Stefano Gianni

Understanding and hijacking the oncogenic signalling adaptor proteins CRKL, Gab2 and Frs2

Stefano GIANNI

RESEARCH AREA: Genetica, biologia e fisiopatologia molecolare-cellulare degli Eucarioti

Receptor tyrosine kinase (RTK) signaling plays key roles in cell physiology and development. Impaired activation of these signaling pathways is critical in the genesis and progression of many types of cancers. RTK signaling pathways are generally activated by growth factor binding to a specific trans-membrane receptor, which activates downstream signaling molecules. These mechanisms require the formation of specific protein complexes mediated by adaptor proteins. Thus, adaptor proteins, while lacking any enzymatic activity provide a critical scaffolding function that facilitates key signaling transduction events and regulates signal specificity and amplification. Among these adaptors, three proteins, namely CRKL, GAB2, and FRS2, represent particularly interesting targets, being recurrently amplified in several types of cancers and essential to cancer cell lines that harbor such amplification. The overexpression of these three proteins is able to transform immortalized human cell lines in in vitro or in vivo models and their knockdown significantly reduces cancer proliferation. Based on these observations, we propose that an effective chemotherapeutic strategy would be that of interfering with CRKL, GAB2 and FRS2 protein-protein interaction network.

To provide a detailed structural characterization of the interactions between CRKL, GAB2 and FRS2 and thier partners, we focused on the interactions between these proteins and p85, Grb2 and SHP2. Therefore we successfully cloned, expressed and purified the N- and C-terminal SH2 domains from p85, the SH3 domain from Grb2 and the N- and C-terminal SH2 domains from SHP2. Subsequently, these constructs were subjected to NMR characterization both in the presence and in the absence of a peptide mimicking GAB2 , which allowed us to assign the structural changes induced by binding. Furthermore, we conducted an extensive biophysical analysis on these proteins domains, which allowed us to describe the mechanism of recognition of these proteins at nearly atomic resolution.

Finally, the research group was also involved in the characterization of the folding of several protein domains, with particular emphasis on the role of intradomain communication in multidomain proteins.

Publications

Nalli M, Puxeddu M, La Regina G, Gianni S, Silvestri R. Emerging Therapeutic Agents for Colorectal Cancer. Molecules. 2021 Dec 9;26(24):7463. doi: 10.3390/molecules26247463.

I.F. 4.412 Pagano L, Malagrinò F, Nardella C, Gianni S, Toto A. Experimental

Characterization of the Interaction between the N-Terminal SH3 Domain of

Crkl and C3G. Int J Mol Sci. 2021 Dec 7;22(24):13174. I.F. 5.924

Nardella C, Malagrinò F, Pagano L, Rinaldo S, Gianni S, Toto A. Determining folding

and binding properties of the C-terminal SH2 domain of SHP2.

Protein Sci. 2021 Dec;30(12):2385-2395. doi: 10.1002/pro.4201. I.F. 6.725

Nardella C, Visconti L, Malagrinò F, Pagano L, Bufano M, Nalli M, Coluccia A, La Regina G, Silvestri R, Gianni S, Toto A. Targeting PDZ domains as potential treatment for viral infections, neurodegeneration and cancer. Biol Direct. 2021 Oct 12;16(1):15. doi: 10.1186/s13062-021-00303-9. I.F. 4.540

Laursen L, Gianni S, Jemth P. Dissecting Inter-domain Cooperativity in the Folding of a Multi Domain Protein. J Mol Biol. 2021 Sep 3;433(18):167148. doi: 10.1016/j.jmb.2021.167148. Epub 2021 Jul 8. I.F. 5.469

Pagano L, Malagrinò F, Visconti L, Troilo F, Pennacchietti V, Nardella C, Toto A, Gianni S. Probing the Effects of Local Frustration in the Folding of a Multidomain Protein. J Mol Biol. 2021 Jul 23;433(15):167087. doi: 10.1016/j.jmb.2021.167087. Epub 2021 Jun 3. I.F. 5.469 Visconti L, Malagrinò F, Troilo F, Pagano L, Toto A, Gianni S. Folding and Misfolding of a PDZ Tandem Repeat. J Mol Biol. 2021 Apr 2;433(7):166862. doi: 10.1016/j.jmb.2021.166862. Epub 2021 Feb 1.I.F. 5.469 Gianni S, Freiberger MI, Jemth P, Ferreiro DU, Wolynes PG, Fuxreiter M. Fuzziness

and Frustration in the Energy Landscape of Protein Folding, Function, and

Assembly. Acc Chem Res. 2021 Mar 2;54(5):1251-1259. doi: 10.1021/acs.accounts.0c00813. Epub 2021 Feb 8.I.F. 22.384 Fusco G, Gianni S. Function, Regulation, and Dysfunction of Intrinsically Disordered Proteins. Life (Basel). 2021 Feb 12;11(2):140. doi: 10.3390/life11020140.I.F. 3.817

Pagano L, Toto A, Malagrinò F, Visconti L, Jemth P, Gianni S. Double Mutant Cycles as a Tool to Address Folding, Binding, and Allostery. Int J Mol Sci. 2021 Jan 15;22(2):828. doi: 10.3390/ijms22020828. I.F. 5.924

Gautier C, Gianni S. Unveiling the Folding Mechanism of PDZ Domains. Methods Mol Biol. 2021;2256:149-156. doi: 10.1007/978-1-0716-1166-1_9. PMID: 34014521 I.F. 1.412

Research Group

Angelo Toto Researchers; Caterina Nardella, Post Doc Daniele Santorelli, Post Doc Livia Pagano, PhD student Awa Diop, PhD student Valeria Pennacchietti, PhD student

Collaborations Per Jemth, Uppsala University

Coronavirus Task force - DrugDesign_SARS2

Search for antiviral small molecules that inhibit ACE2-Spike binding

Roberta Costi, Roberto Di Santo, Antonella Messore Valentina Noemi Madia, Davide Ialongo ,

1. IP Rome,

Introduction The severe and acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a plusstrand RNA virus closely related to SARS and MERS, rapidly spreading worldwide. This virus is responsible for an ongoing global pandemic of coronavirus disease 2019 (COVID-19) causing many casualties in the human population. For these reasons, it is imperative to find drugs able to solve this global health issue as fast as possible. Interaction between the spike (S) protein of SARS-CoV-2 and the cell surface receptor angiotensin-converting enzyme 2 (ACE2) is responsible of the infectivity of the host, allowing the entry of the virus into the host cells. Indeed, ACE2 acts as a ligand-receptor pair that initiates the viral attachment and cellular entry of the virus. In particular, the receptor-binding domain (RBD) of the S protein binds the membrane-distal portion of the ACE2 protein. The S protein forms a homotrimer, which is cleaved shortly after synthesis into two fragments that remain associated non-covalently: S1, which contains the RBD, and S2, which mediates membrane fusion following the binding of Spike to ACE2. Notably, current vaccines induce antibody responses to S protein, and most neutralising antibodies bind to the RBD. Therefore, targeting the binding between the S protein and the ACE2 receptor is a promising approach for virus entry. Inhibitors of the protein-protein interaction between the S protein and human ACE2 are of considerable interest as potential antiviral agents because the interaction between S and ACE2 initiate membrane fusion and virus entry, taking place at an accessible extracellular site. While development of protein-protein interaction inhibitors with small molecules is more challenging than antibodies, small-molecule inhibitors could offer alternatives that are less strain- and mutation-sensitive, suitable for oral or inhaled administration, and more controllable/less immunogenic. Indeed, this strategy has already been successfully applied to inhibit the viral entry of other viruses, as in the caseof two FDA-approved drugs maraviroc and enfuvirtide.

Main results The general approach of our project was to screen as rapidly as possible, with a variety of functional in vitro tests, protein-protein and protein-inhibitor complexes available in the different platforms (including FDA-approved compounds and/or their known metabolites). A flagged version of the recombinant RBD of the SARS-CoV-2 and ACE2 receptor with two different tags constructed by collaborators IP Paris was used for the high-throughput screening (HTS) to identify molecules that interfere with the interaction between the RBD and ACE2. 21 candidates were selected from HTS based on a robust HTRF assay using lanthanide fluorescence for validation in doseresponse experiment and, among them, 8 molecules ware confirmed with an IC50 < 3 µM. Biolayer interferometry (BLI) technology was used as orthogonal assay for hit validation together with microscale thermophoresis (MST) experiments. Two hits were identified from HTRF screen, showing % of inhibition of 51.4 and 25.6 and IC50 of 1 and 2.42 µM vs ACE2 and of 0.44 and 3.58 µM against RBD. Moreover, the two hits showed good affinities to RBD or ACE2 determined by MST with Kd values of 68 and 170 or 52 µM, respectively, and IC50 for protein-protein interaction inhibition of 43 µM as observed with BLI assay (with Ki of 2 vs both RBD and ACE2). In vitro infection assay using the native SARS-CoV-2 virus and VeroE6 cell line showed that the two identified hits possess antiviral activity of 0.29 and 0.42 µM, respectively. When tested against SARS-CoV-2 Delta strain, the compounds reported IC50 of 0.68 and 1.41, respectively. However, the two hits proved to be cytotoxic with CC50 values of 4.9 and 8 µM, respectively. The two hits proved to be cytotoxic also against SARS-CoV-2 Delta strain with CC50 values of 2.7 and 10 µM, respectively. Therefore, in order to reduce the cytotoxic profile of these compounds and to improve their druggability, some analogues were designed. In particular, a structural modification involving the heteroatom linked to the carbon atom of the aromatic core was shifted from nitrogen to oxygen. The heterocyclic compounds were synthesized via a convergent three- or four-steps synthetic route by applying the most modern synthetic approaches. The newly synthesized compounds were purified through flash chromatography and characterized by means of nuclear magnetic resonance, mass spectrometry, and high-performance liquid chromatography. Biological evaluation of the activity of the newly synthesized compounds was carried out in order to deepen their mechanism of action and confirm the better druggability of these molecules in respect to the previously identified hits. The compounds showed good affinities to RBD or ACE2 determined by MST with Kd values of 47.6 and 43.6 µM, respectively, and IC50 for protein-protein interaction inhibition of 100 µM as observed with BLI assay (with Ki of 2). In vitro infection assay using the SARSCoV-2 virus Delta strain and VeroE6 cell line showed that the two identified hits possess antiviral activity of 1.72 and 6.53 µM, respectively. Notably, the newly synthesized compounds proved to be less or no cytotoxic vs both native and Delta strains. Also, experiments with Creoptix confirmed the binding of these molecules to ACE2. Further studies involved the

saturation transfer difference (STD) NMR, one of the most popular ligand-based NMR techniques for the study of protein−ligand interactions. The STD spectrum of RBD showed higher signals for one of the newly synthesized compounds in respect to the analogue Interestingly, the newly synthesized compounds were tested for their activity against phospholipidosis. Indeed, it was recently reported that phospholipidosis was a shared mechanism underlying the antiviral activity of many repurposed drugs, being phospholipidosis correlated with antiviral efficacy. Conversely, drugs that did not induce phospholipidosis were not antiviral. Thus, drugs can provoke phospholipidosis in cells and organs show also antiviral activity. This trend was observed also with SARS-CoV-2. Therefore, the newly synthesized compounds were investigated for their interference with phospholipidosis and, interestingly, one of them demonstrated to induce phospholipidosis.

Perspective The next step in our study will be the identification of the binding pocket of the new best acting compounds through crystallization and, also, the set up of RAPIDWave competition experiments. These data will allow us to elucidate the structure-activity relationships and to optimize the compounds through specific and focused structural modifications.

Publications in compilation

Research Group Roberta Costi

Professor; Roberto Di Santo Professor; Antonella Messore Researcher; Valentina Noemi Madia Post-Doc; Davide Ialongo PhD student.

Collaborations

Felix Rey, Unit of Structural Virology, Institut Pasteur, Paris; Benoit Deprez, Université de Lille, Institut Pasteur de Lille, Lille; Fabrice Agou, Plateforme de Criblage Chemogenomique et Biologique, Institut Pasteur, Paris; Marc Delarue, DSMB, Departement de Biologie Structurale et Chimie, CNRS, Institut Pasteur, Paris; Pierre Lafaye, Plateforme d’Ingenierie des Anticorps, CNRS, Institut Pasteur, Paris.

SCIENTIFIC BOARD RESEARCH PROJECTS

TRANSFORMING GROWTH FACTOR-BETA-INDUCED PROTEIN AS A NOVEL SECRETED IMMUNE CHECK-POINT IN CANCER

VINCENZO BARNABA

RESEARCH AREA: IMMUNOLOGY

Department of Scienze Cliniche Internistiche, Anestesiologiche e Cardiovascolari vincenzo.barnaba@uniroma1.it

We set up a LC/Mass-spectometry (Orbitrap)-based platform to identify the secreted proteome (secretome) in conditioned medium (CM) from fresh tumor and non-tumor surgery samples. By this approach, we selected a multitude of secreted proteins that were upregulated in colorectalcancer (CRC) secretome as compared to the non-tumor, in order to identify those potentially acting as secreted immune check-points (sICs). Their discovery may represent a tremendous resource for tumor specific drug targets, potentially acting as sIC inhibitors in both cold and hot tumors, unlike current IC inhibitors (e.g., IpilumumAb and NivolumAb) causing a partial or no remission in the majority of cold tumors. The transforming growth factor-beta-induced (TGFBI) protein (previously called BIG-H3) was found significantly upregulated in CRC secretome (as compared with the non-tumor), as well as in hepatocellular carcinoma (HCC) secretome. TGFBI is an RGD-containing extracellular matrix protein that binds to type I, II and IV collagens, serves as a ligand recognition sequence for several integrins, and inhibits cell adhesion, including adhesion of mononuclear cells by occupancy of various integrins on endothelial cells, but its role as sIC has not been fully investigated. We first validated by Elisa that TGFBI was overexpressed in CM from CRC or HCC tissue samples (as compared with non-tumor CM), in serum from CRC or HCC patients (as compared with HD sera), and positively correlated with the tumor stage (according to the TNM classification). Interestingly, both flow-cytometry and tissue-confocal microscopy revealed that TGFBI was overexpressed by tumor cells, T cells, monocytes and plasma cells in tumors in a significantly higher extent than in non-tumor, suggesting a massive involvement of the tumor microenvironment (TME) in secreting it. Importantly, the recombinant form of TGFBI, as well as the tumor CM containing high levels of native TGFBI, significantly inhibited various functions (IFN- and TNF- production, GZB and T-bet expression…) of anti-CD3/CD28-activated CD4 and CD8 T cells. In addition, neutralizing anti-TGFBI mAb restored T cell functions in vitro. Finally, we are validating that TGFBI can act as a sIC by using human 3D CRC organoids as a surrogate of animal models in vivo. Human 3D-organoids generated from various tumor tissues allow to determine the interaction

between tumor and immune system, the response (activation, cytokine production, killing…) by autologous CD8 and CD4 T cells derived from cancer patients, the role of sICs in inhibiting anti-tumor T cell response, the role of related sIC inhibitors in unlashing the anti- tumor T cell response. Human-based models, such as human organoids, can offer effective ways “to accelerate transition to a research system that does not involve testing on animals”, as the European Parliament has recently declared (see go.nature.com/3hzprhj).

Publications

Feizi N, Focaccetti C, Pacella I, Tucci G, Rossi A, Costanza M, Pedotti R, Sidney J, Sette A, La Rocca C, Procaccini C, Matarese G, Barnaba V*, Piconese S.. CD8+ T cells specific for cryptic apoptosis-associated epitopes exacerbate experimental autoimmune encephalomyelitis. Cell Death Dis. 2021 Oct 29;12(11):1026 e IF: 8,4

Timperi E, Barnaba V*. CD39 Regulation and Functions in T Cells. Int J Mol Sci. 2021 Jul28;22(15):8068. e IF: 5,9

Veneziani I, Infante P, Ferretti E, Melaiu O, Battistelli C, Lucarini V, Compagnone M, Nicoletti C, Castellano A, Petrini S, Ognibene M, Pezzolo A, Di Marcotullio L, Bei R, Moretta L, Pistoia V, Fruci D, Barnaba V, Locatelli F, Cifaldi L. Nutlin-3a Enhances Natural Killer Cell- Mediated Killing of Neuroblastoma by Restoring p53-Dependent Expression of Ligands for NKG2D and DNAM-1 Receptors. Cancer Immunol Res. 2021 Feb;9(2):170-183. IF: 11,1

Pacella I, Cammarata I, Martire C, Brancaccio G, Gaeta GB, Barnaba V*, Piconese S. CD8+ T cells specific to apoptosis-associated epitopes are expanded in patients with chronic HBV infection and fibrosis. Liver Int. 2021 Mar;41(3):470-481. IF: 5,8

Cossarizza A, Chang HD, Radbruch A, Abrignani S, Addo R, Akdis M, Andrä I, Andreata F, Annunziato F, Arranz E, Bacher P, Bari S, Barnaba V, et al. Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition). Eur J Immunol. 2021 Dec;51(12):2708-3145. IF: 5,5

*corresponding or co-corresponding author

METABOLIC REPROGRAMMING IN PHYSIOLOGY AND PATHOLOGY

FRANCESCA CUTRUZZOLÀ

RESEARCH AREA: GENETICA, BIOLOGIA E FISIOPATOLOGIA MOLECOLARE-CELLULARE DEGLI EUCARIOTI

Department of Biochemical Sciences “A.Rossi Fanelli” francesca.cutruzzola@uniroma1.it

Metabolic reprogramming is a well-known process occurring in cancer as well as in other cell types, like immune cells, proliferating epithelia or during cell differentiation. Our project aims at understanding how reshaping of cellular metabolism is linked to uncontrolled cell growth and maintenance of the proliferation potential in cancer as well as in other pathological states of genetic and sporadic origin. We have focused our attention on serine-glycine one-carbon metabolism (SGOC), a crucial metabolic pathway that fuels the folate and methionine cycles thereby providing cells with the building blocks, as well as the reducing power, necessary to maintain high rates of proliferation. In several cases, an increased activity of the key SGOC enzyme serine hydroxymethyltransferase (SHMT), responsible for reversible conversion of Ser and tetrahydrofolate (THF) into Gly and 5,10-methylene-THF, is observed. All available data suggest that SHMT is a remarkably complex enzyme, for several reasons. First, mitochondrial (SHMT2) and cytosolic (SHMT1/ SHMT2) isoforms of SHMT are known (Figure 1), the latter able to translocate in the nucleus to sustain de novo dTMP synthesis which, in mammals, provides dTTP for DNA replication. SHMT1 is a critical part of the dTMP Synthesis Complex (dTMP-SC) including thymidylate synthase (TYMS) and dihydrofolate reductase (DHFR). Changes in SHMT1 expression directly impact de novo dTMP synthesis by affecting dTMP-SC assembly and its inactivation causes uracil misincorporation and genomic instability. We have recently characterized the intracellular dynamics of the complex in cancer cells by an in situ proximity ligation assay, showing that it is also detected in the cytoplasm (Spizzichino et al., online 2021 granted the cover image of the issue). This result indicates that the role of the thymidylate synthesis complex assembly may go beyond dTMP synthesis. We have also successfully assembled the dTMP synthesis complex in vitro, employing tetrameric SHMT1 and a bifunctional chimeric enzyme comprising human thymidylate synthase and dihydrofolate reductase. We show that the

SHMT1 tetrameric state is required for efficient complex assembly, indicating that this aggregation state is evolutionarily selected in eukaryotes to optimize protein-protein interactions. Lastly, our results regarding the activity of the complete thymidylate cycle in vitro may provide a useful tool with respect to developing drugs targeting the entire complex instead of the individual components.

