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Article ; Online: Canonical fibroblast growth factors in viral infection.

Lottini, Giulia / Plicanti, Erika / Lai, Michele / Quaranta, Paola / Pistello, Mauro / Freer, Giulia

2023 Volume 33, Issue 4, Page(s) e2452

Abstract: Fibroblast growth factors (FGFs) are a family of proteins that play a crucial role in the development and maintenance of various tissues in the body. There are three function-al groups of FGFs: canonical FGFs (cFGFs), intracellularly retained FGFs, and ... ...

Abstract	Fibroblast growth factors (FGFs) are a family of proteins that play a crucial role in the development and maintenance of various tissues in the body. There are three function-al groups of FGFs: canonical FGFs (cFGFs), intracellularly retained FGFs, and metabolic (also called endocrine) FGFs. cFGFs are secreted and act in an autocrine/paracrine fashion to regulate differentiation during foetal development, as well as tissue repair in adults. Recent studies have also begun to unravel the role of cFGFs during viral infections, suggesting that FGF-2 and other canonical FGFs may have an important virus-specific role, also by the regulation of the immune response. Because dysregulation in the FGF pathways is pivotal in cancer development, FGFs are the target of many anticancer drugs. These drugs may be repurposed to treat viral infection, since dysregulation of FGF signalling has been implicated in the pathogenesis of viral infections, such as hepatitis C. Overall, the role of cFGFs during viral infection is an underrepresented area of current research. This review focuses on overviewing the effects of canonical FGFs during infection by different viruses. Many studies highlight that the effects of FGFs during viral infection may be complex and context-dependent. While there is evidence to suggest that FGFs may have a beneficial impact on the immune response and tissue repair during viral infection, further studies are needed to fully understand the mechanisms underlying these effects and to determine in what cases FGFs could be targeted as a therapeutic approach for viral infection.
MeSH term(s)	Humans ; Fibroblast Growth Factors/metabolism ; Antineoplastic Agents ; Neoplasms/metabolism ; Virus Diseases
Chemical Substances	Fibroblast Growth Factors (62031-54-3) ; Antineoplastic Agents
Language	English
Publishing date	2023-04-29
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	1086043-5
ISSN	1099-1654 ; 1052-9276
ISSN (online)	1099-1654
ISSN	1052-9276
DOI	10.1002/rmv.2452
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Article ; Online: Gene silencing of endothelial von Willebrand factor reduces the susceptibility of human endothelial cells to SARS‐CoV‐2 infection

Furini, Giulia / De Carli, A. / Fonnesu, Rossella / Spezia, Pietro Giorgio / Scebba, Francesca / Pistello, Mauro / Lai, Michele / Lionetti, Vincenzo

The FEBS Journal. 2023 Sept., v. 290, no. 17 p.4300-4315

2023

Abstract: Mechanisms underlying vascular endothelial susceptibility to infection by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) are not fully understood. Emerging evidence indicates that patients lacking von Willebrand factor (vWF), an endothelial ...

