@article{%a1.%Y_136,

title = {Developmental and Nutritional Dynamics of Malpighian Tubule Autofluorescence in the Asian Tiger Mosquito Aedes albopictus},

author = {Croce AC and Garbelli A and Moyano A and Soldano S and Tejeda-Guzman C and Missirlis F and Scolari F},

url = {https://www.mdpi.com/1422-0067/25/1/245},

doi = {10.3390/ijms25010245},

year  = {2024},

date = {2024-02-12},

journal = {International journal of molecular sciences},

volume = {25},

issue = {1},

pages = {245},

abstract = {Malpighian tubules (MTs) are arthropod excretory organs crucial for the osmoregulation, detoxification and excretion of xenobiotics and metabolic wastes, which include tryptophan degradation products along the kynurenine (KYN) pathway. Specifically, the toxic intermediate 3-hydroxy kynurenine (3-HK) is metabolized through transamination to xanthurenic acid or in the synthesis of ommochrome pigments. Early investigations in Drosophila larval fat bodies revealed an intracellular autofluorescence (AF) that depended on tryptophan administration. Subsequent observations documented AF changes in the MTs of Drosophila eye-color mutants genetically affecting the conversion of tryptophan to KYN or 3-HK and the intracellular availability of zinc ions. In the present study, the AF properties of the MTs in the Asian tiger mosquito, Aedes albopictus, were characterized in different stages of the insect's life cycle, tryptophan-administered larvae and blood-fed adult females. Confocal imaging and microspectroscopy showed AF changes in the distribution of intracellular, brilliant granules and in the emission spectral shape and amplitude between the proximal and distal segments of MTs across the different samples. The findings suggest AF can serve as a promising marker for investigating the functional status of MTs in response to metabolic alterations, contributing to the use of MTs as a potential research model in biomedicine.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1.%Yb_37,

title = {DEAD-Box RNA Helicases DDX3X and DDX5 as Oncogenes or Oncosuppressors: A Network Perspective},

author = {Secchi M and Lodola C and Garbelli A and Bione S and Maga G},

url = {https://www.mdpi.com/2072-6694/14/15/3820},

doi = {10.3390/cancers14153820},

year  = {2022},

date = {2022-08-18},

journal = {Cancers (Basel)},

volume = {14},

issue = {15},

pages = {3820},

abstract = {RNA helicases of the DEAD-box family are involved in several metabolic pathways, from transcription and translation to cell proliferation, innate immunity and stress response. Given their multiple roles, it is not surprising that their deregulation or mutation is linked to different pathological conditions, including cancer. However, while in some cases the loss of function of a given DEAD-box helicase promotes tumor transformation, indicating an oncosuppressive role, in other contexts the overexpression of the same enzyme favors cancer progression, thus acting as a typical oncogene. The roles of two well-characterized members of this family, DDX3X and DDX5, as both oncogenes and oncosuppressors have been documented in several cancer types. Understanding the interplay of the different cellular contexts, as defined by the molecular interaction networks of DDX3X and DDX5 in different tumors, with the cancer-specific roles played by these proteins could help to explain their apparently conflicting roles as cancer drivers or suppressors.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_131,

title = {A Role for Human DNA Polymerase lambda in Alternative Lengthening of Telomeres},

author = {Mentegari E and Bertoletti F and Kissova M and Zucca E and Galli S and Tagliavini G and Garbelli A and Maffia A and Bione S and Ferrari E and {d'Adda di Fagagna F} and Francia S and Sabbioneda S and Chen LY and Lingner J and Bergoglio V and Hoffmann JS and Hubscher U and Crespan E and Maga G},

url = {https://www.mdpi.com/1422-0067/22/5/2365},

doi = {10.3390/ijms22052365},

year  = {2021},

date = {2021-03-09},

journal = {International journal of molecular sciences},

volume = {22},

number = {5},

pages = {2365},

abstract = {Telomerase negative cancer cell types use the Alternative Lengthening of Telomeres (ALT) pathway to elongate telomeres ends. Here, we show that silencing human DNA polymerase (Pol lambda) in ALT cells represses ALT activity and induces telomeric stress. In addition, replication stress in the absence of Pol lambda, strongly affects the survival of ALT cells. In vitro, Pol lambda can promote annealing of even a single G-rich telomeric repeat to its complementary strand and use it to prime DNA synthesis. The noncoding telomeric repeat containing RNA TERRA and replication protein A negatively regulate this activity, while the Protection of Telomeres protein 1 (POT1)/TPP1 heterodimer stimulates Pol lambda. Pol lambda associates with telomeres and colocalizes with TPP1 in cells. In summary, our data suggest a role of Pol lambda in the maintenance of telomeres by the ALT mechanism.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Yb_60,

