76 results on '"Rosita Accardi"'
Search Results
2. Epigenetic Alteration of the Cancer-Related Gene TGFBI in B Cells Infected with Epstein–Barr Virus and Exposed to Aflatoxin B1: Potential Role in Burkitt Lymphoma Development
- Author
-
Francesca Manara, Antonin Jay, Grace Akinyi Odongo, Fabrice Mure, Mohamed Ali Maroui, Audrey Diederichs, Cecilia Sirand, Cyrille Cuenin, Massimo Granai, Lucia Mundo, Hector Hernandez-Vargas, Stefano Lazzi, Rita Khoueiry, Henri Gruffat, Zdenko Herceg, and Rosita Accardi
- Subjects
Burkitt lymphoma ,EBV ,AFB1 ,DNA methylation ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Burkitt lymphoma (BL) is a malignant B cell neoplasm that accounts for almost half of pediatric cancers in sub-Saharan African countries. Although the BL endemic prevalence is attributable to the combination of Epstein–Barr virus (EBV) infection with malaria and environmental carcinogens exposure, such as the food contaminant aflatoxin B1 (AFB1), the molecular determinants underlying the pathogenesis are not fully understood. Consistent with the role of epigenetic mechanisms at the interface between the genome and environment, AFB1 and EBV impact the methylome of respectively leukocytes and B cells specifically. Here, we conducted a thorough investigation of common epigenomic changes following EBV or AFB1 exposure in B cells. Genome-wide DNA methylation profiling identified an EBV–AFB1 common signature within the TGFBI locus, which encodes for a putative tumor suppressor often altered in cancer. Subsequent mechanistic analyses confirmed a DNA-methylation-dependent transcriptional silencing of TGFBI involving the recruitment of DNMT1 methyltransferase that is associated with an activation of the NF-κB pathway. Our results reveal a potential common mechanism of B cell transformation shared by the main risk factors of endemic BL (EBV and AFB1), suggesting a key determinant of disease that could allow the development of more efficient targeted therapeutic strategies.
- Published
- 2022
- Full Text
- View/download PDF
3. Transforming Properties of Beta-3 Human Papillomavirus E6 and E7 Proteins
- Author
-
Lucia Minoni, Maria Carmen Romero-Medina, Assunta Venuti, Cécilia Sirand, Alexis Robitaille, Gennaro Altamura, Florence Le Calvez-Kelm, Daniele Viarisio, Katia Zanier, Martin Müller, Rosita Accardi, and Massimo Tommasino
- Subjects
beta-3 and mucosal HPV types ,p53 and pRb regulated pathways ,keratinocyte immortalization ,Microbiology ,QR1-502 - Abstract
ABSTRACT The beta human papillomaviruses (HPVs) are subdivided into 5 species (beta-1 to beta-5), and they were first identified in the skin. However, the beta-3 species appears to be more highly represented in the mucosal epithelia than in the skin. Functional studies have also highlighted that beta-3 HPV49 shares some functional similarities with mucosal high-risk (HR) HPV16. Here, we describe the characterization of the in vitro transforming properties of the entire beta-3 species, which includes three additional HPV types: HPV75, HPV76, and HPV115. HPV49, HPV75, and HPV76 E6 and E7 (E6/E7), but not HPV115 E6 and E7, efficiently inactivate the p53 and pRb pathways and immortalize or extend the life span of human foreskin keratinocytes (HFKs). As observed for HR HPV16, cell cycle deregulation mediated by beta-3 HPV E6/E7 expression leads to p16INK4a accumulation, whereas no p16INK4a was detected in beta-2 HPV38 E6/E7 HFKs. As shown for HPV49 E6, HPV75 and HPV76 E6s degrade p53 by an E6AP/proteasome-mediated mechanism. Comparative analysis of cellular gene expression patterns of HFKs containing E6 and E7 from HR HPV16, beta-3 HPV types, and beta-2 HPV38 further highlights the functional similarities of HR HPV16 and beta-3 HPV49, HPV75, and HPV76. The expression profiles of these four HPV HFKs show some similarities and diverge substantially from those of beta-3 HPV115 E6/E7 and beta-2 HPV38 E6/E7 HFKs. In summary, our data show that beta-3 HPV types share some mechanisms with HR HPV types and pave the way for additional studies aiming to evaluate their potential role in human pathologies. IMPORTANCE Human papillomaviruses are currently classified in different genera. Mucosal HPVs belonging to the alpha genus have been clearly associated with carcinogenesis of the mucosal epithelium at different sites. Beta HPV types have been classified as cutaneous. Although findings indicate that some beta HPVs from species 1 and 2 play a role, together with UV irradiation, in skin cancer, very little is known about the transforming properties of most of the beta HPVs. This report shows the transforming activity of E6 and E7 from beta-3 HPV types. Moreover, it highlights that beta-3 HPVs share some biological properties more extensively with mucosal high-risk HPV16 than with beta-2 HPV38. This report provides new paradigms for a better understanding of the biology of the different HPV types and their possible association with lesions at mucosal and/or cutaneous epithelia.
- Published
- 2020
- Full Text
- View/download PDF
4. Human papillomavirus type 38 alters wild-type p53 activity to promote cell proliferation via the downregulation of integrin alpha 1 expression.
- Author
-
Maria Carmen Romero-Medina, Assunta Venuti, Giusi Melita, Alexis Robitaille, Maria Grazia Ceraolo, Laura Pacini, Cecilia Sirand, Daniele Viarisio, Valerio Taverniti, Purnima Gupta, Mariafrancesca Scalise, Cesare Indiveri, Rosita Accardi, and Massimo Tommasino
- Subjects
Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
Tumor suppressors can exert pro-proliferation functions in specific contexts. In the beta human papillomavirus type 38 (HPV38) experimental model, the viral proteins E6 and E7 promote accumulation of a wild-type (WT) p53 form in human keratinocytes (HKs), promoting cellular proliferation. Inactivation of p53 by different means strongly decreases the proliferation of HPV38 E6/E7 HKs. This p53 form is phosphorylated at S392 by the double-stranded RNA-dependent protein kinase PKR, which is highly activated by HPV38. PKR-mediated S392 p53 phosphorylation promotes the formation of a p53/DNMT1 complex, which inhibits expression of integrin alpha 1 (ITGA1), a repressor of epidermal growth factor receptor (EGFR) signaling. Ectopic expression of ITGA1 in HPV38 E6/E7 HKs promotes EGFR degradation, inhibition of cellular proliferation, and cellular death. Itga1 expression was also inhibited in the skin of HPV38 transgenic mice that have an elevated susceptibility to UV-induced skin carcinogenesis. In summary, these findings reveal the existence of a specific WT p53 form that displays pro-proliferation properties.
- Published
- 2020
- Full Text
- View/download PDF
5. Viral driven epigenetic events alter the expression of cancer-related genes in Epstein-Barr-virus naturally infected Burkitt lymphoma cell lines
- Author
-
Hector Hernandez-Vargas, Henri Gruffat, Marie Pierre Cros, Audrey Diederichs, Cécilia Sirand, Romina C. Vargas-Ayala, Antonin Jay, Geoffroy Durand, Florence Le Calvez-Kelm, Zdenko Herceg, Evelyne Manet, Christopher P. Wild, Massimo Tommasino, and Rosita Accardi
- Subjects
Medicine ,Science - Abstract
Abstract Epstein-Barr virus (EBV) was identified as the first human virus to be associated with a human malignancy, Burkitt’s lymphoma (BL), a pediatric cancer endemic in sub-Saharan Africa. The exact mechanism of how EBV contributes to the process of lymphomagenesis is not fully understood. Recent studies have highlighted a genetic difference between endemic (EBV+) and sporadic (EBV−) BL, with the endemic variant showing a lower somatic mutation load, which suggests the involvement of an alternative virally-driven process of transformation in the pathogenesis of endemic BL. We tested the hypothesis that a global change in DNA methylation may be induced by infection with EBV, possibly thereby accounting for the lower mutation load observed in endemic BL. Our comparative analysis of the methylation profiles of a panel of BL derived cell lines, naturally infected or not with EBV, revealed that the presence of the virus is associated with a specific pattern of DNA methylation resulting in altered expression of cellular genes with a known or potential role in lymphomagenesis. These included ID3, a gene often found to be mutated in sporadic BL. In summary this study provides evidence that EBV may contribute to the pathogenesis of BL through an epigenetic mechanism.
- Published
- 2017
- Full Text
- View/download PDF
6. Unique DNA methylation signature in HPV-positive head and neck squamous cell carcinomas
- Author
-
Davide Degli Esposti, Athena Sklias, Sheila C. Lima, Stéphanie Beghelli-de la Forest Divonne, Vincent Cahais, Nora Fernandez-Jimenez, Marie-Pierre Cros, Szilvia Ecsedi, Cyrille Cuenin, Liacine Bouaoun, Graham Byrnes, Rosita Accardi, Anne Sudaka, Valérie Giordanengo, Hector Hernandez-Vargas, Luis Felipe Ribeiro Pinto, Ellen Van Obberghen-Schilling, and Zdenko Herceg
- Subjects
Head and neck squamous cell carcinomas ,HPV ,Differentially methylated regions ,CpG shores ,Predictive models ,Medicine ,Genetics ,QH426-470 - Abstract
Abstract Background Head and neck squamous cell carcinomas (HNSCCs) represent a heterogeneous group of cancers for which human papilloma virus (HPV) infection is an emerging risk factor. Previous studies showed promoter hypermethylation in HPV(+) oropharyngeal cancers, but only few consistent target genes have been so far described, and the evidence of a functional impact on gene expression is still limited. Methods We performed global and stratified pooled analyses of epigenome-wide data in HNSCCs based on the Illumina HumanMethylation450 bead-array data in order to identify tissue-specific components and common viral epigenetic targets in HPV-associated tumours. Results We identified novel differentially methylated CpGs and regions associated with viral infection that are independent of the anatomic site. In particular, most hypomethylated regions were characterized by a marked loss of CpG island boundaries, which showed significant correlations with expression of neighbouring genes. Moreover, a subset of only five CpGs in a few hypomethylated regions predicted HPV status with a high level of specificity in different cohorts. Finally, this signature was a better predictor of survival compared with HPV status determined by viral gene expression by RNA sequencing in The Cancer Genome Atlas cohort. Conclusions We identified a novel epigenetic signature of HPV infection in HNSCCs which is independent of the anatomic site, is functionally correlated with gene expression and may be leveraged for improved stratification of prognosis in HNSCCs.
- Published
- 2017
- Full Text
- View/download PDF
7. Beta HPV38 oncoproteins act with a hit-and-run mechanism in ultraviolet radiation-induced skin carcinogenesis in mice.
- Author
-
Daniele Viarisio, Karin Müller-Decker, Rosita Accardi, Alexis Robitaille, Matthias Dürst, Katrin Beer, Lars Jansen, Christa Flechtenmacher, Matthias Bozza, Richard Harbottle, Catherine Voegele, Maude Ardin, Jiri Zavadil, Sandra Caldeira, Lutz Gissmann, and Massimo Tommasino
- Subjects
Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
Cutaneous beta human papillomavirus (HPV) types are suspected to be involved, together with ultraviolet (UV) radiation, in the development of non-melanoma skin cancer (NMSC). Studies in in vitro and in vivo experimental models have highlighted the transforming properties of beta HPV E6 and E7 oncoproteins. However, epidemiological findings indicate that beta HPV types may be required only at an initial stage of carcinogenesis, and may become dispensable after full establishment of NMSC. Here, we further investigate the potential role of beta HPVs in NMSC using a Cre-loxP-based transgenic (Tg) mouse model that expresses beta HPV38 E6 and E7 oncogenes in the basal layer of the skin epidermis and is highly susceptible to UV-induced carcinogenesis. Using whole-exome sequencing, we show that, in contrast to WT animals, when exposed to chronic UV irradiation K14 HPV38 E6/E7 Tg mice accumulate a large number of UV-induced DNA mutations, which increase proportionally with the severity of the skin lesions. The mutation pattern detected in the Tg skin lesions closely resembles that detected in human NMSC, with the highest mutation rate in p53 and Notch genes. Using the Cre-lox recombination system, we observed that deletion of the viral oncogenes after development of UV-induced skin lesions did not affect the tumour growth. Together, these findings support the concept that beta HPV types act only at an initial stage of carcinogenesis, by potentiating the deleterious effects of UV radiation.
- Published
- 2018
- Full Text
- View/download PDF
8. Epidermal Growth Factor Receptor-Dependent Mutual Amplification between Netrin-1 and the Hepatitis C Virus.
- Author
-
Marie-Laure Plissonnier, Thomas Lahlali, Maud Michelet, Fanny Lebossé, Jessica Cottarel, Melanie Beer, Grégory Neveu, David Durantel, Birke Bartosch, Rosita Accardi, Sophie Clément, Andrea Paradisi, Mojgan Devouassoux-Shisheboran, Shirit Einav, Patrick Mehlen, Fabien Zoulim, and Romain Parent
- Subjects
Biology (General) ,QH301-705.5 - Abstract
Hepatitis C virus (HCV) is an oncogenic virus associated with the onset of hepatocellular carcinoma (HCC). The present study investigated the possible link between HCV infection and Netrin-1, a ligand for dependence receptors that sustains tumorigenesis, in particular in inflammation-associated tumors. We show that Netrin-1 expression is significantly elevated in HCV+ liver biopsies compared to hepatitis B virus (HBV+) and uninfected samples. Furthermore, Netrin-1 was upregulated in all histological stages of HCV+ hepatic lesions, from minimal liver fibrosis to cirrhosis and HCC, compared to histologically matched HCV- tissues. Both cirrhosis and HCV contributed to the induction of Netrin-1 expression, whereas anti-HCV treatment resulted in a reduction of Netrin-1 expression. In vitro, HCV increased the level and translation of Netrin-1 in a NS5A-La-related protein 1 (LARP1)-dependent fashion. Knockdown and forced expression experiments identified the receptor uncoordinated receptor-5 (UNC5A) as an antagonist of the Netrin-1 signal, though it did not affect the death of HCV-infected cells. Netrin-1 enhanced infectivity of HCV particles and promoted viral entry by increasing the activation and decreasing the recycling of the epidermal growth factor receptor (EGFR), a protein that is dysregulated in HCC. Netrin-1 and HCV are, therefore, reciprocal inducers in vitro and in patients, as seen from the increase in viral morphogenesis and viral entry, both phenomena converging toward an increase in the level of infectivity of HCV virions. This functional association involving a cancer-related virus and Netrin-1 argues for evaluating the implication of UNC5 receptor ligands in other oncogenic microbial species.
