Attenuation of Soft-Tissue Sarcomas Resistance to the Cytotoxic Action of TNF-α by Restoring p53 Function

BACKGROUND: Isolated limb perfusion with TNF-α and melphalan is used with remarkable efficiency to treat unresectable limb sarcomas. Here we tested the ability of TNF-α to directly induce apoptosis of sarcoma cells. In addition, we investigated the impact of p53 in the regulation of such effect. METHODOLOGY/PRINCIPAL FINDINGS: We first analysed the ability of TNF-α to induce apoptosis in freshly isolated tumour cells. For this purpose, sarcoma tumours (n = 8) treated ex vivo with TNF-α were processed for TUNEL staining. It revealed substantial endothelial cell apoptosis and levels of tumour cell apoptosis that varied from low to high. In order to investigate the role of p53 in TNF-α-induced cell death, human sarcoma cell lines (n = 9) with different TP53 and MDM2 status were studied for their sensitivity to TNF-α. TP53(Wt) cell lines were sensitive to TNF-α unless MDM2 was over-expressed. However, TP53(Mut) and TP53(Null) cell lines were resistant. TP53 suppression in TP53(Wt) cell lines abrogated TNF-α sensitivity and TP53 overexpression in TP53(Null) cell lines restored it. The use of small molecules that restore p53 activity, such as CP-31398 or Nutlin-3a, in association with TNF-α, potentiated the cell death of respectively TP53(Mut) and TP53(Wt)/MDM2(Ampl). In particular, CP-31398 was able to induce p53 as well as some of its apoptotic target genes in TP53(Mut) cells. In TP53(Wt)/MDM2(Ampl) cells, Nutlin-3a effects were associated with a decrease of TNF-α-induced NF-κB-DNA binding and correlated with a differential regulation of pro- and anti-apoptotic genes such as TP53BP2, GADD45, TGF-β1 and FAIM. CONCLUSION/SIGNIFICANCE: More effective therapeutic approaches are critically needed for the treatment of unresectable limb sarcomas. Our results show that restoring p53 activity in sarcoma cells correlated with increased sensitivity to TNF-α, suggesting that this strategy may be an important determinant of TNF-α-based sarcomas treatment

Analyse du rôle de la voie p53 dans la réponse des sarcomes des tissus mous au traitement par TNF-alpha