Abstract	Mechanisms underlying vascular endothelial susceptibility to infection by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) are not fully understood. Emerging evidence indicates that patients lacking von Willebrand factor (vWF), an endothelial hallmark, are less severely affected by SARS‐CoV‐2 infection, yet the precise role of endothelial vWF in modulating coronavirus entry into endothelial cells is unknown. In the present study, we demonstrated that effective gene silencing by short interfering RNA (siRNA) for vWF expression in resting human umbilical vein endothelial cells (HUVECs) significantly reduced by 56% the cellular levels of SARS‐CoV‐2 genomic RNA. Similar reduction in intracellular SARS‐CoV‐2 genomic RNA levels was observed in non‐activated HUVECs treated with siRNA targeting angiotensin‐converting enzyme 2 (ACE2), the cellular gateway to coronavirus. By integrating quantitative information from real‐time PCR and high‐resolution confocal imaging, we demonstrated that ACE2 gene expression and its plasma membrane localization in HUVECs were both markedly reduced after treatment with siRNA anti‐vWF or anti‐ACE2. Conversely, siRNA anti‐ACE2 did not reduce endothelial vWF gene expression and protein levels. Finally, SARS‐CoV‐2 infection of viable HUVECs was enhanced by overexpression of vWF, which increased ACE2 levels. Of note, we found a similar increase in interferon‐β mRNA levels following transfection with untargeted, anti‐vWF or anti‐ACE2 siRNA and pcDNA3.1‐WT‐VWF. We envision that siRNA targeting endothelial vWF will protect against productive endothelial infection by SARS‐CoV‐2 through downregulation of ACE2 expression and might serve as a novel tool to induce disease resistance by modulating the regulatory role of vWF on ACE2 expression.
Keywords	RNA ; Severe acute respiratory syndrome coronavirus 2 ; blood coagulation factors ; disease resistance ; gene expression ; genes ; genomics ; humans ; peptidyl-dipeptidase A ; plasma membrane ; quantitative polymerase chain reaction ; transfection
Language	English
Dates of publication	2023-09
Size	p. 4300-4315.
Publishing place	John Wiley & Sons, Ltd
Document type	Article ; Online
Note	JOURNAL ARTICLE
ZDB-ID	2173655-8
ISSN	1742-4658 ; 1742-464X
ISSN (online)	1742-4658
ISSN	1742-464X
DOI	10.1111/febs.16808
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Article ; Online: Human T-Cell Leukemia Virus Type 1 Oncogenesis between Active Expression and Latency: A Possible Source for the Development of Therapeutic Targets.

Marino-Merlo, Francesca / Grelli, Sandro / Mastino, Antonio / Lai, Michele / Ferrari, Paola / Nicolini, Andrea / Pistello, Mauro / Macchi, Beatrice

International journal of molecular sciences

2023 Volume 24, Issue 19

Abstract: The human T-cell leukemia virus type 1 (HTLV-1) is the only known human oncogenic retrovirus. HTLV-1 can cause a type of cancer called adult T-cell leukemia/lymphoma (ATL). The virus is transmitted through the body fluids of infected individuals, ... ...

Abstract	The human T-cell leukemia virus type 1 (HTLV-1) is the only known human oncogenic retrovirus. HTLV-1 can cause a type of cancer called adult T-cell leukemia/lymphoma (ATL). The virus is transmitted through the body fluids of infected individuals, primarily breast milk, blood, and semen. At least 5-10 million people in the world are infected with HTLV-1. In addition to ATL, HTLV-1 infection can also cause HTLV-I-associated myelopathy (HAM/TSP). ATL is characterized by a low viral expression and poor prognosis. The oncogenic mechanism triggered by HTLV-1 is extremely complex and the molecular pathways are not fully understood. However, viral regulatory proteins Tax and HTLV-1 bZIP factor (HBZ) have been shown to play key roles in the transformation of HTLV-1-infected T cells. Moreover, several studies have shown that the final fate of HTLV-1-infected transformed Tcell clones is the result of a complex interplay of HTLV-1 oncogenic protein expression with cellular transcription factors that subvert the cell cycle and disrupt regulated cell death, thereby exerting their transforming effects. This review provides updated information on the mechanisms underlying the transforming action of HTLV-1 and highlights potential therapeutic targets to combat ATL.
MeSH term(s)	Adult ; Female ; Humans ; Human T-lymphotropic virus 1/metabolism ; Retroviridae Proteins/genetics ; Retroviridae Proteins/metabolism ; Basic-Leucine Zipper Transcription Factors/genetics ; Basic-Leucine Zipper Transcription Factors/metabolism ; Leukemia-Lymphoma, Adult T-Cell ; Carcinogenesis ; Cell Transformation, Neoplastic/genetics
Chemical Substances	Retroviridae Proteins ; Basic-Leucine Zipper Transcription Factors
Language	English
Publishing date	2023-09-30
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms241914807
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Article ; Online: Gene silencing of endothelial von Willebrand factor reduces the susceptibility of human endothelial cells to SARS-CoV-2 infection.

