Neural Stem Cell Compartments in a Mouse Model of Globoid Cell Leukodystrophy: Implication for Therapeutic Strategies

Globoid cell leukodystrophy (GLD) is a rare genetic lysosomal disorder due to deficiency in the B-galactocerebrosidase (GALC) enzyme. GLD affects mainly children; the prognosis is severe, leading to death few years after the diagnosis. Demyelination is believed to occur as a consequence of Psychosine accumulation and neuroinflammation. Clinical observations in GLD babies suggest that GALC deficiency might affect myelination before overt storage and inflammation, but this issue has been poorly addressed in pre-clinical studies. Similarly, little is known regarding the effect of GALC deficiency on neural stem cell (NSC) in the neurogenic niches during CNS development. The goal of this study was to extensively characterize the role of GALC in regulating the function of NSC niches during the disease progression in Twitcher (Twi) mice, a relevant GLD model. By morphological and functional analysis we showed altered cellular organization and loss of proliferating neuroblasts in the subventricular zone (SVZ) niche of Twi mice as a function of disease progression. These data were confirmed by in vitro experiments showing decreased numbers of primary neurospheres generated from Twi NSC/progenitors. Both defects were rescued to normal levels in symptomatic Twi mice chronically treated with anti-inflammatory drugs. These results, as well as the up-regulation of several inflammatory molecules observed in Twi brains starting from the early symptomatic stage, suggested a major contribution by neuroinflammation at the late stages of the disease. However, these data did not rule out a direct contribution of GALC deficiency nor do they exclude a role of GALC in maintaining a functional niche during CNS development. Indeed, our results indicate decreased proliferation and maturation of NSC/progenitors derived from asymptomatic Twi mice, suggesting that GALC deficiency might lead to neurogenic impairment independently from CNS inflammation. These results improve our understanding of the pathogenic mechanisms of GLD with important implications for therapy

Insights on peptide topology in the computational design of protein ligands: the example of lysozyme binding peptides

Herein, we compared the ability of linear and cyclic peptides generated in silico to target different protein sites: internal pockets and solvent-exposed sites. We selected human lysozyme (HuL) as a model target protein combined with the computational evolution of linear and cyclic peptides. The sequence evolution of these peptides was based on the PARCE algorithm. The generated peptides were screened based on their aqueous solubility and HuL binding affinity. The latter was evaluated by means of scoring functions and atomistic molecular dynamics (MD) trajectories in water, which allowed prediction of the structural features of the protein-peptide complexes. The computational results demonstrated that cyclic peptides constitute the optimal choice for solvent exposed sites, while both linear and cyclic peptides are capable of targeting the HuL pocket effectively. The most promising binders found in silico were investigated experimentally by surface plasmon resonance (SPR), nuclear magnetic resonance (NMR), and electrospray ionization mass spectrometry (ESI-MS) techniques. All tested peptides displayed dissociation constants in the micromolar range, as assessed by SPR; however, both NMR and ESI-MS suggested multiple binding modes, at least for the pocket binding peptides. A detailed NMR analysis confirmed that both linear and cyclic pocket peptides correctly target the binding site they were designed for

Estrogen-dependent downregulation of hypoxia-inducible factor (HIF)-2α in invasive breast cancer cells

The involvement of estrogen (E2) and hypoxia in tumor progression is well established. Hypoxia has been reported to activate and degrade estrogen receptor alpha (ERα) in breast cancer cells. Furthermore, E2 has been shown to regulate hypoxia-inducible factor (HIF)-1α protein, but its role in HIF-2α regulation remains largely unexplored. In this study, we found that both HIF-2α mRNA and protein were down-regulated in ER positive but not ER negative breast cancer cells upon treatment with E2. The analysis of 690 samples derived from 608 mixed and 82 triple-negative breast cancer patients revealed that high nuclear HIF-2α tumor levels are associated with a worse prognosis specifically in human epidermal growth factor receptor 2 (HER2) and hormone receptor positive patients. Consistently, ERα/HER2 positive breast cancer cells displayed less pronounced downregulation of HIF-2α by E2. Experiments using a histone deacetylase inhibitor indicate that the E2 mediated decrease in HIF-2α mRNA is due to transcriptional repression. A functional estrogen response element (ERE) was identified in the first intron of the gene encoding HIF-2α (EPAS1), suggesting transcriptional co-repressor recruitment by ERα. Our results demonstrate a novel modulation of HIF-2α in breast cancer cells, explaining the opposing regulation between HIF-1α and HIF-2α in hormone-responsive breast cancer

“Non sono speciale devo solo pisciare”. Incursioni grafiche per decostruire il binarismo cis-eteronormativo a partire dai bagni pubblici

A tuttx è capitato di utilizzare un bagno pubblico e trovarsi di fronte una scelta: una figurina in pantalone e una con la gonna. Per molte persone immedesimarsi è immediato. Il progetto “Gender free toilet. Just use it” vuole scardinare questo automatismo, partendo dalle esperienze di quelle persone che, invece, di fronte alle due porte si interrogano, si fermano e, spesso, si forzano a scegliere una delle due opzioni. Per mettere in discussione il binarismo cis-eteronormativo il progetto che qui presentiamo, e che proprio in questi giorni ha fatto la sua uscita pubblica sui bagni dello Short Theater festival di Roma, iniziando ad ammiccare da un numero sempre maggiore di porte, vorrebbe sostituire in ogni luogo pubblico ai bagni divisi per genere, uno o più bagni gender free, ovvero attraversabili e utilizzabili da tutte le persone, siano esse trans*, non binarie o cisgender. Come? Affiggendo sulle porte un adesivo, risultato di una riflessione collettiva: abbiamo infatti chiesto a persone amiche di pensare a quante volte fosse capitato loro di dover utilizzare un bagno pubblico e di non trovare sulla porta un simbolo che fosse rappresentativo e che lx facesse sentire a proprio agio. Il risultato è stata una raccolta di idee e forme di autorappresentazione a cui è stata data forma grafica. Partire dalla modifica, anche visiva, degli spazi pubblici, di quegli spazi che quotidianamente attraversiamo e che contribuiscono, come nel caso dei bagni, a dar forma e forza all’invisibilizzazione e all’oppressione di quelle soggettività che vengono percepite come “fuori norma” e “fuori luogo”, è per noi un modo di prendere posizione semplice, ma di grande impatto per il superamento del binarismo di genere. Per trasformare gli ambienti e renderli più sicuri e fruibili per tutte le soggettività che non rientrano nella dicotomia maschile-femminile; per cambiare la società, com’è organizzata e, quindi, anche il nostro modo di pensarla

HUGenomics: A support to personalized medicine research

In the coming years, human genome research will likely transform medical practices. Genome-wide association studies (GWAS) are an example of the research effort made to allowing scientists to identify genes involved in human disease, reaction to treatments or symptom severity. Indeed, the unique genetic profile of an individual and the knowledge of molecular basis of diseases are leading to the development of personalized medicines and therapies, but the exponential growth of available genomic data requires a computational effort that may limit the progress of personalized medicine. Within this context, we propose the development of a novel hardware and software integrated system, named HUGenomics. The framework aims at becoming an advanced support for personalized medicine research. Thanks to more efficient algorithms and data integration from different biological sources, HUGenomics aims at simplifying the interpretation of biological information and facilitating genomic research process by means of both computational and data visualization tools