- Published
- 2016
- Full Text
- View/download PDF
9. Epstein-Barr virus down-regulates tumor suppressor DOK1 expression.
- Author
-
Maha Siouda, Cecilia Frecha, Rosita Accardi, Jiping Yue, Cyrille Cuenin, Henri Gruffat, Evelyne Manet, Zdenko Herceg, Bakary S Sylla, and Massimo Tommasino
- Subjects
Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
The DOK1 tumor suppressor gene encodes an adapter protein that acts as a negative regulator of several signaling pathways. We have previously reported that DOK1 expression is up-regulated upon cellular stress, via the transcription factor E2F1, and down-regulated in a variety of human malignancies due to aberrant hypermethylation of its promoter. Here we show that Epstein Barr virus (EBV) infection of primary human B-cells leads to the down-regulation of DOK1 gene expression via the viral oncoprotein LMP1. LMP1 alone induces recruitment to the DOK1 promoter of at least two independent inhibitory complexes, one containing E2F1/pRB/DNMT1 and another containing at least EZH2. These events result in tri-methylation of histone H3 at lysine 27 (H3K27me3) of the DOK1 promoter and gene expression silencing. We also present evidence that the presence of additional EBV proteins leads to further repression of DOK1 expression with an additional mechanism. Indeed, EBV infection of B-cells induces DNA methylation at the DOK1 promoter region including the E2F1 responsive elements that, in turn, lose the ability to interact with E2F complexes. Treatment of EBV-infected B-cell-lines with the methyl-transferase inhibitor 5-aza-2'-deoxycytidine rescues DOK1 expression. In summary, our data show the deregulation of DOK1 gene expression by EBV and provide novel insights into the regulation of the DOK1 tumor suppressor in viral-related carcinogenesis.
- Published
- 2014
- Full Text
- View/download PDF
10. Oncogenic human papillomaviruses activate the tumor-associated lens epithelial-derived growth factor (LEDGF) gene.
- Author
-
Jenny Leitz, Miriam Reuschenbach, Claudia Lohrey, Anja Honegger, Rosita Accardi, Massimo Tommasino, Manuel Llano, Magnus von Knebel Doeberitz, Karin Hoppe-Seyler, and Felix Hoppe-Seyler
- Subjects
Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
The expression of the human papillomavirus (HPV) E6/E7 oncogenes is crucial for HPV-induced malignant cell transformation. The identification of cellular targets attacked by the HPV oncogenes is critical for our understanding of the molecular mechanisms of HPV-associated carcinogenesis and may open novel therapeutic opportunities. Here, we identify the Lens Epithelial-Derived Growth Factor (LEDGF) gene as a novel cellular target gene for the HPV oncogenes. Elevated LEDGF expression has been recently linked to human carcinogenesis and can protect tumor cells towards different forms of cellular stress. We show that intracellular LEDGF mRNA and protein levels in HPV-positive cancer cells are critically dependent on the maintenance of viral oncogene expression. Ectopic E6/E7 expression stimulates LEDGF transcription in primary keratinocytes, at least in part via activation of the LEDGF promoter. Repression of endogenous LEDGF expression by RNA interference results in an increased sensitivity of HPV-positive cancer cells towards genotoxic agents. Immunohistochemical analyses of cervical tissue specimens reveal a highly significant increase of LEDGF protein levels in HPV-positive lesions compared to histologically normal cervical epithelium. Taken together, these results indicate that the E6/E7-dependent maintenance of intracellular LEDGF expression is critical for protecting HPV-positive cancer cells against various forms of cellular stress, including DNA damage. This could support tumor cell survival and contribute to the therapeutic resistance of cervical cancers towards genotoxic treatment strategies in the clinic.
- Published
- 2014
- Full Text
- View/download PDF
11. Epstein - Barr virus transforming protein LMP-1 alters B cells gene expression by promoting accumulation of the oncoprotein ΔNp73α.
- Author
-
Rosita Accardi, Ikbal Fathallah, Henri Gruffat, Giuseppe Mariggiò, Florence Le Calvez-Kelm, Catherine Voegele, Birke Bartosch, Hector Hernandez-Vargas, James McKay, Bakary S Sylla, Evelyne Manet, and Massimo Tommasino
- Subjects
Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
Many studies have proved that oncogenic viruses develop redundant mechanisms to alter the functions of the tumor suppressor p53. Here we show that Epstein-Barr virus (EBV), via the oncoprotein LMP-1, induces the expression of ΔNp73α, a strong antagonist of p53. This phenomenon is mediated by the LMP-1 dependent activation of c-Jun NH2-terminal kinase 1 (JNK-1) which in turn favours the recruitment of p73 to ΔNp73α promoter. A specific chemical inhibitor of JNK-1 or silencing JNK-1 expression strongly down-regulated ΔNp73α mRNA levels in LMP-1-containing cells. Accordingly, LMP-1 mutants deficient to activate JNK-1 did not induce ΔNp73α accumulation. The recruitment of p73 to the ΔNp73α promoter correlated with the displacement of the histone-lysine N-methyltransferase EZH2 which is part of the transcriptional repressive polycomb 2 complex. Inhibition of ΔNp73α expression in lymphoblastoid cells (LCLs) led to the stimulation of apoptosis and up-regulation of a large number of cellular genes as determined by whole transcriptome shotgun sequencing (RNA-seq). In particular, the expression of genes encoding products known to play anti-proliferative/pro-apoptotic functions, as well as genes known to be deregulated in different B cells malignancy, was altered by ΔNp73α down-regulation. Together, these findings reveal a novel EBV mechanism that appears to play an important role in the transformation of primary B cells.
- Published
- 2013
- Full Text
- View/download PDF
12. Interferon-β induces cellular senescence in cutaneous human papilloma virus-transformed human keratinocytes by affecting p53 transactivating activity.
- Author
-
Maria V Chiantore, Serena Vannucchi, Rosita Accardi, Massimo Tommasino, Zulema A Percario, Gabriele Vaccari, Elisabetta Affabris, Gianna Fiorucci, and Giovanna Romeo
- Subjects
Medicine ,Science - Abstract
Interferon (IFN)-β inhibits cell proliferation and affects cell cycle in keratinocytes transformed by both mucosal high risk Human Papilloma Virus (HPV) and cutaneous HPV E6 and E7 proteins. In particular, upon longer IFN-β treatments, cutaneous HPV38 expressing cells undergo senescence. IFN-β appears to induce senescence by upregulating the expression of the tumor suppressor PML, a well known IFN-induced gene. Indeed, experiments in gene silencing via specific siRNAs have shown that PML is essential in the execution of the senescence programme and that both p53 and p21 pathways are involved. IFN-β treatment leads to a modulation of p53 phosphorylation and acetylation status and a reduction in the expression of the p53 dominant negative ΔNp73. These effects allow the recovery of p53 transactivating activity of target genes involved in the control of cell proliferation. Taken together, these studies suggest that signaling through the IFN pathway might play an important role in cellular senescence. This additional understanding of IFN antitumor action and mechanisms influencing tumor responsiveness or resistance appears useful in aiding further promising development of biomolecular strategies in the IFN therapy of cancer.
- Published
- 2012
- Full Text
- View/download PDF
13. HPV16 E7-dependent transformation activates NHE1 through a PKA-RhoA-induced inhibition of p38alpha.
- Author
-
Rosa A Cardone, Giovanni Busco, Maria R Greco, Antonia Bellizzi, Rosita Accardi, Antonella Cafarelli, Stefania Monterisi, Pierluigi Carratù, Valeria Casavola, Angelo Paradiso, Massimo Tommasino, and Stephan J Reshkin
- Subjects
Medicine ,Science - Abstract
BACKGROUND:Neoplastic transformation originates from a large number of different genetic alterations. Despite this genetic variability, a common phenotype to transformed cells is cellular alkalinization. We have previously shown in human keratinocytes and a cell line in which transformation can be turned on and followed by the inducible expression of the E7 oncogene of human papillomavirus type 16 (HPV16), that intracellular alkalinization is an early and essential physiological event driven by the up-regulation of the Na/(+)H(+) exchanger isoform 1 (NHE1) and is necessary for the development of other transformed phenotypes and the in vivo tumor formation in nude mice. METHODOLOGY:Here, we utilize these model systems to elucidate the dynamic sequence of alterations of the upstream signal transduction systems leading to the transformation-dependent activation of NHE1. PRINCIPAL FINDINGS:We observe that a down-regulation of p38 MAPK activity is a fundamental step in the ability of the oncogene to transform the cell. Further, using pharmacological agents and transient transfections with dominant interfering, constitutively active, phosphorylation negative mutants and siRNA strategy to modify specific upstream signal transduction components that link HPV16 E7 oncogenic signals to up-regulation of the NHE1, we demonstrate that the stimulation of NHE1 activity is driven by an early rise in cellular cAMP resulting in the down-stream inhibition of p38 MAPK via the PKA-dependent phosphorylation of the small G-protein, RhoA, and its subsequent inhibition. CONCLUSIONS:All together these data significantly improve our knowledge concerning the basic cellular alterations involved in oncogene-driven neoplastic transformation.
- Published
- 2008
- Full Text
- View/download PDF
14. Supplementary Figure 3 from Novel ß-HPV49 Transgenic Mouse Model of Upper Digestive Tract Cancer
- Author
-
Massimo Tommasino, Lutz Gissmann, Christa Flechtenmacher, Rosita Accardi, Birgit Aengeneyndt, Ulrich Kloz, Paola Zanna, Karin Müller-Decker, and Daniele Viarisio
- Abstract
Alignment of E6 sequences from HPV16, HPV49, and HPV38 types.
- Published
- 2023
15. Supplementary Figure 2 from Novel ß-HPV49 Transgenic Mouse Model of Upper Digestive Tract Cancer
- Author
-
Massimo Tommasino, Lutz Gissmann, Christa Flechtenmacher, Rosita Accardi, Birgit Aengeneyndt, Ulrich Kloz, Paola Zanna, Karin Müller-Decker, and Daniele Viarisio
- Abstract
E6/E7 gene copy number and E6/E7 transcript levels in the different HPV Tg animal models.
- Published
- 2023
16. Supplementary Figure 1 from Novel ß-HPV49 Transgenic Mouse Model of Upper Digestive Tract Cancer
- Author
-
Massimo Tommasino, Lutz Gissmann, Christa Flechtenmacher, Rosita Accardi, Birgit Aengeneyndt, Ulrich Kloz, Paola Zanna, Karin Müller-Decker, and Daniele Viarisio
- Abstract
Tumor burden in WT and HPV E6/E7-Tg mouse lines of HPV type 38 line 187 and type 49 line 2 after DMBA/TPA treatment
- Published
- 2023
17. Data from Novel ß-HPV49 Transgenic Mouse Model of Upper Digestive Tract Cancer
- Author
-
Massimo Tommasino, Lutz Gissmann, Christa Flechtenmacher, Rosita Accardi, Birgit Aengeneyndt, Ulrich Kloz, Paola Zanna, Karin Müller-Decker, and Daniele Viarisio
- Abstract
The beta genus of human papillomaviruses (ß-HPV) includes approximately 50 different viral types that are subdivided into five species (ß-1 through ß-5). Nonmelanoma cancers may involve some ß-1 and ß-2 HPV types, but the biology of most ß-HPV types and their possible connections to human disease are still little characterized. In this study, we studied the effects of ß-3 type HPV49 in a novel transgenic (Tg) mouse model, using a cytokeratin K14 promoter to drive expression of the E6 and E7 genes from this virus in the basal skin epidermis and the mucosal epithelia of the digestive tract (K14 HPV49 E6/E7-Tg mice). Viral oncogene expression only marginally increased cellular proliferation in the epidermis of Tg animals, compared with wild-type littermates, and we observed no spontaneous tumor formation during their entire lifespan. However, we found that K14 HPV49 E6/E7-Tg mice were highly susceptible to upper digestive tract carcinogenesis upon initiation with 4-nitroquinoline 1-oxide (4NQO). This was a selective effect, as the same mice did not exhibit any skin lesions after chronic UV irradiation. Opposite results were observed in an analogous Tg model expressing the ß-2 HPV38 E6 and E7 oncogenes at the same anatomic sites. While these mice were highly susceptible to UV-induced skin carcinogenesis, as previously shown, they were little affected by 4NQO treatment. Overall, our findings highlight important differences in the biologic properties of certain ß-type HPV that affect their impact on carcinogenesis in an anatomic site-specific manner. Cancer Res; 76(14); 4216–25. ©2016 AACR.
- Published
- 2023
18. Supplementary Table 2 from Novel ß-HPV49 Transgenic Mouse Model of Upper Digestive Tract Cancer
- Author
-
Massimo Tommasino, Lutz Gissmann, Christa Flechtenmacher, Rosita Accardi, Birgit Aengeneyndt, Ulrich Kloz, Paola Zanna, Karin Müller-Decker, and Daniele Viarisio
- Abstract
Analysis of enriched pathways
- Published
- 2023
19. Legend supplementary Figures from Novel ß-HPV49 Transgenic Mouse Model of Upper Digestive Tract Cancer
- Author
-
Massimo Tommasino, Lutz Gissmann, Christa Flechtenmacher, Rosita Accardi, Birgit Aengeneyndt, Ulrich Kloz, Paola Zanna, Karin Müller-Decker, and Daniele Viarisio
- Abstract
Legends of supplementary Figures 1-3
- Published
- 2023
20. Supplementary Materials and Methods from Novel ß-HPV49 Transgenic Mouse Model of Upper Digestive Tract Cancer
- Author
-
Massimo Tommasino, Lutz Gissmann, Christa Flechtenmacher, Rosita Accardi, Birgit Aengeneyndt, Ulrich Kloz, Paola Zanna, Karin Müller-Decker, and Daniele Viarisio
- Abstract
Additional information of methods used in the study
- Published
- 2023
21. Supplementary Table 1 from Novel ß-HPV49 Transgenic Mouse Model of Upper Digestive Tract Cancer
- Author
-
Massimo Tommasino, Lutz Gissmann, Christa Flechtenmacher, Rosita Accardi, Birgit Aengeneyndt, Ulrich Kloz, Paola Zanna, Karin Müller-Decker, and Daniele Viarisio
- Abstract
Sequences of the different primers used for PCR and RT-PCR analyses.