In vitro, we have also shown that cytosolic SHMT1 is able to control the migration of lung cancer cells, suggesting a possible explanation for the formation of brain metastasis and clarified the molecular mechanism underlying this process (Cutruzzolà et al., 2021 and Paone et al., 2021). These results are currently being validated in mouse models, to verify the therapeutic potential of selected molecules in controlling metastasis. The study of metabolic remodeling was also carried out in other pathological settings, such as neurodegenerative diseases (Zuliani et al., 2021). In parallel, we have continued our characterization of the SHMT protein/nucleic acids (DNA/RNA) interactions, by determining the structure of SHMT1/RNA complex by cryo-electron microscopy (CryoEM) and analyzing the effect of RNA-SHMT1 interaction on serine/glycine intracellular levels by a modeling approach (in collaboration with GG Tartaglia). We are also pursuing the analysis of the effect of SHMT-modulating molecules in other proliferative states, such as psoriasis (in collaboration with E. Candi) and the relevance of SHMT isoforms in metformin treatment in cancer (in collab. with J.A. Menendez).

Pubblicazioni

Cutruzzolà F, Bouzidi A, Liberati FR, Spizzichino S, Boumis G, Macone A, Rinaldo S, Giardina G, Paone A. The Emerging Role of Amino Acids of the Brain Microenvironment in the Process of Metastasis Formation. Cancers (Basel). 2021 Jun 9;13(12):2891. doi: 10.3390/cancers13122891. IF 6,639.

Fiorillo, A., Battistoni, A., Ammendola, S., Secli, V., Rinaldo, S., Cutruzzola, F., Demitri, N., Ilari, A. Structure and metal-binding properties of PA4063, a novel player in periplasmic zinc trafficking by Pseudomonas aeruginosa Acta Crystallographica Section D: Structural Biology, 2021, 77, 1401-1410. IF 7,652.

Mantoni F, Scribani Rossi C, Paiardini A, Di Matteo A, Cappellacci L, Petrelli R, Ricciutelli M, Paone A, Cutruzzolà F, Giardina G, Rinaldo S. Studying GGDEF

Domain in the Act: Minimize Conformational Frustration to Prevent Artefacts.

Life (Basel). 2021 Jan 6;11(1):31. IF 3,817.

Paone A, Bouzidi A, Rinaldo S, Giardina G, Cutruzzolà F. Importance of amino acids in brain parenchyma invasion by cancer cells. Oncoscience. 2021 Mar 31;8:47-49.

Spizzichino S, Boi D, Boumis G, Lucchi R, Liberati FR, Capelli D, Montanari R, Pochetti G, Piacentini R, Parisi G, Paone A, Rinaldo S, Contestabile R, Tramonti A, Paiardini A, Giardina G, Cutruzzolà F. Cytosolic localization and in vitro assembly of human de novo thymidylate synthesis complex. FEBS J. 2022 Mar;289(6):1625-1649. IF 5,542.

Zuliani, I., Lanzillotta, C., Tramutola, A., Barone, E., Perluigi, M., Rinaldo, S., Paone, A., Cutruzzolà, F., Bellanti, F., Spinelli, M., Natale, F., Fusco, S., Grassi, C., Di Domenico, F. High-fat diet leads to reduced protein o-glcnacylation and mitochondrial

defects promoting the development of Alzheimer’s disease

signatures. International Journal of Molecular Sciences, 2021, 22 (7), 3746. IF 5,924.

Research Group

Sharon Spizzichino Post-doctoral fellow Francesca Liberati PhD student Federica di Fonzo PhD Student

Collaborations

Gian Gaetano Tartaglia, Sapienza University of Rome (Italy) Javier A. Menendez, Catalan Institute of Oncology (Spain) Eleonora Candi, Univ. Roma Tor Vergata (Italy) Alessandra Baracca Univ. Bologna (Italy)

REDOX-MODULATING COMPOUNDS AS POTENTIAL ANTIVIRALS IN THE TREATMENT OF RESPIRATORY VIRUS INFECTIONS

ANNA TERESA PALAMARA

RESEARCH AREA: INFECTIOUS AGENT AND ASSOCIATED DISEASES

Department of Public Health and Infectious Diseases annateresa.palamara@uniroma1.it

The generation of Reactive Oxygen Species (ROS) is a key event during respiratory virus infections, and it is responsible for the regulation of different steps of virus life cycle. With regard to coronaviruses (CoVs), oxidative stress regulates the virus/host cell binding. In particular, in SARS-CoV-2 model, oxidation of the cysteine residues on the ACE2 peptidase domain and on the RBD of Spike protein, favours viral entry. Based on this evidence, the first period of the project was aimed at exploring the efficacy of thiol compounds against two respiratory viruses, CoV and influenza virus (IV), by evaluating different steps of viral replication. In collaboration with Celio Military Hospital, an aliquot of SARS-CoV-2 was pre-incubated for 1 h at 37° C with different thiol compounds: a GSH-precursor, its dithiol derivative or the N- butanoyl GSH derivative. The mixture was used to infect ACE2-expressing cell lines. In parallel, the same compounds were also added after the viral adsorption of SARS-CoV-2 or influenza virus for 24 h. We found that all the reducing agents were able to impair the infectivity of SARSCoV-2. In particular, the pre-incubation of the virus with both GSH precursors caused a strong decrease in the number of plaques suggesting that reducing conditions strongly impair virus entry. The antiviral effect was confirmed by the western blot analysis of the viral proteins, Spike (S) and Nucleocapsid (N), in lysates obtained from the cells infected with the virus pre-incubated with thiol molecules. We next investigated whether the thiol molecules inhibited intracellular virus life cycle’s steps. The plaque assay showed that all the thiols impaired viral replication, although with differentefficiencies. Treatment with GSH precursors led to over 90% reduction in virus production. Similarly, viral protein expression analysis revealed that thiols were able to interfere with viral replication and confirmed the highest inhibition mediated by the two GSH precursors. The latter also interfered with IV entry. Indeed, alveolar epithelial cells infected with a mixture of IV and GSH-precursor showed a lower expression of viral proteins compared to untreated conditions. Moreover, the same compounds, when added for 24 h post-infection, were able to increase the antioxidant response and inhibit IV replication. In conclusion, these results indicate that redox-modulating compounds can act

by two possible mechanisms: by preventing the binding between viruses and host receptors or by interfering with viral replication by restoring the antioxidant response in host cell. Further studies are in progress to clarify the effect of reducing compounds during the replication of SARS-CoV-2 and their properties in regulating redox-sensitive pathways important in the inflammatory response. Parallel studies of the present project were focused on the potential role of herpes simplex virus- 1 (HSV-1), a neurotropic DNA virus able to establish life-long latent infection in humans followed by periodic reactivations, in neurodegenerative processes related to Alzheimer’s disease (AD), the most common form of dementia in the elderly. In particular, we exploited in vitro and in vivo models of recurrent HSV-1 infection to evaluate whether repeated cycles of virus latency/reactivation may accelerate brain aging. We found that HSV-1 harboring neurons may accumulate markers of senescence such as H4K16 hyperacetylation and HIRA protein, together with the increase in Sin3/HDAC1, which may accelerate neuronal aging. Overall these results strongly suggest that recurrent HSV-1 infection can accelerate pathological aging of neurons thus contributing to neurodegeneration. Moreover, in collaboration with Università Cattolica del Sacro Cuore di Roma (Prof. Grassi teamat the Neuroscience Department), we explored the role played by astrocytes in HSV-1 infection of neurons. We found that HSV-1-infected astrocytes release ATP that contributes to the Ca2+- dependent activation of GSK-3β that is required for HSV-1 entry and replication in neurons. These data indicate that at least in vitro astrocytes promote HSV-1 infection in neurons, thus suggesting a critical role of astrocytes in neuronal function and their response to pathogenic stimuli, including viral infection.

Publications

De Santis, R., Luca, V., Näslund, J., Ehmann, R. K., De Angelis, M., Lundmark, E., Nencioni, L., Faggioni, G., Fillo, S., Amatore, D., Regalbuto, E., Molinari, F., Petralito, G., Wölfel, R., Stefanelli, P., Rezza, G., Palamara, A. T., Antwerpen, M., Forsman, M., & Lista, F. Rapid inactivation of SARSCoV-2 with LED irradiation of visible spectrum wavelengths. Journal of photochemistry and photobiology. 2021. 8, 100082. https://doi.org/10.1016/j.jpap.2021.100082 (IF. 2020: 6.252)

Fiorentino F, De Angelis M, Menna M, Rovere A, Caccuri AM, D'Acunzo F, Palamara AT, Nencioni L, Rotili D, Mai A. Anti-influenza A virus activity and structure-activity relationship of a series of nitrobenzoxadiazole derivatives. J

Enzyme Inhib Med Chem. 2021. 36(1):2128- 2138. doi: 10.1080/14756366.2021.1982932. PMID: 34583607; PMCID: PMC8480593. (IF 2020:5.051)

De Angelis, M., Della-Morte, D., Buttinelli, G., Di Martino, A., Pacifici, F., Checconi, P., Nencioni, L. Protective role of combined polyphenols and micronutrients against influenza a virus and SARS-CoV-2 infection in vitro. Biomedicines. 2021. 9(11) doi:10.3390/biomedicines9111721. (IF 2020: 6.081)

Baldelli S, Limongi D, Coni C, Ciccarone F, Ciotti M, Checconi P, Palamara AT, Ciriolo MR. BK Polyomavirus Activates HSF1 Stimulating Human Kidney Hek293 Cell Proliferation. Oxid Med Cell Longev. 2021. 2021:9176993. doi: 10.1155/2021/9176993. PMID: 34845419; PMCID:PMC8627348. (IF 2020: 6.543)

Chiappini, E., Santamaria, F., Marseglia, G. L., Marchisio, P., Galli, L., Cutrera, R., de Martino,M., Antonini, S., Becherucci, P., Biasci, P., Bortone, B., Bottero, S., Caldarelli, V., Cardinale, F., Gattinara, G. C., Ciarcià, M., Ciofi, D., D'Elios, S., Di Mauro, G., Doria, M… Villani, A. Prevention of recurrent respiratory infections: Inter-society Consensus. Italian journal of pediatrics. 2021. 47(1), 211. https://doi.org/10.1186/s13052-021-01150-0. (IF 2020: 2.638)

Bizzarri BM, Fanelli A, Botta L, De Angelis M, Palamara AT, Nencioni L, Saladino R. Aminomalononitrile inspired prebiotic chemistry as a novel multicomponent tool for the synthesis of imidazole and purine derivatives with anti-influenza A virus activity. RSC Adv. 2021. 8;11(48):30020-30029. doi: 10.1039/d1ra05240c. PMID: 35480240; PMCID: PMC9040849. (IF 2020: 3.361)

Prezioso C, Grimaldi A, Landi D, Nicoletti CG, Brazzini G, Piacentini F, Passerini S, LimongiD, Ciotti M, Palamara AT, Marfia GA, Pietropaolo V. Risk Assessment of Progressive Multifocal Leukoencephalopathy in Multiple Sclerosis Patients during 1 Year of Ocrelizumab Treatment. Viruses. 2021. 25;13(9):1684. doi: 10.3390/v13091684. PMID: 34578264; PMCID: PMC8473394.(IF 2020: 5.048)

Iebba, V., Zanotta, N., Campisciano, G., Zerbato, V., Di Bella, S., Cason, C., Luzzati, R., Confalonieri, M., Palamara, A. T., & Comar, M. Profiling of Oral Microbiota and Cytokines inCOVID-19 Patients. Frontiers in microbiology. 2021. 12, 671813. https://doi.org/10.3389/fmicb.2021.671813 (IF 2020. 5.640)

Pompilio A, Scribano D, Sarshar M, Di Bonaventura G, Palamara AT, Ambrosi

C. Gram- Negative Bacteria Holding Together in a Biofilm: The Acinetobacter baumannii Way. Microorganisms. 2021. 22;9(7):1353. doi: 10.3390/microorganisms9071353. PMID: 34206680; PMCID: PMC8304980. (IF 2020: 4.128)

Napoletani, G., Protto, V., Marcocci, M. E., Nencioni, L., Palamara, A. T., & De Chiara, G.. Recurrent Herpes Simplex Virus Type 1 (HSV-1) Infection Modulates Neuronal Aging Marks inIn Vitro and In Vivo Models. International journal of molecular sciences. 2021. 22(12), 6279. https://doi.org/10.3390/ijms22126279 (IF 2020: 5.923)

Sarshar M, Behzadi P, Scribano D, Palamara AT, Ambrosi C. Acinetobacter baumannii: An Ancient Commensal with Weapons of a Pathogen. Pathogens. 2021. 10(4):387. doi:10.3390/pathogens10040387 (IF 2020: 3.492) Fraternale, A., Zara, C., De Angelis, M., Nencioni, L., Palamara, A. T., Retini, M., Di Mambro, T., Magnani, M., & Crinelli, R. Intracellular RedoxModulated Pathways as Targets for Effective Approaches in the Treatment of Viral Infection. International journal of molecular sciences. 2021. 22(7), 3603. https://doi.org/10.3390/ijms22073603 (IF 2020: 5.923)

Madia VN, De Angelis M, De Vita D, Messore A, De Leo A, Ialongo D, Tudino V, Saccoliti F, De Chiara G, Garzoli S, Scipione L, Palamara AT, Di Santo R, Nencioni L, Costi R. Investigation of Commiphora myrrha (Nees) Engl. Oil and Its Main Components for Antiviral Activity. Pharmaceuticals (Basel). 2021. 9;14(3):243. doi: 10.3390/ph14030243. PMID: 33803165; PMCID: PMC7999460. (IF 2020: 5.863)

Ricordi, C., Pacifici, F., Lanzoni, G., Palamara, A. T., Garaci, E., & DellaMorte, D. Dietary and Protective Factors to Halt or Mitigate Progression of Autoimmunity, COVID-19 and Its Associated Metabolic Diseases. International journal of molecular sciences. 2021. 22(6), 3134. https://doi.org/10.3390/ijms22063134 (IF 2020: 5.923) Ambrosi, C., Prezioso, C., Checconi, P., Scribano, D., Sarshar, M., Capannari, M., Tomino, C., Fini, M., Garaci, E., Palamara, A. T., De Chiara, G., & Limongi, D. SARS-CoV-2: Comparative analysis of different RNA extraction methods. Journal of virological methods. 2021. 287, 114008. https://doi.org/10.1016/j.jviromet.2020.114008 (IF 2020: 2.014)

Li Puma DD, Marcocci ME, Lazzarino G, De Chiara G, Tavazzi B, Palamara AT, Piacentini R, Grassi C. Ca2+ -dependent release of ATP from astrocytes affects herpes simplex virus type 1 infection of neurons. Glia. 2021. 69(1):201215. doi: 10.1002/glia.23895. Epub 2020. PMID: 32818313. (IF 2020: 7.452)

De Angelis, M., Casciaro, B., Genovese, A., Brancaccio, D., Marcocci, M. E., Novellino, E., Carotenuto, A., Palamara, A. T., Mangoni, M. L., & Nencioni, L. Temporin G, an amphibian antimicrobial peptide against influenza and parainfluenza respiratory viruses: Insights into biological activity and mechanism of action. FASEB journal. 2021. 35(2), e21358. https://doi.org/10.1096/fj.202001885RR (IF 2020: 5.191)

Research Group

Marta De AngelisWalter ToscanelliLucia Nencioni Virginia Protto Maria Elena Marcocci

Collaborations

Giovanna De Chiara, Institute of Translational Pharmacology, NationalResearch Council, Rome Maria Luisa Mangoni, Dept. Biochemistry, Sapienza University of Rome Mauro Magnani, Dept. Biomolecular Sciences, University of Urbino Carlo Bo Claudio Grassi, Dept. of Neuroscience, Cattolica University of Rome David Della Morte; Dept. of Systems Medicine, University of Rome "Tor Vergata"; Dept. of Human Sciences and Quality of Life Promotion, San Raffaele Roma Open University; Dept. of Neurology and Evelyn F. McKnight Brain Institute, Miller School of Medicine, University of Miami Paola Stefanelli; Dept. of Infectious Diseases, Istituto Superiore di Sanità Florigio Lista, Dept. Scientific Celio Military Hospital

NEW AGENTS AGAINST COLORECTAL CANCER

ROMANO SILVESTRI

RESEARCH AREA: NOVEL THERAPEUTIC INTERVENTIONS

Department of Drug Chemistry and Technologies romano.silvestri@uniroma1.it

Colorectal cancer (CRC) is the third most common cancer and the fourth leading cause of cancer mortality worldwide. Surgical resection potentially provides the only curative option for patients with dysplastic adenocarcinomas in situ. About 35% of these patients develop local recurrence of tumor relapse that is associated with poor survival. Adjuvant systemic chemotherapy and local radiation are first-line treatments actually recommended for patients with primitive tumors at diagnosis, also being the only standard management for patients with metastatic colorectal cancer. In the era of personalized medicine, the identification of key molecular features or pathways that are specific to a certain CRC subtypes may represent potential therapeutic targets, enabling the implementation of tailored therapies and better patient management.

WNT/-catenin signaling pathway plays key roles in regulating tissue homeostasis. This pathway gets upregulated in several human diseases. Interaction with Frizzled receptor triggers downstream events in the presence of a WNT ligand which prompt the translocation of -catenin to the nucleus followed by recruitment of the scaffold protein Dishevelled (DVL) by the WNT ligand/Frizzled complex and inactivation of -catenin destruction complex by WNT ligand/Frizzled/DVL complex. Thus, abberation of the WNT/β-catenin pathway is responsible of cancer, but, specific agents targeting this signaling pathway have not been approved so far. Negative modulators of DVL1 are able to impair the binding to Frizzled receptors, turning off the aberrant activation of the WNT pathway and leading to anti-cancer activity.