title = {Targeting DDX3X Helicase Activity with BA103 Shows Promising Therapeutic Effects in Preclinical Glioblastoma Models},

author = {Brai A and Riva V and Clementi L and Falsitta L and Zamperini C and Sinigiani V and Festuccia C and Sabetta S and Aiello D and Roselli C and Garbelli A and Trivisani CI and Maccari L and Bugli F and Sanguinetti M and Calandro P and Chiariello M and Quaranta P and Botta L and Angelucci A and Maga G and Botta M},

url = {https://www.mdpi.com/2072-6694/13/21/5569},

doi = {10.3390/cancers13215569},

year  = {2021},

date = {2021-12-06},

journal = {Cancers (Basel)},

volume = {13},

number = {21},

pages = {5569},

abstract = {DDX3X is an ATP-dependent RNA helicase that has recently attracted interest for its involvement in viral replication and oncogenic progression. Starting from hit compounds previously identified by our group, we have designed and synthesized a new series of DDX3X inhibitors that effectively blocked its helicase activity. These new compounds were able to inhibit the proliferation of cell lines from different cancer types, also in DDX3X low-expressing cancer cell lines. According to the absorption, distribution, metabolism, elimination properties, and antitumoral activity, compound BA103 was chosen to be further investigated in glioblastoma models. BA103 determined a significant reduction in the proliferation and migration of U87 and U251 cells, downregulating the oncogenic protein β-catenin. An in vivo evaluation demonstrated that BA103 was able to reach the brain and reduce the tumor growth in xenograft and orthotopic models without evident side effects. This study represents the first demonstration that DDX3X-targeted small molecules are feasible and promising drugs also in glioblastoma.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_429,

title = {DDX3X Inhibitors, an Effective Way to Overcome HIV-1 Resistance Targeting Host Proteins},

author = {Brai A and Riva V and Saladini F and Zamperini C and Trivisani CI and Garbelli A and Pennisi C and Giannini A and Boccuto A and Bugli F and Martini M and Sanguinetti M and Zazzi M and Dreassi E and Botta M and Maga G},

url = {https://www.sciencedirect.com/science/article/pii/S0223523420302889},

doi = {10.1016/j.ejmech.2020.112319},

year  = {2020},

date = {2020-01-01},

journal = {European journal of medicinal chemistry},

volume = {200},

pages = {112319},

abstract = {The huge resources that had gone into Human Immunodeficiency virus (HIV) research led to the development of potent antivirals able to suppress viral load in the majority of treated patients, thus dramatically increasing the life expectancy of people living with HIV. However, life-long treatments could result in the emergence of drug-resistant viruses that can progressively reduce the number of therapeutic options, facilitating the progression of the disease. In this scenario, we previously demonstrated that inhibitors of the human DDX3X helicase can represent an innovative approach for the simultaneous treatment of HIV and other viral infections such as Hepatitis c virus (HCV). We reported herein 6b, a novel DDX3X inhibitor that thanks to its distinct target of action is effective against HIV-1 strains resistant to currently approved drugs. Its improved in vitro ADME properties allowed us to perform preliminary in vivo studies in mice, which highlighted optimal biocompatibility and an improved bioavailability. These results represent a significant advancement in the development of DDX3X inhibitors as a novel class of broad spectrum and safe anti-HIV-1 drugs.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_428,

title = {Exploring the Implication of DDX3X in DENV Infection: Discovery of the First-in-Class DDX3X Fluorescent Inhibitor},

author = {Brai A and Boccuto A and Monti M and Marchi S and Vicenti I and Saladini F and Trivisani CI and Pollutri A and Trombetta CM and Montomoli E and Riva V and Garbelli A and Nola EM and Zazzi M and Maga G and Dreassi E and Botta M},

url = {https://pubs.acs.org/doi/10.1021/acsmedchemlett.9b00681},

doi = {10.1021/acsmedchemlett.9b00681},

year  = {2020},

date = {2020-01-01},

journal = {ACS medicinal chemistry letters},

volume = {11},

number = {5},

pages = {956-962},

abstract = {In the absence of effective drugs or vaccines for the treatment of the five Dengue Virus serotypes, the search for novel antiviral drugs is of primary importance for the scientific community. In this context, drug repurposing represents the most used strategy; however, the study of host targets is now attracting attention since it allows identification of broad-spectrum drugs endowed with high genetic barrier. In the last ten years our research group identified several small molecules DDX3X inhibitors and proved their efficacy against different viruses including novel emerging ones. Herein, starting from a screening of our compounds, we designed and synthesized novel derivatives with potent activity and high selectivity. Finally, we synthesized a fluorescent inhibitor that allowed us to study DDX3X cellular localization during DENV infection in vitro. Immunofluorescence analysis showed that our inhibitor colocalized with DDX3X, promoting the reduction of infected cells and recovering the number of viable cells.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_472,