Introduction: Although named for its antitumor properties, TNF-a is implicatedin a wide spectrum of diseases including chronic inflammation, autoimmunity andcancer. It is used for the loco regional treatment of limb’s sarcoma. P53 is an antioncogenewhose mutation is associated with tumour development. Experimental datademonstrated that TNF-a cytotoxic activity and p53 status are related.Material and methods: Our objective was to study by immunohistochemistrythe p53 status in 110 sarcoma patients treated by isolated limb perfusion with TNF-a. Then, we studied by confocal microscopy in 8 freshly obtained sarcoma tumours,the localization of apoptosis. Finally, in 9 sarcoma cell lines with different p53 status,we tested the p53/TNF-a relationship by abrogating p53 with a sh-RNA and by usingsmall molecules known to restore p53 functions. To better understand the mechanismsof resistance to TNF-a, we measured by EMSA the NF-kB-binding to DNA and withRT-PCR, we explored the regulation of some apoptosis related genes.Results: We demonstrated a relationship between p53 status and thehistological response to TNF-a use in humans. TNF-a induced apoptosis was presentin endothelial cells as well as in the tumour cells. TNF-a cytotoxicity was dependent onthe p53 status since in the cell lines studied, p53 abrogation reduced it and p53restoration allowed it to increase. Moreover, a potentiation of TNF-a cytotoxic effectwas observed when it was combined to CP-31398 or Nutlin-3a. The killing magnitudewas therefore related to the decrease in the NF-kB-binding to DNA when Nutlin-3awas added. A gene expression increase for RIPK2, TP53BP2 and GADD45 and adecrease for TGF-b1 and FAIM was observed if the combined treatment wassynergistic on tumour death cells.Conclusions: These results suggest that the use of compounds able to restorep53 activity could reverse the soft tissue sarcoma’s resistance to TNF-a treatment.Clinical studies should be performed in order to utilize this approach and to use it inthe context of sarcomas that are particularly resistant to conventional treatments.Introduction : Le TNF-a, impliqué dans l’inflammation et la défense de l’hôte, aaussi des propriétés anti-tumorales et est utilisé dans le traitement local des sarcomesdes membres. P53 est un anti-oncogène dont la mutation est associée audéveloppement de tumeurs. Des données expérimentales ont démontré une relationentre activité anti-tumorale de TNF-a et le statut de p53. Matériel et méthodes : Notre objectif a été d’étudier en immunohistochimie lestatut de p53 chez 110 patients atteints de sarcomes et traités par TNF-a. Ensuite,dans 8 sarcomes cultivés ex-vivo, nous avons étudié la localisation de l’apoptoseinduite par le TNF-a par microscopie confocale. Puis, dans 9 lignées de sarcomeshumains, nous avons testé la relation p53/TNF-a en abrogeant p53 grâce à un sh-RNA, ou en utilisant des petites molécules telles que CP-31398 ou Nutlin-3a aptes àrestaurer p53. Enfin, pour mieux comprendre les mécanismes de résistance au TNF-a,nous avons mesuré par méthode EMSA la liaison de NF-kB à l’ADN et recherché parRT-PCR quels gènes de l’apoptose étaient différentiellement régulés.Résultats : Le statut muté de p53 corrèle avec la réponse histologique autraitement par TNF-a chez l’homme. L’apoptose induite par le TNF-a est trouvée aussibien au niveau de la cellule endothéliale que de la cellule tumorale. De plus, laréponse au TNF-a est modulée par le statut de p53 puisque dans les lignées étudiées,l’abrogation de p53 supprime celle-ci alors que la réparation d’une activité p53 permetde l’augmenter. Enfin, une potentialisation de l’apoptose induite par TNF-a estobservée lorsqu’il est associé avec CP-31398 ou Nutlin-3a et elle corrèle avec ladiminution de la liaison du NF-kB à l’ADN. Une augmentation de l’expression desgènes RIPK2, TP53BP2 et GADD45 et une diminution de l’expression de TGF-b1 etFAIM est observée lorsque l’association de Nutlin-3a et TNF-a est synergique sur lamort cellulaire.Conclusions : Ces résultats suggèrent que l’utilisation de molécules capablesde restaurer l’activité de p53 peut inverser la résistance des sarcomes des tissusmous au traitement par TNF-a. Dans ce contexte, des études cliniques pourraientexploiter cette approche pour l’utiliser dans le cadre des sarcomes particulièrementrésistants aux traitements conventionnels ainsi que dans d’autres tumeurs

Role of p53 pathway in the response of soft tissue sarcomas to TNF-alpha treatment