In the “canonical” WNT/-catenin pathway, the binding of morphogen WNTs to Frizzled receptors (FZDs) recruits the PDZ family member DVL at the plasma membrane. DVL is a 736 amino acid long scaffold protein made of three consecutive domains, namely DIX, PDZ and DEP. The DIX domain is constituted by 80–85 amino acids and contributes to DVL oligomerization. The 80–90 amino acid long PDZ domain binds to: (i) a PDZ binding motif (a conserved KTXXXW signaling sequence) allocated at the C-terminal tail of the receptors; (ii) a discontinuous domain spanning through the intracellular loops of the receptors. Finally, the 90–100 amino acid long DEP domain binds to the C-terminal tail of FZD and to its intracellular loops, and it is involved in the intracellular transduction of the WNT/-catenin signaling. The WNT–FZD–DVL complex inactivates the destruction complex formed by Glucose Synthase Kinase 3 (GSK-3), Axin, and Adenomatous Polyposis Coli (APC), and leads to the intracellular accumulation and nuclear translocation of -catenin.

The interaction between proteins is a fundamental event for cellular life that is generally mediated by specialized protein domains or modules. PDZ domains are the largest class of protein–protein interaction modules, involved in several cellular pathways such as signal transduction, cell–cell junctions, cell polarity and adhesion, and protein trafficking. Dysregulation of PDZ domain function often causes the onset of pathologies, thus making this family of domains an interesting pharmaceutical target. Functional features of PDZ domains and their involvement in the cellular and molecular pathways are at the basis of different human pathologies, and strategies to hamper the interaction of PDZ domains with their ligands have been analyzed.

Structure-based virtual screening studiesand molecular dynamic simulations have led to identify hit compounds with promising selective DVL1 binding inhibition. In the in vitro assays, these compounds inhibited the growth of HCT116 cells expressing wild-type APC with micromolar EC50 values and caused high level of ROS production. These results highlight such compounds for the development of new therapeutic agents against WNT-dependent colon cancer. The interactions of prototypic inhibitors of β-catenin have been investigated with the specific aim to understand the structural basis of the binding and improve the inhibition. In fact, to our knowledge, the effect of -catenin sulfonamido inhibitors has been not exhaustively explored, with the exception of very few examples. DVL1 and β-catenin inhibitors may be synergistically employed to get sustained inhibition of CRC.

Publications

Nardella C, Visconti L, Malagrinò F, Pagano L, Bufano M, Nalli M, Coluccia A, La Regina G, Silvestri R, Gianni S, Toto A. Targeting PDZ domains as potential treatment for viral infections, neurodegeneration and cancer. Biology Direct 2021, 16, 15, 1-21. https://doi.org/10.1186/s13062-021-00303-9. IF = 4.78.

Passirani, C, Vessières A, La Regina G, Link W, Silvestri R. Modulating undruggable targets to overcome cancer therapy resistance. Drug Research Updates, accepted 1 Dec 2021. https://doi.org/10.1016/j.drup.2021.100788. IF = 18.50.

Nalli M, Puxeddu M, La Regina G, Gianni S, Silvestri R. Emerging therapeutic agents for colorectal cancer. Molecules 2021, 26, 7463. https://doi.org/10.3390/molecules26247463. IF = 4.41.

Li S, Xu A, Li Y, Tan C, La Regina G, Silvestri R, Wang H, Qi W. RS4651 suppresses lung fibroblast activation via the TGF-β1/SMAD signalling pathway. Eur. J. Pharmacology 2021, 903, 174135. https://doi.org/10.1016/j.ejphar.2021.174135. IF = 4.43

Liu T, Wu J, Han C, Gong Z, La Regina G, Chen J, Dou F, Silvestri R, Chen C, Yu Z.

RS-5645 attenuates inflammatory cytokine storm induced by SARS-CoV-2 spike

protein and LPS by modulating pulmonary microbiota. International Journal of Biological Sciences 2021, 17, 3305-3319. doi:10.7150/ijbs.63329. IF = 6.8.

Research Group Romano Silvestri, Giuseppe La

Regina, Antonio Coluccia: Researchers. Michela Puxeddu, Jessica Sebastiani, Marianna Bufano: PhD Students.

Collaborations Francesca Cutruzzolà, Stefano Gianni,

Biochemical Sciences, Sapienza University Gianluca Canettieri, Molecular Medicine, Sapienza University Mariano Stornaiuolo, Dept. Pharmacy, University of Naples “Federico II” Wolfgang Link, CSIC-UAM, Madrid, Spain Catherine Passirani, MINT, Univ. Angers, UMR INSERM, France Te Liu, Shanghai University, China

STUDY OF MOLECULAR MECHANISMS CONTROLLING EMT/MET DYNAMICS IN CELL DIFFERENTIATION AND TRANSFORMATION

MARCO TRIPODI

Department of Molecular Medicine marco.tripodi@uniroma1.it

Background: Our research focuses on molecular mechanisms controlling epithelial cell dynamics in physiology and pathology. Notably, we previously unveiled a molecular circuitry of reciprocal direct transcriptional repression between Snail, master transcriptional factor of the Epithelial to Mesenchymal Transition (EMT), and HNF4α, master of epithelial differentiation and of the Mesenchymal to Epithelial transition (MET) process, whose balance is responsible for different cell physio-pathological outcomes (i.e. stemness vs differentiation; EMT vs MET; epithelial tumor progression vs tumor suppression). This circuitry has been further integrated by means of specific microRNAs and other co-regulators (Cicchini et al., JCP 2006; Santangelo et al., Hepatology 2011; Garibaldi et al., Cell Death and Diff. 2011; Cicchini et al., BBA gene reg. mech. 2015; Noce et al., Cell Death and Dis. 2019). Notably, we highlighted the role in EMT of HOTAIR, lncRNA largely correlated to tumor progression, metastasis and poor prognosis in several epithelial cancers. HOTAIR is recruited by Snail to specific genomic sites and it is required for Snail repressive activity by acting as a scaffold for EZH2, responsible for the H3K27 trimethylation. In other words, HOTAIR is required for the Snail-mediated repression of epithelial genes required in tumor progression (Battistelli et al., Oncogene 2017). On the other hand, HNF4α was found to directly repress HOTAIR transcription by causing the release of a chromatin loop on its regulatory elements. Therefore, an enhancer blocking activity is attributed for the first time to HNF4α repressive activity and the knowledge about the role of HNF4α as a transcriptional repressor of mesenchymal genes in EMT/MET dynamics is integrated by identifying a new mechanism (Battistelli et al., Cell Death and Diff. 2019).

Annual Report The evidence that Snail was able to convey EZH2 to target sites by means of a direct interaction with HOTAIR in both murine and human cells suggests that specific tridimensional structures of HOTAIR molecules are conserved between these two species and able to act as functional domains (the human HOTAIR shares sequence similarity (55% identity) with its mouse homologue while EZH2 and Snail are very well conserved in the two species (92.8% and 99.3% sequence identity, respectively)). Results by theoretical methods identified potential protein binding regions of murine and human HOTAIR RNAs. Therefore, we designed a HOTAIR deletion mutant form, named HOTAIR-sbid, including the putative Snail-binding domain but depleted of the EZH2-binding domain. HOTAIR-sbid acts as a dominant negative of the

endogenous HOTAIR wild type HOTAIR function in regulating Snail activity. In both murine and human tumor cells, HOTAIR-sbid impairs the ability of HOTAIR to bind Snail and, in turn, trigger H3K27me3/EZH2-mediated repression of Snail epithelial target genes. Notably, HOTAIR-sbid expression was proven to reduce cellular motility, invasiveness, anchorage-independent growth, and responsiveness to TGFβ-induced EMT. Thus, this HOTAIR mutant could represent an innovative tool for an innovative RNA-based gene therapy approach to counteract epithelial tumor metastasis.

Publications

Cecilia Battistelli; Sabrina Garbo; Veronica Riccioni; Claudia Montaldo; Laura Santangelo; Andrea Vandelli; Raffaele Strippoli; Gian Gaetano Tartaglia; Marco Tripodi; Carla Cicchini. Design and Functional Validation of a Mutant Variant of the LncRNA HOTAIR to Counteract Snail Function in Epithelial-to-Mesenchymal Transition. Cancer Research 2021 N.1 Vol.81 p.103-113 IF: 12.701

Montaldo C, Terri M, Riccioni V, Battistelli C, Bordoni V, D'Offizi G, Prado MG, Trionfetti F, Vescovo T, Tartaglia E, Strippoli R, Agrati C, Tripodi M. Fibrogenic signals persist in DAA-treated HCV patients after sustained virological response. J Hepatol. 2021 Dec;75(6):1301-1311. doi: 10.1016/j.jhep.2021.07.003. Epub 2021 Jul 13.PMID: 34271004 Free article.

Pleural Mesothelial Cells Modulate the Inflammatory/Profibrotic Response During SARS-CoV-2 Infection. Matusali G, Trionfetti F, Bordoni V, Nardacci R, Falasca L, Colombo D, Terri M, Montaldo C, Castilletti C, Mariotti D, Del Nonno F, Capobianchi MR, Agrati C, Tripodi M, Strippoli R. Front Mol Biosci. 2021 Nov 26;8:752616. doi: 10.3389/fmolb.2021.752616. eCollection 2021. PMID: 34901152 Free PMC article.

The RNA editing enzyme ADAR2 restricts L1 mobility. Frassinelli L, Orecchini E, Al-Wardat S, Tripodi M, Mancone C, Doria M, Galardi S, Ciafrè SA, Michienzi A. RNA Biol. 2021 Oct 15;18(sup1):75-87. doi: 10.1080/15476286.2021.1940020. Epub 2021 Jul 5. PMID: 34224323 Free PMC article.

Mechanisms of Peritoneal Fibrosis: Focus on Immune Cells-Peritoneal Stroma Interactions. Terri M, Trionfetti F, Montaldo C, Cordani M, Tripodi M, Lopez-Cabrera M, Strippoli R. Front Immunol. 2021 Mar 29;12:607204. doi: 10.3389/fimmu.2021.607204. eCollection 2021.

PMID: 33854496

Garbo S, Tripodi M, Battistelli C. A novel RNA-based approach to counteract EMT. Oncoscience. 2021 May 7;8:53-54. doi: 10.18632/oncoscience.532. eCollection 2021. PMID: 33997108 Free PMC article. No abstract available.

Research Group

Laura Amicone. Associate Professor Cecilia Battistelli Researcher Carla Cicchini Researcher Sabrina Garbo. PhD student Alessandra Marchetti Researcher Valeria Noce. Postdoc fellow Veronica Riccioni PhD student Raffaele Strippoli. Researcher Michela Terri PhD student

Collaborations

Gian Gaetano Tartaglia, Centre for Genomic Regulation (CRG), The Barcelona Institute for Science and Technology and Universitat Pompeu Fabra (UPF), Barcelona, Spain; Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, Genoa, Italy; Department of Biology ‘Charles Darwin’, Sapienza University of Rome, Rome, Italy; Institucio Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain.

KALEIDOSCOPICAL MICROGLIA IN PHYSIO-PATHOLOGICAL CONDITIONS

CRISTINA LIMATOLA1 1. IP Rome, Department of Physiology and Pharmacology Sapienza University of Rome

The growing interest in the role of microglia in the progression of many neurodegenerative diseases is developing in an ever-expedited manner, in part thanks to emergent new tools for studying the morphological and functional features of the CNS. Microglia are the resident immune cells of the central nervous system (CNS) and sustain normal brain functions continuously monitoring cerebral parenchyma to detect neuronal activities and alteration of homeostatic processes. Depending on the brain region, they represent from 5 to 12% of total cell population (1), and continuously monitor the surrounding parenchyma to sense alteration of brain functions (2, 3), controlling neuronal excitability, synaptic activity, neurogenesis, and clearance of apoptotic cells in the healthy adult brain (4). The discovery of specific biomarkers of the microglia phenotype could find application in wide range of human diseases and creates opportunities for the discovery and development of tailored therapeutic interventions. Among these, new pathways have been identified able to modulate microglia phenotype, such as the ion channels. Microglia express many ion channels: H+ channels, Na+ channels, voltage-gated Ca2+ channels, Ca2+ -release-activated Ca2+ channels, voltage-dependent and voltage-independent Cl− channels, voltage-gated and Ca2+ -activated K+ channels, inward rectifier K channels and more recently, the twopore domain channel THIK-1 (K2P13.1) (5). Particularly, we demonstrated that the KCa3.1 microglial channel play a central role in the progression of the neurodegenerative disease Amyotrophic Lateral Sclerosis (ALS); in fact, its inhibition attenuates the proinflammatory phenotype of hypothalamic microglia, increasing food intake in ALS mouse models (hSOD1G93A and TDPA315T mice). These results reveal a new regulatory role for KCa3.1 to counteract weight loss in ALS (6). Concerning brain pathologies, microglia have a pivotal role in supporting the development and growth of the most aggressive brain tumor Glioblastoma (GBM) (7,8). We reported that microglia-derived extravesicles (EVs) affect GBM growth. Our findings indicated that EVs carry messages to cancer cells that modify GBM cell metabolism, reducing lactate, nitric oxide (NO), and glutamate (Glu) release, increasing the expression of Glu transporter Glt-1 on astrocytes. The in vivo benefit of microgliaderived sEVs results in a significantly reduced tumor mass and an increased survival of glioma-bearing mice, depending on miR-124 (7). Furthermore, we take advantage of engineered microglia, modified with recombinant adeno-associated virus serotype 2 (rAAV2) carrying IL-15 (rAAV2-IL-15), to force the production of IL-15, to counteract GBM (8). We demonstrated that the intranasal delivery of rAAV2-IL-15 microglia enhanced NK cell recruitment and pro-inflammatory microglial phenotype in tumor mass of GBM-bearing mice, ultimately counteracted tumor growth. This approach has a

high potential for clinical translatability, highlighting the therapeutic efficacy of forced IL-15 production in microglia: the delivery of engineered rAAV2-IL-15 microglia to boost the immune response paves the way to design a new perspective therapy for glioma patients (8). Lastly, we focused the attention on the role of microglia in sleep. Sleep is a naturally occurring physiological state defined by behavioral criteria that include the absence of voluntary movement and increase in arousal threshold (9). Our data demonstrate that almost complete microglial depletion in wold type mice increases the duration of NREM sleep and reduces the hippocampal excitatory neurotransmission (10). The fractalkine receptor CX3CR1 plays a relevant role in these effects, because cx3cr1GFP/GFP mice recapitulate what found in PLX5622-treated mice. Furthermore, during the light phase, microglia express lower levels of cx3cr1 and a reduction of cx3cr1 expression is also observed when cultured microglial cells are stimulated by ATP, a purinergic molecule released during sleep. Our findings suggest that microglia participate in the regulation of sleep, adapting their cx3cr1 expression in response to the light/dark phase, and modulating synaptic activity in a phase-dependent manner (10). Altogether, our results identify a central role of microglia in orchestrating CNS functions in healthy brain and in neurodegenerative diseases.

References:

1. Lawson LJ, Perry VH, Dri P, Gordon S. Heterogeneity in the distribution and morphology of microglia in the normal adult mouse brain. Neuroscience. (1990) 39:151–70. 10.1016/0306-4522(90)90229-W 2. Davalos D, Grutzendler J, Yang G, Kim JV, Zuo Y, Jung S, et al. ATP mediates rapid microglial response to local brain injury in vivo. Nat Neurosci. (2005) 8:752–8. 10.1038/nn1472 3. Nimmerjahn A, Kirchhoff F, Helmchen F. Resting microglial cells are highly dynamic surveillants of brain parenchyma in vivo. Science. (2005) 308:1314–8. 10.1126/science.1110647 4. Lenz KM, McCarthy MM. A starring role for microglia in brain sex differences. Neuroscientist. (2015) 21:306–21. 10.1177/1073858414536468 5. Cocozza G, Garofalo S, Capitani R, D'Alessandro G, Limatola C. Microglial Potassium Channels: From Homeostasis to Neurodegeneration. Biomolecules. 2021 Nov 26;11(12):1774. doi: 10.3390/biom11121774 6. Cocozza G, Garofalo S, Morotti M, Chece G, Grimaldi A, Lecce M, Scavizzi F, Menghini R, Casagrande V, Federici M, Raspa M, Wulff H, Limatola C. The feeding behaviour of Amyotrophic Lateral Sclerosis mouse models is modulated by the Ca2+ -activated KCa 3.1 channels. Br J Pharmacol. 2021 Dec;178(24):48914906. doi: 10.1111/bph.15665. 7. Serpe C, Monaco L, Relucenti M, Iovino L, Familiari P, Scavizzi F, Raspa M, Familiari G, Civiero L, D'Agnano I, Limatola C, Catalano M. Microglia-Derived Small Extracellular Vesicles Reduce Glioma Growth by Modifying Tumor Cell

Metabolism and Enhancing Glutamate Clearance through miR-124. Cells. 2021

Aug 12;10(8):2066. doi: 10.3390/cells10082066. 8. Mormino A, Bernardini G, Cocozza G, Corbi N, Passananti C, Santoni A,

Limatola C, Garofalo S. Enriched Environment Cues Suggest a New Strategy to

Counteract Glioma: Engineered rAAV2-IL-15 Microglia Modulate the Tumor

Microenvironment. Front Immunol. 2021 Sep 6;12:730128. doi: 10.3389/fimmu.2021.730128. 9. Garofalo S, Picard K, Limatola C, Nadjar A, Pascual O, Tremblay MÈ. Role of

Glia in the Regulation of Sleep in Health and Disease. Compr Physiol. 2020 Mar 12;10(2):687-712. doi: 10.1002/cphy.c190022. 10. Corsi G, Picard K, di Castro MA, Garofalo S, Tucci F, Chece G, Del Percio C, Golia

MT, Raspa M, Scavizzi F, Decoeur F, Lauro C, Rigamonti M, Iannello F,

Ragozzino DA, Russo E, Bernardini G, Nadjar A, Tremblay ME, Babiloni C,

Maggi L, Limatola C. Microglia modulate hippocampal synaptic transmission and sleep duration along the light/dark cycle. Glia. 2022 Jan;70(1):89-105. doi: 10.1002/glia.24090.