- Published
- 2023
22. Transforming Properties of Beta-3 Human Papillomavirus E6 and E7 Proteins
- Author
-
Assunta Venuti, Maria Carmen Romero-Medina, Rosita Accardi, Florence Le Calvez-Kelm, Alexis Robitaille, Cecilia Sirand, Martin Müller, Lucia Minoni, Daniele Viarisio, Katia Zanier, Gennaro Altamura, Massimo Tommasino, Centre International de Recherche contre le Cancer - International Agency for Research on Cancer (CIRC - IARC), Organisation Mondiale de la Santé / World Health Organization Office (OMS / WHO), Centre international de Recherche sur le Cancer (CIRC), Biotechnologie et signalisation cellulaire (BSC), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut de recherche de l'Ecole de biotechnologie de Strasbourg (IREBS), Minoni, L., Romero-Medina, M. C., Venuti, A., Sirand, C., Robitaille, A., Altamura, G., Calvez-Kelm, F. L., Viarisio, D., Zanier, K., Muller, M., Accardi, R., and Tommasino, M.
- Subjects
0301 basic medicine ,Keratinocytes ,Male ,[SDV]Life Sciences [q-bio] ,viruses ,Papillomavirus E7 Proteins ,lcsh:QR1-502 ,Alphapapillomavirus ,medicine.disease_cause ,Sciences du Vivant [q-bio]/Cancer ,lcsh:Microbiology ,Foreskin ,Mice ,0302 clinical medicine ,Gene expression ,NIH 3T3 Cell ,Cells, Cultured ,Skin ,Human papillomavirus 16 ,Alphapapillomaviru ,virus diseases ,QR1-502 ,female genital diseases and pregnancy complications ,3. Good health ,medicine.anatomical_structure ,Papillomavirus E7 Protein ,030220 oncology & carcinogenesis ,beta-3 and mucosal hpv types ,Keratinocyte ,beta-3 and mucosal HPV type ,Human ,Research Article ,keratinocyte immortalization ,Alpha (ethology) ,Biology ,Microbiology ,Host-Microbe Biology ,p53 and pRb regulated pathway ,03 medical and health sciences ,medicine ,Animals ,Humans ,p53 and prb regulated pathways ,Beta (finance) ,Molecular Biology ,Epithelial Cell ,Mucous Membrane ,Animal ,Mechanism (biology) ,Epithelial Cells ,Oncogene Proteins, Viral ,medicine.disease ,In vitro ,030104 developmental biology ,Cancer research ,NIH 3T3 Cells ,Skin cancer ,Carcinogenesis - Abstract
Human papillomaviruses are currently classified in different genera. Mucosal HPVs belonging to the alpha genus have been clearly associated with carcinogenesis of the mucosal epithelium at different sites. Beta HPV types have been classified as cutaneous. Although findings indicate that some beta HPVs from species 1 and 2 play a role, together with UV irradiation, in skin cancer, very little is known about the transforming properties of most of the beta HPVs. This report shows the transforming activity of E6 and E7 from beta-3 HPV types. Moreover, it highlights that beta-3 HPVs share some biological properties more extensively with mucosal high-risk HPV16 than with beta-2 HPV38. This report provides new paradigms for a better understanding of the biology of the different HPV types and their possible association with lesions at mucosal and/or cutaneous epithelia., The beta human papillomaviruses (HPVs) are subdivided into 5 species (beta-1 to beta-5), and they were first identified in the skin. However, the beta-3 species appears to be more highly represented in the mucosal epithelia than in the skin. Functional studies have also highlighted that beta-3 HPV49 shares some functional similarities with mucosal high-risk (HR) HPV16. Here, we describe the characterization of the in vitro transforming properties of the entire beta-3 species, which includes three additional HPV types: HPV75, HPV76, and HPV115. HPV49, HPV75, and HPV76 E6 and E7 (E6/E7), but not HPV115 E6 and E7, efficiently inactivate the p53 and pRb pathways and immortalize or extend the life span of human foreskin keratinocytes (HFKs). As observed for HR HPV16, cell cycle deregulation mediated by beta-3 HPV E6/E7 expression leads to p16INK4a accumulation, whereas no p16INK4a was detected in beta-2 HPV38 E6/E7 HFKs. As shown for HPV49 E6, HPV75 and HPV76 E6s degrade p53 by an E6AP/proteasome-mediated mechanism. Comparative analysis of cellular gene expression patterns of HFKs containing E6 and E7 from HR HPV16, beta-3 HPV types, and beta-2 HPV38 further highlights the functional similarities of HR HPV16 and beta-3 HPV49, HPV75, and HPV76. The expression profiles of these four HPV HFKs show some similarities and diverge substantially from those of beta-3 HPV115 E6/E7 and beta-2 HPV38 E6/E7 HFKs. In summary, our data show that beta-3 HPV types share some mechanisms with HR HPV types and pave the way for additional studies aiming to evaluate their potential role in human pathologies. IMPORTANCE Human papillomaviruses are currently classified in different genera. Mucosal HPVs belonging to the alpha genus have been clearly associated with carcinogenesis of the mucosal epithelium at different sites. Beta HPV types have been classified as cutaneous. Although findings indicate that some beta HPVs from species 1 and 2 play a role, together with UV irradiation, in skin cancer, very little is known about the transforming properties of most of the beta HPVs. This report shows the transforming activity of E6 and E7 from beta-3 HPV types. Moreover, it highlights that beta-3 HPVs share some biological properties more extensively with mucosal high-risk HPV16 than with beta-2 HPV38. This report provides new paradigms for a better understanding of the biology of the different HPV types and their possible association with lesions at mucosal and/or cutaneous epithelia.
- Published
- 2020
23. Human papillomavirus type 38 alters wild-type p53 activity to promote cell proliferation via the downregulation of integrin alpha 1 expression
- Author
-
Valerio Taverniti, Rosita Accardi, Alexis Robitaille, Cecilia Sirand, Giusi Melita, Daniele Viarisio, Assunta Venuti, Maria Grazia Ceraolo, Mariafrancesca Scalise, Laura Pacini, Massimo Tommasino, Maria Carmen Romero-Medina, Cesare Indiveri, and Purnima Gupta
- Subjects
Keratinocytes ,Small interfering RNA ,Integrins ,Skin Neoplasms ,Protein Extraction ,Papillomavirus E7 Proteins ,Gene Expression ,Biochemistry ,Suppressor Genes ,Mice ,Epidermal growth factor receptor ,Biology (General) ,Post-Translational Modification ,Phosphorylation ,Papillomaviridae ,Cells, Cultured ,Extraction Techniques ,0303 health sciences ,biology ,Chemistry ,030302 biochemistry & molecular biology ,Cell biology ,Precipitation Techniques ,Extracellular Matrix ,Nucleic acids ,Cellular Structures and Organelles ,Research Article ,QH301-705.5 ,Tumor Suppressor Genes ,Immunology ,Repressor ,Down-Regulation ,Mice, Transgenic ,Research and Analysis Methods ,Transfection ,Microbiology ,03 medical and health sciences ,Downregulation and upregulation ,Gene Types ,Virology ,Genetics ,Cell Adhesion ,Animals ,Humans ,Immunoprecipitation ,Molecular Biology Techniques ,Non-coding RNA ,Molecular Biology ,030304 developmental biology ,Cell Proliferation ,Papillomavirus Infections ,Wild type ,Membrane Proteins ,Biology and Life Sciences ,Proteins ,Oncogene Proteins, Viral ,Cell Biology ,RC581-607 ,Protein kinase R ,Gene regulation ,Repressor Proteins ,biology.protein ,RNA ,Parasitology ,Ectopic expression ,Immunologic diseases. Allergy ,Tumor Suppressor Protein p53 ,Carrier Proteins - Abstract
Tumor suppressors can exert pro-proliferation functions in specific contexts. In the beta human papillomavirus type 38 (HPV38) experimental model, the viral proteins E6 and E7 promote accumulation of a wild-type (WT) p53 form in human keratinocytes (HKs), promoting cellular proliferation. Inactivation of p53 by different means strongly decreases the proliferation of HPV38 E6/E7 HKs. This p53 form is phosphorylated at S392 by the double-stranded RNA-dependent protein kinase PKR, which is highly activated by HPV38. PKR-mediated S392 p53 phosphorylation promotes the formation of a p53/DNMT1 complex, which inhibits expression of integrin alpha 1 (ITGA1), a repressor of epidermal growth factor receptor (EGFR) signaling. Ectopic expression of ITGA1 in HPV38 E6/E7 HKs promotes EGFR degradation, inhibition of cellular proliferation, and cellular death. Itga1 expression was also inhibited in the skin of HPV38 transgenic mice that have an elevated susceptibility to UV-induced skin carcinogenesis. In summary, these findings reveal the existence of a specific WT p53 form that displays pro-proliferation properties., Author summary This study shows that beta HPV38 can convert p53 functions from a tumor suppressor to an oncoprotein via the formation of a transcriptionally repressive complex, which in turn represses ITGA1 expression, promoting cellular proliferation and UV-induced skin carcinogenesis.
- Published
- 2020
24. Interplay between the Epigenetic Enzyme Lysine (K)-Specific Demethylase 2B and Epstein-Barr Virus Infection
- Author
-
Mohamed Ali Maroui, Maria Carmen Romero-Medina, Marie Pierre Cros, Florence Le Calvez-Kelm, Romina C. Vargas-Ayala, Lucia Mundo, Geoffroy Durand, Antonin Jay, Hector Hernandez-Vargas, Maria Grazia Ceraolo, Alexis Robitaille, Cecilia Sirand, Lorenzo Leoncini, Evelyne Manet, Francesca Manara, Zdenko Herceg, Cyrille Cuenin, Henri Gruffat, Audrey Diederichs, Rosita Accardi, Centre International de Recherche contre le Cancer - International Agency for Research on Cancer (CIRC - IARC), Organisation Mondiale de la Santé / World Health Organization Office (OMS / WHO), Herpesvirus oncogènes – Oncogenic Herpesviruses, Centre International de Recherche en Infectiologie - UMR (CIRI), Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche en Cancérologie de Lyon (UNICANCER/CRCL), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Department of Biotechnology Chemistry and Pharmacy (University of Siena), University of Siena, Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and Università degli Studi di Siena = University of Siena (UNISI)
- Subjects
0301 basic medicine ,Male ,Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Jumonji Domain-Containing Histone Demethylases ,KDM2B ,Epigenesis, Genetic ,0302 clinical medicine ,[SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases ,hemic and lymphatic diseases ,Burkitt lymphomas ,EBV ,Epigenetic ,Child ,ComputingMilieux_MISCELLANEOUS ,B-Lymphocytes ,biology ,Middle Aged ,Burkitt Lymphoma ,Chromatin ,3. Good health ,Virus-Cell Interactions ,030220 oncology & carcinogenesis ,Child, Preschool ,DNA methylation ,[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,Female ,Oncovirus ,Adult ,Chromatin Immunoprecipitation ,Adolescent ,Immunology ,Down-Regulation ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Microbiology ,Cell Line ,03 medical and health sciences ,Young Adult ,Virology ,Humans ,Epigenetics ,F-Box Proteins ,[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,Epigenome ,DNA Methylation ,030104 developmental biology ,Gene Expression Regulation ,Insect Science ,Cancer research ,biology.protein ,Demethylase ,Chromatin immunoprecipitation - Abstract
The histone modifier lysine (K)-specific demethylase 2B (KDM2B) plays a role in the differentiation of hematopoietic cells, and its expression appears to be deregulated in certain cancers of hematological and lymphoid origins. We have previously found that the KDM2B gene is differentially methylated in cell lines derived from Epstein-Barr virus (EBV)-associated endemic Burkitt lymphoma (eBL) compared with that in EBV-negative sporadic Burkitt lymphoma-derived cells. However, whether KDM2B plays a role in eBL development has not been previously investigated. Oncogenic viruses have been shown to hijack the host cell epigenome to complete their life cycle and to promote the transformation process by perturbing cell chromatin organization. Here, we investigated whether EBV alters KDM2B levels to enable its life cycle and promote B-cell transformation. We show that infection of B cells with EBV leads to downregulation of KDM2B levels. We also show that LMP1, one of the main EBV transforming proteins, induces increased DNMT1 recruitment to the KDM2B gene and augments its methylation. By altering KDM2B levels and performing chromatin immunoprecipitation in EBV-infected B cells, we show that KDM2B is recruited to the EBV gene promoters and inhibits their expression. Furthermore, forced KDM2B expression in immortalized B cells led to altered mRNA levels of some differentiation-related genes. Our data show that EBV deregulates KDM2B levels through an epigenetic mechanism and provide evidence for a role of KDM2B in regulating virus and host cell gene expression, warranting further investigations to assess the role of KDM2B in the process of EBV-mediated lymphomagenesis. IMPORTANCE In Africa, Epstein-Barr virus infection is associated with endemic Burkitt lymphoma, a pediatric cancer. The molecular events leading to its development are poorly understood compared with those leading to sporadic Burkitt lymphoma. In a previous study, by analyzing the DNA methylation changes in endemic compared with sporadic Burkitt lymphoma cell lines, we identified several differential methylated genomic positions in the proximity of genes with a potential role in cancer, and among them was the KDM2B gene. KDM2B encodes a histone H3 demethylase already shown to be involved in some hematological disorders. However, whether KDM2B plays a role in the development of Epstein-Barr virus-mediated lymphoma has not been investigated before. In this study, we show that Epstein-Barr virus deregulates KDM2B expression and describe the underlying mechanisms. We also reveal a role of the demethylase in controlling viral and B-cell gene expression, thus highlighting a novel interaction between the virus and the cellular epigenome.