Furini, Giulia / De Carli, Alessandro / Fonnesu, Rossella / Spezia, Pietro Giorgio / Scebba, Francesca / Pistello, Mauro / Lai, Michele / Lionetti, Vincenzo

The FEBS journal

2023 Volume 290, Issue 17, Page(s) 4300–4315

Abstract: Mechanisms underlying vascular endothelial susceptibility to infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are not fully understood. Emerging evidence indicates that patients lacking von Willebrand factor (vWF), an endothelial ...

Abstract	Mechanisms underlying vascular endothelial susceptibility to infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are not fully understood. Emerging evidence indicates that patients lacking von Willebrand factor (vWF), an endothelial hallmark, are less severely affected by SARS-CoV-2 infection, yet the precise role of endothelial vWF in modulating coronavirus entry into endothelial cells is unknown. In the present study, we demonstrated that effective gene silencing by short interfering RNA (siRNA) for vWF expression in resting human umbilical vein endothelial cells (HUVECs) significantly reduced by 56% the cellular levels of SARS-CoV-2 genomic RNA. Similar reduction in intracellular SARS-CoV-2 genomic RNA levels was observed in non-activated HUVECs treated with siRNA targeting angiotensin-converting enzyme 2 (ACE2), the cellular gateway to coronavirus. By integrating quantitative information from real-time PCR and high-resolution confocal imaging, we demonstrated that ACE2 gene expression and its plasma membrane localization in HUVECs were both markedly reduced after treatment with siRNA anti-vWF or anti-ACE2. Conversely, siRNA anti-ACE2 did not reduce endothelial vWF gene expression and protein levels. Finally, SARS-CoV-2 infection of viable HUVECs was enhanced by overexpression of vWF, which increased ACE2 levels. Of note, we found a similar increase in interferon-β mRNA levels following transfection with untargeted, anti-vWF or anti-ACE2 siRNA and pcDNA3.1-WT-VWF. We envision that siRNA targeting endothelial vWF will protect against productive endothelial infection by SARS-CoV-2 through downregulation of ACE2 expression and might serve as a novel tool to induce disease resistance by modulating the regulatory role of vWF on ACE2 expression.
MeSH term(s)	Humans ; COVID-19/genetics ; SARS-CoV-2/genetics ; SARS-CoV-2/metabolism ; von Willebrand Factor/genetics ; von Willebrand Factor/metabolism ; Endothelial Cells/metabolism ; Peptidyl-Dipeptidase A/genetics ; Peptidyl-Dipeptidase A/metabolism ; RNA, Small Interfering/genetics ; Gene Silencing
Chemical Substances	von Willebrand Factor ; Peptidyl-Dipeptidase A (EC 3.4.15.1) ; RNA, Small Interfering
Language	English
Publishing date	2023-05-10
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2173655-8
ISSN	1742-4658 ; 1742-464X
ISSN (online)	1742-4658
ISSN	1742-464X
DOI	10.1111/febs.16808
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Article: A low-cost simple test for weekly detection of Mycoplasma hyorhinis and arginini contaminations in cell cultures and viral preparations

Lai, Michele / Iacono, Elena / Spezia, Pietro Giorgio / Lottini, Giulia / La Rocca, Veronica / Quaranta, Paola / Pistello, Mauro / Freer, Giulia

Journal of virological methods. 2022 Jan., v. 299

2022

Abstract: Mollicutes (Mycoplasma and Acholeplasma) are parasitic bacteria that adhere to cellular surfaces, naturally resistant to many antibiotics and extremely small. They are often found as contaminants in cultured cells, where they go unnoticed. They may be ... ...