Introduction : Le TNF-a, impliqué dans l’inflammation et la défense de l’hôte, aaussi des propriétés anti-tumorales et est utilisé dans le traitement local des sarcomesdes membres. P53 est un anti-oncogène dont la mutation est associée audéveloppement de tumeurs. Des données expérimentales ont démontré une relationentre activité anti-tumorale de TNF-a et le statut de p53. Matériel et méthodes : Notre objectif a été d’étudier en immunohistochimie lestatut de p53 chez 110 patients atteints de sarcomes et traités par TNF-a. Ensuite,dans 8 sarcomes cultivés ex-vivo, nous avons étudié la localisation de l’apoptoseinduite par le TNF-a par microscopie confocale. Puis, dans 9 lignées de sarcomeshumains, nous avons testé la relation p53/TNF-a en abrogeant p53 grâce à un sh-RNA, ou en utilisant des petites molécules telles que CP-31398 ou Nutlin-3a aptes àrestaurer p53. Enfin, pour mieux comprendre les mécanismes de résistance au TNF-a,nous avons mesuré par méthode EMSA la liaison de NF-kB à l’ADN et recherché parRT-PCR quels gènes de l’apoptose étaient différentiellement régulés.Résultats : Le statut muté de p53 corrèle avec la réponse histologique autraitement par TNF-a chez l’homme. L’apoptose induite par le TNF-a est trouvée aussibien au niveau de la cellule endothéliale que de la cellule tumorale. De plus, laréponse au TNF-a est modulée par le statut de p53 puisque dans les lignées étudiées,l’abrogation de p53 supprime celle-ci alors que la réparation d’une activité p53 permetde l’augmenter. Enfin, une potentialisation de l’apoptose induite par TNF-a estobservée lorsqu’il est associé avec CP-31398 ou Nutlin-3a et elle corrèle avec ladiminution de la liaison du NF-kB à l’ADN. Une augmentation de l’expression desgènes RIPK2, TP53BP2 et GADD45 et une diminution de l’expression de TGF-b1 etFAIM est observée lorsque l’association de Nutlin-3a et TNF-a est synergique sur lamort cellulaire.Conclusions : Ces résultats suggèrent que l’utilisation de molécules capablesde restaurer l’activité de p53 peut inverser la résistance des sarcomes des tissusmous au traitement par TNF-a. Dans ce contexte, des études cliniques pourraientexploiter cette approche pour l’utiliser dans le cadre des sarcomes particulièrementrésistants aux traitements conventionnels ainsi que dans d’autres tumeurs.Introduction: Although named for its antitumor properties, TNF-a is implicatedin a wide spectrum of diseases including chronic inflammation, autoimmunity andcancer. It is used for the loco regional treatment of limb’s sarcoma. P53 is an antioncogenewhose mutation is associated with tumour development. Experimental datademonstrated that TNF-a cytotoxic activity and p53 status are related.Material and methods: Our objective was to study by immunohistochemistrythe p53 status in 110 sarcoma patients treated by isolated limb perfusion with TNF-a. Then, we studied by confocal microscopy in 8 freshly obtained sarcoma tumours,the localization of apoptosis. Finally, in 9 sarcoma cell lines with different p53 status,we tested the p53/TNF-a relationship by abrogating p53 with a sh-RNA and by usingsmall molecules known to restore p53 functions. To better understand the mechanismsof resistance to TNF-a, we measured by EMSA the NF-kB-binding to DNA and withRT-PCR, we explored the regulation of some apoptosis related genes.Results: We demonstrated a relationship between p53 status and thehistological response to TNF-a use in humans. TNF-a induced apoptosis was presentin endothelial cells as well as in the tumour cells. TNF-a cytotoxicity was dependent onthe p53 status since in the cell lines studied, p53 abrogation reduced it and p53restoration allowed it to increase. Moreover, a potentiation of TNF-a cytotoxic effectwas observed when it was combined to CP-31398 or Nutlin-3a. The killing magnitudewas therefore related to the decrease in the NF-kB-binding to DNA when Nutlin-3awas added. A gene expression increase for RIPK2, TP53BP2 and GADD45 and adecrease for TGF-b1 and FAIM was observed if the combined treatment wassynergistic on tumour death cells.Conclusions: These results suggest that the use of compounds able to restorep53 activity could reverse the soft tissue sarcoma’s resistance to TNF-a treatment.Clinical studies should be performed in order to utilize this approach and to use it inthe context of sarcomas that are particularly resistant to conventional treatments

Implications of Perioperative Polypharmacy in Adolescents Undergoing Bariatric Surgery: a Single-Center Experience.

Adolescents seeking bariatric surgery may present with pre-existing psychiatric diagnoses for which they use chronic medications. To heighten awareness concerning perioperative polypharmacy in adolescents with extreme obesity, we conducted a retrospective review of patients undergoing laparoscopic sleeve gastrectomy between February 2010 and May 2017 at Children’s National Health System (CNHS). A total of 167 adolescent patients had pre-existing psychiatric diagnoses which included depression (50%), anxiety (23%), ADHD (23%), and binge eating disorder (11%). Medications prescribed to treat these diagnoses included selective serotonin reuptake inhibitors (SSRIs) and serotonin-norepinephrine reuptake inhibitors (SNRIs). Additionally, all patients were given fentanyl, ondansetron, morphine and acetaminophen perioperatively. Although no life threatening symptoms of drug-drug interactions (DDIs) were appreciated, the combined use of many different potent drugs in these patients warrants attention

