31 research outputs found
UMI, a novel RNF168 ubiquitin binding domain involved in the DNA damage signaling pathway
No full textUbiquitination regulates important cellular processes, including the DNA damage response (DDR) and DNA repair. The complexity of the ubiquitin-mediated signals is decoded by ubiquitin receptors, which contain protein modules named ubiquitin binding domains (UBDs). We previously identified a new ubiquitin ligase, RNF168, involved in DDR and endowed with two UBDs named MIU (motif interacting with ubiquitin). Here we have provided the identification of a novel UBD, the UMI (UIM- and MIU-related UBD), present in RNF168, and characterized the interaction surface with ubiquitin, centered on two Leu residues. We have demonstrated that integrity of the UMI, in addition to the MIUs, is necessary for the proper localization of RNF168 and for ubiquitination of nuclear proteins, including histone H2A. Finally, we have shown that simultaneous inactivation of UMI and MIUs prevents the recruitment to DDR foci of the crucial downstream mediator 53BP1
Human papillomavirus type 16 E6 and E7 oncoproteins interact with the nuclear p53-binding protein 1 in an in vitro reconstructed 3D epithelium: new insights for the virus-induced DNA damage response
No full textAbstract Background Despite vaccination and screening measures, anogenital cancer, mainly promoted by HPV16 oncoproteins, still represents the fourth tumor and the second cause of death among women. Cell replication fidelity is the result of the host DNA damage response (DDR). Unlike many DNA viruses that promote their life cycle through the DDR inactivation, HR-HPVs encourage cells proliferation despite the DDR turned on. Why and how it occurs has been only partially elucidated. During HPV16 infection, E6 links and degrades p53 via the binding to the E6AP LXXLL sequence; unfortunately, E6 direct role in the DDR response has not clearly identified yet. Similarly, E7 increases DDR by competing with E2F1-pRb interaction, thus leading to the inactivation of pRb, and promotion, E2F1 mediated, of DDR genes translation, by binding to the pRb-like proteins CBP/p300 and p107, that also harbour LXXLL sequence, and via the interaction and activation of several DDR proteins. Methods To gain information regarding E6 and E7 contribution in DDR activation, we produced an in vitro 3D HPV16-E6E7 infected epithelium, already consolidated study model for HPVs, and validated it by assessing H&E staining and BrdU, HPV16 DNA, E6E7 proteins and γH2A.X/53BP1 double-strand break (DSBs) sensors expression; then we made an immuno-colocalization of E6 and E7 with cyclin E2 and B1. Since 53BP1, like E6 and E7, also binds p53 and pRb, we supposed their possible direct binding. To explore this hypothesis, we performed a double immunofluorescence of E6 and E7 with 53BP1, a sequence analysis of 53BP1 within its BRCT2 domain and then an in situ PLA within CaSki, E6E7HPV16 NHEKs and the 3D model. Results The in vitro epithelium resembled the histology and the events typical of in vivo infected tissues. E6E7HPV16 were both expressed in basal and differentiated strata and induced H2A.X phosphorylation and 53BP1 increment into nuclear foci. After highlighting E6 and E7 co-expression with 53BP1 and a LKVLL sequence within the 53BP1 BRCT2 domain, we demonstrated the bindings via the PLA technique. Conclusions Our results reinforce E6 and E7 role in cellular function control providing potentially new insights into the activity of this tumor virus
Prevalence and impact of COVID-19 sequelae on treatment pathways and survival of patients with cancer who recovered from SARS-Cov-2 infection: results from the OnCovid registry.
