Archive departmental seminars 2019/2020

02 October 2019 Silvia Garavaglia, Alberto Minassi (UNIUPO – DSF)

ALDHs are NAD-dependent enzymes capable of oxidizing aldehydes to their respective acids. In mammals, 19 genes encoding different ALDHs have been identified with a different cytosolic and tissue distribution, and capable of acting on specific substrates. Recent studies have shown a marked expression of ALDH1A3 in HGG glioma CSCs, suggesting for this isoenzyme a primary role in the development of this tumor. Through a molecular and rational approach, biochemical, biocrystalline and organic synthesis studies have made it possible to isolate a small library of compounds with both inhibitory and fluorescent action, capable of selectively binding to the ALDH1A3 enzyme. These molecules can be used both with a pharmacological approach as possible scaffolds for the development of new drugs in the treatment of gliomas, and for the development of fluorescent probes, specific for HGG-CSCs, to be used in the operative phase, for optimum visualisation of the tumour cells during tumour resection

16 ottobre 2019 Daniela Lalli (UNIUPO – DISIT)

The atomic description of cellular processes is an essential prerequisite for the comprehension of life and represents one of the main goals of scientific research. NMR spectroscopy is a powerful technique able to provide atomic level insights into the structure, dynamics and interactions of biomolecules in native environments and in real time. This has rendered it a powerful tool for structural biology highly complementary to X-ray crystallography and Cryo-electron microscopy. Here I provide an overview of the advanced solution and solid-state NMR methods and their applications to study a wide variety of biological processes spanning from Protein-Ligand to Protein-Protein and Protein-Metal Ion interactions. 

06 novembre 2019 Morris Losurdo (UNIUPO – DSF) 

The critical role of neuroinflammation in promoting and accelerating the pathogenic process in Alzheimer’s disease (AD), raising a need to target the cerebral innate immune cells as potential therapeutic strategy to slow down the disease progression. In this scenario, Mesenchymal Stem Cells (MSCs) have aroused great interest thank to their immunomodulatory properties, which have been largely ascribed to the release of extracellular vesicles (EVs), namely exosomes and microvesicles. Here I show, in a triple transgenic model of AD (3xTg mice), the immunomodulatory and neuroprotective effects of intranasally administered EVs derived from cytokine-preconditioned MSCs. MSC-EVs reached the brain, where they dampened the activation of microglia cells and increased dendritic spine density. MSC-EVs polarized in vitro murine primary microglia toward an anti-inflammatory phenotype, suggesting that the neuroprotective effects observed in transgenic mice could result from a positive modulation of the inflammatory status. The possibility of administering MSC-EVs through a non-invasive route and the demonstration of their anti-inflammatory efficacy, might accelerate the chance of a translational exploitation of MSC-EVs in AD.

20 November 2019 Gianluca Papeo (Nerviano Medical Sciences)

The seminar addresses a new method of treating oncological disease that exploits the genetic instability of cancer cells. In particular, the seminar looks at the therapeutic approach intended for cancers characterised by defects in the mechanisms responsible for DNA repair. The goal is to amplify the fragility of tumour DNA through the use of drugs designed to further hinder repair, thus inducing the death of the neoplastic cell. The progenitors of these types of drugs are inhibitors of poly (ADPribose) -polymerase 1 (PARP-1), a key enzyme in restoring damaged DNA. This enzyme exerts its role by building polymers on suitable target proteins, using NAD + as the monomer unit. After a brief introduction on DNA repair mechanisms and the role played by PARP-1, the central topic of the seminar will concern the history of the discovery of PARP-1 inhibitors, their pharmacological similarities and differences, the industrial syntheses of these drugs. and the therapeutic contexts for which they have been approved and placed on the market.

