Valproic acid neuroprotection in 6-OHDA lesioned rat, a model for parkinson's disease

Background: Valproic acid (VPA) a long standing anti-epileptic and anti-manic drug has been recently investigated as a neuroprotective molecule, in relation to its action as an inhibitor of histone deacetylases (HDACs), favoring relaxed configuration of chromatin and thus promoting gene transcription. Methods: In the present study, chronic administration of VPA added to the diet, was tested for neuroprotection in a rat model of Parkinson's disease. The model consists of multiple injections of the dopaminergic toxin, 6-hydroxydopamine (6-OHDA), unilaterally in the striatum with consequent degeneration of the dopaminergic neurons originating the nigro-striatal pathway. This model of neurodegeneration is widely used as a reliable animal model for Parkinson's disease (PD). Results: Chronic VPA administration significantly reduced degeneration of dopaminergic neurons in the substantia nigra, and of dopaminergic terminals in the striatum, in rats subjected to the unilateral lesion of the nigrostriatal pathway. VPA treatment was also able to increase α-synuclein expression in the substantia nigra and to counteract the lesion-dependent decrease of the protein in the substantia nigra itself and in the striatum. Conclusions: Present data, which follow previous results obtained in the rotenone rat model of nigrostriatal degeneration, allow to propose VPA as a treatment to be tested for its effectiveness in other animal models of parkinsonism, in view of possible translation to patients

Ribosome-inactivating proteins and their immunotoxins for cancer therapy: insights into the mechanism of cell death

Ribosome-inactivating proteins (RIPs) are a family of plant toxic enzymes that permanently damage ribosomes and possibly other cellular substrates, thus causing cell death involving different and still not completely understood pathways. The high cytotoxic activity showed by many RIPs makes them ideal candidates for the production of immunotoxins (ITs), chimeric proteins designed for the selective elimination of unwanted or malignant cells. Saporin-S6, a type 1 RIP extracted from Saponaria officinalis L. seeds, has been extensively employed to construct anticancer conjugates because of its high enzymatic activity, stability and resistance to conjugation procedures, resulting in the efficient killing of target cells. Here we investigated the anticancer properties of two saporin-based ITs, anti-CD20 RTX/S6 and anti-CD22 OM124/S6, designed for the experimental treatment of B-cell NHLs. Both ITs showed high cytotoxicity towards CD20-positive B-cells, and their antitumor efficacy was enhanced synergistically by a combined treatment with proteasome inhibitors or fludarabine. Furthermore, the two ITs showed differencies in potency and ability to activate effector caspases, and a different behavior in the presence of the ROS scavenger catalase. Taken together, these results suggest that the different carriers employed to target saporin might influence saporin intracellular routing and saporin-induced cell death mechanisms. We also investigated the early cellular response to stenodactylin, a recently discovered highly toxic type 2 RIP representing an interesting candidate for the design and production of a new IT for the experimental treatment of cancer

Immunoconjugates for Osteosarcoma Therapy: Preclinical Experiences and Future Perspectives

Osteosarcoma (OS) is an aggressive osteoid-producing tumor of mesenchymal origin, which represents the most common primary bone malignancy. It is characterized by a complex and frequently uncertain etiology. The current standard care for high-grade OS treatment is neoadjuvant chemotherapy, followed by surgery and post-operative chemotherapy. In order to ameliorate survival rates of patients, new therapeutic approaches have been evaluated, mainly immunotherapy with antibody-drug conjugates or immunoconjugates. These molecules consist of a carrier (frequently an antibody) joined by a linker to a toxic moiety (drug, radionuclide, or toxin). Although several clinical trials with immunoconjugates have been conducted, mainly in hematological tumors, their potential as therapeutic agents is relatively under-explored in many types of cancer. In this review, we report the immunoconjugates directed against OS surface antigens, considering the in vitro and in vivo studies. To date, several attempts have been made in preclinical settings, reporting encouraging results and demonstrating the validity of the idea. The clinical experience with glembatumumab vedotin may provide new insights into the real efficacy of antibody-drug conjugates for OS therapy, possibly giving more information about patient selection. Moreover, new opportunities could arise from the ongoing clinical trials in OS patients with unconjugated antibodies that could represent future candidates as carrier moieties of immunoconjugates

Apoptosis and necroptosis induced by stenodactylin in neuroblastoma cells can be completely prevented through caspase inhibition plus catalase or necrostatin-1