Publications

Cocozza G, Garofalo S, Capitani R, D'Alessandro G, Limatola C. Microglial Potassium Channels: From Homeostasis to Neurodegeneration. Biomolecules. 2021 Nov 26;11(12):1774. doi: 10.3390/biom11121774. (I.F. 4.569) Cordella F, Sanchini C, Rosito M, Ferrucci L, Pediconi N, Cortese B, Guerrieri F, Pascucci GR, Antonangeli F, Peruzzi G, Giubettini M, Basilico B, Pagani F, Grimaldi A, D'Alessandro G, Limatola C, Ragozzino D, Di Angelantonio S. Antibiotics Treatment Modulates Microglia-Synapses Interaction. Cells. 2021 Oct 4;10(10):2648. doi: 10.3390/cells10102648. (I.F. 6.600) Cocozza G, Garofalo S, Morotti M, Chece G, Grimaldi A, Lecce M, Scavizzi F, Menghini R, Casagrande V, Federici M, Raspa M, Wulff H, Limatola C. The feeding behaviour of Amyotrophic Lateral Sclerosis mouse models is modulated by the Ca2+ -activated KCa 3.1 channels. Br J Pharmacol. 2021 Dec;178(24):4891-4906. doi: 10.1111/bph.15665. (I.F. 8.739) Basilico B, Ferrucci L, Ratano P, Golia MT, Grimaldi A, Rosito M, Ferretti V, Reverte I, Sanchini C, Marrone MC, Giubettini M, De Turris V, Salerno D, Garofalo S, St-Pierre MK, Carrier M, Renzi M, Pagani F, Modi B, Raspa M, Scavizzi F, Gross CT, Marinelli S, Tremblay MÈ, Caprioli D, Maggi L, Limatola C, Di Angelantonio S, Ragozzino D. Microglia control glutamatergic synapses in the adult mouse hippocampus. Glia. 2022 Jan;70(1):173-195. doi: 10.1002/glia.24101. (I.F. 7.452) Mormino A, Bernardini G, Cocozza G, Corbi N, Passananti C, Santoni A, Limatola C, Garofalo S. Enriched Environment Cues Suggest a New Strategy to Counteract Glioma:

Engineered rAAV2-IL-15 Microglia Modulate the Tumor Microenvironment. Front Immunol. 2021 Sep 6;12:730128. doi: 10.3389/fimmu.2021.730128. (I.F. 7.561) Serpe C, Monaco L, Relucenti M, Iovino L, Familiari P, Scavizzi F, Raspa M, Familiari G, Civiero L, D'Agnano I, Limatola C, Catalano M. Microglia-Derived Small Extracellular Vesicles Reduce Glioma Growth by Modifying Tumor Cell Metabolism and Enhancing Glutamate Clearance through miR-124. Cells. 2021 Aug 12;10(8):2066. doi: 10.3390/cells10082066. (I.F. 6.600) Corsi G, Picard K, di Castro MA, Garofalo S, Tucci F, Chece G, Del Percio C, Golia MT, Raspa M, Scavizzi F, Decoeur F, Lauro C, Rigamonti M, Iannello F, Ragozzino DA, Russo E, Bernardini G, Nadjar A, Tremblay ME, Babiloni C, Maggi L, Limatola C. Microglia modulate hippocampal synaptic transmission and sleep duration along the light/dark cycle. Glia. 2022 Jan;70(1):89-105. doi: 10.1002/glia.24090. (I.F. 7.452) Picard K, Bisht K, Poggini S, Garofalo S, Golia MT, Basilico B, Abdallah F, Ciano Albanese N, Amrein I, Vernoux N, Sharma K, Hui CW, C Savage J, Limatola C, Ragozzino D, Maggi L, Branchi I, Tremblay MÈ. Microglial-glucocorticoid receptor depletion alters the response of hippocampal microglia and neurons in a chronic unpredictable mild stress paradigm in female mice. Brain Behav Immun. 2021 Oct;97:423-439. doi: 10.1016/j.bbi.2021.07.022. (I.F. 7.217) Mormino A, Cocozza G, Fontemaggi G, Valente S, Esposito V, Santoro A, Bernardini G, Santoni A, Fazi F, Mai A, Limatola C, Garofalo S. Histone-deacetylase 8 drives the immune response and the growth of glioma. Glia. 2021 Nov;69(11):2682-2698. doi: 10.1002/glia.24065. Epub 2021 Jul 26. (I.F. 7.452) Naziris N, Pippa N, Sereti E, Chrysostomou V, Kędzierska M, Kajdanek J, Ionov M, Miłowska K, Balcerzak Ł, Garofalo S, Limatola C, Pispas S, Dimas K, Bryszewska M, Demetzos C. Chimeric Stimuli-Responsive Liposomes as Nanocarriers for the Delivery of the Anti-Glioma Agent TRAM-34. Int J Mol Sci. 2021 Jun 10;22(12):6271. doi: 10.3390/ijms22126271. (I.F. 4.556) D'Alessandro G, Lauro C, Quaglio D, Ghirga F, Botta B, Trettel F, Limatola C. NeuroSignals from Gut Microbiota: Perspectives for Brain Glioma. Cancers (Basel). 2021 Jun 4;13(11):2810. doi: 10.3390/cancers13112810. (I.F. 6.639) Poggini S, Matte Bon G, Golia MT, Ciano Albanese N, Viglione A, Poleggi A, Limatola C, Maggi L, Branchi I. Selecting antidepressants according to a drug-by-environment interaction: A comparison of fluoxetine and minocycline effects in mice living either in enriched or stressful conditions. Behav Brain Res. 2021 Jun 25;408:113256. doi: 10.1016/j.bbr.2021. (I.F. 3.332)

Research Group

Myriam Catalano Giuseppina D’Alessandro Flavia Trettel Laura Maggi Stefano Garofalo Maria Amalia Di Castro Germana Cocozza Collaborations

Angela Santoni Giovanni Bernardini Richard M Ransohoff Marie Eve Tremblay Fiorenzo Conti Ling Peng Eleonora Aronica

PUBLICATIONS

(derived by studies funded by Istituto Pasteur Italia in the year 2021)

1 During the COVID-19 pandemic where has respiratory syncytial virus gone? Di Mattia G, Nenna R, Mancino E, Rizzo V, Pierangeli A, Villani A, Midulla F. Pediatr Pulmonol. 2021 Oct;56(10):3106-3109. doi: 10.1002/ppul.25582. Epub 2021 Jul 26. PMID: 34273135 2 Urethane-induced lung carcinogenesis. Sozio F, Schioppa T, Sozzani S, Del Prete A. Methods Cell Biol. 2021;163:45-57. doi: 10.1016/bs.mcb.2020.09.005. Epub 2020 Oct 23. PMID: 33785168 3 Microglia control glutamatergic synapses in the adult mouse hippocampus. Basilico B, Ferrucci L, Ratano P, Golia MT, Grimaldi A, Rosito M, Ferretti V, Reverte I, Sanchini C, Marrone MC, Giubettini M, De Turris V, Salerno D, Garofalo S, St-Pierre MK, Carrier M, Renzi M, Pagani F, Modi B, Raspa M, Scavizzi F, Gross CT, Marinelli S, Tremblay MÈ, Caprioli D, Maggi L, Limatola C, Di Angelantonio S, Ragozzino D. Glia. 2022 Jan;70(1):173-195. doi: 10.1002/glia.24101. Epub 2021 Oct 18. PMID: 34661306 4 New insight into the catalytic -dependent and -independent roles of METTL3 in sustaining aberrant translation in chronic myeloid leukemia. Ianniello Z, Sorci M, Ceci Ginistrelli L, Iaiza A, Marchioni M, Tito C, Capuano E, Masciarelli S, Ottone T, Attrotto C, Rizzo M, Franceschini L, de Pretis S, Voso MT, Pelizzola M, Fazi F, Fatica A. Cell Death Dis. 2021 Sep 24;12(10):870. doi: 10.1038/s41419-021-04169-7. PMID: 34561421 5 Microglia modulate hippocampal synaptic transmission and sleep duration along the light/dark cycle. Corsi G, Picard K, di Castro MA, Garofalo S, Tucci F, Chece G, Del Percio C, Golia MT, Raspa M, Scavizzi F, Decoeur F, Lauro C, Rigamonti M, Iannello F, Ragozzino DA, Russo E, Bernardini G, Nadjar A, Tremblay ME, Babiloni C, Maggi L, Limatola C. Glia. 2022 Jan;70(1):89-105. doi: 10.1002/glia.24090. Epub 2021 Sep 6. PMID: 34487590 Microglia, the brain's resident macrophages, actively contribute to the homeostasis of cerebral parenchyma by sensing neuronal activity and supporting synaptic remodeling and plasticity. ... 6 Type I Interferons in COVID-19 Pathogenesis. Palermo E, Di Carlo D, Sgarbanti M, Hiscott J. Biology (Basel). 2021 Aug 26;10(9):829. doi: 10.3390/biology10090829. PMID: 34571706 7 Ion Channels in Glioma Malignancy. Catacuzzeno L, Sforna L, Esposito V, Limatola C, Franciolini F. Rev Physiol Biochem Pharmacol. 2021;181:223-267. doi: 10.1007/112_2020_44. PMID: 32930879

8 Antibiotics Treatment Modulates Microglia-Synapses Interaction. Cordella F, Sanchini C, Rosito M, Ferrucci L, Pediconi N, Cortese B, Guerrieri F, Pascucci GR, Antonangeli F, Peruzzi G, Giubettini M, Basilico B, Pagani F, Grimaldi A, D'Alessandro G, Limatola C, Ragozzino D, Di Angelantonio S. Cells. 2021 Oct 4;10(10):2648. doi: 10.3390/cells10102648. PMID: 34685628 9 Inoculum effect of antimicrobial peptides. Loffredo MR, Savini F, Bobone S, Casciaro B, Franzyk H, Mangoni ML, Stella L. Proc Natl Acad Sci U S A. 2021 May 25;118(21):e2014364118. doi: 10.1073/pnas.2014364118. PMID: 34021080 10 Microglial Potassium Channels: From Homeostasis to Neurodegeneration. Cocozza G, Garofalo S, Capitani R, D'Alessandro G, Limatola C. Biomolecules. 2021 Nov 26;11(12):1774. doi: 10.3390/biom11121774. PMID: 34944418 Free PMC article. Review. Among these, recent studies highlight the pivotal role of the potassium channels in regulating microglial functions in physiological and pathological conditions such as Alzheimer's Disease, Parkinson's Disease, and Amyotrophic Lateral Sclerosis. ... 11 CD39 Regulation and Functions in T Cells. Timperi E, Barnaba V. Int J Mol Sci. 2021 Jul 28;22(15):8068. doi: 10.3390/ijms22158068. PMID: 34360833 12 Prevention of recurrent respiratory infections : Inter-society Consensus. Chiappini E, Santamaria F, Marseglia GL, Marchisio P, Galli L, Cutrera R, de Martino M, Antonini S, Becherucci P, Biasci P, Bortone B, Bottero S, Caldarelli V, Cardinale F, Gattinara GC, Ciarcià M, Ciofi D, D'Elios S, Di Mauro G, Doria M, Indinnimeo L, Lo Vecchio A, Macrì F, Mattina R, Miniello VL, Del Giudice MM, Morbin G, Motisi MA, Novelli A, Palamara AT, Panatta ML, Pasinato A, Peroni D, Perruccio K, Piacentini G, Pifferi M, Pignataro L, Sitzia E, Tersigni C, Torretta S, Trambusti I, Trippella G, Valentini D, Valentini S, Varricchio A, Verga MC, Vicini C, Zecca M, Villani A. Ital J Pediatr. 2021 Oct 25;47(1):211. doi: 10.1186/s13052-021-01150-0. PMID: 34696778 13 Editorial: TGF-β as a Key Regulator of NK and ILCs Development and Functions. Bottino C, Walzer T, Santoni A, Castriconi R. Front Immunol. 2021 Jan 19;11:631712. doi: 10.3389/fimmu.2020.631712. eCollection 2020. PMID: 33542726 14 Editorial: Cellular Stress and Inflammation: How the Immune System Drives Tissue Homeostasis. Antonangeli F, Grimsholm O, Rossi MN, Velotti F. Front Immunol. 2021 Mar 18;12:668876. doi: 10.3389/fimmu.2021.668876. eCollection 2021. PMID: 33828567 15 Naringenin is a powerful inhibitor of SARS-CoV-2 infection in vitro. Clementi N, Scagnolari C, D'Amore A, Palombi F, Criscuolo E, Frasca F, Pierangeli A, Mancini N, Antonelli G, Clementi M, Carpaneto A, Filippini A. Pharmacol Res. 2021 Jan;163:105255. doi: 10.1016/j.phrs.2020.105255. Epub 2020 Oct 20. PMID: 33096221

16 NK Cell Anti-Tumor Surveillance in a Myeloid Cell-Shaped Environment. Russo E, Laffranchi M, Tomaipitinca L, Del Prete A, Santoni A, Sozzani S, Bernardini G. Front Immunol. 2021 Dec 17;12:787116. doi: 10.3389/fimmu.2021.787116. eCollection 2021. PMID: 34975880 17 Neuro-Signals from Gut Microbiota: Perspectives for Brain Glioma. D'Alessandro G, Lauro C, Quaglio D, Ghirga F, Botta B, Trettel F, Limatola C. Cancers (Basel). 2021 Jun 4;13(11):2810. doi: 10.3390/cancers13112810. PMID: 34199968 18 Fighting HIV-1 Persistence: At the Crossroads of "Shoc-K and B-Lock". Acchioni C, Palermo E, Sandini S, Acchioni M, Hiscott J, Sgarbanti M. Pathogens. 2021 Nov 20;10(11):1517. doi: 10.3390/pathogens10111517. PMID: 34832672 19 Mutational and immunogenetic landscape of HCV-associated B-cell lymphoproliferative disorders. Defrancesco I, Visentini M, Zibellini S, Minafò YA, Rattotti S, Ferretti VV, Rizzo E, Varettoni M, Frigeni M, Pulsoni A, Casato M, Colantuono S, Rossi M, Candido C, Zerbi C, Bergamini F, Cristinelli C, Fabbri N, Merli M, Zuccaro V, Bruno R, Paulli M, Arcaini L. Am J Hematol. 2021 Jun 1;96(6):E210-E214. doi: 10.1002/ajh.26167. Epub 2021 Apr 9. PMID: 33755245 20 New Insights into Curcumin- and Resveratrol-Mediated Anti-Cancer Effects. Arena A, Romeo MA, Benedetti R, Masuelli L, Bei R, Gilardini Montani MS, Cirone M. Pharmaceuticals (Basel). 2021 Oct 22;14(11):1068. doi: 10.3390/ph14111068. PMID: 34832850 21 Editorial: Control of Regulatory T Cell Stability, Plasticity, and Function in Health and Disease. Piconese S, Walker LSK, Dominguez-Villar M. Front Immunol. 2021 Jan 27;11:611591. doi: 10.3389/fimmu.2020.611591. eCollection 2020. PMID: 33584690 22 SARS-CoV-2-associated ssRNAs activate inflammation and immunity via TLR7/8. Salvi V, Nguyen HO, Sozio F, Schioppa T, Gaudenzi C, Laffranchi M, Scapini P, Passari M, Barbazza I, Tiberio L, Tamassia N, Garlanda C, Del Prete A, Cassatella MA, Mantovani A, Sozzani S, Bosisio D. JCI Insight. 2021 Sep 22;6(18):e150542. doi: 10.1172/jci.insight.150542. PMID: 34375313 23 The RNA editing enzyme ADAR2 restricts L1 mobility. Frassinelli L, Orecchini E, Al-Wardat S, Tripodi M, Mancone C, Doria M, Galardi S, Ciafrè SA, Michienzi A. RNA Biol. 2021 Oct 15;18(sup1):75-87. doi: 10.1080/15476286.2021.1940020. Epub 2021 Jul 5. PMID: 34224323 24 Reduced RNA turnover as a driver of cellular senescence. Mullani N, Porozhan Y, Mangelinck A, Rachez C, Costallat M, Batsché E, Goodhardt M, Cenci G, Mann C, Muchardt C. Life Sci Alliance. 2021 Jan 14;4(3):e202000809. doi: 10.26508/lsa.202000809. Print 2021 Mar. PMID: 33446491

25 Determinants of therapeutic lag in multiple sclerosis. Roos I, Leray E, Frascoli F, Casey R, Brown JWL, Horakova D, Havrdova EK, Debouverie M, Trojano M, Patti F, Izquierdo G, Eichau S, Edan G, Prat A, Girard M, Duquette P, Onofrj M, Lugaresi A, Grammond P, Ciron J, Ruet A, Ozakbas S, De Seze J, Louapre C, Zephir H, Sá MJ, Sola P, Ferraro D, Labauge P, Defer G, Bergamaschi R, Lebrun-Frenay C, Boz C, Cartechini E, Moreau T, Laplaud D, Lechner-Scott J, Grand'Maison F, Gerlach O, Terzi M, Granella F, Alroughani R, Iuliano G, Van Pesch V, Van Wijmeersch B, Spitaleri D, Soysal A, Berger E, Prevost J, Aguera-Morales E, McCombe P, Castillo Triviño T, Clavelou P, Pelletier J, Turkoglu R, Stankoff B, Gout O, Thouvenot E, Heinzlef O, Sidhom Y, Gouider R, Csepany T, Bourre B, Al Khedr A, Casez O, Cabre P, Montcuquet A, Wahab A, Camdessanche JP, Maurousset A, Patry I, Hankiewicz K, Pottier C, Maubeuge N, Labeyrie C, Nifle C, Coles A, Malpas CB, Vukusic S, Butzkueven H, Kalincik T. Mult Scler. 2021 Oct;27(12):1838-1851. doi: 10.1177/1352458520981300. Epub 2021 Jan 11. PMID: 33423618 26 Knowns and Unknowns of Vitamin B6 Metabolism in Escherichia coli. Tramonti A, Nardella C, di Salvo ML, Barile A, D'Alessio F, de Crécy-Lagard V, Contestabile R. EcoSal Plus. 2021 Apr;9(2):10.1128/ecosalplus.ESP-0004-2021. doi: 10.1128/ecosalplus.ESP-00042021. PMID: 33787481 27 RNA Flow Cytometry for the Study of T Cell Metabolism. Rossi A, Pacella I, Piconese S. Int J Mol Sci. 2021 Apr 9;22(8):3906. doi: 10.3390/ijms22083906. PMID: 33918901 28 First-in-Class Cyclic Temporin L Analogue: Design, Synthesis, and Antimicrobial Assessment. Bellavita R, Casciaro B, Di Maro S, Brancaccio D, Carotenuto A, Falanga A, Cappiello F, Buommino E, Galdiero S, Novellino E, Grossmann TN, Mangoni ML, Merlino F, Grieco P. J Med Chem. 2021 Aug 12;64(15):11675-11694. doi: 10.1021/acs.jmedchem.1c01033. Epub 2021 Jul 23. PMID: 34296619 29 Drug Delivery Systems for Hedgehog Inhibitors in the Treatment of SHH-Medulloblastoma. Caimano M, Lospinoso Severini L, Loricchio E, Infante P, Di Marcotullio L. Front Chem. 2021 Jun 7;9:688108. doi: 10.3389/fchem.2021.688108. eCollection 2021. PMID: 34164380 30 CCRL2 Modulates Physiological and Pathological Angiogenesis During Retinal Development. Ben Dhaou C, Del Prete A, Sozzani S, Parmentier M. Front Cell Dev Biol. 2021 Dec 23;9:808455. doi: 10.3389/fcell.2021.808455. eCollection 2021. PMID: 35004698 31 The innovative potential of selenium-containing agents for fighting cancer and viral infections. Ali W, Benedetti R, Handzlik J, Zwergel C, Battistelli C. Drug Discov Today. 2021 Jan;26(1):256263. doi: 10.1016/j.drudis.2020.10.014. Epub 2020 Oct 23. PMID: 33164821 32 Editorial: The RNA Revolution in Embryonic Development and Cell Differentiation in Health and Disease. Rosa A, Ciaudo C, Sumazin P, Fazi F. Front Cell Dev Biol. 2021 Sep 14;9:715341. doi: 10.3389/fcell.2021.715341. eCollection 2021. PMID: 34595170