Abstract	Mollicutes (Mycoplasma and Acholeplasma) are parasitic bacteria that adhere to cellular surfaces, naturally resistant to many antibiotics and extremely small. They are often found as contaminants in cultured cells, where they go unnoticed. They may be present in viral stocks because they are present in supernatants of cells where cultured viruses are released. The best way to keep laboratories free of Mycoplasma is to discard infected cultures, but, as judged by the very common finding of Mycoplasma-contaminated cultures in many laboratories, this is not done as often as it should be. A possible reason is that most procedures recommended take as long as performing a simple experiment and many laboratories delay testing to save money and time. Indeed, many methods exist to detect Mycoplasma infection of cell lines, but they take at least a couple of hours of hands-on work, if not more. Here we describe a procedure to screen viral stocks and tissue cultures for Mycoplasma presence. It relies on isolation of Mycoplasma on ordinary horse blood agar directly from exhausted tissue culture supernatants and does not require experienced personnel or expensive equipment. It only requires minutes of hands-on work, and, for this, it may be useful for weekly screening of cultures. It yields semiquantitative results in roughly 5 days, which is the time that usually passes between one subculture passage of cells in vitro to another. Because of its simplicity, it may be useful for detecting Mycoplasma in viral stocks and for frequent screening of cultures in research laboratories.
Keywords	Acholeplasma ; Mycoplasma hyorhinis ; culture media ; equipment ; horses ; human resources ; mycoplasmosis ; tissue culture
Language	English
Dates of publication	2022-01
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	8013-5
ISSN	1879-0984 ; 0166-0934
ISSN (online)	1879-0984
ISSN	0166-0934
DOI	10.1016/j.jviromet.2021.114327
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Article: Evolution of viruses and the emergence of SARS-CoV-2 variants.

Freer, Giulia / Lai, Michele / Quaranta, Paola / Spezia, Pietro Giorgio / Pistello, Mauro

The new microbiologica

2021 Volume 44, Issue 4, Page(s) 191–204

Abstract: Life implies adaptation. This is one of the fundamental principles that has permitted most living species to survive through ages in an ever-changing environment. Spontaneously occurring events have shaped also virus populations and their fitness. Thanks ...

Abstract	Life implies adaptation. This is one of the fundamental principles that has permitted most living species to survive through ages in an ever-changing environment. Spontaneously occurring events have shaped also virus populations and their fitness. Thanks to their plasticity, viruses have thrived in extremely dissimilar conditions. Unsurprisingly, SARS-CoV-2, the etiological agent of COVID-19, is no exception. Thanks to an unprecedented rate of molecular tracing and sequence scrutiny, the virus was followed in all its changes and shown to evolve in such a way as to possibly determine subsequent waves of infection after the first global and massive outbreak. This review illustrates the major modifications occurred to the virus since its discovery. We describe the potential advantages that these changes conveyed as regards SARS-CoV-2 transmissibility, resistance to host innate and adaptive barriers and molecular diagnosis.
MeSH term(s)	COVID-19 ; Humans ; SARS-CoV-2 ; Spike Glycoprotein, Coronavirus
Chemical Substances	Spike Glycoprotein, Coronavirus
Language	English
Publishing date	2021-12-19
Publishing country	Italy
Document type	Journal Article ; Review
ZDB-ID	756168-4
ISSN	1121-7138 ; 0391-5352
ISSN	1121-7138 ; 0391-5352
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Article ; Online: Design, synthesis, and biological screening of a series of 4'-fluoro-benzotriazole-acrylonitrile derivatives as microtubule-destabilising agents (MDAs).

Riu, Federico / Ibba, Roberta / Zoroddu, Stefano / Sestito, Simona / Lai, Michele / Piras, Sandra / Sanna, Luca / Bordoni, Valentina / Bagella, Luigi / Carta, Antonio

Journal of enzyme inhibition and medicinal chemistry

2022 Volume 37, Issue 1, Page(s) 2223–2240

Abstract: Introduction: ...

