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Article ; Online: The Intertwined Role of 8-oxodG and G4 in Transcription Regulation.

Gorini, Francesca / Ambrosio, Susanna / Lania, Luigi / Majello, Barbara / Amente, Stefano

International journal of molecular sciences

2023 Volume 24, Issue 3

Abstract: The guanine base in nucleic acids is, among the other bases, the most susceptible to being converted into 8-Oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) when exposed to reactive oxygen species. In double-helix DNA, 8-oxodG can pair with adenine; hence, it ...

Abstract	The guanine base in nucleic acids is, among the other bases, the most susceptible to being converted into 8-Oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) when exposed to reactive oxygen species. In double-helix DNA, 8-oxodG can pair with adenine; hence, it may cause a G > T (C > A) mutation; it is frequently referred to as a form of DNA damage and promptly corrected by DNA repair mechanisms. Moreover, 8-oxodG has recently been redefined as an epigenetic factor that impacts transcriptional regulatory elements and other epigenetic modifications. It has been proposed that 8-oxodG exerts epigenetic control through interplay with the G-quadruplex (G4), a non-canonical DNA structure, in transcription regulatory regions. In this review, we focused on the epigenetic roles of 8-oxodG and the G4 and explored their interplay at the genomic level.
MeSH term(s)	8-Hydroxy-2'-Deoxyguanosine ; Deoxyguanosine ; DNA Damage ; DNA Repair ; DNA/chemistry
Chemical Substances	8-Hydroxy-2'-Deoxyguanosine (88847-89-6) ; Deoxyguanosine (G9481N71RO) ; DNA (9007-49-2)
Language	English
Publishing date	2023-01-19
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/ijms24032031
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Article: Autophagy Roles in Genome Maintenance.

Ambrosio, Susanna / Majello, Barbara

Cancers

2020 Volume 12, Issue 7

Abstract: In recent years, a considerable correlation has emerged between autophagy and genome integrity. A range of mechanisms appear to be involved where autophagy participates in preventing genomic instability, as well as in DNA damage response and cell fate ... ...

Abstract	In recent years, a considerable correlation has emerged between autophagy and genome integrity. A range of mechanisms appear to be involved where autophagy participates in preventing genomic instability, as well as in DNA damage response and cell fate decision. These initial findings have attracted particular attention in the context of malignancy; however, the crosstalk between autophagy and DNA damage response is just beginning to be explored and key questions remain that need to be addressed, to move this area of research forward and illuminate the overall consequence of targeting this process in human therapies. Here we present current knowledge on the complex crosstalk between autophagy and genome integrity and discuss its implications for cancer cell survival and response to therapy.
Language	English
Publishing date	2020-07-04
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2527080-1
ISSN	2072-6694
ISSN	2072-6694
DOI	10.3390/cancers12071793
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Article ; Online: OxiDIP-Seq for Genome-wide Mapping of Damaged DNA Containing 8-Oxo-2'-Deoxyguanosine.

Gorini, Francesca / Scala, Giovanni / Ambrosio, Susanna / Majello, Barbara / Amente, Stefano

Bio-protocol

2022 Volume 12, Issue 21

Abstract: 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) is considered to be a premutagenic DNA lesion generated by 2'-deoxyguanosine (dG) oxidation due to reactive oxygen species (ROS). In recent years, the 8-oxodG distribution in human, mouse, and yeast genomes ... ...

Abstract	8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) is considered to be a premutagenic DNA lesion generated by 2'-deoxyguanosine (dG) oxidation due to reactive oxygen species (ROS). In recent years, the 8-oxodG distribution in human, mouse, and yeast genomes has been underlined using various next-generation sequencing (NGS)-based strategies. The present study reports the OxiDIP-Seq protocol, which combines specific 8-oxodG immuno-precipitation of single-stranded DNA with NGS, and the pipeline analysis that allows the genome-wide 8-oxodG distribution in mammalian cells. The development of this OxiDIP-Seq method increases knowledge on the oxidative DNA damage/repair field, providing a high-resolution map of 8-oxodG in human cells.
Language	English
Publishing date	2022-11-05
Publishing country	United States
Document type	Journal Article
ZDB-ID	2833269-6
ISSN	2331-8325 ; 2331-8325
ISSN (online)	2331-8325
ISSN	2331-8325
DOI	10.21769/BioProtoc.4540
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Article: Targeting Histone Demethylase LSD1/KDM1a in Neurodegenerative Diseases.