33 Mechanisms of Peritoneal Fibrosis: Focus on Immune Cells-Peritoneal Stroma Interactions. Terri M, Trionfetti F, Montaldo C, Cordani M, Tripodi M, Lopez-Cabrera M, Strippoli R. Front Immunol. 2021 Mar 29;12:607204. doi: 10.3389/fimmu.2021.607204. eCollection 2021. PMID: 33854496 34 METTL3-dependent MALAT1 delocalization drives c-Myc induction in thymic epithelial tumors. Iaiza A, Tito C, Ianniello Z, Ganci F, Laquintana V, Gallo E, Sacconi A, Masciarelli S, De Angelis L, Aversa S, Diso D, Anile M, Petrozza V, Facciolo F, Melis E, Pescarmona E, Venuta F, Marino M, Blandino G, Fontemaggi G, Fatica A, Fazi F. Clin Epigenetics. 2021 Sep 16;13(1):173. doi: 10.1186/s13148-021-01159-6. PMID: 34530916 35 BK Polyomavirus Activates HSF1 Stimulating Human Kidney Hek293 Cell Proliferation. Baldelli S, Limongi D, Coni C, Ciccarone F, Ciotti M, Checconi P, Palamara AT, Ciriolo MR. Oxid Med Cell Longev. 2021 Nov 20;2021:9176993. doi: 10.1155/2021/9176993. eCollection 2021. PMID: 34845419 36 Gram-Negative Bacteria Holding Together in a Biofilm: The Acinetobacter baumannii Way. Pompilio A, Scribano D, Sarshar M, Di Bonaventura G, Palamara AT, Ambrosi C. Microorganisms. 2021 Jun 22;9(7):1353. doi: 10.3390/microorganisms9071353. PMID: 34206680 37 m6A RNA methylation and beyond - The epigenetic machinery and potential treatment options. Garbo S, Zwergel C, Battistelli C. Drug Discov Today. 2021 Nov;26(11):2559-2574. doi: 10.1016/j.drudis.2021.06.004. Epub 2021 Jun 12. PMID: 34126238 38 Measuring and Modelling Nonlinear Elasticity of Ex Vivo Mouse Muscles. Rizzuto E, De Luca R, Musarò A, Del Prete Z. J Healthc Eng. 2021 Nov 17;2021:5579232. doi: 10.1155/2021/5579232. eCollection 2021. PMID: 34840699 Free PMC article. Elastography is a noninvasive imaging technique that provides information on soft tissue stiffness. Young's modulus is typically used to characterize soft tissues' response to the applied force, as soft tissues are often considered linear elastic, isotropic, and quasi-inco … 39 Multi-Dimensional Gene Regulation in Innate and Adaptive Lymphocytes: A View From Regulomes. Fernando N, Sciumè G, O'Shea JJ, Shih HY. Front Immunol. 2021 Mar 25;12:655590. doi: 10.3389/fimmu.2021.655590. eCollection 2021. PMID: 33841440. 40 Histone-deacetylase 8 drives the immune response and the growth of glioma. Mormino A, Cocozza G, Fontemaggi G, Valente S, Esposito V, Santoro A, Bernardini G, Santoni A, Fazi F, Mai A, Limatola C, Garofalo S. Glia. 2021 Nov;69(11):2682-2698. doi: 10.1002/glia.24065. Epub 2021 Jul 26. PMID: 34310727. 41 Importance of amino acids in brain parenchyma invasion by cancer cells. Paone A, Bouzidi A, Rinaldo S, Giardina G, Cutruzzolà F. Oncoscience. 2021 Mar 31;8:47-49. doi: 10.18632/oncoscience.530. eCollection 2021. PMID: 33884287

42 Design and Synthesis of New Withaferin A Inspired Hedgehog Pathway Inhibitors. Bonandi E, Mori M, Infante P, Basili I, Di Marcotullio L, Calcaterra A, Catti F, Botta B, Passarella D. Chemistry. 2021 Jun 4;27(32):8350-8357. doi: 10.1002/chem.202100315. Epub 2021 May 4. PMID: 33811701 43 Laccase-Catalyzed 1,4-Dioxane-Mediated Synthesis of Belladine N-Oxides with Anti-Influenza A Virus Activity. Zippilli C, Botta L, Bizzarri BM, Nencioni L, De Angelis M, Protto V, Giorgi G, Baratto MC, Pogni R, Saladino R. Int J Mol Sci. 2021 Jan 29;22(3):1337. doi: 10.3390/ijms22031337. PMID: 33572794 44 Stress Responses in Down Syndrome Neurodegeneration: State of the Art and Therapeutic Molecules. Lanzillotta C, Di Domenico F. Biomolecules. 2021 Feb 11;11(2):266. doi: 10.3390/biom11020266. PMID: 33670211 Down syndrome (DS) is the most common genomic disorder characterized by the increased incidence of developing early Alzheimer's disease (AD). In DS, the triplication of genes on chromosome 21 is intimately associated with the increase of AD pathological hallmarks and with … 45 NK cell and ILC heterogeneity in colorectal cancer. New perspectives from high dimensional data. Pietropaolo G, Scarno G, Stabile H, Grimaldi A, Gismondi A, Santoni A, Sciumè G. Mol Aspects Med. 2021 Aug;80:100967. doi: 10.1016/j.mam.2021.100967. Epub 2021 May 1. PMID: 33941383 46 ER Stress, UPR Activation and the Inflammatory Response to Viral Infection. Cirone M. Viruses. 2021 Apr 29;13(5):798. doi: 10.3390/v13050798. PMID: 33946891 47 Age-Related Alterations at Neuromuscular Junction: Role of Oxidative Stress and Epigenetic Modifications. Dobrowolny G, Barbiera A, Sica G, Scicchitano BM. Cells. 2021 May 24;10(6):1307. doi: 10.3390/cells10061307. PMID: 34074012 48 The Emerging Role of Amino Acids of the Brain Microenvironment in the Process of Metastasis Formation. Cutruzzolà F, Bouzidi A, Liberati FR, Spizzichino S, Boumis G, Macone A, Rinaldo S, Giardina G, Paone A. Cancers (Basel). 2021 Jun 9;13(12):2891. doi: 10.3390/cancers13122891. PMID: 34207731 49 Harnessing the Activation of RIG-I Like Receptors to Inhibit Glioblastoma Tumorigenesis. Bufalieri F, Basili I, Di Marcotullio L, Infante P. Front Mol Neurosci. 2021 Jul 8;14:710171. doi: 10.3389/fnmol.2021.710171. eCollection 2021. PMID: 34305530 50 Circulating myomiRs in Muscle Denervation: From Surgical to ALS Pathological Condition. Casola I, Scicchitano BM, Lepore E, Mandillo S, Golini E, Nicoletti C, Barberi L, Dobrowolny G, Musarò A. Cells. 2021 Aug 10;10(8):2043. doi: 10.3390/cells10082043. PMID: 34440812

51 Fibrogenic signals persist in DAA-treated HCV patients after sustained virological response. Montaldo C, Terri M, Riccioni V, Battistelli C, Bordoni V, D'Offizi G, Prado MG, Trionfetti F, Vescovo T, Tartaglia E, Strippoli R, Agrati C, Tripodi M. J Hepatol. 2021 Dec;75(6):1301-1311. doi: 10.1016/j.jhep.2021.07.003. Epub 2021 Jul 13. PMID: 34271004 52 Anti-influenza A virus activity and structure-activity relationship of a series of nitrobenzoxadiazole derivatives. Fiorentino F, De Angelis M, Menna M, Rovere A, Caccuri AM, D'Acunzo F, Palamara AT, Nencioni L, Rotili D, Mai A. J Enzyme Inhib Med Chem. 2021 Dec;36(1):2128-2138. doi: 10.1080/14756366.2021.1982932. PMID: 34583607 53 Studying GGDEF Domain in the Act: Minimize Conformational Frustration to Prevent Artefacts. Mantoni F, Scribani Rossi C, Paiardini A, Di Matteo A, Cappellacci L, Petrelli R, Ricciutelli M, Paone A, Cutruzzolà F, Giardina G, Rinaldo S. Life (Basel). 2021 Jan 6;11(1):31. doi: 10.3390/life11010031. PMID: 33418960 54 Editorial: Secondary Metabolites and Peptides as Unique Natural Reservoirs of New Therapeutic Leads for Treatment of Cancer and Microbial Infections. Mangoni ML, Bhunia A, Botta B, Ghirga F. Front Chem. 2021 Aug 12;9:748180. doi: 10.3389/fchem.2021.748180. eCollection 2021. PMID: 34458241 55 A Novel, Easy Assay Method for Human Cysteine Sulfinic Acid Decarboxylase. Tramonti A, Contestabile R, Florio R, Nardella C, Barile A, Di Salvo ML. Life (Basel). 2021 May 14;11(5):438. doi: 10.3390/life11050438. PMID: 34068845 56 Intracellular Redox-Modulated Pathways as Targets for Effective Approaches in the Treatment of Viral Infection. Fraternale A, Zara C, De Angelis M, Nencioni L, Palamara AT, Retini M, Di Mambro T, Magnani M, Crinelli R. Int J Mol Sci. 2021 Mar 30;22(7):3603. doi: 10.3390/ijms22073603. PMID: 33808471 57 KI and WU Polyomavirus in Respiratory Samples of SARS-CoV-2 Infected Patients. Prezioso C, Moens U, Oliveto G, Brazzini G, Piacentini F, Frasca F, Viscido A, Scordio M, Guerrizio G, Rodio DM, Pierangeli A, d'Ettorre G, Turriziani O, Antonelli G, Scagnolari C, Pietropaolo V. Microorganisms. 2021 Jun 9;9(6):1259. doi: 10.3390/microorganisms9061259. PMID: 34207902 58 CD8+ T cells specific for cryptic apoptosis-associated epitopes exacerbate experimental autoimmune encephalomyelitis. Feizi N, Focaccetti C, Pacella I, Tucci G, Rossi A, Costanza M, Pedotti R, Sidney J, Sette A, La Rocca C, Procaccini C, Matarese G, Barnaba V, Piconese S. Cell Death Dis. 2021 Oct 29;12(11):1026. doi: 10.1038/s41419-021-04310-6. PMID: 34716313

59 ALS-related FUS mutations alter axon growth in motoneurons and affect HuD/ELAVL4 and FMRP activity. Garone MG, Birsa N, Rosito M, Salaris F, Mochi M, de Turris V, Nair RR, Cunningham TJ, Fisher EMC, Morlando M, Fratta P, Rosa A. Commun Biol. 2021 Sep 1;4(1):1025. doi: 10.1038/s42003-021-02538-8. PMID: 34471224 60 Circulating microRNAs from the Molecular Mechanisms to Clinical Biomarkers: A Focus on the Clear Cell Renal Cell Carcinoma. Tito C, De Falco E, Rosa P, Iaiza A, Fazi F, Petrozza V, Calogero A. Genes (Basel). 2021 Jul 28;12(8):1154. doi: 10.3390/genes12081154. PMID: 34440329 61 Potential IFNγ Modulation of Inflammasome Pathway in Chlamydia trachomatis Infected Synovial Cells. Filardo S, Di Pietro M, Frasca F, Diaco F, Scordio M, Antonelli G, Scagnolari C, Sessa R. Life (Basel). 2021 Dec 7;11(12):1359. doi: 10.3390/life11121359. PMID: 34947890 62 Emerging Single-Cell Technological Approaches to Investigate Chromatin Dynamics and Centromere Regulation in Human Health and Disease. Leo L, Colonna Romano N. Int J Mol Sci. 2021 Aug 16;22(16):8809. doi: 10.3390/ijms22168809. PMID: 34445507 63 Regulation of Ribosome Function by RNA Modifications in Hematopoietic Development and Leukemia: It Is Not Only a Matter of m6A. Fazi F, Fatica A. Int J Mol Sci. 2021 Apr 30;22(9):4755. doi: 10.3390/ijms22094755. PMID: 33946178 64 The Dysregulation of OGT/OGA Cycle Mediates Tau and APP Neuropathology in Down Syndrome. Zuliani I, Lanzillotta C, Tramutola A, Francioso A, Pagnotta S, Barone E, Perluigi M, Di Domenico F. Neurotherapeutics. 2021 Jan;18(1):340-363. doi: 10.1007/s13311-020-00978-4. Epub 2020 Nov 30. PMID: 33258073 Protein O-GlcNAcylation is a nutrient-related post-translational modification that, since its discovery some 30 years ago, has been associated with the development of neurodegenerative diseases. As reported in Alzheimer's disease (AD), flaws in the cerebral glucose uptake … 65 The role of RNA-binding and ribosomal proteins as specific RNA translation regulators in cellular differentiation and carcinogenesis. Ceci M, Fazi F, Romano N. Biochim Biophys Acta Mol Basis Dis. 2021 Apr 1;1867(4):166046. doi: 10.1016/j.bbadis.2020.166046. Epub 2020 Dec 28. PMID: 33383105 66 Engineered extracellular vesicle decoy receptor-mediated modulation of the IL6 trans-signalling pathway in muscle.

Conceição M, Forcina L, Wiklander OPB, Gupta D, Nordin JZ, Vrellaku B, McClorey G, Mäger I, Gӧrgens A, Lundin P, Musarò A, Wood MJA, Andaloussi SE, Roberts TC. Biomaterials. 2021 Jan;266:120435. doi: 10.1016/j.biomaterials.2020.120435. Epub 2020 Oct 5. PMID: 33049461 67 Chronic PERK induction promotes Alzheimer-like neuropathology in Down syndrome: Insights for therapeutic intervention. Lanzillotta C, Zuliani I, Tramutola A, Barone E, Blarzino C, Folgiero V, Caforio M, Valentini D, Villani A, Locatelli F, Butterfield DA, Head E, Perluigi M, Abisambra JF, Di Domenico F. Prog Neurobiol. 2021 Jan;196:101892. doi: 10.1016/j.pneurobio.2020.101892. Epub 2020 Aug 11. PMID: 32795489 68 Granzyme A and CD160 expression delineates ILC1 with graded functions in the mouse liver. Di Censo C, Marotel M, Mattiola I, Müller L, Scarno G, Pietropaolo G, Peruzzi G, Laffranchi M, Mazej J, Hasim MS, Asif S, Russo E, Tomaipitinca L, Stabile H, Lee SH, Vian L, Gadina M, Gismondi A, Shih HY, Mikami Y, Capuano C, Bernardini G, Bonelli M, Sozzani S, Diefenbach A, Ardolino M, Santoni A, Sciumè G. Eur J Immunol. 2021 Nov;51(11):2568-2575. doi: 10.1002/eji.202149209. Epub 2021 Aug 19. PMID: 34347289 69 Expression of CCRL2 Inhibits Tumor Growth by Concentrating Chemerin and Inhibiting Neoangiogenesis. Al Delbany D, Robert V, Dubois-Vedrenne I, Del Prete A, Vernimmen M, Radi A, Lefort A, Libert F, Wittamer V, Sozzani S, Parmentier M. Cancers (Basel). 2021 Oct 5;13(19):5000. doi: 10.3390/cancers13195000. PMID: 34638484 70 Chimeric Stimuli-Responsive Liposomes as Nanocarriers for the Delivery of the Anti-Glioma Agent TRAM-34. Naziris N, Pippa N, Sereti E, Chrysostomou V, Kędzierska M, Kajdanek J, Ionov M, Miłowska K, Balcerzak Ł, Garofalo S, Limatola C, Pispas S, Dimas K, Bryszewska M, Demetzos C. Int J Mol Sci. 2021 Jun 10;22(12):6271. doi: 10.3390/ijms22126271. PMID: 34200955 71 New insights into the evolution of human Y chromosome palindromes through mutation and gene conversion. Bonito M, D'Atanasio E, Ravasini F, Cariati S, Finocchio A, Novelletto A, Trombetta B, Cruciani F. Hum Mol Genet. 2021 Nov 16;30(23):2272-2285. doi: 10.1093/hmg/ddab189. PMID: 34244762 72 Self-assembling ferritin-dendrimer nanoparticles for targeted delivery of nucleic acids to myeloid leukemia cells. Palombarini F, Masciarelli S, Incocciati A, Liccardo F, Di Fabio E, Iazzetti A, Fabrizi G, Fazi F, Macone A, Bonamore A, Boffi A. J Nanobiotechnology. 2021 Jun 9;19(1):172. doi: 10.1186/s12951021-00921-5. PMID: 34107976 73 Cancer cells dysregulate PI3K/AKT/mTOR pathway activation to ensure their survival and proliferation: mimicking them is a smart strategy of gammaherpesviruses. Cirone M. Crit Rev Biochem Mol Biol. 2021 Oct;56(5):500-509. doi: 10.1080/10409238.2021.1934811. Epub 2021 Jun 15. PMID: 34130564