Abstract Background Stenodactylin is a highly toxic plant lectin purified from the caudex of Adenia stenodactyla , with molecular structure, intracellular routing and enzyme activity similar to those of ricin, a well-known type 2 ribosome-inactivating protein. However, in contrast with ricin, stenodactylin is retrogradely transported not only in peripheral nerves but also in the central nervous system. Purpose Stenodactylin properties make it a potential candidate for application in neurobiology and in experimental therapies against cancer. Thus, it is necessary to better clarify the toxic activity of this compound. Study design We investigated the mechanism of stenodactylin-induced cell death in the neuroblastoma-derived cell line, NB100, evaluating the implications of different death pathways and the involvement of oxidative stress. Methods Stenodactylin cytotoxicity was determined by evaluating protein synthesis and other viability parameters. Cell death pathways and oxidative stress were analysed through flow cytometry and microscopy. Inhibitors of apoptosis, oxidative stress and necroptosis were tested to evaluate their protective effect against stenodactylin cytotoxicity. Results Stenodactylin efficiently blocked protein synthesis and reduced the viability of neuroblastoma cells at an extremely low concentration and over a short time (1 pM, 24 h). Stenodactylin induced the strong and rapid activation of apoptosis and the production of free radicals. Here, for the first time, a complete and long lasting protection from the lethal effect induced by a toxic type 2 ribosome-inactivating protein has been obtained by combining the caspase inhibitor Z-VAD-fmk, to either the hydrogen peroxide scavenger catalase or the necroptotic inhibitor necrostatin-1. Conclusion In respect to stenodactylin cytotoxicity, our results: (i) confirm the high toxicity to nervous cells, (ii) indicate that multiple cell death pathways can be induced, (iii) show that apoptosis is the main death pathway, (iv) demonstrate the involvement of necroptosis and (v) oxidative stress

Web tools to fight pandemics : the COVID-19 experience

The current outbreak of COVID-19 has generated an unprecedented scientific response worldwide, with the generation of vast amounts of publicly available epidemiological, biological and clinical data. Bioinformatics scientists have quickly produced online methods to provide non-computational users with the opportunity of analyzing such data. In this review, we report the results of this effort, by cataloguing the currently most popular web tools for COVID-19 research and analysis. Our focus was driven on tools drawing data from the fields of epidemiology, genomics, interactomics and pharmacology, in order to provide a meaningful depiction of the current state of the art of COVID-19 online resources

Morphological analysis of the Hippocampal region of aged rats, role of Clasmatodendrosis

Clasmatodendrosis is a phenomenon first described by Alzheimer in 1910, which was observed in aged nervous system and in the course of neurodegenerative diseases. It consists in the loss of astrocytic distal processes. The occurrence of clasmatodendrosis is frequently associated with an increase of autofluorescent aggregates in different cell types of nervous tissue. In this study we designed a calibrated excitation/emission method of spectral unmixing aimed to discriminate the fluorescence emitted by commercial fluorochrome-conjugated antibodies from the autofluorescent signal, by using confocal microscopy and multiphoton fluorescence lifetime imaging techniques. By this method, the immunolabeled GFAP localization in the CA1 Hippocampal region of aged rats was analyzed. Autofluorescent debris showed a strong positivity to GFAP labeling, suggesting that the detached fragments of clasmatodendrotic astrocytes might take part in the generation of these structures. By 3D confocal analysis we found that these aggregates, were located on neuronal cell surfaces, as well as inside the soma and that, in addition, the presence of autofluorescent aggregates seemed to be related with increased adhesion phenomena among neurons. These data were compared with those obtained in control adult rats and in rats infused with lipopolysaccharide (LPS) in the 4th ventricle to induce a chronic inflammatory state. The presence of autofluorescent aggregates was detected in LPS rats and also in control rats, even if they appeared smaller and with a lesser intensity as compared with the aged rats. These findings suggest that clasmatodendrosis is a process involving the interaction of neurons and astrocytes in a prolonged timespan of life. Its severity increases with aging or under inflammatory and/or neurodegenerative diseases. In conclusion, our results seem to suggest that clasmatodendrosis can affect neuron functionality not only due to a decreased astrocyte activity, but also by direct interaction of the detached astrocytic fragments with neuron somata

Detection of subtype-specific breast cancer surface protein biomarkers via a novel transcriptomics approach

Background: Cell-surface proteins have been widely used as diagnostic and prognostic markers in cancer research and as targets for the development of anticancer agents. So far, very few attempts have been made to characterize the surfaceome of patients with breast cancer, particularly in relation with the current molecular breast cancer (BRCA) classification. In this view, we developed a new computational method to infer cell-surface protein activities from transcriptomics data, termed ‘SURFACER’. Methods: Gene expression data from GTEx were used to build a normal breast network model as input to infer differential cell-surface proteins activity in BRCA tissue samples retrieved from TCGA versus normal samples. Data were stratified according to the PAM50 transcriptional subtypes (Luminal A, Luminal B, HER2 and Basal), while unsupervised clustering techniques were applied to define BRCA subtypes according to cell-surface proteins activity. Results: Our approach led to the identification of 213 PAM50 subtypes-specific deregulated surface genes and the definition of five BRCA subtypes, whose prognostic value was assessed by survival analysis, identifying a cell-surface activity configuration at increased risk. The value of the SURFACER method in BRCA genotyping was tested by evaluating the performance of 11 different machine learning classification algorithms. Conclusions: BRCA patients can be stratified into five surface activity-specific groups having the potential to identify subtype-specific actionable targets to design tailored targeted therapies or for diagnostic purposes. SURFACER-defined subtypes show also a prognostic value, identifying surface-activity profiles at higher risk