Ambrosio, Susanna / Majello, Barbara

Journal of experimental neuroscience

2018 Volume 12, Page(s) 1179069518765743

Abstract: The autophagy-lysosome pathway sustains cellular homeostasis and is a protective mechanism against neurodegenerative diseases. Recent findings highlight the role of the histone demethylases LSD1/LDM1A as a pivotal regulator of autophagy process, by ... ...

Abstract	The autophagy-lysosome pathway sustains cellular homeostasis and is a protective mechanism against neurodegenerative diseases. Recent findings highlight the role of the histone demethylases LSD1/LDM1A as a pivotal regulator of autophagy process, by controlling the mTORC1 cascade, in neuroblastoma cells. LSD1 binds to the promoter region of the
Language	English
Publishing date	2018-03-20
Publishing country	United States
Document type	Journal Article
ZDB-ID	2659991-0
ISSN	1179-0695
ISSN	1179-0695
DOI	10.1177/1179069518765743
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Article ; Online: Accumulation of 8-oxodG within the human mitochondrial genome positively associates with transcription.

Scala, Giovanni / Ambrosio, Susanna / Menna, Margherita / Gorini, Francesca / Caiazza, Carmen / Cooke, Marcus S / Majello, Barbara / Amente, Stefano

NAR genomics and bioinformatics

2023 Volume 5, Issue 4, Page(s) lqad100

Abstract: Mitochondrial DNA (mtDNA) can be subject to internal and environmental stressors that lead to oxidatively generated damage and the formation of 8-oxo-7,8-dihydro-2'-deoxyguanine (8-oxodG). The accumulation of 8-oxodG has been linked to degenerative ... ...

Abstract	Mitochondrial DNA (mtDNA) can be subject to internal and environmental stressors that lead to oxidatively generated damage and the formation of 8-oxo-7,8-dihydro-2'-deoxyguanine (8-oxodG). The accumulation of 8-oxodG has been linked to degenerative diseases and aging, as well as cancer. Despite the well-described implications of 8-oxodG in mtDNA for mitochondrial function, there have been no reports of mapping of 8-oxodG across the mitochondrial genome. To address this, we used OxiDIP-Seq and mapped 8-oxodG levels in the mitochondrial genome of human MCF10A cells. Our findings indicated that, under steady-state conditions, 8-oxodG is non-uniformly distributed along the mitochondrial genome, and that the longer non-coding region appeared to be more protected from 8-oxodG accumulation compared with the coding region. However, when the cells have been exposed to oxidative stress, 8-oxodG preferentially accumulated in the coding region which is highly transcribed as H1 transcript. Our data suggest that 8-oxodG accumulation in the mitochondrial genome is positively associated with mitochondrial transcription.
Language	English
Publishing date	2023-11-06
Publishing country	England
Document type	Journal Article
ISSN	2631-9268
ISSN (online)	2631-9268
DOI	10.1093/nargab/lqad100
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Article ; Online: Histone methyl-transferases and demethylases in the autophagy regulatory network: the emerging role of KDM1A/LSD1 demethylase.

Ambrosio, Susanna / Ballabio, Andrea / Majello, Barbara

Autophagy

2018 Volume 15, Issue 2, Page(s) 187–196

Abstract: Macroautophagy/autophagy is a physiological mechanism that is essential for the maintenance of cellular homeostasis and stress adaptation. Defective autophagy is associated with many human diseases, including cancer and neurodegenerative disorders. The ... ...