04 December 2019 Silvia Fallarini, Tracey Pirali (UNIUPO – DSF)

Indoleamine 2,3-dioxygenase 1 (IDO1) is one of the enzymes responsible for the oxidative degradation of tryptophan in the kynurenin pathway. Alterations in the physiological levels of IDO1 are associated with numerous pathologies including cancer, where it participates in the determination of the immunosuppressive microenvironment. None of the IDO1 inhibitors tested in clinical trials have to date been approved for clinical use, suggesting that the identification of an IDO1 inhibitor still represents an open challenge. Evaluation of the inhibitory activity of approximately 50 purchasable and drug-like compounds identified through virtual screening led to the identification of a new and potent IDO1 inhibitor (IC50 = 16 nM), which does not resemble any reported inhibitor and shows a modality of peculiar bond. The SAR analysis of this compound made it possible to identify an equally active analogue. Both were found to be selective, active in different tumour cell lines, capable of reducing the inhibitory activity exerted by immunosuppressive monocytes. The inhibitor with the best metabolic profile was also able to reduce kynurenine levels in vivo.

Project funded by Local Research Call

18 December 2019 Carla Distasi (UNIUPO – DSF)

Oxaliplatin (OHP) is an antineoplastic drug used for the treatment of solid tumors and in particular colorectal cancer. Complications affecting the peripheral nervous system accompanied by neuropathic pain are the factor that limits its use in the clinic. The neurotoxic profile of OHP is singular as it produces an acute neuropathy which develops in a short time (hours) and which manifests itself with paresthesia and severe hypersalgesia in the cold. OHP essentially acts at the level of the cell body of the neurons of the dorsal root ganglia where it alters cell functions and neuronal excitability. OHP in fact modifies the expression and activity of different types of ion channels but the mechanisms underlying these effects are not fully understood. From the study of the expression and activity of some ion channels (purinergic receptors, TRP and K2P channels), we have shown that oxaliplatin modifies the excitability of sensory neurons by significantly altering the homeostasis of intracellular pH.

Project funded by Local Research Call

08 gennaio 2020 Beatrice Riva (UNIUPO – DSF)

STIM and ORAI proteins play a fundamental role in calcium signalling, allowing for calcium influx through the plasma membrane upon depletion of intracellular stores in a process known as store-operated Ca2+-entry (SOCE). Gain-of-function mutations on STIM1 or Orai1 are responsible for a cluster of rare diseases (tubular aggregate myopathy, Stormorken syndrome and York platelet syndrome), multi-organ disorders mainly characterized by muscle disturbances, haematopoietic dyscrasias and platelet dysfunction. In this project, we have characterized some of the mutations that lead to these disorders in vitro and have completely characterized an animal model that bears one of the responsible mutations (STIM1 I115F). Furthermore, in collaboration with the medicinal chemistry group, we have discovered two separate series of inhibitors of SOCE that mitigate in vitro and in vivo the effect of the defect. The main aims of this project are to establish whether these disorders are amenable to pharmacological treatment with SOCE inhibitors, and to improve the knowledge on these neglected rare diseases providing both indications of mechanisms underlying the phenotype and possible surrogate biomarkers to follow the disorders.

05 February 2020 Erika Del Grosso (UNIUPO - DSF)

One of the successful strategies to counter the spread of malaria in endemic areas is today represented by the containment of the vector insect population (the Anopheles mosquito). A vital metabolic pathway in both humans and insects is represented by the "kynurenine pathway", through which the tryptophan in excess of the needs of protein synthesis (and the production of serotonin and melatonin), is enzymatically demolished. In adult insects, an excess of 3-hydroxyquinurenine (3-HK), an intermediate metabolite of the pathway, is cytotoxic and its detoxification is totally dependent on the metabolic capacity of the insect to transform it through direct transamination; this occurs by the enzyme 3-hydroxyquinurenine transaminase (3-HKT). The complete biochemical and structural characterization of the Anopheles 3-HKT enzyme and the development of new highly selective and specific coupled analytical methods allowed us to study the inhibitory activity of new synthetic molecules on the Anopheles 3-HKT enzyme, thus allowing us to identify new inhibitors.

