SINEUP Non-coding RNA Targeting GDNF Rescues Motor Deficits and Neurodegeneration in a Mouse Model of Parkinson's Disease.

International audience; Glial cell-derived neurotrophic factor (GDNF) has a potent action in promoting the survival of dopamine (DA) neurons. Several studies indicate that increasing GDNF levels may be beneficial for the treatment of Parkinson's disease (PD) by reducing neurodegeneration of DA neurons. Despite a plethora of preclinical studies showing GDNF efficacy in PD animal models, its application in humans remains questionable for its poor efficacy and side effects due to its uncontrolled, ectopic expression. Here we took advantage of SINEUPs, a new class of antisense long non-coding RNA, that promote translation of partially overlapping sense protein-coding mRNAs with no effects on their mRNA levels. By synthesizing a SINEUP targeting Gdnf mRNA, we were able to increase endogenous GDNF protein levels by about 2-fold. Adeno-associated virus (AAV)9-mediated delivery in the striatum of wild-type (WT) mice led to an increase of endogenous GDNF protein for at least 6 months and the potentiation of the DA system's functions while showing no side effects. Furthermore, SINEUP-GDNF was able to ameliorate motor deficits and neurodegeneration of DA neurons in a PD neurochemical mouse model. Our data indicate that SINEUP-GDNF could represent a new strategy to increase endogenous GDNF protein levels in a more physiological manner for therapeutic treatments of PD

Influence of Antisynthetase Antibodies Specificities on Antisynthetase Syndrome Clinical Spectrum TimeCourse

Introduction: Increased cardiovascular (CV) morbidity and mortality is observed in inflammatory joint diseases (IJDs) such as rheumatoid arthritis, ankylosing spondylitis, and psoriatic arthritis. However, the management of CV disease in these conditions is far from being well established.Areas covered: This review summarizes the main epidemiologic, pathophysiological, and clinical risk factors of CV disease associated with IJDs. Less common aspects on early diagnosis and risk stratification of the CV disease in these conditions are also discussed. In Europe, the most commonly used risk algorithm in patients with IJDs is the modified SCORE index based on the revised recommendations proposed by the EULAR task force in 2017.Expert opinion: Early identification of IJD patients at high risk of CV disease is essential. It should include the use of complementary noninvasive imaging techniques. A multidisciplinary approach aimed to improve heart-healthy habits, including strict control of classic CV risk factors is crucial. Adequate management of the underlying IJD is also of main importance since the reduction of disease activity decreases the risk of CV events. Non-steroidal anti-inflammatory drugs may have a lesser harmful effect in IJD than in the general population, due to their anti-inflammatory effects along with other potential beneficial effects.This research was partially funded by FOREUM—Foundation for Research in Rheumatolog