- Published
- 2019
25. Merkel Cell Polyomavirus Downregulates N-myc Downstream-Regulated Gene 1, Leading to Cellular Proliferation and Migration
- Author
-
Laura Pacini, Lise Brault, Naveed Shahzad, Hector Hernandez-Vargas, Alexis Robitaille, Purnima Gupta, Tarik Gheit, Alexis Harold, Geoffroy Durand, Maria Carmen Romero-Medina, Masahiro Shuda, Rosita Accardi, Assunta Venuti, Florence Le Calvez-Kelm, Massimo Tommasino, and Valerio Taverniti
- Subjects
Gene Expression Regulation, Viral ,Keratinocytes ,Skin Neoplasms ,Tumor suppressor gene ,Carcinogenesis ,Immunology ,Merkel cell polyomavirus ,Down-Regulation ,Cell Cycle Proteins ,medicine.disease_cause ,Microbiology ,Cell Line ,03 medical and health sciences ,0302 clinical medicine ,Cyclin D1 ,Cell Movement ,Virology ,medicine ,Humans ,Antigens, Viral, Tumor ,030304 developmental biology ,Cell Proliferation ,Skin ,0303 health sciences ,Gene knockdown ,Polyomavirus Infections ,biology ,Merkel cell carcinoma ,Cell growth ,Intracellular Signaling Peptides and Proteins ,biology.organism_classification ,medicine.disease ,Virus-Cell Interactions ,Carcinoma, Merkel Cell ,Gene Expression Regulation, Neoplastic ,Tumor Virus Infections ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Insect Science ,Cancer research ,Merkel cell ,Transcriptome - Abstract
Merkel cell polyomavirus (MCPyV) is the first human polyomavirus etiologically associated with Merkel cell carcinoma (MCC), a rare and aggressive form of skin cancer. Similar to other polyomaviruses, MCPyV encodes early T antigen genes, viral oncogenes required for MCC tumor growth. To identify the unique oncogenic properties of MCPyV, we analyzed the gene expression profiles in human spontaneously immortalized keratinocytes (NIKs) expressing the early genes from six distinct human polyomaviruses (PyVs), including MCPyV. A comparison of the gene expression profiles revealed 28 genes specifically deregulated by MCPyV. In particular, the MCPyV early gene downregulated the expression of the tumor suppressor gene N-myc downstream-regulated gene 1 (NDRG1) in MCPyV gene-expressing NIKs and hTERT-MCPyV gene-expressing human keratinocytes (HK) compared to their expression in the controls. In MCPyV-positive MCC cells, the expression of NDRG1 was downregulated by the MCPyV early gene, as T antigen knockdown rescued the level of NDRG1. In addition, NDRG1 overexpression in hTERT-MCPyV gene-expressing HK or MCC cells resulted in a decrease in the number of cells in S phase and cell proliferation inhibition. Moreover, a decrease in wound healing capacity in hTERT-MCPyV gene-expressing HK was observed. Further analysis revealed that NDRG1 exerts its biological effect in Merkel cell lines by regulating the expression of the cyclin-dependent kinase 2 (CDK2) and cyclin D1 proteins. Overall, NDRG1 plays an important role in MCPyV-induced cellular proliferation. IMPORTANCE Merkel cell carcinoma was first described in 1972 as a neuroendocrine tumor of skin, most cases of which were reported in 2008 to be caused by a PyV named Merkel cell polyomavirus (MCPyV), the first PyV linked to human cancer. Thereafter, numerous studies have been conducted to understand the etiology of this virus-induced carcinogenesis. However, it is still a new field, and much work is needed to understand the molecular pathogenesis of MCC. In the current work, we sought to identify the host genes specifically deregulated by MCPyV, as opposed to other PyVs, in order to better understand the relevance of the genes analyzed on the biological impact and progression of the disease. These findings open newer avenues for targeted drug therapies, thereby providing hope for the management of patients suffering from this highly aggressive cancer.
- Published
- 2019
26. Transforming properties of Felis catus papillomavirus type 2 E6 and E7 putative oncogenes in vitro and their transcriptional activity in feline squamous cell carcinoma in vivo
- Author
-
Annunziata Corteggio, Giovanna Maria Pierantoni, Andrea Conte, Massimo Tommasino, Gennaro Altamura, Giuseppe Borzacchiello, Laura Pacini, Rosita Accardi, Altamura, Gennaro, Corteggio, Annunziata, Pacini, Laura, Conte, Andrea, Pierantoni, GIOVANNA MARIA, Tommasino, Massimo, Accardi, Rosita, and Borzacchiello, Giuseppe
- Subjects
Gene Expression Regulation, Viral ,Transcriptional Activation ,0301 basic medicine ,Felis catus papillomaviru ,040301 veterinary sciences ,Papillomavirus E7 Proteins ,Veterinary medicine ,Cell ,Cyclin A ,Oncogenic viruse ,Virus ,Cell Line ,0403 veterinary science ,Mice ,03 medical and health sciences ,Downregulation and upregulation ,Virology ,medicine ,Animals ,RNA, Messenger ,Papillomaviridae ,Gene ,E7 ,E6 ,Cyclin-dependent kinase 1 ,biology ,Papillomavirus Infections ,Oncogene Proteins, Viral ,Feline oncology ,04 agricultural and veterinary sciences ,Molecular biology ,Veterinary virology ,In vitro ,030104 developmental biology ,medicine.anatomical_structure ,Carcinoma, Squamous Cell ,Cats ,biology.protein ,Carrier Proteins ,Feline SCC ,Oncovirus ,Protein Binding ,Signal Transduction - Abstract
Felis catus papillomavirus type 2 (FcaPV2) DNA is found in feline cutaneous squamous cell carcinomas (SCCs); however, its biological properties are still uncharacterized. In this study, we successfully expressed FcaPV2 E6 and E7 putative oncogenes in feline epithelial cells and demonstrated that FcaPV2 E6 binds to p53, impairing its protein level. In addition, E6 and E7 inhibited ultraviolet B (UVB)-triggered accumulation of p53, p21 and pro-apoptotic markers such as Cleaved Caspase3, Bax and Bak, suggesting a synergistic action of the virus with UV exposure in tumour pathogenesis. Furthermore, FcaPV2 E7 bound to feline pRb and impaired pRb levels, resulting in upregulation of the downstream pro-proliferative genes Cyclin A and Cdc2. Importantly, we demonstrated mRNA expression of FcaPV2 E2, E6 and E7 in feline SCC samples, strengthening the hypothesis of a causative role in the development of feline SCC.
- Published
- 2016
27. Downregulation of Toll-Like Receptor 9 Expression by Beta Human Papillomavirus 38 and Implications for Cell Cycle Control
- Author
-
Claudia Savini, Laura Pacini, Uzma Hasan, Rosita Accardi, Raffaella Ghittoni, Jérôme Lamartine, Massimo Tommasino, and Djamel Saidj
- Subjects
Keratinocytes ,Papillomavirus E7 Proteins ,Immunology ,Down-Regulation ,Biology ,Methylation ,Microbiology ,Cell Line ,Histones ,Immune system ,RNA interference ,Virology ,Humans ,Gene silencing ,Enhancer of Zeste Homolog 2 Protein ,RNA, Messenger ,RNA, Small Interfering ,Kinase activity ,Promoter Regions, Genetic ,Papillomaviridae ,Cell Proliferation ,Innate immune system ,Cyclin-Dependent Kinase 2 ,Papillomavirus Infections ,Polycomb Repressive Complex 2 ,Cell Cycle Checkpoints ,Cell cycle ,Molecular biology ,I-kappa B Kinase ,Virus-Cell Interactions ,Cell biology ,p21-Activated Kinases ,Toll-Like Receptor 9 ,Insect Science ,RNA, Viral ,RNA Interference ,Ectopic expression ,Cyclin-Dependent Kinase Inhibitor p27 - Abstract
Innate immunity is the first line of host defense against infections. Many oncogenic viruses can deregulate several immune-related pathways to guarantee the persistence of the infection. Here, we show that the cutaneous human papillomavirus 38 (HPV38) E6 and E7 oncoproteins suppress the expression of the double-stranded DNA sensor Toll-like receptor 9 (TLR9) in human foreskin keratinocytes (HFK), a key mediator of the antiviral innate immune host response. In particular, HPV38 E7 induces TLR9 mRNA downregulation by promoting accumulation of ΔNp73α, an antagonist of p53 and p73. Inhibition of ΔNp73α expression by antisense oligonucleotide in HPV38 E6/E7 HFK strongly rescues mRNA levels of TLR9, highlighting a key role of ΔNp73α in this event. Chromatin immunoprecipitation experiments showed that ΔNp73α is part of a negative transcriptional regulatory complex with IκB kinase beta (IKKβ) that binds to a NF-κB responsive element within the TLR9 promoter. In addition, the Polycomb protein enhancer of zeste homolog 2 (EZH2), responsible for gene expression silencing, is also recruited into the complex, leading to histone 3 trimethylation at lysine 27 (H3K27me3) in the same region of the TLR9 promoter. Ectopic expression of TLR9 in HPV38 E6/E7 cells resulted in an accumulation of the cell cycle inhibitors p21 WAF1 and p27 Kip1 , decreased CDK2-associated kinase activity, and inhibition of cellular proliferation. In summary, our data show that HPV38, similarly to other viruses with well-known oncogenic activity, can downregulate TLR9 expression. In addition, they highlight a new role for TLR9 in cell cycle regulation. IMPORTANCE The mucosal high-risk HPV types have been clearly associated with human carcinogenesis. Emerging lines of evidence suggest the involvement of certain cutaneous HPV types in development of skin squamous cell carcinoma, although this association is still under debate. Oncogenic viruses have evolved different strategies to hijack the host immune system in order to guarantee the persistence of the infection. Their capability to evade the immune system is as important as their ability to promote cellular transformation. Therefore, understanding the viral mechanisms involved in viral persistence is a valid tool to evaluate their potential role in human carcinogenesis. Here, we show that E6 and E7 oncoproteins from the cutaneous HPV38 downregulate the expression of the double-stranded DNA sensor TLR9 of innate immunity. We also present evidence that the HPV38-mediated downregulation of TLR9 expression, in addition to its potential impact on the innate immune response, is linked to cell cycle deregulation.
- Published
- 2015
28. Interplay between the epigenetic enzyme lysine (K)-specific demethylase 2B and Epstein-Barr virus infection
- Author
-
Hector Hernandez-Vargas, Mohamed Ali Maroui, Lorenzo Leoncini, Marie Pierre Cros, Henri Gruffat, Evelyne Manet, Geoffroy Durand, Antonin Jay, Alexis Robitaille, Cecilia Sirand, Maria Grazia Ceraolo, Maria Carmen Romero-Medina, Florence Le Calvez-Kelm, Lucia Mundo, Cyrille Cuenin, Romina C. Vargas-Ayala, Zdenko Herceg, Audrey Diederichs, and Rosita Accardi
- Subjects
biology ,hemic and lymphatic diseases ,DNMT1 ,biology.protein ,Cancer research ,Demethylase ,KDM2B ,Epigenome ,Epigenetics ,Chromatin immunoprecipitation ,Oncovirus ,Chromatin - Abstract
Histone modifier lysine (K)-specific demethylase 2B(KDM2B) plays a role in hematopoietic cells differentiation and its expression appears to be deregulated in certain cancers of hematological and lymphoid origins. We have previously found that KDM2B gene is differentially methylated in cell lines derived from the Epstein-Barr virus (EBV) associated endemic Burkitt’s lymphomas (eBL) compared to EBV negative sporadic BL cells. However, whether KDM2B plays a role in eBL development has never been previously demonstrated. Oncogenic viruses have been shown to hijack the host cell epigenome to complete their life cycle and to promote the transformation process by perturbing cell chromatin organization. Here we investigated whether EBV would alter KDM2B levels to enable its life cycle and promote B-cells transformation. We show that infection of B-cells with EBV leads to down-regulation of KDM2B levels. We also show that LMP1, one of the main EBV transforming proteins, induces increased DNMT1 recruitment to KDM2B gene and augments its methylation. By altering KDM2B levels and performing chromatin immunoprecipitation in EBV infected B-cells, we were able to show that KDM2B is recruited to the EBV gene promoters and inhibits their expression. Furthermore, forced KDM2B expression in immortalized B-cells led to altered mRNA levels of some differentiation-related genes. Our data show that EBV deregulates KDM2B levels through an epigenetic mechanism and provide evidence for a role of KDM2B in regulating virus and host cell gene expression, warranting further investigations to assess the role of KDM2B in the process of EBV-mediated lymphomagenesis.IMPORTANCE. In Africa, Epstein-Barr virus infection is associated with endemic Burkitt lymphoma, a pediatric cancer. The molecular events leading to its development are poorly understood compared to the sporadic Burkitt lymphoma. In a previous study, by analyzing the DNA methylation changes in endemic compared to sporadic Burkitt lymphomas cell lines, we identified several differential methylated genomic positions in proximity of genes with a potential role in cancer, among them the KDM2B gene. KDM2B encodes a histone H3 demethylase already shown to be involved in some hematological disorders. However, whether KDM2B plays a role in the development of Epstein-Barr virus-mediated lymphoma has never been investigated before. In this study we show that Epstein-Barr virus deregulates KDM2B expression and describe the underlying mechanisms. We also reveal a role of the demethylase in controlling viral and B-cells genes expression, thus highlighting a novel interaction between the virus and the cellular epigenome.
- Published
- 2018
29. Human Papillomavirus E6 and E7 oncoproteins affect the cell microenvironment by classical secretion and extracellular vesicles delivery of inflammatory mediators
- Author
-
Giovanna Romeo, Maria Simona Zangrillo, Rosita Accardi, Giorgio Mangino, Massimo Tommasino, Maria Vincenza Chiantore, Gianna Fiorucci, and Marco Iuliano
- Subjects
0301 basic medicine ,Chemokine ,inflammatory mediators ,Papillomavirus E7 Proteins ,Immunology ,Inflammation ,medicine.disease_cause ,Biochemistry ,Malignant transformation ,Cell Line ,03 medical and health sciences ,Extracellular Vesicles ,0302 clinical medicine ,Immune system ,human papilloma virus ,medicine ,Extracellular ,Immunology and Allergy ,Humans ,Gene Silencing ,RNA, Messenger ,Molecular Biology ,Human Papilloma Virus ,Tumor virology ,Inflammatory mediators ,Extracellular vesicles ,Tumor microenvironment ,biology ,Microvesicle ,Hematology ,Oncogene Proteins, Viral ,tumor virology ,Repressor Proteins ,030104 developmental biology ,Cellular Microenvironment ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,medicine.symptom ,Inflammation Mediators ,Carcinogenesis - Abstract
The connection between chronic inflammation and risk of cancer has been supported by several studies. The development of cancer might be a process driven by the presence of a specific combination of inflammatory mediators, including cytokines, chemokines and enzymes, in the tumor microenvironment. Virus-induced tumors, like HPV-induced Squamous Cell Carcinomas, represent a paradigmatic example of the interplay between inflammation, as integral part of the innate antiviral response, and malignant transformation. Here, the role of inflammatory microenvironment in the HPV-induced carcinogenesis is addressed, with a specific focus on the involvement of the immune molecules as well as their delivery through the microvesicle cargo possibly correlated to the different HPV genotype. The expression of the inflammatory mediators in HPV positive cells has been analyzed in primary human foreskin keratinocytes and keratinocytes transduced by E6 and E7 from mucosal HPV-16 or cutaneous HPV-38 genotypes. HPV E6 and E7 proteins can modulate the expression of immune mediators in HPV-infected cells and can affect the levels of immune molecules, mainly chemokines, in the extracellular milieu. HPV-16 E6 and E7 oncoproteins have been silenced to confirm the specificity of the modulation of the inflammatory microenvironment. Our results suggest that the expression of HPV oncoproteins allows the modification of the tumor milieu through the synthesis and release of specific pro-inflammatory cytokines and chemokines, affecting the efficacy of the immune response. The microenvironment can also be conditioned by an altered mRNA cargo delivered by extracellular vesicles, thereby efficiently affecting the surrounding cells with possible implication for tumorigenesis and tumor diagnosis.