No full textretrospective, registry study David J Pinato, Josep Tabernero, Mark Bower, Lorenza Scotti, Meera Patel, Emeline Colomba, Saoirse Dolly, Angela Loizidou, John Chester, Uma Mukherjee, Alberto Zambelli, Alessia Dalla Pria, Juan Aguilar-Company, Diego Ottaviani, Amani Chowdhury, Eve Merry, Ramon Salazar, Alexia Bertuzzi, Joan Brunet, Matteo Lambertini, Marco Tagliamento, Anna Pous, Ailsa Sita-Lumsden, Krishnie Srikandarajah, Johann Colomba, Fanny Pommeret, Elia Segui, Daniele Generali, Salvatore Grisanti, Paolo Pedrazzoli, Gianpiero Rizzo, Michela Libertini, Charlotte Moss, Joanne S Evans, Beth Russell, Nadia Harbeck, Bruno Vincenzi, Federica Biello, Rossella Bertulli, Raquel Lilian, Sabrina Rossi, Maria Carmen Carmona-Garcia, Carlo Tondini, Laura Fox, Alice Baggi, Vittoria Fotia, Alessandro Parisi, Giampero Porzio, Maristella Saponara, Claudia Andrea Cruz, David Garcia-Illescas, Eudald Felip, Ariadna Roque Lloveras, Rachel Sharkey, Elisa Roldan, Roxana Reyes, Irina Earnshaw, Daniela Ferrante, Javier Marco-Hernandez, Isabel Ruiz-Camps, Gianluca Gaidano, Andrea Patriarca, Riccardo Bruna, Anna Sureda, Clara Martinez-Vila, Ana Sanchez de Torre, Luca Cantini, Marco Filetti, Lorenza Rimassa, Lorenzo Chiudinelli, Michela Franchi, Marco Krengli, Armando Santoro, Aleix Prat, Mieke Van Hemelrijck, Nikolaos Diamantis, Thomas Newsom-Davis, Alessandra Gennari, Alessio Cortellini, on behalf of the OnCovid study group
sj-docx-1-tam-10.1177_17588359231225028 – Supplemental material for Impact of SARS-CoV-2 vaccines and recent chemotherapy on COVID-19 morbidity and mortality in patients with soft tissue sarcoma: an analysis from the OnCovid registry
No full textSupplemental material, sj-docx-1-tam-10.1177_17588359231225028 for Impact of SARS-CoV-2 vaccines and recent chemotherapy on COVID-19 morbidity and mortality in patients with soft tissue sarcoma: an analysis from the OnCovid registry by Bruno Vincenzi, Alessio Cortellini, Alessandro Mazzocca, Sarah Orlando, Davide Romandini, Juan Aguilar-Company, Isabel Ruiz-Camps, Claudia Valverde Morales, Simeon Eremiev-Eremiev, Carlo Tondini, Joan Brunet, Rossella Bertulli, Salvatore Provenzano, Mark Bower, Daniele Generali, Ramon Salazar, Anna Sureda, Aleix Prat, Michalarea Vasiliki, Mieke Van Hemelrijck, Ailsa Sita-Lumsden, Alexia Bertuzzi, Sabrina Rossi, Amanda Jackson, Federica Grosso, Alvin J. X. Lee, Cian Murphy, Katherine Belessiotis, Uma Mukherjee, Fanny Pommeret, Angela Loizidou, Gianluca Gaidano, Gino M. Dettorre, Salvatore Grisanti, Marco Tucci, Claudia A. M. Fulgenzi, Alessandra Gennari, Andrea Napolitano and David J. Pinato in Therapeutic Advances in Medical Oncology</p
RNF168, a new RING finger, MIU-containing protein that modifies chromatin by ubiquitination of histones H2A and H2AX
No full textBACKGROUND
Modulation of chromatin structure has emerged as a critical molecular device to control gene expression. Histones undergo different post-translational modifications that increase chromatin accessibility to a number of regulatory factors. Among them, histone ubiquitination appears relevant in nuclear processes that govern gene silencing, either by inhibiting or activating transcription, and maintain genome stability, acting as scaffold to properly organize the DNA damage response. Thus, it is of paramount importance the identification and the characterization of new ubiquitin ligases that address histones.
RESULTS
We identified and characterized RNF168, a new chromatin-associated RING finger protein. We demonstrated that RNF168 is endowed with ubiquitin ligase activity both in vitro and in vivo, which targets histones H2A and H2AX, but not H2B, forming K63 polyubiquitin chains. We previously described the presence within RNF168 sequence of two MIU domains, responsible for the binding to ubiquitinated proteins. Here we showed that inactivation of the MIUs impairs ubiquitin binding ability in vitro and reduces chromatin association of RNF168 in vivo. Moreover, upon formation of DNA double strand breaks induced by chemical and physical agents, RNF168 is recruited to the DNA damage foci, where it co-localizes with gammaH2AX and 53BP1. The localization of RNF168 at the site of damage highly increases the local concentration of ubiquitinated proteins and determines the prolonged ubiquitination signal.