A regulatory crosstalk in Down syndrome: competing mRNA-miRNA network

Down syndrome (DS) - also known as Trisomy 21 - is a genetic disorder caused by an extra copy of all or part of human chromosome 21 (HSA21). DS is a complex genetic condition characterized by over 80 clinically different phenotypes of variable penetrance and expressivity. Individuals with DS show alterations - both structural and functional - affecting distinct organs and systems, which suggest that a perturbation of embryogenesis occurs in individuals with Down syndrome, due to Trisomy 21. Large-scale gene expression studies have revealed a more complex scenario, highlighting a global transcriptional deregulation, extended also to genes mapping outside the DS Critical Region (DSCR) on HSA21, as well as on the other chromosomes. In this context, it is clear that direct and/or indirect interactions between gene products of HSA21 and those from the other chromosomes can better explain the complexity of the clinical manifestations of the disorder. Recent studies have pointed out a previously unexpected role of non-coding RNA (ncRNAs), such as pseudogenes and lincRNAs, on gene expression regulation. In particular, a new crosstalk mediated by the competition for the binding to specific miRNAs has been demonstrated between messenger RNAs and ncRNAs. Such a regulatory crosstalk represents a new and interesting "RNA language" through which different mRNAs regulate each other by competing for miRNAs' availability. Given these recent findings and considering that Trisomy 21-induced gene imbalance perturbs the entire transcriptome and occurs throughout the embryogenesis, the above-described regulatory crosstalk may be altered by HSA21 trisomy in DS fetuses. The pathological overexpression of HSA21 genes may perturb gene expression during embryogenesis through the alteration of the crosstalk mediated by the interaction with specific miRNAs. In turn, it would cause typical multisystem clinical manifestations of Down syndrome. Thus, the aim of this PhD project is to deeply explore the global gene deregulation, and the potential role of microRNAs' "sponges" played by HSA21 genes that are overexpressed in DS during embryonic development. Taking advantage of publicly available mRNA and miRNA expression datasets of human embryos (4-6 weeks), a regulatory miRNA/mRNA network in these crucial weeks of the human embryonic development was established. A significant fraction of genes within the network belong to developmental-related pathways. Afterwards, HSA21 genes overexpressed in DS embryos that belong to this network were identified using RNA-Seq datasets from DS and euploid matched chorionic villi. HUNK gene was computationally selected as the best candidate to be an HSA21-derived miRNAs' sponge. Experimental studies confirmed that the overexpression of its 3'UTR induces an increased expression of genes involved in embryonic development, including BCL2, CLIC5, EPHA5, ERBB4, HIPK2, MECP2, ONECUT2, and WNT5A, and a reduction of the expression of correlated miRNAs, including miR-17, miR-20a, miR-20b, miR-128 and miR-200c. Although further studies are still in progress, the results of both the computational and the experimental studies strongly suggest that the overexpression of HSA21 genes may perturb the physiologic regulatory miRNA/mRNA network during DS embryos' development. This would explain, at least in part, the multisystemic nature of the alterations the typically occur in individuals with the syndrome

The Olive Oil Sector: A Comparison between Consumers and “experts” Choices by the Sensory Analysis

AbstractThe analysis is focused on olive oil, given its importance in the present competitive scenario and also for the renewed and growing interest that this product has in nutrition, health and wellbeing. It is in the interests of the different categories of olive oil producers to highlight the value of the specific attributes of their products, through the certification systems, geographical indications or organic farming. We have analysed consumer liking in order to understand what sensory attributes guide the choice, because this can help managers to develop marketing strategies focused on consumers’ demands. This study was conducted to identify and define sensory characteristics of five Italian olive oils and to link these differences to consumer and “experts” (the chefs) preferences through the application of preference mapping. This study confirms the hypothesis that experts give more importance to intrinsic attributes than “novices”, and also that the chefs are more aware than consumers on the EU certification systems and geographical indications or organic farming

The development of new G-quadruplex stabilising ligands using click chemistry

EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Novel Transcription Factor Variants through RNA-Sequencing: The Importance of Being “Alternative”

Alternative splicing is a pervasive mechanism of RNA maturation in higher eukaryotes, which increases proteomic diversity and biological complexity. It has a key regulatory role in several physiological and pathological states. The diffusion of Next Generation Sequencing, particularly of RNA-Sequencing, has exponentially empowered the identification of novel transcripts revealing that more than 95% of human genes undergo alternative splicing. The highest rate of alternative splicing occurs in transcription factors encoding genes, mostly in Krüppel-associated box domains of zinc finger proteins. Since these molecules are responsible for gene expression, alternative splicing is a crucial mechanism to “regulate the regulators”. Indeed, different transcription factors isoforms may have different or even opposite functions. In this work, through a targeted re-analysis of our previously published RNA-Sequencing datasets, we identified nine novel transcripts in seven transcription factors genes. In silico analysis, combined with RT-PCR, cloning and Sanger sequencing, allowed us to experimentally validate these new variants. Through computational approaches we also predicted their novel structural and functional properties. Our findings indicate that alternative splicing is a major determinant of transcription factor diversity, confirming that accurate analysis of RNA-Sequencing data can reliably lead to the identification of novel transcripts, with potentially new functions

Short interspersed DNA elements and miRNAs: a novel hidden gene regulation layer in zebrafish?