title = {Novel alternative ribonucleotide excision repair pathways in human cells by DDX3X and specialized DNA polymerases. },

author = {Riva V and Garbelli A and Casiraghi F and Arena F and Trivisani CI and Gagliardi A and Bini L and Schroeder M and Maffia A and Sabbioneda S and Maga G},

url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7672437/},

doi = {10.1093/nar/gkaa948},

year  = {2020},

date = {2020-01-01},

journal = {Nucleic acids research},

volume = {48},

number = {20},

pages = {11551-11565},

abstract = {Removal of ribonucleotides (rNMPs) incorporated into the genome by the ribonucleotide excision repair (RER) is essential to avoid genetic instability. In eukaryotes, the RNaseH2 is the only known enzyme able to incise 5' of the rNMP, starting the RER process, which is subsequently carried out by replicative DNA polymerases (Pols) delta or epsilon, together with Flap endonuclease 1 (Fen-1) and DNA ligase 1. Here, we show that the DEAD-box RNA helicase DDX3X has RNaseH2-like activity and can support fully reconstituted in vitro RER reactions, not only with Pol δ but also with the repair Pols beta and lambda. Silencing of DDX3X causes accumulation of rNMPs in the cellular genome. These results support the existence of alternative RER pathways conferring high flexibility to human cells in responding to the threat posed by rNMPs incorporation.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_427,

title = {Novel Insights into the Biochemical Mechanism of CK1epsilon and its Functional Interplay with DDX3X},

author = {Bono B and Franco G and Riva V and Garbelli A and Maga G},

url = {https://www.mdpi.com/1422-0067/21/17/6449},

doi = {10.3390/ijms21176449},

year  = {2020},

date = {2020-01-01},

journal = {International journal of molecular sciences},

volume = {21},

number = {17},

pages = {E6449},

abstract = {Casein Kinase 1 epsilon (CK1epsilon) is a member of the serine (Ser)/threonine (Thr) CK1 family, known to have crucial roles in several biological scenarios and, ever more frequently, in pathological contexts, such as cancer. Recently, the human DEAD-box RNA helicase 3 X-linked (DDX3X), involved in cancer proliferation and viral infections, has been identified as one of CK1epsilon substrates and its positive regulator in the Wnt/beta-catenin network. However, the way by which these two proteins influence each other has not been fully clarified. In order to further investigate their interplay, we defined the kinetic parameters of CK1epsilon towards its substrates: ATP, casein, Dvl2 and DDX3X. CK1epsilon affinity for ATP depends on the nature of the substrate: increasing of casein concentrations led to an increase of KmATP, while increasing DDX3X reduced it. In literature, DDX3X is described to act as an allosteric activator of CK1epsilon. However, when we performed kinase reactions combining DDX3X and casein, we did not find a positive effect of DDX3X on casein phosphorylation by CK1epsilon, while both substrates were phosphorylated in a competitive manner. Moreover, CK1epsilon positively stimulates DDX3X ATPase activity. Our data provide a more detailed kinetic characterization on the functional interplay of these two proteins.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_5555,

title = {Unique Domain for a Unique Target: Selective Inhibitors of Host Cell DDX3X to Fight Emerging Viruses},

author = {Riva V and Garbelli A and Brai A and Casiraghi F and Fazi R and Trivisani CI and Boccuto A and Saladini F and Vicenti I and Martelli F and Zazzi M and Giannecchini S and Dreassi E and Botta M and Maga G},