74 Host - Bacterial Pathogen Communication: The Wily Role of the Multidrug Efflux Pumps of the MFS Family. Pasqua M, Bonaccorsi di Patti MC, Fanelli G, Utsumi R, Eguchi Y, Trirocco R, Prosseda G, Grossi M, Colonna B. Front Mol Biosci. 2021 Jul 26;8:723274. doi: 10.3389/fmolb.2021.723274. eCollection 2021. PMID: 34381818 75 Investigation of Commiphora myrrha (Nees) Engl. Oil and Its Main Components for Antiviral Activity. Madia VN, De Angelis M, De Vita D, Messore A, De Leo A, Ialongo D, Tudino V, Saccoliti F, De Chiara G, Garzoli S, Scipione L, Palamara AT, Di Santo R, Nencioni L, Costi R. Pharmaceuticals (Basel). 2021 Mar 9;14(3):243. doi: 10.3390/ph14030243. PMID: 33803165 76 SARS-CoV-2 Entry Genes Expression in Relation with Interferon Response in Cystic Fibrosis Patients. Bitossi C, Frasca F, Viscido A, Oliveto G, Scordio M, Belloni L, Cimino G, Pietropaolo V, Gentile M, d'Ettorre G, Midulla F, Trancassini M, Antonelli G, Pierangeli A, Scagnolari C. Microorganisms. 2021 Jan 3;9(1):93. doi: 10.3390/microorganisms9010093. PMID: 33401565 77 Antiviral Potential of the Antimicrobial Drug Atovaquone against SARS-CoV-2 and Emerging Variants of Concern. Carter-Timofte ME, Arulanandam R, Kurmasheva N, Fu K, Laroche G, Taha Z, van der Horst D, Cassin L, van der Sluis RM, Palermo E, Di Carlo D, Jacobs D, Maznyi G, Azad T, Singaravelu R, Ren F, Hansen AL, Idorn M, Holm CK, Jakobsen MR, van Grevenynghe J, Hiscott J, Paludan SR, Bell JC, Seguin J, Sabourin LA, Côté M, Diallo JS, Alain T, Olagnier D. ACS Infect Dis. 2021 Nov 12;7(11):3034-3051. doi: 10.1021/acsinfecdis.1c00278. Epub 2021 Oct 18. PMID: 34658235 78 Identification and characterization of the pyridoxal 5'-phosphate allosteric site in Escherichia coli pyridoxine 5'-phosphate oxidase. Barile A, Battista T, Fiorillo A, di Salvo ML, Malatesta F, Tramonti A, Ilari A, Contestabile R. J Biol Chem. 2021 Jan-Jun;296:100795. doi: 10.1016/j.jbc.2021.100795. Epub 2021 May 18. PMID: 34019876 79 Anticancer effect of AZD2461 PARP inhibitor against colon cancer cells carrying wt or dysfunctional p53. Romeo MA, Gilardini Montani MS, Benedetti R, Arena A, Maretto M, Bassetti E, Caiazzo R, D'Orazi G, Cirone M. Exp Cell Res. 2021 Nov 15;408(2):112879. doi: 10.1016/j.yexcr.2021.112879. Epub 2021 Oct 13. PMID: 34653407 80 Glabrescione B delivery by self-assembling micelles efficiently inhibits tumor growth in preclinical models of Hedgehog-dependent medulloblastoma. Infante P, Malfanti A, Quaglio D, Balducci S, De Martin S, Bufalieri F, Mastrotto F, Basili I, Garofalo M, Lospinoso Severini L, Mori M, Manni I, Moretti M, Nicoletti C, Piaggio G, Caliceti P, Botta B, Ghirga F, Salmaso S, Di Marcotullio L. Cancer Lett. 2021 Feb 28;499:220-231. doi: 10.1016/j.canlet.2020.11.028. Epub 2020 Nov 26. PMID: 33249196

81 The ATPase SRCAP is associated with the mitotic apparatus, uncovering novel molecular aspects of Floating-Harbor syndrome. Messina G, Prozzillo Y, Delle Monache F, Santopietro MV, Atterrato MT, Dimitri P. BMC Biol. 2021 Sep 2;19(1):184. doi: 10.1186/s12915-021-01109-x. PMID: 34474679 82 CXCR4 antagonism sensitizes cancer cells to novel indole-based MDM2/4 inhibitors in glioblastoma multiforme. Daniele S, La Pietra V, Piccarducci R, Pietrobono D, Cavallini C, D'Amore VM, Cerofolini L, Giuntini S, Russomanno P, Puxeddu M, Nalli M, Pedrini M, Fragai M, Luchinat C, Novellino E, Taliani S, La Regina G, Silvestri R, Martini C, Marinelli L. Eur J Pharmacol. 2021 Apr 15;897:173936. doi: 10.1016/j.ejphar.2021.173936. Epub 2021 Feb 10. PMID: 33581134 83 Seizure-mediated iron accumulation and dysregulated iron metabolism after status epilepticus and in temporal lobe epilepsy. Zimmer TS, David B, Broekaart DWM, Schidlowski M, Ruffolo G, Korotkov A, van der Wel NN, van Rijen PC, Mühlebner A, van Hecke W, Baayen JC, Idema S, François L, van Eyll J, Dedeurwaerdere S, Kessels HW, Surges R, Rüber T, Gorter JA, Mills JD, van Vliet EA, Aronica E. Acta Neuropathol. 2021 Oct;142(4):729-759. doi: 10.1007/s00401-021-02348-6. Epub 2021 Jul 22. PMID: 34292399 84 Ca2+ -dependent release of ATP from astrocytes affects herpes simplex virus type 1 infection of neurons. Li Puma DD, Marcocci ME, Lazzarino G, De Chiara G, Tavazzi B, Palamara AT, Piacentini R, Grassi C. Glia. 2021 Jan;69(1):201-215. doi: 10.1002/glia.23895. Epub 2020 Aug 20. PMID: 32818313 85 Risk Assessment of Progressive Multifocal Leukoencephalopathy in Multiple Sclerosis Patients during 1 Year of Ocrelizumab Treatment. Prezioso C, Grimaldi A, Landi D, Nicoletti CG, Brazzini G, Piacentini F, Passerini S, Limongi D, Ciotti M, Palamara AT, Marfia GA, Pietropaolo V. Viruses. 2021 Aug 25;13(9):1684. doi: 10.3390/v13091684. PMID: 34578264 CONCLUSIONS: Ocrelizumab in JCPyV-DNA positive patients is safe and did not determine PML cases. Combined monitoring of ocrelizumab's effects on JCPyV pathogenicity and on host immunity might offer a complete insight towards predicting PML risk.... 86 Beneficial effects of a combination of natural product activators of autophagy on endothelial cells and platelets. Carnevale R, Nocella C, Schiavon S, Cammisotto V, Cotugno M, Forte M, Valenti V, Marchitti S, Vecchio D, Biondi Zoccai G, Rubattu S, Martinelli O, Pignatelli P, Violi F, Volpe M, Versaci F, Frati L, Frati G, Sciarretta S. Br J Pharmacol. 2021 May;178(10):2146-2159. doi: 10.1111/bph.15399. Epub 2021 Apr 24. PMID: 33512008 87 T Cell Memory in Infection, Cancer, and Autoimmunity. Barnaba V. Front Immunol. 2022 Jan 3;12:811968. doi: 10.3389/fimmu.2021.811968. eCollection 2021. PMID: 35069600

88 Role of UPR Sensor Activation in Cell Death-Survival Decision of Colon Cancer Cells Stressed by DPE Treatment. Benedetti R, Gilardini Montani MS, Romeo MA, Arena A, Santarelli R, D'Orazi G, Cirone M. Biomedicines. 2021 Sep 18;9(9):1262. doi: 10.3390/biomedicines9091262. PMID: 34572447 89 Chronic cancer and non-cancer pain and opioid-induced hyperalgesia share common mechanisms: neuroinflammation and central sensitization. Santoni A, Mercadante S, Arcuri E. Minerva Anestesiol. 2021 Feb;87(2):210-222. doi: 10.23736/S0375-9393.20.14822-3. Epub 2020 Dec 10. PMID: 33300326 90 Differential induction of type I and III interferon genes in the upper respiratory tract of patients with coronavirus disease 2019 (COVID-19). Scagnolari C, Pierangeli A, Frasca F, Bitossi C, Viscido A, Oliveto G, Scordio M, Mazzuti L, Di Carlo D, Gentile M, Solimini A, Ceccarelli G, Pugliese F, Mastroianni CM, d'Ettorre G, Turriziani O, Antonelli G; Virology and Infectious Diseases COVID-19 Study Group of Sapienza University. Virus Res. 2021 Apr 2;295:198283. doi: 10.1016/j.virusres.2020.198283. Epub 2021 Jan 5. PMID: 33418027 91 Proprotein Convertase Subtilisin Kexin Type 9 Inhibitors Reduce Platelet Activation Modulating ox-LDL Pathways. Cammisotto V, Baratta F, Castellani V, Bartimoccia S, Nocella C, D'Erasmo L, Cocomello N, Barale C, Scicali R, Di Pino A, Piro S, Del Ben M, Arca M, Russo I, Purrello F, Carnevale R, Violi F, Pastori D, Pignatelli P. Int J Mol Sci. 2021 Jul 3;22(13):7193. doi: 10.3390/ijms22137193. PMID: 34281247 92 Sustained Systemic Levels of IL-6 Impinge Early Muscle Growth and Induce Muscle Atrophy and Wasting in Adulthood. Pelosi L, Berardinelli MG, Forcina L, Ascenzi F, Rizzuto E, Sandri M, De Benedetti F, Scicchitano BM, Musarò A. Cells. 2021 Jul 18;10(7):1816. doi: 10.3390/cells10071816. PMID: 34359985 93 IRE1 Alpha/XBP1 Axis Sustains Primary Effusion Lymphoma Cell Survival by Promoting Cytokine Release and STAT3 Activation. Gonnella R, Gilardini Montani MS, Guttieri L, Romeo MA, Santarelli R, Cirone M. Biomedicines. 2021 Jan 27;9(2):118. doi: 10.3390/biomedicines9020118. PMID: 33513694 Free PMC article. Primary Effusion Lymphoma (PEL) is a highly aggressive B cell lymphoma associated with Kaposi's Sarcoma-associated Herpesvirus (KSHV). It is characterized by a high level of basal Endoplasmic Reticulum (ER) stress, Unfolded Protein Response (UPR) activation and constitutiv … 94 A New Smoothened Antagonist Bearing the Purine Scaffold Shows Antitumour Activity In Vitro and In Vivo. Zárate AM, Espinosa-Bustos C, Guerrero S, Fierro A, Oyarzún-Ampuero F, Quest AFG, Di Marcotullio L, Loricchio E, Caimano M, Calcaterra A, González-Quiroz M, Aguirre A, Meléndez J, Salas CO. Int J Mol Sci. 2021 Aug 4;22(16):8372. doi: 10.3390/ijms22168372. PMID: 34445078

95 Modulation of OMV Production by the Lysis Module of the DLP12 Defective Prophage of Escherichia coli K12. Pasqua M, Zennaro A, Trirocco R, Fanelli G, Micheli G, Grossi M, Colonna B, Prosseda G. Microorganisms. 2021 Feb 12;9(2):369. doi: 10.3390/microorganisms9020369. PMID: 33673345 96 Design and Functional Validation of a Mutant Variant of the LncRNA HOTAIR to Counteract Snail Function in Epithelial-to-Mesenchymal Transition. Battistelli C, Garbo S, Riccioni V, Montaldo C, Santangelo L, Vandelli A, Strippoli R, Tartaglia GG, Tripodi M, Cicchini C. Cancer Res. 2021 Jan 1;81(1):103-113. doi: 10.1158/0008-5472.CAN20-1764. Epub 2020 Nov 6. PMID: 33158813 97 CD8+ T cells specific to apoptosis-associated epitopes are expanded in patients with chronic HBV infection and fibrosis. Pacella I, Cammarata I, Martire C, Brancaccio G, Gaeta GB, Barnaba V, Piconese S. Liver Int. 2021 Mar;41(3):470-481. doi: 10.1111/liv.14720. Epub 2020 Nov 25. PMID: 33159402 98 Enriched Environment Cues Suggest a New Strategy to Counteract Glioma: Engineered rAAV2IL-15 Microglia Modulate the Tumor Microenvironment. Mormino A, Bernardini G, Cocozza G, Corbi N, Passananti C, Santoni A, Limatola C, Garofalo S. Front Immunol. 2021 Sep 6;12:730128. doi: 10.3389/fimmu.2021.730128. eCollection 2021. PMID: 34552593 99 Fenretinide Beneficial Effects on Amyotrophic Lateral Sclerosis-associated SOD1G93A Mutant Protein Toxicity: In Vitro and In Vivo Evidences. Orienti I, Armida M, Dobrowolny G, Pepponi R, Sollazzini G, Pezzola A, Casola I, Musarò A, Popoli P, Potenza RL. Neuroscience. 2021 Oct 1;473:1-12. doi: 10.1016/j.neuroscience.2021.07.033. Epub 2021 Aug 5. PMID: 34363869 100 Essential oils from Artemisia species inhibit biofilm formation and the virulence of Escherichia coli EPEC 2348/69. Mathlouthi A, Saadaoui N, Pennacchietti E, De Biase D, Ben-Attia M. Biofouling. 2021 Feb;37(2):174-183. doi: 10.1080/08927014.2021.1886278. Epub 2021 Feb 15. PMID: 33588649 101 ACE2 expression is related to the interferon response in airway epithelial cells but is that functional for SARS-CoV-2 entry? Scagnolari C, Bitossi C, Viscido A, Frasca F, Oliveto G, Scordio M, Petrarca L, Mancino E, Nenna R, Riva E, De Vito C, Midulla F, Antonelli G, Pierangeli A. Cytokine. 2021 Apr;140:155430. doi: 10.1016/j.cyto.2021.155430. Epub 2021 Jan 15. PMID: 33508651 102 Immunohistopathological response against anisakid nematode larvae and a coccidian in Micromesistius poutassou from NE Atlantic waters. Sayyaf Dezfuli B, Simoni E, Bosi G, Palomba M, Mattiucci S, Giulietti L, Bao M, Levsen A, Cipriani P. J Helminthol. 2021 Mar 10;95:e14. doi: 10.1017/S0022149X20000942. PMID: 33750484

103 Hypoxia Enhances the Expression of RNASET2 in Human Monocyte-Derived Dendritic Cells: Role of PI3K/AKT Pathway. Monaci S, Coppola F, Giuntini G, Roncoroni R, Acquati F, Sozzani S, Carraro F, Naldini A. Int J Mol Sci. 2021 Jul 15;22(14):7564. doi: 10.3390/ijms22147564. PMID: 34299186 104 System-oriented optimization of multi-target 2,6-diaminopurine derivatives: Easily accessible broad-spectrum antivirals active against flaviviruses, influenza virus and SARS-CoV-2. Vicenti I, Martina MG, Boccuto A, De Angelis M, Giavarini G, Dragoni F, Marchi S, Trombetta CM, Crespan E, Maga G, Eydoux C, Decroly E, Montomoli E, Nencioni L, Zazzi M, Radi M. Eur J Med Chem. 2021 Nov 15;224:113683. doi: 10.1016/j.ejmech.2021.113683. Epub 2021 Jul 5. PMID: 34273661 105 Control of replication stress and mitosis in colorectal cancer stem cells through the interplay of PARP1, MRE11 and RAD51. Manic G, Musella M, Corradi F, Sistigu A, Vitale S, Soliman Abdel Rehim S, Mattiello L, Malacaria E, Galassi C, Signore M, Pallocca M, Scalera S, Goeman F, De Nicola F, Guarracino A, Pennisi R, Antonangeli F, Sperati F, Baiocchi M, Biffoni M, Fanciulli M, Maugeri-Saccà M, Franchitto A, Pichierri P, De Maria R, Vitale I. Cell Death Differ. 2021 Jul;28(7):2060-2082. doi: 10.1038/s41418-020-00733-4. Epub 2021 Feb 2. PMID: 33531658 106 The cross-talk between STAT1/STAT3 and ROS up-regulates PD-L1 and promotes the release of pro-inflammatory/immune suppressive cytokines in primary monocytes infected by HHV-6B. Romeo MA, Gilardini Montani MS, Benedetti R, Giambelli L, D'Aprile R, Gaeta A, Faggioni A, Cirone M. Virus Res. 2021 Jan 15;292:198231. doi: 10.1016/j.virusres.2020.198231. Epub 2020 Nov 15. PMID: 33207265 107 Bartonella henselae Persistence within Mesenchymal Stromal Cells Enhances Endothelial Cell Activation and Infectibility That Amplifies the Angiogenic Process. Scutera S, Mitola S, Sparti R, Salvi V, Grillo E, Piersigilli G, Bugatti M, Alotto D, Schioppa T, Sozzani S, Musso T. Infect Immun. 2021 Jul 15;89(8):e0014121. doi: 10.1128/IAI.00141-21. Epub 2021 Jul 15. PMID: 34031126 108 PGE2 Released by Pancreatic Cancer Cells Undergoing ER Stress Transfers the Stress to DCs Impairing Their Immune Function. Gilardini Montani MS, Benedetti R, Piconese S, Pulcinelli FM, Timperio AM, Romeo MA, Masuelli L, Mattei M, Bei R, D'Orazi G, Cirone M. Mol Cancer Ther. 2021 May;20(5):934-945. doi: 10.1158/1535-7163.MCT-20-0699. Epub 2021 Feb 25. PMID: 33632872 109 In Vivo Silencing of Genes Coding for dTip60 Chromatin Remodeling Complex Subunits Affects Polytene Chromosome Organization and Proper Development in Drosophila melanogaster. Prozzillo Y, Cuticone S, Ferreri D, Fattorini G, Messina G, Dimitri P. Int J Mol Sci. 2021 Apr 26;22(9):4525. doi: 10.3390/ijms22094525. PMID: 33926075

