4 research outputs found
Dysregulations of DNA and mRNA epigenetic modifications in breast cancer
Cette thèse est axée en deux parties :une première partie explorant la dérégulation d’une modification épigénétique localisée sur l’ADN tandis que la seconde porte sur une modification des ARN messagers. Ces deux modifications sont étudiées dans le cadre du cancer du sein. La première partie concernant l’hydroxymethylation de l’ADN décrit l’influence de l’infiltration immune sur l’expression de l’enzyme TET1, responsable de l’ajout de cette modification. En effet, notre étude met en évidence une anti-corrélation entre l’expression de TET1 et l’infiltration immune (via IHC et analyses in silico). De plus, nous démontrons que l’activation de la voie de signalisation NF-B dans le cancer du sein « basal-like » réduit l’expression de TET1. Finalement, nous avons pu mettre en évidence, grâce à plusieurs expériences dont de l’immunoprécipitation que cette régulation se déroulait (au moins en partie) via liaison d’NF-B au promoteur de TET1. Dans le but d’élargir notre model à d’autres cancers, nous avons pu également observer cette anti-corrélation et l’effet de NF-B dans les cancers de la thyroïde, du poumon et de la peau. Avec le développement récent de l’immunothérapie, ce lien entre modification épigénétique et immunité pourrait s’avérer intéressant à exploiter. La seconde partie décrit quant à elle la dérégulation de la modification m6A dans le cancer du sein en général et non plus dans un sous-type particulier. Nous avons non seulement été en mesure de décrire une accumulation de la marque dans les tissus cancéreux mais également de proposer une cause et une conséquence à cette dérégulation. En effet, cette augmentation de la présence d’m6A (observée par spectrométrie de masse) serait causée par la baisse d’expression d’une enzyme responsable de sa suppression (l’enzyme FTO). Cette dérégulation, en agissant directement sur les ARNm de la voie de signalisation WNT comme le montre notre m6A-seq, active d’avantage la voie. Cette suractivation des WNT entraine alors une transition épithélio-mésenchymateuse des cellules, étape importante dans le développement de métastases. Ainsi, les patientes présentant une tumeur à basse expression de FTO pourraient être plus sujettes à des récurrences tumorales. Nous avons également observé cette même régulation dans d’autres cancers comme celui de la prostate, à la fois in vitro, in vivo et in silico. De plus, notre recherche met également en évidence une plus grande sensitivité de ces tumeurs à un inhibiteur des WNT. En effet, nos expériences de xénogreffes soulignent que des tumeurs avec peu de FTO, traitées avec l’inhibiteur des WNT ICRT-3, sont bien moins actives et grandes que les tumeurs contrôle et développent moins de métastases. Mesurer la quantité d’FTO tumorale pourrait donc en théorie permettre une meilleure discrimination des patientes pour de tels traitements. Ces deux projets ont pour but d’améliorer notre compréhension de l’épigénétique dans le cancer du sein et ainsi proposer de potentielles implications thérapeutiques.This thesis is divided in two parts: the first part explores the dysregulation of an epigenetic modification localized on DNA and the second part is centred on a mRNA modification. These two marks are being studied in the context of breast cancer (BC). The first part describes the influence of immune cell infiltration on the expression of the TET1 enzyme, which is responsible for the addition of hydroxymethylation to DNA. Our study indeed first describes an anti-correlation between TET1 expression and immune infiltration (through both IHC and in silico analysis). In addition, we show that NF-B pathway activation in « basal-like » breast cancer cells downregulates TET1 expression. Finally, we highlighted through several experiments including immunoprecipitation, a TET1 promoter binding of NF-B. In an effort to generalize our data to other cancers, we also investigated this anticorrelation and the NF-B regulation in several cancers. Our results indicate that this immune related regulation also occurs in lung, thyroid and skin cancers. With the increasing use of immunotherapy as anti-cancer treatment, this link between an epigenetic modification and immunity might be interesting to explore. The second part focuses on the dysregulation of the m6A modification in breast cancer in general. We describe an accumulation of m6A in cancerous tissues and we further unravel cause and consequence of this dysregulation. Indeed, we show that this increase of m6A observed by mass spectrometry correlates with the