url = {https://pubs.acs.org/doi/10.1021/acs.jmedchem.0c01039},

doi = {10.1021/acs.jmedchem.0c01039},

year  = {2020},

date = {2020-12-17},

urldate = {2020-12-17},

journal = {Journal of medicinal chemistry},

volume = {63},

number = {17},

pages = {9876-9877},

abstract = {•	Emerging viruses like dengue, West Nile, chikungunya, and Zika can cause widespread viral epidemics. Developing novel drugs or vaccines against specific targets for each virus is a difficult task. As obligate parasites, all viruses exploit common cellular pathways, providing the possibility to develop broad-spectrum antiviral agents targeting host factors. The human DEAD-box RNA helicase DDX3X is an essential cofactor for viral replication but dispensable for cell viability. Herein, we exploited the presence of a unique structural motif of DDX3X not shared by other cellular enzymes to develop a theoretical model to aid in the design of a novel class of highly selective inhibitors acting against such specific targets, thus limiting off-targeting effects. High-throughput virtual screening led us to identify hit compound 5, endowed with promising antienzymatic activity. To improve its aqueous solubility, 5 and its two enantiomers were synthesized and converted into their corresponding acetate salts (compounds 11, 12, and 13). In vitro mutagenesis and biochemical and cellular assays further confirmed that the developed molecules were selective for DDX3X and were able to suppress replication of West Nile and dengue viruses in infected cells in the micromolar range while showing no toxicity for uninfected cells. These results provide proof of principle for a novel strategy in developing highly selective and broad-spectrum antiviral molecules active against emerging and dangerous viral pathogens. This study paves the way for the development of larger focused libraries targeting such domain to expand SAR studies and fully characterize their mode of interaction.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y%o,

title = {DDX3X Helicase Inhibitors as a New Strategy To Fight the West Nile Virus Infection.},

author = {Brai A and Martelli F and Riva V and Garbelli A and Fazi R and Zamperini C and Pollutri A and Falsitta L and Ronzini S and Maccari L and Maga G and Giannecchini S and Botta M},

url = {https://pubs.acs.org/doi/10.1021/acs.jmedchem.8b01403},

doi = {10.1021/acs.jmedchem.8b01403},

year  = {2019},

date = {2019-02-17},

journal = {Journal of medicinal chemistry},

volume = {62},

number = {5},

pages = {2333-2347},

abstract = {Increased frequency of arbovirus outbreaks in the last 10 years represents an important emergence for global health. Climate warming, extensive urbanization of tropical regions, and human migration flows facilitate the expansion of anthropophilic mosquitos and the emerging or re-emerging of new viral infections. Only recently the human adenosinetriphosphatase/RNA helicase X-linked DEAD-box polypeptide 3 (DDX3X) emerged as a novel therapeutic target in the fight against infectious diseases. Herein, starting from our previous studies, a new family of DDX3X inhibitors was designed, synthesized, validated on the target enzyme, and evaluated against the West Nile virus (WNV) infection. Time of addition experiments after virus infection indicated that the compounds exerted their antiviral activities after the entry process, likely at the protein translation step of WNV replication. Finally, the most interesting compounds were then analyzed for their in vitro pharmacokinetic parameters, revealing favorable absorption, distribution, metabolism, and excretion values. The good safety profile together with a good activity against WNV for which no treatments are currently available, make this new class of molecules a good starting point for further in vivo studies.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y%p,

title = {Synthesis and Antiviral Activity of Novel 1,3,4-Thiadiazole Inhibitors of DDX3X.},

author = {Brai A and Ronzini S and Riva V and Botta L and Zamperini C and Borgini M and Trivisani CI and Garbelli A and Pennisi C and Boccuto A and Saladini F and Zazzi M and Maga G and Botta M},

url = {https://www.mdpi.com/1420-3049/24/21/3988},

doi = {10.3390/molecules24213988},

year  = {2019},

date = {2019-11-04},

journal = {Molecules},

volume = {24},

number = {21},

pages = { pii: E3988},

abstract = {The human ATPase/RNA helicase X-linked DEAD-box polypeptide 3 (DDX3X) emerged as a novel therapeutic target in the fight against both infectious diseases and cancer. Herein, a new family of DDX3X inhibitors was designed, synthesized, and tested for its inhibitory action on the ATPase activity of the enzyme. The potential use of the most promising derivatives it has been investigated by evaluating their anti-HIV-1 effects, revealing inhibitory activities in the low micromolar range. A preliminary ADME analysis demonstrated high metabolic stability and good aqueous solubility. The promising biological profile, together with the suitable in vitro pharmacokinetic properties, make these novel compounds a very good starting point for further development.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_219,

title = {How to win the HIV-1 drug resistance hurdle race: running faster or jumping higher?},