CONCLUSION
The RING finger protein RNF168 is a new ubiquitin ligase that functions as chromatin modifier, through histone ubiquitination. We hypothesize a dual function for RNF168. In normal condition RNF168 modifies chromatin structure by modulating ubiquitination of histone H2A. Upon DNA lesions, RNF168 is recruited to DNA damage response foci where it contributes to increase the amount of ubiquitinated proteins, thereby facilitating the downstream signalling cascade
Políticas sociales en cercanía : infancia y juventud en contexto de pobreza y vulnerabilidad social
No full textPresentación / Susana Mallo -- Introducción / Martín Couto y Betty Weisz-- El proceso de construcción de la Estrategia Nacional para la Infancia y la Adolescencia (ENIA): ideas, filtros institucionales y capacidades estatales en juego / Mauro Casa González y Belén Villegas Plá -- La infancia primero: una prioridad del gobierno de Mujica. ¿Y la igualdad de género? Análisis del caso de Uruguay Crece Contigo / Silvana Balsa Ruella -- Cuidados infantiles y vulnerabilidad. Sobre estrategias y percepciones/ Ana Casteluccio, Lorena Caffaro y Valeria Santana-- La protección de la primera infancia en Uruguay. Significados y sentidos de una estrategia integral: la experiencia del Programa Uruguay Crece Contigo / Cristian Pinato y Natalie Robaina -- Trayectorias ocultas tras el concepto “ni-ni”. ¿Qué tienen estos jóvenes para decir? / Leonel Rivero -- Jóvenes en Red: ¿construyendo ciudadanía?/ Alejandra Triñanes y Lucía Curiel -- La mirada desde los jóvenes. Jóvenes en Red en el departamento de Cerro Largo/ Milena Delgado Larrama -- Las nuevas políticas sociales de articulación en el territorio. El caso de Cercanías en la región norte de Uruguay/Sofía Angulo Benítez y Agustina Marques -- Estrategia de cercanía: modalidades, vínculos y sujetos. Los programas prioritarios en la zona de la Ruta 5 - Canelones / Natalia Amarillo Lema, Graciela Fagúndez Núñez Moraes y Sabrina Jassid Hernánde
Políticas sociales en cercanía : infancia y juventud en contexto de pobreza y vulnerabilidad social
No full textPresentación / Susana Mallo -- Introducción / Martín Couto y Betty Weisz-- El proceso de construcción de la Estrategia Nacional para la Infancia y la Adolescencia (ENIA): ideas, filtros institucionales y capacidades estatales en juego / Mauro Casa González y Belén Villegas Plá -- La infancia primero: una prioridad del gobierno de Mujica. ¿Y la igualdad de género? Análisis del caso de Uruguay Crece Contigo / Silvana Balsa Ruella -- Cuidados infantiles y vulnerabilidad. Sobre estrategias y percepciones/ Ana Casteluccio, Lorena Caffaro y Valeria Santana-- La protección de la primera infancia en Uruguay. Significados y sentidos de una estrategia integral: la experiencia del Programa Uruguay Crece Contigo / Cristian Pinato y Natalie Robaina -- Trayectorias ocultas tras el concepto “ni-ni”. ¿Qué tienen estos jóvenes para decir? / Leonel Rivero -- Jóvenes en Red: ¿construyendo ciudadanía?/ Alejandra Triñanes y Lucía Curiel -- La mirada desde los jóvenes. Jóvenes en Red en el departamento de Cerro Largo/ Milena Delgado Larrama -- Las nuevas políticas sociales de articulación en el territorio. El caso de Cercanías en la región norte de Uruguay/Sofía Angulo Benítez y Agustina Marques -- Estrategia de cercanía: modalidades, vínculos y sujetos. Los programas prioritarios en la zona de la Ruta 5 - Canelones / Natalia Amarillo Lema, Graciela Fagúndez Núñez Moraes y Sabrina Jassid Hernánde
Capsaicin 8% patch for treprostinil subcutaneous infusion site pain in pulmonary hypertension patients
No full textBackgroundTreprostinil sodium improves haemodynamics and symptoms in pulmonary arterial hypertension (PAH) patients, but its subcutaneous (s.c.) administration can produce severe local site pain, and lead to discontinuation of vital treatment. Treprostinil is a prostacyclin analogue which stimulates prostacyclin receptors in skin nociceptor terminals, resulting in pain and cutaneous hypersensitivity, for which current pain remedies have limited effect. Capsaicin 8% patch relieves neuropathic pain for 3 months after a single 60 min cutaneous application; we investigated whether its pre-application can reduce s.c. trepostinil-induced pain.MethodsA single-centre, double-blind, randomized, placebo-controlled, crossover study was conducted to assess the safety and efficacy of a single capsaicin 8% patch pre-application for s.c. treprostinil pain in 11 PAH patients, relative to control patch with low-dose capsaicin 0.075% cream.ResultsThe primary efficacy endpoint, mean difference between the two treatment arms in an 11-point numerical pain rating scale from baseline to 2 weeks after patch applications, was significantly lower on the capsaicin 8% patch treatment arm [P=0.01, mean difference=-1.47 units, 95% credible interval (CI): -2. 