Antagonism of the prokineticin system prevents and reverses allodynia and inflammation in a mouse model of diabetes

Neuropathic pain is a severe diabetes complication and its treatment is not satisfactory. It is associated with neuroinflammation-related events that participate in pain generation and chronicization. Prokineticins are a new family of chemokines that has emerged as critical players in immune system, inflammation and pain. We investigated the role of prokineticins and their receptors as modulators of neuropathic pain and inflammatory responses in experimental diabetes. In streptozotocin-induced-diabetes in mice, the time course expression of prokineticin and its receptors was evaluated in spinal cord and sciatic nerves, and correlated with mechanical allodynia. Spinal cord and sciatic nerve pro- and anti-inflammatory cytokines were measured as protein and mRNA, and spinal cord GluR subunits expression studied. The effect of preventive and therapeutic treatment with the prokineticin receptor antagonist PC1 on behavioural and biochemical parameters was evaluated. Peripheral immune activation was assessed measuring macrophage and T-helper cytokine production. An up-regulation of the Prokineticin system was present in spinal cord and nerves of diabetic mice, and correlated with allodynia. Therapeutic PC1 reversed allodynia while preventive treatment blocked its development. PC1 normalized prokineticin levels and prevented the up-regulation of GluN2B subunits in the spinal cord. The antagonist restored the pro-/anti-inflammatory cytokine balance altered in spinal cord and nerves and also reduced peripheral immune system activation in diabetic mice, decreasing macrophage proinflammatory cytokines and the T-helper 1 phenotype. The prokineticin system contributes to altered sensitivity in diabetic neuropathy and its inhibition blocked both allodynia and inflammatory events underlying disease

NF-κB/Rel-Mediated Regulation of the Neural Fate in Drosophila

Two distinct roles are described for Dorsal, Dif and Relish, the three NF-κB/Rel proteins of Drosophila, in the development of the peripheral nervous system. First, these factors regulate transcription of scute during the singling out of sensory organ precursors from clusters of cells expressing the proneural genes achaete and scute. This effect is possibly mediated through binding sites for NF-κB/Rel proteins in a regulatory module of the scute gene required for maintenance of scute expression in precursors as well as repression in cells surrounding precursors. Second, genetic evidence suggests that the receptor Toll-8, Relish, Dif and Dorsal, and the caspase Dredd pathway are active over the entire imaginal disc epithelium, but Toll-8 expression is excluded from sensory organ precursors. Relish promotes rapid turnover of transcripts of the target genes scute and asense through an indirect, post-transcriptional mechanism. We propose that this buffering of gene expression levels serves to keep the neuro-epithelium constantly poised for neurogenesis

Evolution de la régulation du complexe achaete-scute et évolution du patron des soies chez les Diptères

STRASBOURG-Sc. et Techniques (674822102) / SudocSudocFranceF

Mutual Exclusion of Sensory Bristles and Tendons on the Notum of Dipteran Flies

AbstractBackground: Genes of the achaete-scute complex encode transcription factors whose activity regulates the development of neural cells. The spatially restricted expression of achaete-scute on the mesonotum of higher flies governs the development and positioning of the large sensory bristles. On the scutum the bristles are arranged into conserved patterns, based on an ancestral arrangement of four longitudinal rows. This pattern appears to date back to the origin of cyclorraphous flies about 100–140 million years ago. The origin of the four-row bauplan, which is independent of body size, and the reasons for its conservation, are not known.Results: We report that tendons for attachment of the indirect flight muscles are invariably located between the bristle rows of the scutum throughout the Diptera. Tendon development depends on the activity of a transcription factor encoded by the gene stripe. In Drosophila, stripe and achaete-scute have separate expression domains, leading to spatial segregation of tendon precursors and bristle precursors. Furthermore the products of these genes act antagonistically: ectopic sr expression prevents bristle development and ectopic sc expression prevents normal muscle attachment. The product of stripe acts downstream of Achaete-Scute and interferes with the development of bristle precursors.Conclusions: The pattern of flight muscles has changed little throughout the Diptera and we argue that the sites of muscle attachment may have constrained the positioning of bristles during the course of evolution. This could account for the pattern of four bristle rows on the scutum

Engineering Geological and Geophysical Investigations to Characterise the Unstable Rock Slope of the Sopu Promontory (Gozo, Malta)

Different engineering geological and geophysical investigations were performed at the Sopu promontory in the island of Gozo (Malta), involved in an impressive lateral spreading process due to the superimposition of a stiff limestone (ULC) on a ductile clay (BC). The applied techniques include: traditional geological and engineering geological surveys, unmanned aerial vehicles (UAV) survey, electrical resistivity tomography (ERT) survey, ground-penetrating radar (GPR) investigations, single-station seismic ambient noise measurements, and array seismic ambient noise measurements. The integration of the obtained results allowed us to reconstruct a subsoil model of the promontory that includes features related to the local geology of the slope and to the landslide process, as well as to define a conceptual model that describes the main evolution phases of the expansion process. The presence of back-tilted rock blocks with no features of polarization of Rayleigh waves evidenced the different failure mechanism of the rigid UCL plateau at the Sopu promontory with respect to the Selmun promontory, located in the close island of Malta, where the lateral spreading due to the same geological setting tends to produce unstable rock blocks with a toppling mechanism. This result encourages further future observations and analyses of this topic