110 High-Fat Diet Leads to Reduced Protein O-GlcNAcylation and Mitochondrial Defects Promoting the Development of Alzheimer's Disease Signatures. Zuliani I, Lanzillotta C, Tramutola A, Barone E, Perluigi M, Rinaldo S, Paone A, Cutruzzolà F, Bellanti F, Spinelli M, Natale F, Fusco S, Grassi C, Di Domenico F. Int J Mol Sci. 2021 Apr 3;22(7):3746. doi: 10.3390/ijms22073746. PMID: 33916835 111 p53-R273H Sustains ROS, Pro-Inflammatory Cytokine Release and mTOR Activation While Reducing Autophagy, Mitophagy and UCP2 Expression, Effects Prevented by wtp53. Romeo MA, Gilardini Montani MS, Benedetti R, Arena A, D'Orazi G, Cirone M. Biomolecules. 2021 Feb 24;11(3):344. doi: 10.3390/biom11030344. PMID: 33668399 112 Microglia-Derived Small Extracellular Vesicles Reduce Glioma Growth by Modifying Tumor Cell Metabolism and Enhancing Glutamate Clearance through miR-124. Serpe C, Monaco L, Relucenti M, Iovino L, Familiari P, Scavizzi F, Raspa M, Familiari G, Civiero L, D'Agnano I, Limatola C, Catalano M. Cells. 2021 Aug 12;10(8):2066. doi: 10.3390/cells10082066. PMID: 34440835 113 Lovastatin reduces PEL cell survival by phosphorylating ERK1/2 that blocks the autophagic flux and engages a cross-talk with p53 to activate p21. Santarelli R, Pompili C, Gilardini Montani MS, Romeo MA, Gonnella R, D'Orazi G, Cirone M. IUBMB Life. 2021 Jul;73(7):968-977. doi: 10.1002/iub.2503. Epub 2021 May 21. PMID: 33987937 114 Alteration of type I interferon response is associated with subclinical atherosclerosis in virologically suppressed HIV-1-infected male patients. Santinelli L, De Girolamo G, Borrazzo C, Vassalini P, Pinacchio C, Cavallari EN, Statzu M, Frasca F, Scordio M, Bitossi C, Viscido A, Ceccarelli G, Mancone M, Mastroianni CM, Antonelli G, d'Ettorre G, Scagnolari C. J Med Virol. 2021 Aug;93(8):4930-4938. doi: 10.1002/jmv.27028. Epub 2021 May 3. PMID: 33913525 115 Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28Mediated Inflammatory Signals in the Absence of MHC Class II Molecules. Kunkl M, Amormino C, Caristi S, Tedeschi V, Fiorillo MT, Levy R, Popugailo A, Kaempfer R, Tuosto L. Front Immunol. 2021 Aug 13;12:723689. doi: 10.3389/fimmu.2021.723689. eCollection 2021. PMID: 34489975 116 Temporin G, an amphibian antimicrobial peptide against influenza and parainfluenza respiratory viruses: Insights into biological activity and mechanism of action. De Angelis M, Casciaro B, Genovese A, Brancaccio D, Marcocci ME, Novellino E, Carotenuto A, Palamara AT, Mangoni ML, Nencioni L. FASEB J. 2021 Feb;35(2):e21358. doi: 10.1096/fj.202001885RR. PMID: 33538061 117 SMA-miRs (miR-181a-5p, -324-5p, and -451a) are overexpressed in spinal muscular atrophy skeletal muscle and serum samples. Abiusi E, Infante P, Cagnoli C, Lospinoso Severini L, Pane M, Coratti G, Pera MC, D'Amico A, Diano F, Novelli A, Spartano S, Fiori S, Baranello G, Moroni I, Mora M, Pasanisi MB, Pocino K, Le Pera L, D'Amico D, Travaglini L, Ria F, Bruno C, Locatelli D, Bertini ES, Morandi LO,

Mercuri E, Di Marcotullio L, Tiziano FD. Elife. 2021 Sep 20;10:e68054. doi: 10.7554/eLife.68054. PMID: 34542403 118 Functional Inactivation of Drosophila GCK Orthologs Causes Genomic Instability and Oxidative Stress in a Fly Model of MODY-2. Mascolo E, Liguori F, Stufera Mecarelli L, Amoroso N, Merigliano C, Amadio S, Volonté C, Contestabile R, Tramonti A, Vernì F. Int J Mol Sci. 2021 Jan 18;22(2):918. doi: 10.3390/ijms22020918. PMID: 33477627 119 Recurrent Herpes Simplex Virus Type 1 (HSV-1) Infection Modulates Neuronal Aging Marks in In Vitro and In Vivo Models. Napoletani G, Protto V, Marcocci ME, Nencioni L, Palamara AT, De Chiara G. Int J Mol Sci. 2021 Jun 11;22(12):6279. doi: 10.3390/ijms22126279. PMID: 34208020 120 A longitudinal study defined circulating microRNAs as reliable biomarkers for disease prognosis and progression in ALS human patients. Dobrowolny G, Martone J, Lepore E, Casola I, Petrucci A, Inghilleri M, Morlando M, Colantoni A, Scicchitano BM, Calvo A, Bisogni G, Chiò A, Sabatelli M, Bozzoni I, Musarò A. Cell Death Discov. 2021 Jan 11;7(1):4. doi: 10.1038/s41420-020-00397-6. PMID: 33431881. 121 Esc peptides as novel potentiators of defective cystic fibrosis transmembrane conductance regulator: an unprecedented property of antimicrobial peptides. Ferrera L, Cappiello F, Loffredo MR, Puglisi E, Casciaro B, Botta B, Galietta LJV, Mori M, Mangoni ML. Cell Mol Life Sci. 2021 Dec 31;79(1):67. doi: 10.1007/s00018-021-04030-2. PMID: 34971429 122 Hepatitis C virus-associated indolent B-cell lymphomas: A review on the role of the new direct antiviral agents therapy. Mazzaro C, Dal Maso L, Visentini M, Ermacora A, Andreone P, Gattei V, Pozzato G. Hematol Oncol. 2021 Oct;39(4):439-447. doi: 10.1002/hon.2862. Epub 2021 Mar 26. PMID: 34477233 123 Antipseudomonal and Immunomodulatory Properties of Esc Peptides: Promising Features for Treatment of Chronic Infectious Diseases and Inflammation. Cappiello F, Carnicelli V, Casciaro B, Mangoni ML. Int J Mol Sci. 2021 Jan 8;22(2):557. doi: 10.3390/ijms22020557. PMID: 33429882 124 Novel genotyping approaches to easily detect genomic admixture between the major Afrotropical malaria vector species, Anopheles coluzzii and An. gambiae. Caputo B, Pichler V, Bottà G, De Marco C, Hubbart C, Perugini E, Pinto J, Rockett KA, Miles A, Della Torre A. Mol Ecol Resour. 2021 Jul;21(5):1504-1516. doi: 10.1111/1755-0998.13359. Epub 2021 Apr 1. PMID: 33590707

125 Sequence Read Depth Analysis of a Monophyletic Cluster of Y Chromosomes Characterized by Structural Rearrangements in the AZFc Region Resulting in DYS448 Deletion and DYF387S1 Duplication. Ravasini F, D'Atanasio E, Bonito M, Bonucci B, Della Rocca C, Berti A, Trombetta B, Cruciani F. Front Genet. 2021 Apr 16;12:669405. doi: 10.3389/fgene.2021.669405. eCollection 2021. PMID: 33936180 126 Genetic identification of Anisakis spp. (Nematoda: Anisakidae) from cetaceans of the Southwestern Atlantic Ocean: ecological and zoogeographical implications. Irigoitia MM, Palomba M, Braicovich PE, Lanfranchi AL, Denuncio PE, Gana JCM, Mattiucci S, Timi JT. Parasitol Res. 2021 May;120(5):1699-1711. doi: 10.1007/s00436-021-07088-w. Epub 2021 Mar 6. PMID: 33674924 127 Self or Non-Self? It Is also a Matter of RNA Recognition and Editing by ADAR1. Tassinari V, Cerboni C, Soriani A. Biology (Basel). 2022 Apr 8;11(4):568. doi: 10.3390/biology11040568. PMID: 35453767 128 Aminomalononitrile inspired prebiotic chemistry as a novel multicomponent tool for the synthesis of imidazole and purine derivatives with anti-influenza A virus activity. Bizzarri BM, Fanelli A, Botta L, De Angelis M, Palamara AT, Nencioni L, Saladino R. RSC Adv. 2021 Sep 8;11(48):30020-30029. doi: 10.1039/d1ra05240c. eCollection 2021 Sep 6. PMID: 35480240 129 The PDE4 Inhibitor Tanimilast Blunts Proinflammatory Dendritic Cell Activation by SARS-CoV2 ssRNAs. Nguyen HO, Schioppa T, Tiberio L, Facchinetti F, Villetti G, Civelli M, Del Prete A, Sozio F, Gaudenzi C, Passari M, Barbazza I, Sozzani S, Salvi V, Bosisio D. Front Immunol. 2022 Jan 24;12:797390. doi: 10.3389/fimmu.2021.797390. eCollection 2021. PMID: 35140709 130 Cytosolic localization and in vitro assembly of human de novo thymidylate synthesis complex. Spizzichino S, Boi D, Boumis G, Lucchi R, Liberati FR, Capelli D, Montanari R, Pochetti G, Piacentini R, Parisi G, Paone A, Rinaldo S, Contestabile R, Tramonti A, Paiardini A, Giardina G, Cutruzzolà F. FEBS J. 2022 Mar;289(6):1625-1649. doi: 10.1111/febs.16248. Epub 2021 Nov 12. PMID: 34694685 131 An overview on tumor treating fields (TTFields) technology as a new potential subsidiary biophysical treatment for COVID-19. Farmani AR, Mahdavinezhad F, Scagnolari C, Kouhestani M, Mohammadi S, Ai J, Shoormeij MH, Rezaei N. Drug Deliv Transl Res. 2022 Jul;12(7):1605-1615. doi: 10.1007/s13346-021-010675. Epub 2021 Sep 20. PMID: 34542840 132 Influenza Virus Down-Modulates G6PD Expression and Activity to Induce Oxidative Stress and Promote Its Replication. De Angelis M, Amatore D, Checconi P, Zevini A, Fraternale A, Magnani M, Hiscott J, De Chiara G, Palamara AT, Nencioni L. Front Cell Infect Microbiol. 2022 Jan 6;11:804976. doi: 10.3389/fcimb.2021.804976. eCollection 2021. PMID: 35071051

133 DNA damage triggers an interplay between wtp53 and c-Myc affecting lymphoma cell proliferation and Kaposi sarcoma herpesvirus replication. Arena A, Gilardini Montani MS, Romeo MA, Benedetti R, Gaeta A, Cirone M. Biochim Biophys Acta Mol Cell Res. 2022 Jan;1869(1):119168. doi: 10.1016/j.bbamcr.2021.119168. Epub 2021 Oct 30. PMID: 34728235 134 First COVID-19 lockdown resulted in most respiratory viruses disappearing among hospitalised children, with the exception of rhinoviruses. Nenna R, Matera L, Pierangeli A, Oliveto G, Viscido A, Petrarca L, La Regina DP, Mancino E, Di Mattia G, Villani A, Midulla F. Acta Paediatr. 2022 Jul;111(7):1399-1403. doi: 10.1111/apa.16326. Epub 2022 Mar 16. PMID: 35266576 135 Dietary and Protective Factors to Halt or Mitigate Progression of Autoimmunity, COVID-19 and Its Associated Metabolic Diseases. Ricordi C, Pacifici F, Lanzoni G, Palamara AT, Garaci E, Della-Morte D. Int J Mol Sci. 2021 Mar 19;22(6):3134. doi: 10.3390/ijms22063134. PMID: 33808574 136 Neuroserpin: structure, function, physiology and pathology. D'Acunto E, Fra A, Visentin C, Manno M, Ricagno S, Galliciotti G, Miranda E. Cell Mol Life Sci. 2021 Oct;78(19-20):6409-6430. doi: 10.1007/s00018-021-03907-6. Epub 2021 Aug 17. PMID: 34405255 137 The Regulatory Activity of Noncoding RNAs in ILCs. Grimaldi A, Pietropaolo G, Stabile H, Kosta A, Capuano C, Gismondi A, Santoni A, Sciumè G, Fionda C. Cells. 2021 Oct 14;10(10):2742. doi: 10.3390/cells10102742. PMID: 34685721 138 Function, Regulation, and Dysfunction of Intrinsically Disordered Proteins. Fusco G, Gianni S. Life (Basel). 2021 Feb 12;11(2):140. doi: 10.3390/life11020140. PMID: 33673214 139 Anticancer innovative therapy congress: Highlights from the 10th anniversary edition. De Santis F, Fucà G, Schadendorf D, Mantovani A, Magnani L, Lisanti M, Pettitt S, Bellone M, Del Sal G, Minucci S, Eggermont A, Bruzzi P, Bicciato S, Conte P, Noberini R, Hiscott J, De Braud F, Del Vecchio M, Di Nicola M. Cytokine Growth Factor Rev. 2021 Jun;59:1-8. doi: 10.1016/j.cytogfr.2021.02.001. Epub 2021 Feb 14. PMID: 33610464 140 Drug Delivery Systems for Hedgehog Inhibitors in the Treatment of SHH-Medulloblastoma. Caimano M, Lospinoso Severini L, Loricchio E, Infante P, Di Marcotullio L. Front Chem. 2021 Jun 7;9:688108. doi: 10.3389/fchem.2021.688108. eCollection 2021. PMID: 34164380 141 Fuzziness and Frustration in the Energy Landscape of Protein Folding, Function, and Assembly. Gianni S, Freiberger MI, Jemth P, Ferreiro DU, Wolynes PG, Fuxreiter M. Acc Chem Res. 2021 Mar 2;54(5):1251-1259. doi: 10.1021/acs.accounts.0c00813. Epub 2021 Feb 8. PMID: 33550810

142 Oncogenic pathways activated by pro-inflammatory cytokines promote mutant p53 stability: clue for novel anticancer therapies. D'Orazi G, Cordani M, Cirone M. Cell Mol Life Sci. 2021 Mar;78(5):1853-1860. doi: 10.1007/s00018-020-03677-7. Epub 2020 Oct 17. PMID: 33070220 143 Targeting PDZ domains as potential treatment for viral infections, neurodegeneration and cancer. Nardella C, Visconti L, Malagrinò F, Pagano L, Bufano M, Nalli M, Coluccia A, La Regina G, Silvestri R, Gianni S, Toto A. Biol Direct. 2021 Oct 12;16(1):15. doi: 10.1186/s13062-021-00303-9. PMID: 34641953 144 Understanding Drivers of Ocular Fibrosis: Current and Future Therapeutic Perspectives. Mallone F, Costi R, Marenco M, Plateroti R, Minni A, Attanasio G, Artico M, Lambiase A. Int J Mol Sci. 2021 Oct 29;22(21):11748. doi: 10.3390/ijms222111748. PMID: 34769176 145 NK cell surveillance of hematological malignancies. Therapeutic implications and regulation by chemokine receptors. Tomaipitinca L, Russo E, Bernardini G. Mol Aspects Med. 2021 Aug;80:100968. doi: 10.1016/j.mam.2021.100968. Epub 2021 May 24. PMID: 34045078 146 An Unbalanced Synaptic Transmission: Cause or Consequence of the Amyloid Oligomers Neurotoxicity? Sciaccaluga M, Megaro A, Bellomo G, Ruffolo G, Romoli M, Palma E, Costa C. Int J Mol Sci. 2021 Jun 1;22(11):5991. doi: 10.3390/ijms22115991. PMID: 34206089 147 Double Mutant Cycles as a Tool to Address Folding, Binding, and Allostery. Pagano L, Toto A, Malagrinò F, Visconti L, Jemth P, Gianni S. Int J Mol Sci. 2021 Jan 15;22(2):828. doi: 10.3390/ijms22020828. PMID: 33467625 148 Unveiling the Folding Mechanism of PDZ Domains. Gautier C, Gianni S. Methods Mol Biol. 2021;2256:149-156. doi: 10.1007/978-1-0716-1166-1_9. PMID: 34014521 149 Exciplex Formation in Lipid-bound Escherichia coli Flavohemoglobin. Marcelli A, Patrizi B, Bonamore A, Boffi A, Becucci M, Foggi P. Chemphyschem. 2021 Jun 4;22(11):1134-1140. doi: 10.1002/cphc.202100019. Epub 2021 Apr 28. PMID: 33794073 150 Recent Advances in Recovery of Lycopene from Tomato Waste: A Potent Antioxidant with Endless Benefits. Madia VN, De Vita D, Ialongo D, Tudino V, De Leo A, Scipione L, Di Santo R, Costi R, Messore A. Molecules. 2021 Jul 26;26(15):4495. doi: 10.3390/molecules26154495. PMID: 34361654 151 Acetylcholinesterase inhibitors for the treatment of Alzheimer's disease - a patent review (2016present). Bortolami M, Rocco D, Messore A, Di Santo R, Costi R, Madia VN, Scipione L, Pandolfi F. Expert Opin Ther Pat. 2021 May;31(5):399-420. doi: 10.1080/13543776.2021.1874344. Epub 2021 Jan 14. PMID: 33428491

152 Folding and Misfolding of a PDZ Tandem Repeat. Visconti L, Malagrinò F, Troilo F, Pagano L, Toto A, Gianni S. J Mol Biol. 2021 Apr 2;433(7):166862. doi: 10.1016/j.jmb.2021.166862. Epub 2021 Feb 1. PMID: 33539879 153 Small-molecule Inhibitors of HIV-1 Reverse Transcriptase-Associated Ribonuclease H Function: Challenges and Recent Developments. Madia VN, Messore A, De Leo A, Tudino V, Pindinello I, Saccoliti F, De Vita D, Scipione L, Costi R, Di Santo R. Curr Med Chem. 2021;28(30):6146-6178. doi: 10.2174/0929867328666210322164557. PMID: 34225606 154 ERAP1 as an emerging therapeutic target for medulloblastoma. Bufalieri F, Fruci D, Di Marcotullio L. Trends Cancer. 2022 Jan;8(1):4-8. doi: 10.1016/j.trecan.2021.09.005. Epub 2021 Oct 20. PMID: 34686465 155 Role of Aiolos and Ikaros in the Antitumor and Immunomodulatory Activity of IMiDs in Multiple Myeloma: Better to Lose Than to Find Them. Cippitelli M, Stabile H, Kosta A, Petillo S, Gismondi A, Santoni A, Fionda C. Int J Mol Sci. 2021 Jan 22;22(3):1103. doi: 10.3390/ijms22031103. PMID: 33499314 156 Probing the Effects of Local Frustration in the Folding of a Multidomain Protein. Pagano L, Malagrinò F, Visconti L, Troilo F, Pennacchietti V, Nardella C, Toto A, Gianni S. J Mol Biol. 2021 Jul 23;433(15):167087. doi: 10.1016/j.jmb.2021.167087. Epub 2021 Jun 3. PMID: 34089717 157 Cereblon regulates NK cell cytotoxicity and migration via Rac1 activation. Fionda C, Stabile H, Molfetta R, Kosta A, Peruzzi G, Ruggeri S, Zingoni A, Capuano C, Soriani A, Paolini R, Gismondi A, Cippitelli M, Santoni A. Eur J Immunol. 2021 Nov;51(11):2607-2617. doi: 10.1002/eji.202149269. Epub 2021 Sep 18. PMID: 34392531 158 Dissecting Inter-domain Cooperativity in the Folding of a Multi Domain Protein. Laursen L, Gianni S, Jemth P. J Mol Biol. 2021 Sep 3;433(18):167148. doi: 10.1016/j.jmb.2021.167148. Epub 2021 Jul 8. PMID: 34245784 159 A combination of PARP and CHK1 inhibitors efficiently antagonizes MYCN-driven tumors. Di Giulio S, Colicchia V, Pastorino F, Pedretti F, Fabretti F, Nicolis di Robilant V, Ramponi V, Scafetta G, Moretti M, Licursi V, Belardinilli F, Peruzzi G, Infante P, Goffredo BM, Coppa A, Canettieri G, Bartolazzi A, Ponzoni M, Giannini G, Petroni M. Oncogene. 2021 Oct;40(43):61436152. doi: 10.1038/s41388-021-02003-0. Epub 2021 Sep 10. PMID: 34508175 160 Experimental Characterization of the Interaction between the N-Terminal SH3 Domain of Crkl and C3G. Pagano L, Malagrinò F, Nardella C, Gianni S, Toto A. Int J Mol Sci. 2021 Dec 7;22(24):13174. doi: 10.3390/ijms222413174. PMID: 34947971 Free PMC article. Translational control of polyamine metabolism by CNBP is required for Drosophila locomotor function.