downregulation of an enzyme (FTO), which is responsible for the removal of this mark. In addition, we observed with TCGA data that this downregulation occurs in multiple cancers such as prostate or lung cancers. Our results indicate that this dysregulation activates the WNT pathway through interaction with WNT related mRNAs (identified by our m6A-seq). This overactivation (observed with TOP FOP, IHC and western blot assays) promotes epithelialmesenchymal transition (EMT) which is essential in the development of metastasis. Consistent with this, we observed that patients with low FTO expressing breast tumors are more prone to tumor progression and recurrence. Moreover, we were able to generalize our data to other cancers including prostate cancer both through in vitro, in vivo and in silico data. Finally, our research also shows a greater sensitivity of FTO-low tumors to WNT inhibition. Indeed, our mice xenografts experiments showed that FTO-low tumors treated with the WNT inhibitor ICRT-3 are less metabolically active, smaller than controls and develop less metastasis. Measuring the amount of FTO in tumors could therefore allow for the stratification better of breast cancer patients for WNT inhibition therapy. Both projects aim to improve our understanding of the role of epigenetics in breast cancer and thus propose potential therapeutic implications.Doctorat en Sciences biomédicales et pharmaceutiques (Médecine)info:eu-repo/semantics/nonPublishe
Immunity drives TET1 regulation in cancer through NF-ÎşB
Ten-eleven translocation enzymes (TET1, TET2, and TET3), which induce DNA demethylation and gene regulation by converting 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), are often down-regulated in cancer. We uncover, in basal-like breast cancer (BLBC), genome-wide 5hmC changes related to TET1 regulation. We further demonstrate that TET1 repression is associated with high expression of immune markers and high infiltration by immune cells. We identify in BLBC tissues an anticorrelation between TET1 expression and the major immunoregulator family nuclear factor ÎşB (NF-ÎşB). In vitro and in mice, TET1 is down-regulated in breast cancer cells upon NF-ÎşB activation through binding of p65 to its consensus sequence in the TET1 promoter. We lastly show that these findings extend to other cancer types, including melanoma, lung, and thyroid cancers. Together, our data suggest a novel mode of regulation for TET1 in cancer and highlight a new paradigm in which the immune system can influence cancer cell epigenetics.SCOPUS: ar.jinfo:eu-repo/semantics/publishe
Patient-reported outcomes in terms of swallowing and quality of life after prophylactic versus reactive percutaneous endoscopic gastrostomy tube placement in advanced oropharyngeal cancer patients treated with definitive chemo-radiotherapy: Swall PEG study
Abstract Background Percutaneous endoscopic gastrostomy (PEG) is often used to provide nutritional support in locally advanced head and neck cancer patients undergoing multimodality treatment. However, there is little published data on the impact of prophylactic versus reactive PEG. PEG placement may affect swallowing-related physiology, function, and quality of life. The Swall PEG study is a randomized controlled phase III trial testing the impact of prophylactic versus reactive PEG on patient-reported outcomes in terms of swallowing and quality of life in oropharyngeal cancer patients. Methods Patients with locally advanced oropharyngeal cancer receiving chemo-radiotherapy will be randomized to either the prophylactic or reactive PEG tube group. Randomization will be stratified by human papillomavirus (HPV) status and unilateral versus bilateral positive neck lymph nodes. The primary objective of the study is the patient’s reported outcome in terms of swallowing (MD Anderson Dysphagia Inventory (MDADI)) at 6 months. Secondary objectives include health-related quality of life, dosimetric parameters associated with patient-reported outcomes, chemo-radiation toxicities, PEG tube placement complications, the impact of nutritional status on survival and toxicity outcomes, loco-regional control, overall survival, the impact of HPV and tobacco smoking on survival outcomes and toxicities, and the cost-effectiveness of each treatment strategy. Discussion Findings from this study will enhance clinical evidence regarding nutritional management in oropharyngeal cancer patients treated by concurrent chemo-radiation. Trial registration ClinicalTrials.gov NCT04019548, study protocol version 2.0_08/08/2019. Registered on 15 July 2019info:eu-repo/semantics/publishe