author = {Garbelli A and Riva V and Crespan E and Maga G},

url = {http://www.biochemj.org/content/474/10/1559.long},

doi = {10.1042/BCJ20160772},

year  = {2017},

date = {2017-02-23},

journal = {Biochemical journal},

volume = {474},

number = {10},

pages = {1559-1577},

abstract = {Infections by the human immunodeficiency virus type 1 (HIV-1), the causative agent of the acquired immunodeficiency syndrome (AIDS), are still totaling an appalling 36.7 millions worldwide, with 1.1 million AIDS deaths/year and a similar number of yearly new infections. All this, in spite of the discovery of HIV-1 as the AIDS etiological agent more than 30 years ago and the introduction of an effective combinatorial antiretroviral therapy (cART), able to control disease progression, more than 20 years ago. Although very effective, current cART is plagued by the emergence of drug-resistant viral variants and most of the efforts in the development of novel direct-acting antiviral agents (DAAs) against HIV-1 have been devoted toward the fighting of resistance. In this review, rather than providing a detailed listing of all the drugs and the corresponding resistance mutations, we aim, through relevant examples, at presenting to the general reader the conceptual shift in the approaches that are being taken to overcome the viral resistance hurdle. From the classic 'running faster' strategy, based on the development of novel DAAs active against the mutant viruses selected by the previous drugs and/or presenting to the virus a high genetic barrier toward the development of resilience, to a 'jumping higher' approach, which looks at the cell, rather than the virus, as a source of valuable drug targets, in order to make the cellular environment non-permissive toward the replication of both wild-type and mutated viruses. 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_254,

title = {Human DDX3 protein is a valuable target to develop broad spectrum antiviral agents.},

author = {Brai A and Fazi R and Tintori C and Zamperini C and Bugli F and Sanguinetti M and Stigliano E and Este' J and Badia R and Franco S and Martinez MA and Martinez JP and Meyerhans A and Saladini F and Zazzi M and Garbelli A and Maga G and Botta M},

url = {http://www.pnas.org/content/113/19/5388.long},

doi = {10.1073/pnas.1522987113},

year  = {2016},

date = {2016-05-10},

journal = {Proceedings of the National Academy of Sciences of the United States of America},

volume = {113},

number = {9},

pages = {5388-5393},

abstract = {Targeting a host factor essential for the replication of different viruses but not for the cells offers a higher genetic barrier to the development of resistance, may simplify therapy regimens for coinfections, and facilitates management of emerging viral diseases. DEAD-box polypeptide 3 (DDX3) is a human host factor required for the replication of several DNA and RNA viruses, including some of the most challenging human pathogens currently circulating, such as HIV-1, Hepatitis C virus, Dengue virus, and West Nile virus. Herein, we showed for the first time, to our knowledge, that the inhibition of DDX3 by a small molecule could be successfully exploited for the development of a broad spectrum antiviral agent. In addition to the multiple antiviral activities, hit compound 16d retained full activity against drug-resistant HIV-1 strains in the absence of cellular toxicity. Pharmacokinetics and toxicity studies in rats confirmed a good safety profile and bioavailability of 16d. Thus, DDX3 is here validated as a valuable therapeutic target.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_370,

title = {Homology Model-Based Virtual Screening for the Identification of Human Helicase DDX3 Inhibitors.},

author = {Fazi R and Tintori C and Brai A and Botta L and Selvaraj M and Garbelli A and Maga G and Botta M},

url = {https://pubs.acs.org/doi/10.1021/acs.jcim.5b00419},

doi = {10.1021/acs.jcim.5b00419},

year  = {2015},

date = {2015-11-23},

journal = {Journal of Chemical Information and Modeling},

volume = {55},

number = {11},

pages = {2443-2454},

abstract = {Targeting cellular cofactors instead of viral enzymes represents a new strategy to combat infectious diseases, which should help to overcome the problem of viral resistance. Recently, it has been revealed that the cellular ATPase/RNA helicase X-linked DEAD-box polypeptide 3 (DDX3) is an essential host factor for the replication of several viruses such as HIV, HCV, JEV, Dengue, and West Nile. Accordingly, a drug targeting DDX3 could theoretically inhibit all viruses that are dependent on this host factor. Herein, for the first time, a model of hDDX3 in its closed conformation, which binds the viral RNA was developed by using the homology module of Prime through the Maestro interface of Schrodinger. Next, a structure-based virtual screening protocol was applied to identify DDX3 small molecule inhibitors targeting the RNA binding pocket. As a result, an impressive hit rate of 40% was obtained with the identification of 10 active compounds out of the 25 tested small molecules. The best poses of the active ligands highlighted the crucial residues to be targeted for the inhibition of the helicase activity of DDX3. The obtained results confirm the reliability of the constructed DDX3/RNA model and the proposed computational strategy for investigating novel DDX3 inhibitors.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}