- Published
- 2017
30. Viral driven epigenetic events alter the expression of cancer-related genes in Epstein-Barr-virus naturally infected Burkitt lymphoma cell lines
- Author
-
Marie Pierre Cros, Evelyne Manet, Geoffroy Durand, Antonin Jay, Christopher P. Wild, Florence Le Calvez-Kelm, Audrey Diederichs, Rosita Accardi, Hector Hernandez-Vargas, Henri Gruffat, Romina C. Vargas-Ayala, Cecilia Sirand, Massimo Tommasino, Zdenko Herceg, Centre International de Recherche contre le Cancer - International Agency for Research on Cancer (CIRC - IARC), Organisation Mondiale de la Santé / World Health Organization Office (OMS / WHO), Herpesvirus oncogènes – Oncogenic Herpesviruses, Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre International de Recherche en Infectiologie - UMR (CIRI), Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)
- Subjects
0301 basic medicine ,Herpesvirus 4, Human ,Science ,Down-Regulation ,Biology ,medicine.disease_cause ,Virus ,Article ,Epigenesis, Genetic ,Viral Matrix Proteins ,03 medical and health sciences ,Germline mutation ,hemic and lymphatic diseases ,Cell Line, Tumor ,medicine ,Humans ,Epigenetics ,Gene Silencing ,RNA, Messenger ,Regulation of gene expression ,Mutation ,Multidisciplinary ,DNA Methylation ,[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology ,Pediatric cancer ,Virology ,Epstein–Barr virus ,Burkitt Lymphoma ,3. Good health ,Neoplasm Proteins ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,DNA methylation ,Medicine ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,CpG Islands ,Inhibitor of Differentiation Proteins - Abstract
Epstein-Barr virus (EBV) was identified as the first human virus to be associated with a human malignancy, Burkitt’s lymphoma (BL), a pediatric cancer endemic in sub-Saharan Africa. The exact mechanism of how EBV contributes to the process of lymphomagenesis is not fully understood. Recent studies have highlighted a genetic difference between endemic (EBV+) and sporadic (EBV−) BL, with the endemic variant showing a lower somatic mutation load, which suggests the involvement of an alternative virally-driven process of transformation in the pathogenesis of endemic BL. We tested the hypothesis that a global change in DNA methylation may be induced by infection with EBV, possibly thereby accounting for the lower mutation load observed in endemic BL. Our comparative analysis of the methylation profiles of a panel of BL derived cell lines, naturally infected or not with EBV, revealed that the presence of the virus is associated with a specific pattern of DNA methylation resulting in altered expression of cellular genes with a known or potential role in lymphomagenesis. These included ID3, a gene often found to be mutated in sporadic BL. In summary this study provides evidence that EBV may contribute to the pathogenesis of BL through an epigenetic mechanism.
- Published
- 2017
31. UV Radiation Activates Toll-Like Receptor 9 Expression in Primary Human Keratinocytes, an Event Inhibited by Human Papillomavirus 38 E6 and E7 Oncoproteins
- Author
-
Assunta Venuti, Giusi Melita, Uzma Hasan, Maria Grazia Ceraolo, Laura Pacini, Massimo Tommasino, Rosita Accardi, Centre International de Recherche contre le Cancer - International Agency for Research on Cancer (CIRC - IARC), Organisation Mondiale de la Santé / World Health Organization Office (OMS / WHO), Università degli Studi di Milano = University of Milan (UNIMI), DIBIT, San Raffaele Scientific Institute, Réponse immunitaire innée dans les maladies infectieuses et auto-immunes – Innate immunity in infectious and autoimmune diseases, Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Service d'immunologie [Centre Hospitalier Lyon Sud - HCL], Centre Hospitalier Lyon Sud [CHU - HCL] (CHLS), Hospices Civils de Lyon (HCL)-Hospices Civils de Lyon (HCL), University of Milan, Centre International de Recherche en Infectiologie - UMR (CIRI), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
0301 basic medicine ,HPV38 ,Keratinocytes ,p53 ,Skin Neoplasms ,Papillomavirus E7 Proteins ,medicine.disease_cause ,Cell Transformation ,0302 clinical medicine ,RNA interference ,Cellular stress response ,Gene expression ,RNA, Small Interfering ,Promoter Regions, Genetic ,Papillomaviridae ,Cells, Cultured ,Skin ,Cultured ,Virus-Cell Interactions ,Cell biology ,Cell Transformation, Neoplastic ,[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,RNA Interference ,Ultraviolet Rays ,Cells ,Immunology ,Biology ,Small Interfering ,Microbiology ,primary keratinocytes ,UV radiation ,Promoter Regions ,03 medical and health sciences ,Viral Proteins ,Downregulation and upregulation ,Genetic ,Virology ,medicine ,Gene silencing ,Humans ,Transcription factor ,Cell Proliferation ,Neoplastic ,JNK Mitogen-Activated Protein Kinases ,[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology ,Enzyme Activation ,030104 developmental biology ,Insect Science ,Toll-Like Receptor 9 ,RNA ,Tumor Suppressor Protein p53 ,Carcinogenesis ,030215 immunology - Abstract
Several lines of evidence indicate that cutaneous human papillomavirus (HPV) types belonging to the beta genus of the HPV phylogenetic tree synergize with UV radiation in the development of skin cancer. Accordingly, the E6 and E7 oncoproteins from some beta HPV types are able to deregulate pathways related to immune response and cellular transformation. Toll-like receptor 9 (TLR9), in addition to playing a role in innate immunity, has been shown to be involved in the cellular stress response. Using primary human keratinocytes as experimental models, we have shown that UV irradiation (and other cellular stresses) activates TLR9 expression. This event is closely linked to p53 activation. Silencing the expression of p53 or deleting its encoding gene affected the activation of TLR9 expression after UV irradiation. Using various strategies, we have also shown that the transcription factors p53 and c-Jun are recruited onto a specific region of the TLR9 promoter after UV irradiation. Importantly, the E6 and E7 oncoproteins from beta HPV38, by inducing the accumulation of the p53 antagonist ΔNp73α, prevent the UV-mediated recruitment of these transcription factors onto the TLR9 promoter, with subsequent impairment of TLR9 gene expression. This study provides new insight into the mechanism that mediates TLR9 upregulation in response to cellular stresses. In addition, we show that HPV38 E6 and E7 are able to interfere with this mechanism, providing another explanation for the possible cooperation of beta HPV types with UV radiation in skin carcinogenesis. IMPORTANCE Beta HPV types have been suggested to act as cofactors in UV-induced skin carcinogenesis by altering several cellular mechanisms activated by UV radiation. We show that the expression of TLR9, a sensor of damage-associated molecular patterns produced during cellular stress, is activated by UV radiation in primary human keratinocytes (PHKs). Two transcription factors known to be activated by UV radiation, p53 and c-Jun, play key roles in UV-activated TLR9 expression. The E6 and E7 oncoproteins from beta HPV38 strongly inhibit UV-activated TLR9 expression by preventing the recruitment of p53 and c-Jun to the TLR9 promoter. Our findings provide additional support for the role that beta HPV types play in skin carcinogenesis by preventing activation of specific pathways upon exposure of PHKs to UV radiation.
- Published
- 2017
32. The T Antigen Locus of Merkel Cell Polyomavirus Downregulates Human Toll-Like Receptor 9 Expression
- Author
-
Masahiro Shuda, Hyun Jin Kwun, Yuan Chang, Claudia Zannetti, Patrick S. Moore, Naveed Shahzad, Djamel Saidj, Rosita Accardi, Iris Cornet, Massimo Tommasino, Tarik Gheit, and Uzma Hasan
- Subjects
viruses ,Immunology ,Down-Regulation ,Merkel cell polyomavirus ,chemical and pharmacologic phenomena ,Biology ,medicine.disease_cause ,Microbiology ,Virus ,Merkel Cells ,Transactivation ,Antigen ,Virology ,medicine ,Humans ,Gene silencing ,Antigens, Viral, Tumor ,Promoter Regions, Genetic ,Polyomavirus Infections ,biology.organism_classification ,Virus-Cell Interactions ,Carcinoma, Merkel Cell ,Tumor Virus Infections ,Toll-Like Receptor 9 ,Insect Science ,Host-Pathogen Interactions ,Cancer research ,Carcinogenesis ,Oncovirus - Abstract
Establishment of a chronic infection is a key event in virus-mediated carcinogenesis. Several cancer-associated, double-stranded DNA (dsDNA) viruses act via their oncoproteins to downregulate Toll-like receptor 9 (TLR9), a key receptor in the host innate immune response that senses viral or bacterial dsDNA. A novel oncogenic virus, Merkel cell polyomavirus (MCPyV), has been recently identified that causes up to 80% of Merkel cell carcinomas (MCCs). However, it is not yet known whether this oncogenic virus also disrupts immune-related pathways. We find that MCPyV large T antigen (LT) expression downregulates TLR9 expression in epithelial and MCC-derived cells. Accordingly, silencing of LT expression results in upregulation of mRNA TLR9 levels. In addition, small T antigen (sT) also appears to inhibit TLR9 expression, since inhibition of its expression also resulted in an increase of TLR9 mRNA levels. LT inhibits TLR9 expression by decreasing the mRNA levels of the C/EBPβ transactivator, a positive regulator of the TLR9 promoter. Chromatin immunoprecipitation reveals that C/EBPβ binding at a C/EBPβ response element (RE) in the TLR9 promoter is strongly inhibited by expression of MCPyV early genes and that mutation of the C/EBP RE prevents MCPyV downregulation of TLR9. A survey of BK polyomavirus (BKPyV), JC polyomavirus (JCPyV), KI polyomavirus (KIPyV), MCPyV, simian virus 40 (SV40), and WU polyomavirus (WUPyV) early genes revealed that only BKPyV and MCPyV are potent inhibitors of TLR9 gene expression. MCPyV LT targeting of C/EBP transactivators is likely to play an important role in viral persistence and potentially inhibit host cell immune responses during MCPyV tumorigenesis.
- Published
- 2013
33. Oncoprotein E7 from Beta Human Papillomavirus 38 Induces Formation of an Inhibitory Complex for a Subset of p53-Regulated Promoters
- Author
-
Hector Hernandez-Vargas, Francesca Guarino, Rosita Accardi, Marie-Pierre Cros, Bakary S. Sylla, Massimo Tommasino, and Djamel Saidj
- Subjects
Keratinocytes ,Papillomavirus E7 Proteins ,Fluorescent Antibody Technique ,Mice ,p53 function ,0302 clinical medicine ,Gene expression ,DNA (Cytosine-5-)-Methyltransferases ,NF-kB ,RNA, Small Interfering ,Luciferases ,Promoter Regions, Genetic ,Papillomaviridae ,Cells, Cultured ,protein EZH2 ,IKK-Beta ,p73 ,transforming properties ,0303 health sciences ,Reverse Transcriptase Polymerase Chain Reaction ,EZH2 ,NF-kappa B ,Polycomb Repressive Complex 2 ,Nuclear Proteins ,Virus-Cell Interactions ,I-kappa B Kinase ,3. Good health ,DNA-Binding Proteins ,Histone ,030220 oncology & carcinogenesis ,DNA methylation ,DNA (Cytosine-5-)-Methyltransferase 1 ,Chromatin Immunoprecipitation ,Blotting, Western ,Immunology ,Biology ,Real-Time Polymerase Chain Reaction ,Microbiology ,DNA methyltransferase ,Colony-Forming Units Assay ,03 medical and health sciences ,Virology ,Animals ,Humans ,Immunoprecipitation ,Enhancer of Zeste Homolog 2 Protein ,RNA, Messenger ,Epigenetics ,030304 developmental biology ,Tumor Suppressor Proteins ,Papillomavirus Infections ,Tumor Protein p73 ,Promoter ,Fibroblasts ,Molecular biology ,Insect Science ,biology.protein ,Tumor Suppressor Protein p53 ,Chromatin immunoprecipitation - Abstract
Our previous studies on cutaneous beta human papillomavirus 38 (HPV38) E6 and E7 oncoproteins highlighted a novel activity of IκB kinase beta (IKKβ) in the nucleus of human keratinocytes, where it phosphorylates and stabilizes ΔNp73α, an antagonist of p53/p73 functions. Here, we further characterize the role of the IKKβ nuclear form. We show that IKKβ nuclear translocation and ΔNp73α accumulation are mediated mainly by HPV38 E7 oncoprotein. Chromatin immunoprecipitation (ChIP)/Re-ChIP experiments showed that ΔNp73α and IKKβ are part, together with two epigenetic enzymes DNA methyltransferase 1 (DNMT1) and the enhancer of zeste homolog 2 (EZH2), of a transcriptional regulatory complex that inhibits the expression of some p53-regulated genes, such as PIG3 . Recruitment to the PIG3 promoter of EZH2 and DNMT1 resulted in trimethylation of histone 3 on lysine 27 and in DNA methylation, respectively, both events associated with gene expression silencing. Decreases in the intracellular levels of HPV38 E7 or ΔNp73α strongly affected the recruitment of the inhibitory transcriptional complex to the PIG3 promoter, with consequent restoration of p53-regulated gene expression. Finally, the ΔNp73α/IKKβ/DNMT1/EZH2 complex appears to bind a subset of p53-regulated promoters. In fact, the complex is efficiently recruited to several promoters of genes encoding proteins involved in DNA repair and apoptosis, whereas it does not influence the expression of the prosurvival factor Survivin. In summary, our data show that HPV38 via E7 protein promotes the formation of a multiprotein complex that negatively regulates the expression of several p53-regulated genes.