Dataset related to article "Important functional role of the protein osteopontin in the progression of malignant pleural mesothelioma"

<p>This record contains raw data related to article "Important functional role of the protein osteopontin in the progression of malignant pleural mesothelioma"</p><p><strong>Background: </strong>Malignant Pleural Mesothelioma (MPM) is an aggressive cancer of the mesothelial lining associated with exposure to airborne non-degradable asbestos fibers. Its poor response to currently available treatments prompted us to explore the biological mechanisms involved in its progression. MPM is characterized by chronic non-resolving inflammation; in this study we investigated which inflammatory mediators are mostly expressed in biological tumor samples from MPM patients, with a focus on inflammatory cytokines, chemokines and matrix components.</p><p><strong>Methods: </strong>Expression and quantification of Osteopontin (OPN) was detected in tumor and plasma samples of MPM patients by mRNA, immunohistochemistry and ELISA. The functional role of OPN was investigated in mouse MPM cell lines <i>in vivo</i> using an orthotopic syngeneic mouse model.</p><p><strong>Results: </strong>In patients with MPM, the protein OPN was significantly more expressed in tumors than in normal pleural tissues and predominantly produced by mesothelioma cells; plasma levels were elevated in patients and associated with poor prognosis. However, modulation of OPN levels was not significantly different in a series of 18 MPM patients receiving immunotherapy with durvalumab alone or with pembrolizumab in combination with chemotherapy, some of whom achieved a partial clinical response. Two established murine mesothelioma cell lines: AB1 and AB22 of sarcomatoid and epithelioid histology, respectively, spontaneously produced high levels of OPN. Silencing of the OPN gene (<i>Spp1</i>) dramatically inhibited tumor growth <i>in vivo</i> in an orthotopic model, indicating that OPN has an important promoting role in the proliferation of MPM cells. Treatment of mice with anti-CD44 mAb, blocking a major OPN receptor, significantly reduced tumor growth <i>in vivo</i>.</p><p><strong>Conclusion: </strong>These results demonstrate that OPN is an endogenous growth factor for mesothelial cells and inhibition of its signaling may be helpful to restrain tumor progression <i>in vivo</i>. These findings have translational potential to improve the therapeutic response of human MPM.</p&gt

Oncogenic KRAS-Induced Protein Signature in the Tumor Secretome Identifies Laminin-C2 and Pentraxin-3 as Useful Biomarkers for the Early Diagnosis of Pancreatic Cancer

KRAS mutations characterize pancreatic cell transformation from the earliest stages of carcinogenesis, and are present in >95% of pancreatic ductal adenocarcinoma (PDAC) cases. In search of novel biomarkers for the early diagnosis of PDAC, we identified the proteins secreted by the normal human pancreatic cell line (HPDE) recently transformed by inducing the overexpression of the KRASG12V oncogene. We report a proteomic signature of KRAS-induced secreted proteins, which was confirmed in surgical tumor samples from resected PDAC patients. The putative diagnostic performance of three candidates, Laminin-C2 (LAMC2), Tenascin-C (TNC) and Pentraxin-3 (PTX3), was investigated by ELISA quantification in two cohorts of PDAC patients (n = 200) eligible for surgery. Circulating levels of LAMC2, TNC and PTX3 were significantly higher in PDAC patients compared to the healthy individuals (p < 0.0001). The Receiver Operating Characteristics (ROC) curve showed good sensitivity (1) and specificity (0.63 and 0.85) for LAMC2 and PTX3, respectively, but not for TNC, and patients with high levels of LAMC2 had significantly shorter overall survival (p = 0.0007). High levels of LAMC2 and PTX3 were detected at early stages (I–IIB) and in CA19-9-low PDAC patients. In conclusion, pancreatic tumors release LAMC2 and PTX3, which can be quantified in the systemic circulation, and may be useful in selecting patients for further diagnostic imaging

PC1 does not affect body weight, blood glucose and insulin levels.

<p>Effect of PC1 administration from day 21 to day 35 on body weight (panel A), blood glucose levels (panel B) and insulin levels (panel C). Plasma insulin levels were measured at the end of the PC1 therapeutic treatment, i.e. on day 35 after MLD-STZ. Effect of preventive PC1 administration (from day 0 to 14) on body weight (panel D), blood glucose levels (panel E) and insulin levels (panel F). Plasma insulin levels were measured at the end of the PC1 therapeutic treatment, i.e. on day 14 after MLD-STZ. ***p<0.001 vs vehicle; * p<0.05 vs vehicle</p

Anti-allodynic effect of PKR antagonist (PC1) administration.

<p>Anti-allodynic effect of PC1 administration as a single bolus (A) or as repeated administrations (B, C). A: acute PC1(s.c. 150 μg/ kg) was administered 21 days after MLD-STZ. B: therapeutic PC1 protocol- PC1 was administered (s.c. 150 μg/ kg, twice-daily) for 14 days, from day 21 to 35 after MLD-STZ. C: preventive PC1 treatment-PC1 was administered (s.c. 150 μg /kg, twice-daily) for 14 days, starting from day 0, time point corresponding to the first STZ administration. Data represent mean± SEM of 6 mice / group. Two way ANOVA was used for statistical evaluation, followed by Bonferroni’s test. ***p<0.001 vs vehicle/CTR and CTR + PC1; °°p<0.01, °°°p<0.001 vs STZ.</p