Abstract	Macroautophagy/autophagy is a physiological mechanism that is essential for the maintenance of cellular homeostasis and stress adaptation. Defective autophagy is associated with many human diseases, including cancer and neurodegenerative disorders. The emerging implication of epigenetic events in the control of the autophagic process opens new avenues of investigation and offers the opportunity to develop novel therapeutic strategies in diseases associated with dysfunctional autophagy-lysosomal pathways. Accumulating evidence reveals that several methyltransferases and demethylases are essential regulators of autophagy, and recent studies have led to the identification of the lysine demethylase KDM1A/LSD1 as a promising drug target. KDM1A/LSD1 modulates autophagy at multiple levels, participating in the transcriptional control of several downstream effectors. This review summarizes our current understanding of the role of KDM1A/LSD1 in the autophagy regulatory network.
MeSH term(s)	Animals ; Autophagy/genetics ; Epigenesis, Genetic ; Histone Demethylases/metabolism ; Histone Methyltransferases/metabolism ; Humans ; Models, Biological ; Signal Transduction/genetics
Chemical Substances	Histone Demethylases (EC 1.14.11.-) ; Histone Methyltransferases (EC 2.1.1.-)
Language	English
Publishing date	2018-09-22
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2454135-7
ISSN	1554-8635 ; 1554-8627
ISSN (online)	1554-8635
ISSN	1554-8627
DOI	10.1080/15548627.2018.1520546
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Article ; Online: 8-oxodG accumulation within super-enhancers marks fragile CTCF-mediated chromatin loops.

Scala, Giovanni / Gorini, Francesca / Ambrosio, Susanna / Chiariello, Andrea M / Nicodemi, Mario / Lania, Luigi / Majello, Barbara / Amente, Stefano

Nucleic acids research

2022 Volume 50, Issue 6, Page(s) 3292–3306

Abstract: 8-Oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), a major product of the DNA oxidization process, has been proposed to have an epigenetic function in gene regulation and has been associated with genome instability. NGS-based methodologies are contributing ... ...

Abstract	8-Oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), a major product of the DNA oxidization process, has been proposed to have an epigenetic function in gene regulation and has been associated with genome instability. NGS-based methodologies are contributing to the characterization of the 8-oxodG function in the genome. However, the 8-oxodG epigenetic role at a genomic level and the mechanisms controlling the genomic 8-oxodG accumulation/maintenance have not yet been fully characterized. In this study, we report the identification and characterization of a set of enhancer regions accumulating 8-oxodG in human epithelial cells. We found that these oxidized enhancers are mainly super-enhancers and are associated with bidirectional-transcribed enhancer RNAs and DNA Damage Response activation. Moreover, using ChIA-PET and HiC data, we identified specific CTCF-mediated chromatin loops in which the oxidized enhancer and promoter regions physically associate. Oxidized enhancers and their associated chromatin loops accumulate endogenous double-strand breaks which are in turn repaired by NHEJ pathway through a transcription-dependent mechanism. Our work suggests that 8-oxodG accumulation in enhancers-promoters pairs occurs in a transcription-dependent manner and provides novel mechanistic insights on the intrinsic fragility of chromatin loops containing oxidized enhancers-promoters interactions.
MeSH term(s)	8-Hydroxy-2'-Deoxyguanosine/metabolism ; CCCTC-Binding Factor/metabolism ; Chromatin/genetics ; DNA ; Enhancer Elements, Genetic ; Epigenesis, Genetic ; Genomic Instability ; Humans ; Promoter Regions, Genetic ; Transcription, Genetic
Chemical Substances	CCCTC-Binding Factor ; CTCF protein, human ; Chromatin ; 8-Hydroxy-2'-Deoxyguanosine (88847-89-6) ; DNA (9007-49-2)
Language	English
Publishing date	2022-03-02
Publishing country	England
Document type	Journal Article
ZDB-ID	186809-3
ISSN	1362-4962 ; 1362-4954 ; 0301-5610 ; 0305-1048
ISSN (online)	1362-4962 ; 1362-4954
ISSN	0301-5610 ; 0305-1048
DOI	10.1093/nar/gkac143
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Zs.A 1067: Show issues

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Article ; Online: EGR1 drives cell proliferation by directly stimulating TFEB transcription in response to starvation.