- Published
- 2013
34. Transcriptional Regulation of the Human Tumor Suppressor DOK1 by E2F1
- Author
-
Marion Creveaux, Maha Siouda, Sophie Guillermier, Jiping Yue, Bakary S. Sylla, Ruchi Shukla, Massimo Tommasino, Zdenko Herceg, and Rosita Accardi
- Subjects
Transcriptional Activation ,endocrine system ,Apoptosis ,Biology ,Decitabine ,Response Elements ,Cell Line ,Epigenetics of physical exercise ,Transcription (biology) ,Cell Line, Tumor ,Transcriptional regulation ,Humans ,E2F1 ,Gene silencing ,Gene Silencing ,Molecular Biology ,Cell Proliferation ,Etoposide ,RNA-Binding Proteins ,E2F1 Transcription Factor ,Promoter ,Methyltransferases ,Articles ,Cell Biology ,DNA Methylation ,Chromatin Assembly and Disassembly ,Phosphoproteins ,Antineoplastic Agents, Phytogenic ,DNA-Binding Proteins ,HEK293 Cells ,DNA methylation ,Azacitidine ,Cancer research ,biological phenomena, cell phenomena, and immunity ,DNA Damage - Abstract
The expression of the tumor suppressor DOK1 is repressed in a variety of human tumors as a result of hypermethylation of its promoter region. However, the molecular mechanisms by which DOK1 expression is regulated have been poorly investigated. Here, we show that the expression of DOK1 is regulated mainly by the transcription factor E2F1. We identified three putative E2F1 response elements (EREs) in the DOK1 promoter region. E2F1 had a relatively higher binding affinity for the ERE located between bp -498 and -486 compared with the other two EREs. E2F1 gene silencing strongly inhibited DOK1 expression. E2F1-driven DOK1 transcription occurred in the presence of cellular stresses, such as accumulation of DNA damage induced by etoposide. DOK1 silencing promoted cell proliferation and protected against etoposide-induced apoptosis, indicating that DOK1 acts as a key mediator of cellular stress-induced cell death. Most importantly, we observed that DNA methylation of the DOK1 core promoter region found in head and neck cancer cell lines hampered the recruitment of E2F1 to the DOK1 promoter and compromised DOK1 expression. In summary, our data show that E2F1 is a key factor in DOK1 expression and provide novel insights into the regulation of these events in cancer cells.
- Published
- 2012
35. Unique DNA methylation signature in HPV-positive head and neck squamous cell carcinomas
- Author
-
Athena Sklias, Luis Felipe Ribeiro Pinto, Marie-Pierre Cros, Liacine Bouaoun, Graham Byrnes, Ellen Van Obberghen-Schilling, Anne Sudaka, Szilvia Ecsedi, Hector Hernandez-Vargas, Vincent Cahais, Rosita Accardi, D Degli Esposti, Zdenko Herceg, Valérie Giordanengo, Stéphanie Beghelli-de la Forest Divonne, Sheila Coelho Soares Lima, Nora Fernandez-Jimenez, and Cyrille Cuenin
- Subjects
0301 basic medicine ,Oncology ,Adult ,Male ,medicine.medical_specialty ,HPV ,lcsh:QH426-470 ,lcsh:Medicine ,Differentially methylated regions ,Biology ,Predictive models ,03 medical and health sciences ,Young Adult ,Internal medicine ,Genetics ,medicine ,Humans ,Genetics(clinical) ,Epigenetics ,Promoter Regions, Genetic ,Molecular Biology ,Gene ,Genetics (clinical) ,Aged ,Regulation of gene expression ,Aged, 80 and over ,Genome, Human ,Squamous Cell Carcinoma of Head and Neck ,Research ,lcsh:R ,Papillomavirus Infections ,Head and neck squamous cell carcinomas ,HPV infection ,DNA Methylation ,Middle Aged ,medicine.disease ,lcsh:Genetics ,030104 developmental biology ,CpG site ,Gene Expression Regulation ,Head and Neck Neoplasms ,DNA methylation ,Carcinoma, Squamous Cell ,Molecular Medicine ,Human genome ,CpG Islands ,Female ,CpG shores - Abstract
Background Head and neck squamous cell carcinomas (HNSCCs) represent a heterogeneous group of cancers for which human papilloma virus (HPV) infection is an emerging risk factor. Previous studies showed promoter hypermethylation in HPV(+) oropharyngeal cancers, but only few consistent target genes have been so far described, and the evidence of a functional impact on gene expression is still limited. Methods We performed global and stratified pooled analyses of epigenome-wide data in HNSCCs based on the Illumina HumanMethylation450 bead-array data in order to identify tissue-specific components and common viral epigenetic targets in HPV-associated tumours. Results We identified novel differentially methylated CpGs and regions associated with viral infection that are independent of the anatomic site. In particular, most hypomethylated regions were characterized by a marked loss of CpG island boundaries, which showed significant correlations with expression of neighbouring genes. Moreover, a subset of only five CpGs in a few hypomethylated regions predicted HPV status with a high level of specificity in different cohorts. Finally, this signature was a better predictor of survival compared with HPV status determined by viral gene expression by RNA sequencing in The Cancer Genome Atlas cohort. Conclusions We identified a novel epigenetic signature of HPV infection in HNSCCs which is independent of the anatomic site, is functionally correlated with gene expression and may be leveraged for improved stratification of prognosis in HNSCCs. Electronic supplementary material The online version of this article (doi:10.1186/s13073-017-0419-z) contains supplementary material, which is available to authorized users.
- Published
- 2016
36. Novel ß-HPV49 Transgenic Mouse Model of Upper Digestive Tract Cancer
- Author
-
Karin Müller-Decker, Daniele Viarisio, Massimo Tommasino, Ulrich Kloz, Rosita Accardi, Paola Zanna, Birgit Aengeneyndt, Lutz Gissmann, and Christa Flechtenmacher
- Subjects
0301 basic medicine ,Genetically modified mouse ,Cancer Research ,Pathology ,medicine.medical_specialty ,Skin Neoplasms ,Ultraviolet Rays ,Transgene ,Papillomavirus E7 Proteins ,Viral Oncogene ,Mice, Transgenic ,Biology ,medicine.disease_cause ,Digestive System Neoplasms ,Virus ,03 medical and health sciences ,Cytokeratin ,Mice ,medicine ,Animals ,Papillomaviridae ,Epidermis (botany) ,Cancer ,Oncogene Proteins, Viral ,medicine.disease ,Disease Models, Animal ,030104 developmental biology ,Oncology ,Cancer research ,Carcinoma, Squamous Cell ,Female ,Carcinogenesis - Abstract
The beta genus of human papillomaviruses (ß-HPV) includes approximately 50 different viral types that are subdivided into five species (ß-1 through ß-5). Nonmelanoma cancers may involve some ß-1 and ß-2 HPV types, but the biology of most ß-HPV types and their possible connections to human disease are still little characterized. In this study, we studied the effects of ß-3 type HPV49 in a novel transgenic (Tg) mouse model, using a cytokeratin K14 promoter to drive expression of the E6 and E7 genes from this virus in the basal skin epidermis and the mucosal epithelia of the digestive tract (K14 HPV49 E6/E7-Tg mice). Viral oncogene expression only marginally increased cellular proliferation in the epidermis of Tg animals, compared with wild-type littermates, and we observed no spontaneous tumor formation during their entire lifespan. However, we found that K14 HPV49 E6/E7-Tg mice were highly susceptible to upper digestive tract carcinogenesis upon initiation with 4-nitroquinoline 1-oxide (4NQO). This was a selective effect, as the same mice did not exhibit any skin lesions after chronic UV irradiation. Opposite results were observed in an analogous Tg model expressing the ß-2 HPV38 E6 and E7 oncogenes at the same anatomic sites. While these mice were highly susceptible to UV-induced skin carcinogenesis, as previously shown, they were little affected by 4NQO treatment. Overall, our findings highlight important differences in the biologic properties of certain ß-type HPV that affect their impact on carcinogenesis in an anatomic site-specific manner. Cancer Res; 76(14); 4216–25. ©2016 AACR.
- Published
- 2016
37. IκB Kinase β Promotes Cell Survival by Antagonizing p53 Functions through ΔNp73α Phosphorylation and Stabilization
- Author
-
Jiping Yue, Christine Carreira, Rosita Accardi, Agnese Collino, Tarik Gheit, Lutz Gissmann, Bakary S. Sylla, Ishraq Hussain, Massimo Tommasino, Cesare Indiveri, and Mariafrancesca Scalise
- Subjects
Transcription, Genetic ,Cell Survival ,Immunoblotting ,Gene Expression ,Breast Neoplasms ,IκB kinase ,Biology ,Mice ,RNA interference ,Cell Line, Tumor ,Serine ,Animals ,Humans ,Immunoprecipitation ,Gene silencing ,Phosphorylation ,RNA, Small Interfering ,Nuclear protein ,Molecular Biology ,Cells, Cultured ,Protein Stability ,Reverse Transcriptase Polymerase Chain Reaction ,Tumor Suppressor Proteins ,HEK 293 cells ,NF-kappa B ,I-Kappa-B Kinase ,Nuclear Proteins ,Tumor Protein p73 ,Articles ,Cell Biology ,NFKB1 ,I-kappa B Kinase ,DNA-Binding Proteins ,HEK293 Cells ,Mutation ,Cancer research ,RNA Interference ,Tumor Suppressor Protein p53 - Abstract
ΔNp73α, a dominant-negative inhibitor of p53 and p73, exhibits antiapoptotic and transforming activity in in vitro models and is often found to be upregulated in human cancers. The mechanisms involved in the regulation of ΔNp73α protein levels in normal and cancer cells are poorly characterized. Here, we show that that IκB kinase beta (IKKβ) increases ΔNp73α protein stability independently of its ability to activate NF-κB. IKKβ associates with and phosphorylates ΔNp73α at serine 422 (S422), leading to its accumulation in the nucleus, where it binds and represses several p53-regulated genes. S422A mutation in ΔNp73α abolished IKKβ-mediated stabilization and inhibition of p53-regulated gene expression. Inhibition of IKKβ activity by chemical inhibitors, overexpression of dominant-negative mutants, or gene silencing by siRNA also resulted in ΔNp73α destabilization, which under these conditions was rapidly translocated into the cytoplasm and degraded by a calpain-mediated mechanism. We also present evidence for the IKKβ and ΔNp73α cross talk in cancer-derived cell lines and primary cancers. Our data unveil a new mechanism involved in the regulation of the p73 and p53 network.
- Published
- 2011
38. Beta HPV38 oncoproteins act with a hit-and-run mechanism in ultraviolet radiation-induced skin carcinogenesis in mice
- Author
-
Karin Müller-Decker, Daniele Viarisio, Christa Flechtenmacher, Sandra Caldeira, Rosita Accardi, Catherine Voegele, Matthias Dürst, Alexis Robitaille, Katrin Beer, Lutz Gissmann, Jiri Zavadil, Matthias Bozza, Richard Harbottle, Maude Ardin, Massimo Tommasino, and Lars Jansen
- Subjects
Keratinocytes ,0301 basic medicine ,Neoplasms, Radiation-Induced ,Skin Neoplasms ,Light ,Carcinogenesis ,Gene Identification and Analysis ,Pathology and Laboratory Medicine ,medicine.disease_cause ,Epithelium ,Mice ,0302 clinical medicine ,Animal Cells ,Medicine and Health Sciences ,Biology (General) ,Skin ,Mutation ,Radiation ,Receptors, Notch ,Physics ,Electromagnetic Radiation ,Animal Models ,Recombinant Proteins ,Neoplasm Proteins ,Tumor Burden ,Experimental Organism Systems ,Oncology ,030220 oncology & carcinogenesis ,Physical Sciences ,Female ,Cellular Types ,Anatomy ,Research Article ,QH301-705.5 ,Ultraviolet Rays ,Transgene ,Immunology ,Mouse Models ,Mice, Transgenic ,Biology ,Research and Analysis Methods ,Microbiology ,Viral Proteins ,03 medical and health sciences ,Signs and Symptoms ,Model Organisms ,Germline mutation ,Diagnostic Medicine ,Ultraviolet Radiation ,Virology ,Genetics ,medicine ,Animals ,Betapapillomavirus ,Beta (finance) ,Mutation Detection ,Molecular Biology ,Nuclear Physics ,Epidermis (botany) ,Mutagenesis ,Biology and Life Sciences ,Epithelial Cells ,Cell Biology ,Oncogene Proteins, Viral ,RC581-607 ,Genes, p53 ,medicine.disease ,Biological Tissue ,030104 developmental biology ,Lesions ,Beta Radiation ,Cancer research ,Somatic Mutation ,Parasitology ,Immunologic diseases. Allergy ,Epidermis ,Skin cancer ,Gene Deletion - Abstract
Cutaneous beta human papillomavirus (HPV) types are suspected to be involved, together with ultraviolet (UV) radiation, in the development of non-melanoma skin cancer (NMSC). Studies in in vitro and in vivo experimental models have highlighted the transforming properties of beta HPV E6 and E7 oncoproteins. However, epidemiological findings indicate that beta HPV types may be required only at an initial stage of carcinogenesis, and may become dispensable after full establishment of NMSC. Here, we further investigate the potential role of beta HPVs in NMSC using a Cre-loxP-based transgenic (Tg) mouse model that expresses beta HPV38 E6 and E7 oncogenes in the basal layer of the skin epidermis and is highly susceptible to UV-induced carcinogenesis. Using whole-exome sequencing, we show that, in contrast to WT animals, when exposed to chronic UV irradiation K14 HPV38 E6/E7 Tg mice accumulate a large number of UV-induced DNA mutations, which increase proportionally with the severity of the skin lesions. The mutation pattern detected in the Tg skin lesions closely resembles that detected in human NMSC, with the highest mutation rate in p53 and Notch genes. Using the Cre-lox recombination system, we observed that deletion of the viral oncogenes after development of UV-induced skin lesions did not affect the tumour growth. Together, these findings support the concept that beta HPV types act only at an initial stage of carcinogenesis, by potentiating the deleterious effects of UV radiation., Author summary Many epidemiological and biological findings support the hypothesis that beta HPV types cooperate with UV radiation in the induction of NMSC, the most common form of human cancer. We have previously shown that K14 HPV38 E6/E7 Tg mice, when exposed to long-term UV radiation, developed NMSC, whereas WT animals subjected to identical treatments did not develop any type of skin lesions. Here, we show that the high skin cancer susceptibility of these Tg animals tightly correlates with their tendency to accumulate UV-induced mutations in genes that are frequently mutated in human NMSC. Importantly, deletion of the HPV38 E6 and E7 genes in existing skin lesions did not affect the further growth of the cancer cells. Together, these findings support the model that beta HPV infection is a co-factor in skin carcinogenesis, facilitating the accumulation of the UV-induced DNA mutations.