In this study, we investigated by in silico analysis the possible correlation between microRNAs (miRNAs) and Anamnia V-SINEs (a superfamily of short interspersed nuclear elements), which belong to those retroposon families that have been preserved in vertebrate genomes for millions of years and are actively transcribed because they are embedded in the 3' untranslated region (UTR) of several genes. We report the results of the analysis of the genomic distribution of these mobile elements in zebrafish (Danio rerio) and discuss their involvement in generating miRNA gene loci. The computational study showed that the genes predicted to bear V-SINEs can be targeted by miRNAs with a very high hybridization E-value. Gene ontology analysis indicates that these genes are mainly involved in metabolic, membrane, and cytoplasmic signaling pathways. Nearly all the miRNAs that were predicted to target the V-SINEs of these genes, i.e., miR-338, miR-9, miR-181, miR-724, miR-735, and miR-204, have been validated in similar regulatory roles in mammals. The large number of genes bearing a V-SINE involved in metabolic and cellular processes suggests that V-SINEs may play a role in modulating cell responses to different stimuli and in preserving the metabolic balance during cell proliferation and differentiation. Although they need experimental validation, these preliminary results suggest that in the genome of D. rerio, as in other TE families in vertebrates, the preservation of V-SINE retroposons may also have been favored by their putative role in gene network modulation

Late-onset mucopolysaccharidosis type IIIA mimicking Usher syndrome

: Mucopolysaccharidosis type IIIA (MPS IIIA or Sanfilippo syndrome type A) is an autosomal recessive lysosomal storage disorder caused by pathogenic variants in the SGSH gene encoding N-sulfoglucosamine sulfohydrolase, an enzyme involved in the degradation of heparan sulfate. MPS IIIA is typically characterized by neurocognitive decline and hepatosplenomegaly with childhood onset. Here, we report on a 53-year-old male subject initially diagnosed with Usher syndrome for the concurrence of retinitis pigmentosa and sensorineural hearing loss. Clinical exome sequencing identified biallelic missense variants in SGSH, and biochemical assays showed complete deficiency of sulfamidase activity and increased urinary glycosaminoglycan excretion. Reverse phenotyping revealed left ventricle pseudo-hypertrophy, hepatosplenomegaly, bilateral deep white matter hyperintensities upon brain MRI, and decreased cortical metabolic activity by PET-CT. On neuropsychological testing, the proband presented only partial and isolated verbal memory deficits. This case illustrates the power of unbiased, comprehensive genetic testing for the diagnosis of challenging mild or atypical forms of MPS IIIA

Genetic epidemiology of inherited retinal diseases in a large patient cohort followed at a single center in Italy

Abstract Inherited retinal diseases (IRDs) are the leading cause of vision loss in the working-age population. We performed a retrospective epidemiological study to determine the genetic basis of IRDs in a large Italian cohort (n = 2790) followed at a single referral center. We provided, mainly by next generation sequencing, potentially conclusive molecular diagnosis for 2036 patients (from 1683 unrelated families). We identified a total of 1319 causative sequence variations in 132 genes, including 353 novel variants, and 866 possibly actionable genotypes for therapeutic approaches. ABCA4 was the most frequently mutated gene (n = 535; 26.3% of solved cases), followed by USH2A (n = 228; 11.2%) and RPGR (n = 102; 5.01%). The other 129 genes had a lower contribution to IRD pathogenesis (e.g. CHM 3.5%, RHO 3.5%; MYO7A 3.4%; CRB1 2.7%; RPE65 2%, RP1 1.8%; GUCY2D 1.7%). Seventy-eight genes were mutated in five patients or less. Mitochondrial DNA variants were responsible for 2.1% of cases. Our analysis confirms the complex genetic etiology of IRDs and reveals the high prevalence of ABCA4 and USH2A mutations. This study also uncovers genetic associations with a spectrum of clinical subgroups and highlights a valuable number of cases potentially eligible for clinical trials and, ultimately, for molecular therapies