Guidelines for clothing in the operating theatre, 2021

International audienceOBJECTIVE: To provide guidelines for the choice of items of clothing (except sterile surgical gown) for staff working in the operating theatre. DESIGN: A committee of nine experts from SFAR and the SF2H learned societies was convened. A formal conflict-of-interest policy was developed at the beginning of the process and enforced throughout. Likewise, it did not benefit from any funding from a company marketing a health product (drug or medical device). The authors were required to follow the rules of the GRADE® method (Grading of Recommendations Assessment, Development and Evaluation) to assess the quality of the evidence on which the recommendations were based. METHODS: We aimed to formulate recommendations according to the GRADE® methodology for four different fields: operating theatre suits, operating theatre hats, masks, and shoes/over-shoes. Each question was formulated according to the PICO format (Patient, Intervention, Comparison, Outcome). The literature review and recommendations were formulated according to the GRADE® methodology. RESULTS: The experts’ synthesis work and their application of the GRADE® method resulted in 13 recommendations. As the GRADE® method could not be integrally applied to all questions, some recommendations were formulated as expert opinions. CONCLUSION: Based on strong agreement between experts, we produced 13 recommendations to guide the choice of operating theatre attire

European Society of Anaesthesiology and Intensive Care consensus document on sustainability: 4 scopes to achieve a more sustainable practice

Climate change is a defining issue for our generation. The carbon footprint of clinical practice accounts for 4.7% of European greenhouse gas emissions, with the European Union ranking as the third largest contributor to the global healthcare industry's carbon footprint, after the United States and China. Recognising the importance of urgent action, the European Society of Anaesthesiology and Intensive Care (ESAIC) adopted the Glasgow Declaration on Environmental Sustainability in June 2023. Building on this initiative, the ESAIC Sustainability Committee now presents a consensus document in perioperative sustainability. Acknowledging wider dimensions of sustainability, beyond the environmental one, the document recognizes healthcare professionals as cornerstones for sustainable care, and puts forward recommendations in four main areas: direct emissions, energy, supply chain and waste management, and psychological and self-care of healthcare professionals. Given the urgent need to cut global carbon emissions, and the scarcity of evidence-based literature on perioperative sustainability, our methodology is based on expert opinion recommendations. A total of 90 recommendations were drafted by 13 sustainability experts in anaesthesia in March 2023, then validated by 36 experts from 24 different countries in a two-step Delphi validation process in May and June 2023. To accommodate different possibilities for action in high- versus middle-income countries, an 80% agreement threshold was set to ease implementation of the recommendations Europe-wide. All recommendations surpassed the 80% agreement threshold in the first Delphi round, and 88 recommendations achieved an agreement >90% in the second round. Recommendations include the use of very low fresh gas flow, choice of anaesthetic drug, energy and water preserving measures, "5R" policies including choice of plastics and their disposal, and recommendations to keep a healthy work environment or on the importance of fatigue in clinical practice. Executive summaries of recommendations in areas 1, 2 and 3 are available as cognitive aids that can be made available for quick reference in the operating room.Sin financiación3.6 Q2 JCR 20220.987 Q1 SJR 2023No data IDR 2022UE

Tenue vestimentaire au bloc opératoire 2021

International audienceObjectifÉmettre des recommandations pour le choix des éléments composant la tenue vestimentaire du personnel (hors casaque chirurgicale stérile) exerçant au bloc opératoire.ConceptionUn comité de neuf experts issus de la SFAR et de la SF2H a été constitué. Une politique de déclaration des liens d’intérêts a été appliquée et respectée durant tout le processus de réalisation du référentiel. De même, celui-ci n’a bénéficié d’aucun financement provenant d’une entreprise commercialisant un produit de santé (médicament ou dispositif médical). Le comité devait respecter et suivre la méthode GRADE® (Grading of Recommendations Assessment, Development and Evaluation) pour évaluer la qualité des données factuelles sur lesquelles étaient fondées les recommandations.MéthodesNous avons formulé des recommandations selon la méthodologie GRADE® en identifiant quatre champs différents : tuniques, articles coiffants, masques et chaussures/sur-chaussures. Chaque question a été formulée selon le format PICO (Patients, Intervention, Comparison, Outcome). L’analyse de la littérature et les recommandations ont été formulées selon la méthodologie GRADE®.RésultatsLe travail de synthèse des experts et l’application de la méthode GRADE® ont abouti à 13 recommandations. Pour l’ensemble des questions, la méthode GRADE® ne pouvant pas s’appliquer en totalité, les recommandations ont été formulées sous forme d’avis d’experts.ConclusionÀ partir d’un accord fort entre experts, nous avons pu formuler 13 recommandations sur la tenue vestimentaire au bloc opératoire