- Published
- 2018
39. E7 properties of mucosal human papillomavirus types 26, 53 and 66 correlate with their intermediate risk for cervical cancer development
- Author
-
Rosita Accardi, Uzma Hasan, Majid Touka, Anne-Sophie Gabet, Bakary S. Sylla, Anouk Smet, Mariam Mansour, Angelica Bellopede, and Massimo Tommasino
- Subjects
Keratinocytes ,Papillomavirus E7 Proteins ,Cyclin A ,Uterine Cervical Neoplasms ,Alphapapillomavirus ,Biology ,Retinoblastoma Protein ,Cell Line ,Cell cycle deregulation ,Risk Factors ,Virology ,Cyclin E ,medicine ,Humans ,Cells, Cultured ,E7 ,Cell Line, Transformed ,Cyclin ,Cervical cancer ,HPV26, 53 and 66 ,Cell growth ,Intermediate risk ,Cyclin-Dependent Kinase 2 ,Papillomavirus Infections ,Cyclin-dependent kinase 2 ,G1 Phase ,Retinoblastoma protein ,Fibroblasts ,Cell cycle ,medicine.disease ,In vitro ,Up-Regulation ,Cell Transformation, Neoplastic ,Immunology ,biology.protein ,Female - Abstract
Epidemiological studies have demonstrated that 15 different mucosal human papillomavirus (HPV) types of the genus alpha of the HPV phylogetic tree are classified as high risk for cervical cancer development. Three additional HPV types of the same genus, HPV26, 53 and 66, are classified as probable high-risk types. In this study, we have characterized the biological properties of the E7 oncoproteins from these three HPV types. All of the corresponding E7 proteins were able to associate with retinoblastoma protein (pRb) and up-regulated the expression of several positive cell cycle regulators, i.e. CDK2, cyclin A and cylin E. However, HPV26 E7 appears to be more efficient than HPV53 and 66 E7 in up-regulating the transcription of cyclin A. Unlike E7 from the high-risk type HPV16 protein, HPV26, 53 and 66 did not efficiently promote pRb degradation. In addition, E7 from these viruses was able to promote proliferation of primary human keratinocytes and circumvent G1 arrest imposed by overexpression of p16 INK4a , but with less efficiency than the high-risk HPV16 E7. Together, our data show that in vitro properties of these E7 proteins correlate with the epidemiological classification of HPV26, 53 and 66 as HPV types with an intermediate risk for cervical cancer development.
- Published
- 2007
40. TLR9 Expression and Function Is Abolished by the Cervical Cancer-Associated Human Papillomavirus Type 16
- Author
-
Isabelle Vincent, Mario Sideri, Alexandra Biliato, Rosita Accardi, Lutz Gissmann, Fumihiko Takeshita, Véronique Bouvard, Uzma Hasan, Thomas Iftner, Elizabeth E.M. Bates, Mariam Mansour, Frank Stubenrauch, and Massimo Tommasino
- Subjects
Transcription, Genetic ,Papillomavirus E7 Proteins ,viruses ,Immunology ,Down-Regulation ,Uterine Cervical Neoplasms ,Biology ,Virus ,Immune system ,Transcription (biology) ,Humans ,Immunology and Allergy ,Human papillomavirus 16 ,Innate immune system ,Human papillomavirus 18 ,Papillomavirus Infections ,virus diseases ,TLR9 ,Oncogene Proteins, Viral ,Cell cycle ,Immunity, Innate ,female genital diseases and pregnancy complications ,Repressor Proteins ,Cell Transformation, Neoplastic ,CpG site ,Apoptosis ,Toll-Like Receptor 9 ,Female ,HeLa Cells - Abstract
Cervical cancer development is linked to the persistent infection by high-risk mucosal human papillomaviruses (HPVs) types. The E6 and E7 major oncoproteins from this dsDNA virus play a key role in the deregulation of the cell cycle, apoptosis, and adaptive immune surveillance. In this study, we show for the first time that HPV type 16 (HPV16), the most carcinogenic type among the high-risk subgroup, interferes with innate immunity by affecting the expression of TLRs. Infection of human primary keratinocytes with HPV16 E6 and E7 recombinant retroviruses inhibits TLR9 transcription and hence functional loss of TLR9-regulated pathways. Similar findings were achieved in HPV16-positive cancer-derived cell lines and primary cervical cancers, demonstrating that this event occurs also in an in vivo context. Interestingly, E6 and E7 from the low-risk HPV type 6 are unable to down-regulate the TLR9 promoter. In addition, E6 and E7 from the high-risk HPV type 18, which are known to persist less competently in the host than HPV16, have reduced efficiency compared with HPV16 in inhibiting TLR9 transcription. Furthermore, a CpG motif derived from the HPV16 E6 DNA sequence activated TLR9, indicating this virus is able to initiate innate responses via the receptor it later down-regulates. This study reveals a novel mechanism used by HPV16 to suppress the host immune response by deregulating the TLR9 transcript, providing evidence that abolishing innate responses may be a crucial step involved in the carcinogenic events mediated by HPVs.
- Published
- 2007
41. The mycotoxin aflatoxin B1 stimulates Epstein-Barr virus-induced B-cell transformation in in vitro and in vivo experimental models
- Author
-
Christopher P. Wild, François-Loïc Cosset, Henri Gruffat, Rosita Accardi, Evelyne Manet, Massimo Tommasino, Florence Le Calvez-Kelm, Tarik Gheit, Cecilia Sirand, Floriane Fusil, Hector Hernandez-Vargas, Alexandra Traverse-Glehen, Centre International de Recherche contre le Cancer - International Agency for Research on Cancer (CIRC - IARC), Organisation Mondiale de la Santé / World Health Organization Office (OMS / WHO), Herpesvirus oncogènes – Oncogenic Herpesviruses, Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Virus enveloppés, vecteurs et immunothérapie – Enveloped viruses, Vectors and Immuno-therapy (EVIR), Laboratoire Central d'Anatomie et de Cytologie Pathologiques [Hôpital Edouard Herriot - HCL], Hôpital Edouard Herriot [CHU - HCL], Hospices Civils de Lyon (HCL)-Hospices Civils de Lyon (HCL), Centre International de Recherche en Infectiologie - UMR (CIRI), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
Male ,Cancer Research ,Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Aflatoxin B1 ,Carcinogenesis ,Cells ,Population ,Mice, SCID ,Biology ,Cell Transformation ,SCID ,medicine.disease_cause ,Virus Replication ,Virus ,Mice ,In vivo ,Mice, Inbred NOD ,hemic and lymphatic diseases ,medicine ,Animals ,Humans ,education ,B cell ,Cells, Cultured ,Neoplastic ,education.field_of_study ,B-Lymphocytes ,Cultured ,Herpesvirus 4 ,General Medicine ,Environmental exposure ,Environmental Exposure ,[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology ,Epstein–Barr virus ,Virology ,Burkitt Lymphoma ,3. Good health ,medicine.anatomical_structure ,Cell Transformation, Neoplastic ,Lytic cycle ,[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,Inbred NOD ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,Female ,Virus Activation ,Human ,Signal Transduction - Abstract
International audience; Although Epstein-Barr virus (EBV) infection is widely distributed, certain EBV-driven malignancies are geographically restricted. EBV-associated Burkitt's lymphoma (eBL) is endemic in children living in sub-Saharan Africa. This population is heavily exposed to food contaminated with the mycotoxin aflatoxin B1 (AFB1). Here, we show that exposure to AFB1 in in vitro and in vivo models induces activation of the EBV lytic cycle and increases EBV load, two events that are associated with an increased risk of eBL in vivo. AFB1 treatment leads to the alteration of cellular gene expression, with consequent activations of signaling pathways, e.g. PI3K, that in turn mediate reactivation of the EBV life cycle. Finally, we show that AFB1 triggers EBV-driven cellular transformation both in primary human B cells and in a humanized animal model. In summary, our data provide evidence for a role of AFB1 as a cofactor in EBV-mediated carcinogenesis.
- Published
- 2015
42. Role of human papillomaviruses in carcinogenesis
- Author
-
Susanna Chiocca, Raffaella Ghittoni, Rosita Accardi, and Massimo Tommasino
- Subjects
Cancer Research ,DNA damage ,HPV vaccines ,Review ,Bioinformatics ,medicine.disease_cause ,papillomavirus ,03 medical and health sciences ,0302 clinical medicine ,Medicine ,Gene ,030304 developmental biology ,Cervical cancer ,0303 health sciences ,business.industry ,Cell cycle ,medicine.disease ,3. Good health ,Oncology ,Apoptosis ,030220 oncology & carcinogenesis ,Cancer research ,Skin cancer ,business ,Carcinogenesis ,carcinogenesis - Abstract
The human papillomavirus (HPV) family comprises more than 170 different types that preferentially infect the mucosa of the genitals, upper-respiratory tract, or the skin. The 'high-risk HPV type', a sub-group of mucosal HPVs, is the cause of approximately 5% of all human cancers, which corresponds to one-third of all virus-induced tumours. Within the high-risk group, HPV16 is the most oncogenic type, being responsible for approximatively 50% of all worldwide cervical cancers. Many studies suggest that, in addition to the high-risk mucosal HPV types, certain cutaneous HPVs also have a role in the development of non-melanoma skin cancer (NMSC). Functional studies on the HPV early gene products showed that E6 and E7 play a key role in carcinogenesis. These two proteins use multiple mechanisms to evade host immune surveillance, allowing viral persistence, and to deregulate cell cycle and apoptosis control, thus facilitating the accumulation of DNA damage and ultimately cellular transformation. The demonstration that high-risk HPV types are the etiological agents of cervical cancer allowed the implementation in the clinical routine of novel screening strategies for cervical lesions, as well as the development of a very efficient prophylactic vaccine. Because of these remarkable achievements, there is no doubt that in the coming decades we will witness a dramatic reduction of cervical cancer incidence worldwide.
- Published
- 2015
43. Expression of the Epidermodysplasia Verruciformis-Associated Genes EVER1 and EVER2 Is Activated by Exogenous DNA and Inhibited by LMP1 Oncoprotein from Epstein-Barr Virus
- Author
-
C. Cohen, Djamel Saidj, Ivonne Rubio, Patrick van Uden, Maha Siouda, Rosita Accardi, Massimo Tommasino, Patrick Lomonte, Cecilia Frecha, Sébastien A. Chevalier, Centre de génétique et de physiologie moléculaire et cellulaire (CGPhiMC), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, Université Claude Bernard Lyon 1 (UCBL), Université de Lyon, Institut NeuroMyoGène (INMG), and Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)
- Subjects
Herpesvirus 4, Human ,EBV-LMP1 ,Immunology ,education ,EVER1 ,EVER2 ,Biology ,medicine.disease_cause ,Microbiology ,Virus ,Viral Matrix Proteins ,Ciencias Biológicas ,Virology ,medicine ,Humans ,Gene ,ComputingMilieux_MISCELLANEOUS ,Regulation of gene expression ,B-Lymphocytes ,Viral matrix protein ,Membrane Proteins ,Epidermodysplasia verruciformis ,medicine.disease ,Epstein–Barr virus ,3. Good health ,Virus-Cell Interactions ,Gene Expression Regulation ,Insect Science ,Epidermodysplasia Verruciformis ,Host-Pathogen Interactions ,[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,Ectopic expression ,Signal transduction ,Virología ,CIENCIAS NATURALES Y EXACTAS ,BCELLS - Abstract
EVER1 and EVER2 are mutated in epidermodysplasia verruciformis patients, who are susceptible to human betapapillomavirus (HPV) infection. It is unknown whether their products control the infection of other viruses. Here, we show that the expression of both genes in B cells is activated immediately after Epstein-Barr virus (EBV) infection, whereas at later stages, it is strongly repressed via activation of the NF-κB signaling pathway by latent membrane protein 1 (LMP1). Ectopic expression of EVER1 impairs the ability of EBV to infect B cells. Fil: Frecha, Cecilia Ariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Biología Celular y Neurociencia "Prof. Eduardo de Robertis". Universidad de Buenos Aires. Facultad de Medicina. Instituto de Biología Celular y Neurociencia; Argentina. International Agency For Research On Cancer; Francia Fil: Chevalier, Sébastien A.. International Agency For Research On Cancer; Francia Fil: van Uden, Patrick. International Agency For Research On Cancer; Francia Fil: Rubio, Ivonne. International Agency For Research On Cancer; Francia Fil: Siouda, Maha. International Agency For Research On Cancer; Francia Fil: Saidj, Djamel. International Agency For Research On Cancer; Francia Fil: Cohen, Camille. Université Claude Bernard Lyon 1; Francia. Centre National de la Recherche Scientifique; Francia Fil: Lomonte, Patrick. Centre National de la Recherche Scientifique; Francia. Université Claude Bernard Lyon 1; Francia Fil: Accardi, Rosita. International Agency For Research On Cancer; Francia Fil: Tommasino, Massimo. International Agency For Research On Cancer; Francia
- Published
- 2015
44. The role of microenvironment in tumorigenesis: Focus on dendritic cells in human papillomavirus E6, E7-transformed keratinocytes
- Author
-
Maria Vincenza Chiantore, Marco Iuliano, Giovanna Romeo, Rosita Accardi, Massimo Tommasino, Gianna Fiorucci, and Giorgio Mangino
- Subjects
Tumor microenvironment ,Priming (immunology) ,General Medicine ,Biology ,medicine.disease_cause ,Human Papillomavirus ,dendritic cells ,miRNA ,CTL ,Immune system ,Antigen ,Immunology ,Cancer cell ,medicine ,Cancer research ,Cytotoxic T cell ,tumor microenvironment ,Carcinogenesis ,human papillomavirus - Abstract
The inception of tumor microenvironment (TME), a complex and dynamic system constituted by different types of cells engaged by tumor and extracellular matrix surrounding cancer cells, is a way for them to elude the immune surveillance. Dendritic cells (DCs), as a sentinel, are able to recognize alteration in the microenvironment and predispose the immune system response. The relationship between cancer and virus infection is well documented. High-risk Human Papillomavirus (HR-HPV) has a well-characterized transforming property and has been associated with squamous cell carcinoma of the ano-genital and oral tracts. Transforming ability of HR-HPVs is based on the function of E6 and E7 viral oncoproteins, which interact and inactivate p53 and pRb oncosuppressors, respectively. Recently, it was demonstrated that HPV oncoproteins are also able to affect a number of microRNAs (miRNAs) regulating the expression of genes involved in proliferative control. For these reasons DC-based vaccines targeting oncogenic E6 and E7 are ideal candidates to elicit strong immune responses. Here we summarize significant data about the analysis of TME in HPV-induced tumorigenesis. We also report original results produced by cytotoxic T lymphocyte (CTL) in vitro priming against E6 and E7 HPV16 antigens, performed using human monocyte-derived dendritic cells. Dendritic cells, maturated by the exposition to necrotic or apoptotic keratinocytes expressing both oncoproteins of HPV16, show different expression levels of specific maturation markers. Evidence indicating the ability of necrotic keratinocytes to alter the microRNA profile in DCs, macrophages (MΦ) and Langerhans cells (LCs) compared to prototypical stimuli as bacterial lipopolysaccharide was also provided. We can speculate that, based on transformed cells death pathway (i.e. apoptosis versus necrosis), virus-induced immune alterations might show different results in creating an immunotolerogenic microenvironment during the carcinogenesis process.