Coni S, Falconio FA, Marzullo M, Munafò M, Zuliani B, Mosti F, Fatica A, Ianniello Z, Bordone R, Macone A, Agostinelli E, Perna A, Matkovic T, Sigrist S, Silvestri G, Canettieri G, Ciapponi L. Elife. 2021 Sep 14;10:e69269. doi: 10.7554/eLife.69269. PMID: 34517941 161 Determining folding and binding properties of the C-terminal SH2 domain of SHP2. Nardella C, Malagrinò F, Pagano L, Rinaldo S, Gianni S, Toto A. Protein Sci. 2021 Dec;30(12):2385-2395. doi: 10.1002/pro.4201. Epub 2021 Oct 9. PMID: 34605082 162 Analytical Characterization of an Inulin-Type Fructooligosaccharide from Root-Tubers of Asphodelusramosus L. Madia VN, De Vita D, Messore A, Toniolo C, Tudino V, De Leo A, Pindinello I, Ialongo D, Saccoliti F, D'Ursi AM, Grimaldi M, Ceccobelli P, Scipione L, Di Santo R, Costi R. Pharmaceuticals (Basel). 2021 Mar 19;14(3):278. doi: 10.3390/ph14030278. PMID: 33808608 163 A PCR-RFLP method for genotyping of inversion 2Rc in Anopheles coluzzii. Montanez-Gonzalez R, Vallera AC, Calzetta M, Pichler V, Love RR, Guelbeogo MW, Dabire RK, Pombi M, Costantini C, Simard F, Della Torre A, Besansky NJ. Parasit Vectors. 2021 Mar 22;14(1):174. doi: 10.1186/s13071-021-04657-x. PMID: 33752733 164 Towards a new application of amaranth seed oil as an agent against Candida albicans. De Vita D, Messore A, Toniolo C, Frezza C, Scipione L, Bertea CM, Micera M, Di Sarno V, Madia VN, Pindinello I, Roscilli P, Botto A, Simonetti G, Orekhova A, Manfredini S, Costi R, Di Santo R. Nat Prod Res. 2021 Nov;35(22):4621-4626. doi: 10.1080/14786419.2019.1696335. Epub 2019 Dec 4. PMID: 31795749 165 Rapid inactivation of SARS-CoV-2 with LED irradiation of visible spectrum wavelengths. De Santis R, Luca V, Näslund J, Ehmann RK, De Angelis M, Lundmark E, Nencioni L, Faggioni G, Fillo S, Amatore D, Regalbuto E, Molinari F, Petralito G, Wölfel R, Stefanelli P, Rezza G, Palamara AT, Antwerpen M, Forsman M, Lista F. J Photochem Photobiol. 2021 Dec;8:100082. doi: 10.1016/j.jpap.2021.100082. Epub 2021 Oct 28. PMID: 34729540 166 The Organization of the Golgi Structures during Drosophila Male Meiosis Requires the Citrate Lyase ATPCL. Morciano P, Di Giorgio ML, Tullo L, Cenci G. Int J Mol Sci. 2021 May 27;22(11):5745. doi: 10.3390/ijms22115745. PMID: 34072207 167 Design, Synthesis and Biological Evaluation of New Pyrimidine Derivatives as Anticancer Agents. Madia VN, Nicolai A, Messore A, De Leo A, Ialongo D, Tudino V, Saccoliti F, De Vita D, Scipione L, Artico M, Taurone S, Taglieri L, Di Santo R, Scarpa S, Costi R. Molecules. 2021 Feb 2;26(3):771. doi: 10.3390/molecules26030771. PMID: 33540875 168 Quinolinonyl Non-Diketo Acid Derivatives as Inhibitors of HIV-1 Ribonuclease H and Polymerase Functions of Reverse Transcriptase. Messore A, Corona A, Madia VN, Saccoliti F, Tudino V, De Leo A, Ialongo D, Scipione L, De Vita D, Amendola G, Novellino E, Cosconati S, Métifiot M, Andreola ML, Esposito F, Grandi N,

Tramontano E, Costi R, Di Santo R. J Med Chem. 2021 Jun 24;64(12):8579-8598. doi: 10.1021/acs.jmedchem.1c00535. Epub 2021 Jun 9. PMID: 34106711 169 Functional Gastrointestinal Disorders in Patients With Epilepsy: Reciprocal Influence and Impact on Seizure Occurrence. Avorio F, Cerulli Irelli E, Morano A, Fanella M, Orlando B, Albini M, Basili LM, Ruffolo G, Fattouch J, Manfredi M, Russo E, Striano P, Carabotti M, Giallonardo AT, Severi C, Di Bonaventura C. Front Neurol. 2021 Aug 6;12:705126. doi: 10.3389/fneur.2021.705126. eCollection 2021. PMID: 34421803 170 Protective Role of Combined Polyphenols and Micronutrients against Influenza A Virus and SARS-CoV-2 Infection In Vitro. De Angelis M, Della-Morte D, Buttinelli G, Di Martino A, Pacifici F, Checconi P, Ambrosio L, Stefanelli P, Palamara AT, Garaci E, Ricordi C, Nencioni L. Biomedicines. 2021 Nov 19;9(11):1721. doi: 10.3390/biomedicines9111721. PMID: 34829949 171 Effect of heparanase inhibitor on tissue factor overexpression in platelets and endothelial cells induced by anti-β2-GPI antibodies. Capozzi A, Riitano G, Recalchi S, Manganelli V, Costi R, Saccoliti F, Pulcinelli F, Garofalo T, Misasi R, Longo A, Di Santo R, Sorice M. J Thromb Haemost. 2021 Sep;19(9):2302-2313. doi: 10.1111/jth.15417. Epub 2021 Jul 4. PMID: 34107171 172 Dolutegravir-Based Regimen for Maintenance of Viral Suppression in People Living with HIV: 48-Week Results in Real-Life Setting. Di Carlo D, Falasca F, Palermo E, Mezzaroma I, Fimiani C, Siccardi G, Celani L, Di Campli FM, d'Ettorre G, Antonelli G, Turriziani O. AIDS Res Hum Retroviruses. 2021 Jun;37(6):478-485. doi: 10.1089/AID.2020.0196. Epub 2021 Mar 1. PMID: 33487130 173 Dissecting the Molecular Determinants of GABAA Receptors Current Rundown, a Hallmark of Refractory Human Epilepsy. Cifelli P, Di Angelantonio S, Alfano V, Morano A, De Felice E, Aronica E, Ruffolo G, Palma E. Brain Sci. 2021 Mar 30;11(4):441. doi: 10.3390/brainsci11040441. PMID: 33808090 174 New Pyrimidine and Pyridine Derivatives as Multitarget Cholinesterase Inhibitors: Design, Synthesis, and In Vitro and In Cellulo Evaluation. Bortolami M, Pandolfi F, Tudino V, Messore A, Madia VN, De Vita D, Di Santo R, Costi R, Romeo I, Alcaro S, Colone M, Stringaro A, Espargaró A, Sabatè R, Scipione L. ACS Chem Neurosci. 2021 Nov 3;12(21):4090-4112. doi: 10.1021/acschemneuro.1c00485. Epub 2021 Oct 15. PMID: 34652128 175 Toxicological aspects of cannabinoid, pesticide and metal levels detected in light Cannabis inflorescences grown in Italy. Amendola G, Bocca B, Picardo V, Pelosi P, Battistini B, Ruggieri F, Attard Barbini D, De Vita D, Madia VN, Messore A, Di Santo R, Costi R. Food Chem Toxicol. 2021 Oct;156:112447. doi: 10.1016/j.fct.2021.112447. Epub 2021 Jul 31. PMID: 34343597

176 Anti-Tumoral Effects of a (1H-Pyrrol-1-yl)Methyl-1H-Benzoimidazole Carbamate Ester Derivative on Head and Neck Squamous Carcinoma Cell Lines. Nicolai A, Madia VN, Messore A, De Vita D, De Leo A, Ialongo D, Tudino V, Tortorella E, Scipione L, Taurone S, Pergolizzi T, Artico M, Di Santo R, Costi R, Scarpa S. Pharmaceuticals (Basel). 2021 Jun 12;14(6):564. doi: 10.3390/ph14060564. PMID: 34204738 177 Enzymatic Spermine Metabolites Induce Apoptosis Associated with Increase of p53, caspase-3 and miR-34a in Both Neuroblastoma Cells, SJNKP and the N-Myc-Amplified Form IMR5. Kanamori Y, Finotti A, Di Magno L, Canettieri G, Tahara T, Timeus F, Greco A, Tirassa P, Gasparello J, Fino P, Di Liegro CM, Proia P, Schiera G, Di Liegro I, Gambari R, Agostinelli E. Cells. 2021 Jul 31;10(8):1950. doi: 10.3390/cells10081950. PMID: 34440719 178 Design, synthesis and biological evaluation of a series of iron and copper chelating deferiprone derivatives as new agents active against Candida albicans. Bortolami M, Pandolfi F, Messore A, Rocco D, Feroci M, Di Santo R, De Vita D, Costi R, Cascarino P, Simonetti G, Scipione L. Bioorg Med Chem Lett. 2021 Jun 15;42:128087. doi: 10.1016/j.bmcl.2021.128087. Epub 2021 May 6. PMID: 33964446 179 Outcomes of early repeat sweat testing in infants with cystic fibrosis transmembrane conductance regulator-related metabolic syndrome/CF screen-positive, inconclusive diagnosis. Terlizzi V, Claut L, Colombo C, Tosco A, Castaldo A, Fabrizzi B, Lucarelli M, Cimino G, Carducci C, Dolce D, Biffi A, Bonomi P, Timpano S, Padoan R. Pediatr Pulmonol. 2021 Dec;56(12):37853791. doi: 10.1002/ppul.25683. Epub 2021 Sep 29. PMID: 34549893 180 Selecting antidepressants according to a drug-by-environment interaction: A comparison of fluoxetine and minocycline effects in mice living either in enriched or stressful conditions. Poggini S, Matte Bon G, Golia MT, Ciano Albanese N, Viglione A, Poleggi A, Limatola C, Maggi L, Branchi I. Behav Brain Res. 2021 Jun 25;408:113256. doi: 10.1016/j.bbr.2021.113256. Epub 2021 Mar 26. PMID: 33775780 181 DNA Methylation Patterns Correlate with the Expression of SCNN1A, SCNN1B, and SCNN1G (Epithelial Sodium Channel, ENaC) Genes. Pierandrei S, Truglio G, Ceci F, Del Porto P, Bruno SM, Castellani S, Conese M, Ascenzioni F, Lucarelli M. Int J Mol Sci. 2021 Apr 4;22(7):3754. doi: 10.3390/ijms22073754. PMID: 33916525 182 Investigating the genetic structure of the parasites Anisakis pegreffii and A. berlandi (Nematoda: Anisakidae) in a sympatric area of the southern Pacific Ocean waters using a multilocus genotyping approach: first evidence of their interspecific hybridization. Bello E, Palomba M, Webb SC, Paoletti M, Cipriani P, Nascetti G, Mattiucci S. Infect Genet Evol. 2021 Aug;92:104887. doi: 10.1016/j.meegid.2021.104887. Epub 2021 Apr 30. PMID: 33940197 183 A survey of the prevalence, management and outcome of infants with an inconclusive diagnosis following newborn bloodspot screening for cystic fibrosis (CRMS/CFSPID) in six Italian centres. Terlizzi V, Claut L, Tosco A, Colombo C, Raia V, Fabrizzi B, Lucarelli M, Angeloni A, Cimino G, Castaldo A, Marsiglio L, Timpano S, Cirilli N, Moroni L, Festini F, Piccinini P, Zavataro L,

Bonomi P, Taccetti G, Southern KW, Padoan R. J Cyst Fibros. 2021 Sep;20(5):828-834. doi: 10.1016/j.jcf.2021.03.015. Epub 2021 Apr 18. PMID: 33883100 184 Microglial-glucocorticoid receptor depletion alters the response of hippocampal microglia and neurons in a chronic unpredictable mild stress paradigm in female mice. Picard K, Bisht K, Poggini S, Garofalo S, Golia MT, Basilico B, Abdallah F, Ciano Albanese N, Amrein I, Vernoux N, Sharma K, Hui CW, C Savage J, Limatola C, Ragozzino D, Maggi L, Branchi I, Tremblay MÈ. Brain Behav Immun. 2021 Oct;97:423-439. doi: 10.1016/j.bbi.2021.07.022. Epub 2021 Jul 31. PMID: 34343616 185 Theratyping cystic fibrosis in vitro in ALI culture and organoid models generated from patientderived nasal epithelial conditionally reprogrammed stem cells. Sette G, Lo Cicero S, Blaconà G, Pierandrei S, Bruno SM, Salvati V, Castelli G, Falchi M, Fabrizzi B, Cimino G, De Maria R, Biffoni M, Eramo A, Lucarelli M. Eur Respir J. 2021 Dec 2;58(6):2100908. doi: 10.1183/13993003.00908-2021. Print 2021 Dec. PMID: 34413153 186 Salmonella Typhimurium and Pseudomonas aeruginosa Respond Differently to the Fe Chelator Deferiprone and to Some Novel Deferiprone Derivatives. Ammendola S, Secli V, Pacello F, Bortolami M, Pandolfi F, Messore A, Di Santo R, Scipione L, Battistoni A. Int J Mol Sci. 2021 Sep 23;22(19):10217. doi: 10.3390/ijms221910217. PMID: 34638558 187 Nutlin-3a Enhances Natural Killer Cell-Mediated Killing of Neuroblastoma by Restoring p53Dependent Expression of Ligands for NKG2D and DNAM-1 Receptors. Veneziani I, Infante P, Ferretti E, Melaiu O, Battistelli C, Lucarini V, Compagnone M, Nicoletti C, Castellano A, Petrini S, Ognibene M, Pezzolo A, Di Marcotullio L, Bei R, Moretta L, Pistoia V, Fruci D, Barnaba V, Locatelli F, Cifaldi L. Cancer Immunol Res. 2021 Feb;9(2):170-183. doi: 10.1158/2326-6066.CIR-20-0313. Epub 2020 Dec 10. PMID: 33303573 188 Immunomodulatory effect of NEDD8-activating enzyme inhibition in Multiple Myeloma: upregulation of NKG2D ligands and sensitization to Natural Killer cell recognition. Petillo S, Capuano C, Molfetta R, Fionda C, Mekhloufi A, Pighi C, Antonangeli F, Zingoni A, Soriani A, Petrucci MT, Galandrini R, Paolini R, Santoni A, Cippitelli M. Cell Death Dis. 2021 Sep 4;12(9):836. doi: 10.1038/s41419-021-04104-w. PMID: 34482362 189 Specific Protein 1 and p53 Interplay Modulates the Expression of the KCTD-Containing Cullin3 Adaptor Suppressor of Hedgehog 2. Angrisani A, Di Fiore A, Di Trani CA, Fonte S, Petroni M, Lospinoso Severini L, Bordin F, Belloni L, Ferretti E, Canettieri G, Moretti M, De Smaele E. Front Cell Dev Biol. 2021 Apr 8;9:638508. doi: 10.3389/fcell.2021.638508. eCollection 2021. PMID: 33898425 190 Crystal structure and functional characterization of an oligosaccharide dehydrogenase from Pycnoporus cinnabarinus provides insights into fungal breakdown of lignocellulose. Cerutti G, Gugole E, Montemiglio LC, Turbé-Doan A, Chena D, Navarro D, Lomascolo A, Piumi F, Exertier C, Freda I, Vallone B, Record E, Savino C, Sciara G. Biotechnol Biofuels. 2021 Jul 22;14(1):161. doi: 10.1186/s13068-021-02003-y. PMID: 34294139

191 Structural determinants driving the binding process between PDZ domain of wild type human PALS1 protein and SLiM sequences of SARS-CoV E proteins. Lo Cascio E, Toto A, Babini G, De Maio F, Sanguinetti M, Mordente A, Della Longa S, Arcovito A. Comput Struct Biotechnol J. 2021;19:1838-1847. doi: 10.1016/j.csbj.2021.03.014. Epub 2021 Mar 18. PMID: 33758649

ADMINISTRATIVE BOARD

The Board of Administration is chaired by a President. President

Luigi Frati

Members Angela Santoni (Scientific Director), Vincenzo Barba, Giuseppe Sancetta, Administrative Secretary

Maria Pia Lorenzoni

Auditors

Alessandra De Marco, Carla Vassallo, Adriana Vittazzi

SCIENTIFIC BOARD

The Scientific Council is a board of scientists active in the field of the pasteurian sciences. Scientific Director Angela Santoni (Immunology) Members Vincenzo Barnaba (Molecular Medicine), Francesca Cutruzzolà (Molecular Biology), Stefano Gianni (Biochemistry), Cristina Limatola (Neurosciences), Anna Teresa Palamara (Microbiology and Infectious Diseases), Sergio Pimpinelli (Genetics), Romano Silvestri (Drug Sciences), Marco Tripodi (Cell and Developmental Biology)

STAFF

Sara Atzeni, Elisabetta Canigiani, Francesca Lattanzi, Maria Pia Lorenzoni,

Lynda Romani

ISTITUTO PASTEUR ITALIA – FONDAZIONE CENCI BOLOGNETTI Viale Regina Elena 291 – 00161 – Rome Tel. 0039 06 49255625 mail: pasteuritalia@istitutopasteuritalia.it www.istitutopasteuritalia.it