Abstract	Introduction:
MeSH term(s)	Acrylonitrile/pharmacology ; Antineoplastic Agents/chemistry ; Cell Line, Tumor ; Cell Proliferation ; Colchicine/chemistry ; Drug Screening Assays, Antitumor ; HeLa Cells ; Humans ; Ligands ; Microtubules/metabolism ; Molecular Docking Simulation ; Structure-Activity Relationship ; Triazoles ; Tubulin/metabolism ; Tubulin Modulators
Chemical Substances	Antineoplastic Agents ; Ligands ; Triazoles ; Tubulin ; Tubulin Modulators ; benzotriazole (86110UXM5Y) ; Acrylonitrile (MP1U0D42PE) ; Colchicine (SML2Y3J35T)
Language	English
Publishing date	2022-08-17
Publishing country	England
Document type	Journal Article
ZDB-ID	2082578-X
ISSN	1475-6374 ; 1475-6366
ISSN (online)	1475-6374
ISSN	1475-6366
DOI	10.1080/14756366.2022.2111680
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Article ; Online: Effects of Nutraceuticals on Cisplatin-Induced Cytotoxicity in HEI-OC1 Cells.

Guidotti, Lorenzo / Tomassi, Elena / Marracci, Silvia / Lai, Michele / Lapi, Dominga / Pesi, Rossana / Pucci, Laura / Novellino, Ettore / Albi, Elisabetta / Garcia-Gil, Mercedes

International journal of molecular sciences

2023 Volume 24, Issue 24

Abstract: Cisplatin is a chemotherapeutic drug for the treatment of several solid tumors, whose use is limited by its nephrotoxicity, neurotoxicity, ototoxicity, and development of resistance. The toxicity is caused by DNA cross-linking, increase in reactive ... ...

Abstract	Cisplatin is a chemotherapeutic drug for the treatment of several solid tumors, whose use is limited by its nephrotoxicity, neurotoxicity, ototoxicity, and development of resistance. The toxicity is caused by DNA cross-linking, increase in reactive oxygen species and/or depletion of cell antioxidant defenses. The aim of the work was to study the effect of antioxidant compounds (Lisosan G, Taurisolo
MeSH term(s)	Cisplatin/pharmacology ; Cisplatin/metabolism ; Antioxidants/pharmacology ; Reactive Oxygen Species/metabolism ; Sphingomyelin Phosphodiesterase/metabolism ; Hair Cells, Auditory/metabolism ; Antineoplastic Agents/pharmacology ; Antineoplastic Agents/metabolism ; Apoptosis ; Caspases/metabolism ; Dietary Supplements ; Cell Survival
Chemical Substances	Cisplatin (Q20Q21Q62J) ; erucin (CTE370DL3U) ; Antioxidants ; Reactive Oxygen Species ; Sphingomyelin Phosphodiesterase (EC 3.1.4.12) ; Antineoplastic Agents ; Caspases (EC 3.4.22.-)
Language	English
Publishing date	2023-12-12
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms242417416
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Article ; Online: Anti-inflammatory reprogramming of microglia cells by metabolic modulators to counteract neurodegeneration; a new role for Ranolazine.

Piano, Ilaria / Votta, Arianna / Colucci, Patrizia / Corsi, Francesca / Vitolo, Sara / Cerri, Chiara / Puppi, Dario / Lai, Michele / Maya-Vetencourt, José Fernando / Leigheb, Massimiliano / Gabellini, Chiara / Ferraro, Elisabetta

Scientific reports

2023 Volume 13, Issue 1, Page(s) 20138

Abstract: Microglia chronic activation is a hallmark of several neurodegenerative diseases, including the retinal ones, possibly contributing to their etiopathogenesis. However, some microglia sub-populations have anti-inflammatory and neuroprotective functions, ... ...