Project funded by Local Research Call

12 February 2020 Francesca Consonni (UNIUPO - DSF)

Although the pathological significance, as well as the therapeutic potential, of tumour-associated macrophages (TAMs) heterogeneity is still poorly understood, TAMs reprogramming is viewed as a promising anticancer therapy. Here we show that a distinct subset of TAMs (F4/80highCD115highC3aRhighCD88high) endowed with high rates of heme catabolism by the stressresponsive enzyme heme oxygenase-1 (HMOX1/HO-1) plays a critical role in shaping a prometastatic tumour microenvironment, favoring immunosuppression and angiogenesis. This population originates from F4/80+HO-1+ marrow precursors, accumulates in the blood of tumour bearers, and preferentially localizes at the invasive margin through a mechanism regulated by the NFκB1/CSFR1/C3aR axis. Fittingly, relative expression levels of HO-1 in peripheral monocyte subsets discriminate survival among stage IIIC/IV melanoma patients. Overall, these results identify a distinct cancer-induced HO-1+ myeloid subgroup as novel anti-metastatic target and prognostic blood-marker. 

19 February 2020 Dmitry Lim (UNIUPO - DSF)

Astrocytes are principal homeostatic cells in the CNS. During neuronal activity they generate intracellular calcium (Ca2+) signals which are strongly implicated in astroglial homeostatic functions. How these signals are translated in homeostatic activities of astrocytes is poorly understood. We have generated mice with astrocyte-specific calcineurin (CaN) KO (ACN-KO) which at 1 month of age show (i) neuronal hypo-excitability due to inactivation of astrocytic Na+/K+ ATPase, (ii) deregulation of protein, but not gene, expression in both astrocytes and neurons due to inhibition of astrocytic protein synthesis, (iv) alterations of protein expression related to neurodegeneration and seizures. Strikingly, from 5 months of age ACN-KO mice show increasing risk of epileptic seizures. Deletion of CaN from astrocytes thus re-creates features of neuropathology and predisposes to seizures. Altogether, our data put astroglial CaN centre stage as an activity-dependent regulator of CNS homeostasis.

As part of a FAR-2016 project Ruolo della Calcineurina Astrogliale nella regolazione dell’eccitabilità neuronale mediata della ATPasi Na+/K

24 June 2020 Pietro Picconi & Danilo Mirizzi (Nerviano Medical Sciences). Host: Gian Cesare Tron (UNIUPO-DSF)

Targeted Protein Degradation (TPD) has received a great deal of attention in the past five years, becoming one of the trends in medicinal chemistry. With an ever-increasing number of publications and patents, it has been subject to huge investments by the pharma industry, delivering in 2019 the first two clinical trials dealing with the technology. From a “cute chemical curiosity”, as stated by one of its fathers (Craig Crews, Nature, 2019, 298 Protein-slaying Drugs Could Be the Next Blockbuster Therapies), TPD has become a potential tool in the hands of medicinal chemists for the discovery of new therapeutic agents targeting diseases-causing proteins. Since its mechanism deeply differs from inhibition, it can be complementary to it, even including the possibility to target (and destroy) proteins previously described as undruggable, expanding the weaponry in the fight against human diseases. Among the means to achieve TPD, heterobifunctional molecules (such as PROteolysis TArgeting Chimeras – PROTACs) represent the main class of chemical entities. They comprise a Target Protein Ligand (TPL) and an E3 ligase ligand (E3LL) connected by a spacer (linker). PROTACs achieve their goal by recruiting the TP to the E3L, resulting in its poly-ubiquitylation and successive degradation of the tagged TP by the proteasome. The main features of the technology, alongside the essential tools for TPD-based drug discovery projects will be discussed.

9 July 2020 Maria Luisa Torre (DipSF-UNIPV). Host: Lorena Segale (UNIUPO-DSF)

Mesenchymal Stem Cells (MSCs) play a vital role in regenerative medicine and their safety and efficacy are well established. While it was initially believed that the mechanism of action was linked to their multipotency, today it is shown that they act mainly through a paracrine action, mediated by their secretome. Therefore, with the secretome it is possible to exploit the power of MSCs with all the advantages of acellular therapy. The difficulties in using the secretome in the clinic derive from the fact that today it is not yet a drug: for it to become one, it is necessary to produce it in industrial quantities according to the rules of good manufacturing, to convey it in a pharmaceutical form, and to demonstrate its safety and efficacy. In this context, some challenges that have been faced and overcome will be described: the development of a scalable GMP-compliant technological process for the production, its formulation in a lyophilized product, called lyosecretoma, and some in vitro and in vivo tests. to determine safety and efficacy.

ATEx Project (Id. 637541), Programme of Interreg Cooperation V-A Italia Svizzera