Cesana, Marcella / Tufano, Gennaro / Panariello, Francesco / Zampelli, Nicolina / Ambrosio, Susanna / De Cegli, Rossella / Mutarelli, Margherita / Vaccaro, Lorenzo / Ziller, Micheal J / Cacchiarelli, Davide / Medina, Diego L / Ballabio, Andrea

PLoS biology

2023 Volume 21, Issue 3, Page(s) e3002034

Abstract: The stress-responsive transcription factor EB (TFEB) is a master controller of lysosomal biogenesis and autophagy and plays a major role in several cancer-associated diseases. TFEB is regulated at the posttranslational level by the nutrient-sensitive ... ...

Abstract	The stress-responsive transcription factor EB (TFEB) is a master controller of lysosomal biogenesis and autophagy and plays a major role in several cancer-associated diseases. TFEB is regulated at the posttranslational level by the nutrient-sensitive kinase complex mTORC1. However, little is known about the regulation of TFEB transcription. Here, through integrative genomic approaches, we identify the immediate-early gene EGR1 as a positive transcriptional regulator of TFEB expression in human cells and demonstrate that, in the absence of EGR1, TFEB-mediated transcriptional response to starvation is impaired. Remarkably, both genetic and pharmacological inhibition of EGR1, using the MEK1/2 inhibitor Trametinib, significantly reduced the proliferation of 2D and 3D cultures of cells displaying constitutive activation of TFEB, including those from a patient with Birt-Hogg-Dubé (BHD) syndrome, a TFEB-driven inherited cancer condition. Overall, we uncover an additional layer of TFEB regulation consisting in modulating its transcription via EGR1 and propose that interfering with the EGR1-TFEB axis may represent a therapeutic strategy to counteract constitutive TFEB activation in cancer-associated conditions.
MeSH term(s)	Humans ; Mechanistic Target of Rapamycin Complex 1/genetics ; Mechanistic Target of Rapamycin Complex 1/metabolism ; Autophagy/genetics ; Lysosomes/metabolism ; Cell Proliferation/genetics ; Early Growth Response Protein 1/genetics ; Early Growth Response Protein 1/metabolism ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/metabolism
Chemical Substances	Mechanistic Target of Rapamycin Complex 1 (EC 2.7.11.1) ; EGR1 protein, human ; Early Growth Response Protein 1 ; TFEB protein, human ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors
Language	English
Publishing date	2023-03-08
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2126776-5
ISSN	1545-7885 ; 1544-9173
ISSN (online)	1545-7885
ISSN	1544-9173
DOI	10.1371/journal.pbio.3002034
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Zs.A 6193: Show issues

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Article: Epigenetic regulation of epithelial to mesenchymal transition by the Lysine-specific demethylase LSD1/KDM1A.

Ambrosio, Susanna / Saccà, Carmen D / Majello, Barbara

Biochimica et biophysica acta. Gene regulatory mechanisms

2017 Volume 1860, Issue 9, Page(s) 905–910

Abstract: The Lysine-specific demethylase 1, KDM1A/LSD1, plays a central role in the regulation of Pol II transcription through the removal of the activation mark (mono- and dimethyl lysine 4 of histone H3). LSD1 is often deregulated in human cancers, and it is ... ...