59 to -0.38] in the patients who completed the study per protocol, although intention-to-treat analysis did not show significant difference (P=0.28). Heat pain thresholds were decreased (P=0.027, mean difference=5.43°C, 95% CI: 0.71-10.21) and laser Doppler flux increased (P=0.016, mean difference=370 units, 95% CI: 612 to 127.9) at the application site immediately after capsaicin 8% patch, confirming activity. ConclusionsFurther investigation of the efficacy of capsaicin 8% patch in this indication is warranted.Clinical trial registrationClinicalTrials.gov: NCT01393795. © 2013 © The Author [2013]. Published by Oxford University Press on behalf of the British Journal of Anaesthesia. All rights reserved. For Permissions, please email: [email protected]
A novel ubiquitin mark at the N-terminal tail of histone H2As targeted by RNF168 ubiquitin ligase
No full textUbiquitination of histones plays a critical role in the regulation of several processes within the nucleus, including maintenance of genome stability and transcriptional regulation. The only known ubiquitination site on histones is represented by a conserved Lys residue located at the C terminus of the protein. Here, we describe a novel ubiquitin mark at the N-terminal tail of histone H2As consisting of two Lys residues at positions 13 and 15 (K13/K15). This "bidentate" site is a target of the DNA damage response (DDR) ubiquitin ligases RNF8 and RNF168. Histone mutants lacking the K13/K15 site impair RNF168- and DNA damage-dependent ubiquitination. Conversely, inactivation of the canonical C-terminal site prevents the constitutive monoubiquitination of histone H2As but does not abolish the ubiquitination induced by RNF168. A ubiquitination-defective mutant is obtained by inactivating both the N- and the C-terminal sites, suggesting that these are unique, non-redundant acceptors of ubiquitination on histone H2As. This unprecedented result implies that RNF168 generates a qualitatively different Ub mark on chromatin
Codon-specific KRAS mutations predict survival benefit of trifluridine/tipiracil in metastatic colorectal cancer
No full textGenomics has greatly improved how patients with cancer are being treated; however, clinical-grade genomic biomarkers for chemotherapies are currently lacking. Using whole-genome analysis of 37 patients with metastatic colorectal cancer (mCRC) treated with the chemotherapy trifluridine/tipiracil (FTD/TPI), we identified KRAS codon G12 (KRASG12) mutations as a potential biomarker of resistance. Next, we collected real-world data of 960 patients with mCRC receiving FTD/TPI and validated that KRASG12 mutations were significantly associated with poor survival, also in analyses restricted to the RAS/RAF mutant subgroup. We next analyzed the data of the global, double-blind, placebo-controlled, phase 3 RECOURSE trial (n = 800 patients) and found that KRASG12 mutations (n = 279) were predictive biomarkers for reduced overall survival (OS) benefit of FTD/TPI versus placebo (unadjusted interaction P = 0.0031, adjusted interaction P = 0.015). For patients with KRASG12 mutations in the RECOURSE trial, OS was not prolonged with FTD/TPI versus placebo (n = 279; hazard ratio (HR) = 0.97; 95% confidence interval (CI) = 0.73–1.20; P = 0.85). In contrast, patients with KRASG13 mutant tumors showed significantly improved OS with FTD/TPI versus placebo (n = 60; HR = 0.29; 95% CI = 0.15–0.55; P < 0.001). In isogenic cell lines and patient-derived organoids, KRASG12 mutations were associated with increased resistance to FTD-based genotoxicity. In conclusion, these data show that KRASG12 mutations are biomarkers for reduced OS benefit of FTD/TPI treatment, with potential implications for approximately 28% of patients with mCRC under consideration for treatment with FTD/TPI. Furthermore, our data suggest that genomics-based precision medicine may be possible for a subset of chemotherapies.</p
