central nervous system. The aggressiveness of the disorder is not tightly correlated with the genotype indeed there are many cases reported in literature of patients carrying the same mutation but different pathological symptoms. Typical features of the disease are the unrestrained activation of inflammatory mechanisms and neuronal cell death in neuronopathic conditions, whose molecular pathways are still not fully understood. Recent evidences show that upregulation of the Hippo pathway is associated with neuroinflammation and neuronal cell death, while the necroptosis pathway was found involved in a murine model of Gaucher Disease.