- Published
- 2015
45. Over-expression in Escherichia coli and characterization of two recombinant isoforms of human FAD synthetase
- Author
-
Michele Galluccio, Rosita Accardi, Carmen Brizio, Cesare Indiveri, Maria Barile, Elisabetta Gianazza, Enza Maria Torchetti, Valeria Bafunno, and Robin Wait
- Subjects
Gene isoform ,Molecular Sequence Data ,Biophysics ,medicine.disease_cause ,Riboflavin kinase ,Biochemistry ,Gene Expression Regulation, Enzymologic ,Cofactor ,FLAD1 ,Affinity chromatography ,Settore BIO/10 - Biochimica ,Escherichia coli ,medicine ,Humans ,Amino Acid Sequence ,Molecular Biology ,FMN adenylyl transferase ,chemistry.chemical_classification ,FAD synthetase ,Sequence Homology, Amino Acid ,biology ,Molecular mass ,Gene Expression Regulation, Bacterial ,Cell Biology ,Nucleotidyltransferases ,Molecular biology ,Recombinant Proteins ,Enzyme Activation ,Isoenzymes ,Enzyme ,chemistry ,biology.protein ,FAD synthesis ,Isoforms - Abstract
FAD synthetase (FADS) (EC 2.7.7.2) is a key enzyme in the metabolic pathway that converts riboflavin into the redox cofactor FAD. Two hypothetical human FADSs, which are the products of FLAD1 gene, were over-expressed in Escherichia coli and identified by ESI-MS/MS. Isoform 1 was over-expressed as a T7-tagged protein which had a molecular mass of 63 kDa on SDS–PAGE. Isoform 2 was over-expressed as a 6-His-tagged fusion protein, carrying an extra 84 amino acids at the N-terminal with an apparent molecular mass of 60 kDa on SDS–PAGE. It was purified near to homogeneity from the soluble cell fraction by one-step affinity chromatography. Both isoforms possessed FADS activity and had a strict requirement for MgCl 2 , as demonstrated using both spectrophotometric and chromatographic methods. The purified recombinant isoform 2 showed a specific activity of 6.8 ± 1.3 nmol of FAD synthesized/min/mg protein and exhibited a K M value for FMN of 1.5 ± 0.3 μM. This is the first report on characterization of human FADS, and the first cloning and over-expression of FADS from an organism higher than yeast.
- Published
- 2006
46. A Nuclear Export Signal and Phosphorylation Regulate Dok1 Subcellular Localization and Functions
- Author
-
Yamei Niu, Charlotte Andrieu-Soler, Amélie Thépot, François Roy, Frédéric Saltel, Bakary S. Sylla, Rosita Accardi, Pierre Jurdic, Wen Dong, Nadège Foiselle, Massimo Tommasino, and Anne-Lise Chantegrel
- Subjects
Cytoplasm ,Active Transport, Cell Nucleus ,Biology ,Mice ,chemistry.chemical_compound ,Cell Movement ,Cell Adhesion ,medicine ,Animals ,Humans ,Amino Acids ,Phosphorylation ,Nuclear protein ,Growth Substances ,Phosphotyrosine ,Nuclear export signal ,Molecular Biology ,Cells, Cultured ,Cell Proliferation ,Cell Nucleus ,Nuclear Export Signals ,Swiss 3T3 Cells ,RNA-Binding Proteins ,Tyrosine phosphorylation ,Articles ,Cell Biology ,Fibroblasts ,Phosphoproteins ,Subcellular localization ,I-kappa B Kinase ,Cell biology ,DNA-Binding Proteins ,Protein Transport ,Cell nucleus ,src-Family Kinases ,medicine.anatomical_structure ,Gene Expression Regulation ,chemistry ,Mutation ,NIH 3T3 Cells ,Nuclear lamina ,Nuclear localization sequence - Abstract
Dok1 is believed to be a mainly cytoplasmic adaptor protein which down-regulates mitogen-activated protein kinase activation, inhibits cell proliferation and transformation, and promotes cell spreading and cell migration. Here we show that Dok1 shuttles between the nucleus and cytoplasm. Treatment of cells with leptomycin B (LMB), a specific inhibitor of the nuclear export signal (NES)-dependent receptor CRM1, causes nuclear accumulation of Dok1. We have identified a functional NES (348LLKAKLTDPKED359) that plays a major role in the cytoplasmic localization of Dok1. Src-induced tyrosine phosphorylation prevented the LMB-mediated nuclear accumulation of Dok1. Dok1 cytoplasmic localization is also dependent on IKKbeta. Serum starvation or maintaining cells in suspension favor Dok1 nuclear localization, while serum stimulation, exposure to growth factor, or cell adhesion to a substrate induce cytoplasmic localization. Functionally, nuclear NES-mutant Dok1 had impaired ability to inhibit cell proliferation and to promote cell spreading and cell motility. Taken together, our results provide the first evidence that Dok1 transits through the nucleus and is actively exported into the cytoplasm by the CRM1 nuclear export system. Nuclear export modulated by external stimuli and phosphorylation may be a mechanism by which Dok1 is maintained in the cytoplasm and membrane, thus regulating its signaling functions.
- Published
- 2006
47. Skin Hyperproliferation and Susceptibility to Chemical Carcinogenesis in Transgenic Mice Expressing E6 and E7 of Human Papillomavirus Type 38
- Author
-
Lutz Gissmann, Rosita Accardi, Bakary S. Sylla, Zhao Qi Wang, Wen Dong, Matthias Dürst, Massimo Tommasino, Lars Jansen, Ulrich Kloz, Sandra Caldeira, and Wei Min Tong
- Subjects
Keratinocytes ,Genetically modified mouse ,Cell cycle checkpoint ,Carcinogenicity Tests ,Papillomavirus E7 Proteins ,Transgene ,Immunology ,Mice, Transgenic ,medicine.disease_cause ,Microbiology ,Transformation and Oncogenesis ,Mice ,Virology ,Keratin ,medicine ,Animals ,Papillomaviridae ,Cell Proliferation ,chemistry.chemical_classification ,biology ,Epidermis (botany) ,Cell Transformation, Viral ,biology.organism_classification ,Cell Transformation, Neoplastic ,chemistry ,Insect Science ,Cancer research ,Epidermis ,Carcinogenesis - Abstract
The oncoproteins E6 and E7 of human papillomavirus type 38 (HPV38) display several transforming activities in vitro, including immortalization of primary human keratinocytes. To evaluate the oncogenic activities of the viral proteins in an in vivo model, we generated transgenic mice expressing HPV38 E6 and E7 under the control of the bovine homologue of the human keratin 10 (K10) promoter. Two distinct lines of HPV38 E6/E7-expressing transgenic mice that express the viral genes at different levels were obtained. In both lines, HPV38 E6 and E7 induced cellular proliferation, hyperplasia, and dysplasia in the epidermis. The rate of occurrence of these events was proportional to the levels of HPV38 E6 and E7 expression in the two transgenic lines. Exposure of the epidermis of nontransgenic mice to UV led to p21 WAF1 accumulation and cell cycle arrest. In contrast, keratinocytes from transgenic mice continued to proliferate and were not positive for p21 WAF1 , indicating that cell cycle checkpoints are altered in keratinocytes expressing the viral genes. Although the HPV38 E6/E7-expressing transgenic mice did not develop spontaneous tumors during their life span, two-stage carcinogen treatment led to a high incidence of papillomas, keratoacanthomas, and squamous-cell carcinomas in HPV38 mice compared with nontransgenic animals. Together, these data show that HPV38 E6 and E7 display transforming properties in vivo, providing further support for the role of HPV38 in carcinogenesis.
- Published
- 2005
48. Human Papillomavirus Type 16 E6 Promotes Retinoblastoma Protein Phosphorylation and Cell Cycle Progression
- Author
-
Ilaria Malanchi, Elliot J. Androphy, Rosita Accardi, Joerg Hoheisel, Frank Diehl, Anouk Smet, and Massimo Tommasino
- Subjects
Tumor suppressor gene ,Immunology ,Cyclin A ,Retinoblastoma Protein ,Microbiology ,Transformation and Oncogenesis ,S Phase ,Mice ,Virology ,Animals ,Humans ,Phosphorylation ,E2F ,neoplasms ,Cells, Cultured ,Cell Proliferation ,Cyclin ,Cyclin-dependent kinase 1 ,biology ,Cell Cycle ,Cyclin-dependent kinase 2 ,G1 Phase ,Mouth Mucosa ,Retinoblastoma protein ,3T3 Cells ,Oncogene Proteins, Viral ,Fibroblasts ,Cell cycle ,Cell biology ,Repressor Proteins ,Gene Expression Regulation ,Insect Science ,biology.protein ,Cancer research ,biological phenomena, cell phenomena, and immunity - Abstract
We show that E6 proteins from benign human papillomavirus type 1 (HPV1) and oncogenic HPV16 have the ability to alter the regulation of the G 1 /S transition of the cell cycle in primary human fibroblasts. Overexpression of both viral proteins induces cellular proliferation, retinoblastoma (pRb) phosphorylation, and accumulation of products of genes that are negatively regulated by pRb, such as p16 INK4a , CDC2, E2F-1, and cyclin A. Hyperphosphorylated forms of pRb are present in E6-expressing cells even in the presence of ectopic levels of p16 INK4a . The E6 proteins strongly increased the cyclin A/cyclin-dependent kinase 2 (CDK2) activity, which is involved in pRb phosphorylation. In addition, mRNA and protein levels of the CDK2 inhibitor p21 WAF1/CIP1 were strongly down-regulated in cells expressing E6 proteins. The down-regulation of the p21 WAF1/CIP1 gene appears to be independent of p53 inactivation, since HPV1 E6 and an HPV16 E6 mutant unable to target p53 were fully competent in decreasing p21 WAF1/CIP1 levels. E6 from HPV1 and HPV16 also enabled cells to overcome the G 1 arrest imposed by oncogenic ras . Immunofluorescence staining of cells coexpressing ras and E6 from either HPV16 or HPV1 revealed that antiproliferative (p16 INK4a ) and proliferative (Ki67) markers were coexpressed in the same cells. Together, these data underline a novel activity of E6 that is not mediated by inactivation of p53.
- Published
- 2004
49. p53 mutations are common in human papillomavirus type 38-positive non-melanoma skin cancers
- Author
-
Raffaele Filotico, Massimo Tommasino, Sandra Caldeira, Rosita Accardi, Ingeborg Zehbe, Silvia Franceschi, and Repositório da Universidade de Lisboa
- Subjects
Keratinocytes ,p53 ,Human papillomavirus ,Cancer Research ,E6 protein ,Skin Neoplasms ,Non-melanoma-skin cancer ,Immunoblotting ,medicine.disease_cause ,Polymerase Chain Reaction ,medicine ,Animals ,Codon ,Papillomaviridae ,Gene ,Cells, Cultured ,E6 ,integumentary system ,Reverse Transcriptase Polymerase Chain Reaction ,business.industry ,DNA ,Exons ,Cervical cells ,Fibroblasts ,Genes, p53 ,Coculture Techniques ,Rats ,Retroviridae ,Oncology ,Mutation ,Cancer research ,Carcinogenesis ,business ,Non melanoma - Abstract
Copyright © 2003 Elsevier Ireland Ltd. All rights reserved., In cervical cells, the E6 protein of the oncogenic human papillomavirus (HPV) types inactivates p53, promoting its degradation. Consequently, mutations of the p53 gene are rarely seen in these cancers. Our recent data indicate that the cutaneous HPV38 is involved in skin carcinogenesis. In this study, we have determined the presence of HPV38 and the status of p53 gene in 32 non-melanoma skin cancers. We found that p53 gene is frequently mutated in HPV38-positive skin cancers and that HPV38 E6 does not promote p53 degradation. Thus, different mechanisms appear to be involved in the development of HPV-positive cervical and skin cancers.
- Published
- 2004
50. Frameshift mutation in the Dok1 gene in chronic lymphocytic leukemia
- Author
-
Charles Dumontet, Bakary S. Sylla, Rosita Accardi, Emeline Cros, François Roy, Alexandra Viller, Jocelyne Michelon, Sanghoon Lee, and Carlos M. Galmarini
- Subjects
Cancer Research ,Mutant ,Biology ,medicine.disease_cause ,Frameshift mutation ,chemistry.chemical_compound ,Genetics ,medicine ,Humans ,Nuclear protein ,Frameshift Mutation ,Molecular Biology ,Sequence Deletion ,Mutation ,Wild type ,RNA-Binding Proteins ,Tyrosine phosphorylation ,Phosphoproteins ,Leukemia, Lymphocytic, Chronic, B-Cell ,Molecular biology ,DNA-Binding Proteins ,chemistry ,Mitogen-activated protein kinase ,Cancer research ,biology.protein ,Mitogen-Activated Protein Kinases ,Tyrosine kinase - Abstract
B-cell chronic lymphocytic leukemia (B-CLL) is a malignant disease characterized by an accumulation of monoclonal CD5+ mature B cells, with a high percentage of cells arrested in the G0/G1 phase of the cell cycle, and a particular resistance toward apoptosis-inducing agents. Dok1 (downstream of tyrosine kinases) is an abundant Ras-GTPase-activating protein (Ras-GAP)-associated tyrosine kinase substrate, which negatively regulates cell proliferation, downregulates MAP kinase activation and promotes cell migration. The gene encoding Dok1 maps to human chromosome 2p13, a region previously found to be rearranged in B-CLL. We have screened the Dok1 gene for mutations from 46 individuals with B-CLL using heteroduplex analysis. A four-nucleotide GGCC deletion in the coding region was found in the leukemia cells from one patient. This mutation causes a frameshift leading to protein truncation at the carboxyl-terminus, with the acquisition of a novel amino-acid sequence. In contrast to the wild-type Dok1 protein, which has cytoplasmic/membrane localization, the mutant Dok1 is a nuclear protein containing a functional bipartite nuclear localization signal. Whereas overexpression of wild-type Dok1 inhibited PDGF-induced MAP kinase activation, this inhibition was not observed with the mutant Dok1. Furthermore the mutant Dok1 forms heterodimers with Dok1 wild type and the association can be enhanced by Lck-mediated tyrosine-phosphorylation. This is the first example of a Dok1 mutation in B-CLL and the data suggest that Dok1 might play a role in leukemogenesis.
- Published
- 2004
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.