Abstract	The Lysine-specific demethylase 1, KDM1A/LSD1, plays a central role in the regulation of Pol II transcription through the removal of the activation mark (mono- and dimethyl lysine 4 of histone H3). LSD1 is often deregulated in human cancers, and it is frequently overexpressed in human solid cancers and leukemia. LSD1 regulates the epithelial mesenchymal transition (EMT) in epithelial cells, i.e., the ability to transition into mesenchymal cells, to lose homotypic adhesion and to acquire migratory capacity. From its initial discovery as a component of the Snail complex, multiple studies highlighted the causative role of LSD1 in cell invasiveness and EMT, describing its direct involvement in different molecular processes through the interaction with specific partners. Here we present an overview of the role of LSD1 in the EMT process, summarizing recent findings on its emerging functions in cell migration and invasion in cancer.
MeSH term(s)	Cell Movement/genetics ; Epigenesis, Genetic/genetics ; Epithelial-Mesenchymal Transition/genetics ; Epithelial-Mesenchymal Transition/physiology ; Gene Expression Regulation, Neoplastic/genetics ; Histone Demethylases/metabolism ; Humans ; Lysine/metabolism ; Neoplasms/genetics ; Neoplasms/metabolism ; Neoplasms/pathology
Chemical Substances	Histone Demethylases (EC 1.14.11.-) ; KDM1A protein, human (EC 1.5.-) ; Lysine (K3Z4F929H6)
Language	English
Publishing date	2017-07-15
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	60-7
ISSN	1879-2596 ; 1879-260X ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650 ; 1874-9399 ; 0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439
ISSN (online)	1879-2596 ; 1879-260X ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650
ISSN	1874-9399 ; 0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439
DOI	10.1016/j.bbagrm.2017.07.001
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Article ; Online: TFEB and TFE3 control glucose homeostasis by regulating insulin gene expression.

Pasquier, Adrien / Pastore, Nunzia / D'Orsi, Luca / Colonna, Rita / Esposito, Alessandra / Maffia, Veronica / De Cegli, Rossella / Mutarelli, Margherita / Ambrosio, Susanna / Tufano, Gennaro / Grimaldi, Antonio / Cesana, Marcella / Cacchiarelli, Davide / Delalleau, Nathalie / Napolitano, Gennaro / Ballabio, Andrea

The EMBO journal

2023 Volume 42, Issue 21, Page(s) e113928

Abstract: To fulfill their function, pancreatic beta cells require precise nutrient-sensing mechanisms that control insulin production. Transcription factor EB (TFEB) and its homolog TFE3 have emerged as crucial regulators of the adaptive response of cell ... ...

Abstract	To fulfill their function, pancreatic beta cells require precise nutrient-sensing mechanisms that control insulin production. Transcription factor EB (TFEB) and its homolog TFE3 have emerged as crucial regulators of the adaptive response of cell metabolism to environmental cues. Here, we show that TFEB and TFE3 regulate beta-cell function and insulin gene expression in response to variations in nutrient availability. We found that nutrient deprivation in beta cells promoted TFEB/TFE3 activation, which resulted in suppression of insulin gene expression. TFEB overexpression was sufficient to inhibit insulin transcription, whereas beta cells depleted of both TFEB and TFE3 failed to suppress insulin gene expression in response to amino acid deprivation. Interestingly, ChIP-seq analysis showed binding of TFEB to super-enhancer regions that regulate insulin transcription. Conditional, beta-cell-specific, Tfeb-overexpressing, and Tfeb/Tfe3 double-KO mice showed severe alteration of insulin transcription, secretion, and glucose tolerance, indicating that TFEB and TFE3 are important physiological mediators of pancreatic function. Our findings reveal a nutrient-controlled transcriptional mechanism that regulates insulin production, thus playing a key role in glucose homeostasis at both cellular and organismal levels.
MeSH term(s)	Animals ; Mice ; Autophagy/genetics ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/metabolism ; Gene Expression ; Glucose ; Insulin ; Lysosomes/metabolism
Chemical Substances	Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; Glucose (IY9XDZ35W2) ; Insulin ; Tcfe3 protein, mouse (136896-33-8) ; Tcfeb protein, mouse
Language	English
Publishing date	2023-09-15
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	586044-1
ISSN	1460-2075 ; 0261-4189
ISSN (online)	1460-2075
ISSN	0261-4189
DOI	10.15252/embj.2023113928
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