Abstract	Microglia chronic activation is a hallmark of several neurodegenerative diseases, including the retinal ones, possibly contributing to their etiopathogenesis. However, some microglia sub-populations have anti-inflammatory and neuroprotective functions, thus making arduous deciphering the role of these cells in neurodegeneration. Since it has been proposed that functionally different microglia subsets also rely on different metabolic routes, we hypothesized that modulating microglia metabolism might be a tool to enhance their anti-inflammatory features. This would have a preventive and therapeutic potential in counteracting neurodegenerative diseases. For this purpose, we tested various molecules known to act on cell metabolism, and we revealed the anti-inflammatory effect of the FDA-approved piperazine derivative Ranolazine on microglia cells, while confirming the one of the flavonoids Quercetin and Naringenin, both in vitro and in vivo. We also demonstrated the synergistic anti-inflammatory effect of Quercetin and Idebenone, and the ability of Ranolazine, Quercetin and Naringenin to counteract the neurotoxic effect of LPS-activated microglia on 661W neuronal cells. Overall, these data suggest that using the selected molecules -also in combination therapies- might represent a valuable approach to reduce inflammation and neurodegeneration while avoiding long term side effects of corticosteroids.
MeSH term(s)	Humans ; Microglia/metabolism ; Ranolazine/pharmacology ; Ranolazine/therapeutic use ; Quercetin/therapeutic use ; Anti-Inflammatory Agents/therapeutic use ; Inflammation/pathology ; Neurodegenerative Diseases/drug therapy ; Neurodegenerative Diseases/metabolism ; Lipopolysaccharides/pharmacology ; Neuroprotective Agents/therapeutic use
Chemical Substances	Ranolazine (A6IEZ5M406) ; Quercetin (9IKM0I5T1E) ; Anti-Inflammatory Agents ; Lipopolysaccharides ; Neuroprotective Agents
Language	English
Publishing date	2023-11-17
Publishing country	England
Document type	Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-023-47540-8
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Article ; Online: Acid ceramidase controls apoptosis and increases autophagy in human melanoma cells treated with doxorubicin.

Lai, Michele / Amato, Rachele / La Rocca, Veronica / Bilgin, Mesut / Freer, Giulia / Spezia, Piergiorgio / Quaranta, Paola / Piomelli, Daniele / Pistello, Mauro

Scientific reports

2021 Volume 11, Issue 1, Page(s) 11221

Abstract: Acid ceramidase (AC) is a lysosomal hydrolase encoded by the ASAH1 gene, which cleaves ceramides into sphingosine and fatty acid. AC is expressed at high levels in most human melanoma cell lines and may confer resistance against chemotherapeutic agents. ... ...

Abstract	Acid ceramidase (AC) is a lysosomal hydrolase encoded by the ASAH1 gene, which cleaves ceramides into sphingosine and fatty acid. AC is expressed at high levels in most human melanoma cell lines and may confer resistance against chemotherapeutic agents. One such agent, doxorubicin, was shown to increase ceramide levels in melanoma cells. Ceramides contribute to the regulation of autophagy and apoptosis. Here we investigated the impact of AC ablation via CRISPR-Cas9 gene editing on the response of A375 melanoma cells to doxorubicin. We found that doxorubicin activates the autophagic response in wild-type A375 cells, which effectively resist apoptotic cell death. In striking contrast, doxorubicin fails to stimulate autophagy in A375 AC-null cells, which rapidly undergo apoptosis when exposed to the drug. The present work highlights changes that affect melanoma cells during incubation with doxorubicin, in A375 melanoma cells lacking AC. We found that the remarkable reduction in recovery rate after doxorubicin treatment is strictly associated with the impairment of autophagy, that forces the AC-inhibited cells into apoptotic path.
MeSH term(s)	Acid Ceramidase/genetics ; Acid Ceramidase/metabolism ; Antineoplastic Agents/pharmacology ; Apoptosis/physiology ; Autophagy/physiology ; Cell Line, Tumor ; Ceramides/metabolism ; Doxorubicin/pharmacology ; Humans ; Melanoma/drug therapy ; Melanoma/metabolism ; Melanoma/pathology
Chemical Substances	Antineoplastic Agents ; Ceramides ; Doxorubicin (80168379AG) ; Acid Ceramidase (EC 3.5.1.23)
Language	English
Publishing date	2021-05-27
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-021-90219-1
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