Epigenetic prognostic biomarkers in colorectal cancer

Colorectal cancer is one of the most common diagnosed cancers worldwide, and is the second most important cause of cancer mortality in Europe. The current TNM staging system used at the time of diagnosis is insufficient, as patients with the same tumor stage show wide variations in survival and tumor recurrence. Therefore, there is a need for identification of new biomarkers in colorectal cancer in order to identify high-risk patients and to guide treatment decision-making. In this thesis, epigenetic markers, including DNA methylation and histone modifications were studied in colorectal cancer patients. Several epigenetic clinically prognostic biomarkers were identified in colorectal cancer in this thesis, including both genome-wide and gene-specific patterns of DNA methylation and histone modifications. Knowledge of tumor biology is of key importance in the development of new therapies and the making of informed treatment decisions. Pathway-focused approaches, as presented in this thesis, provide information regarding possible synergistic interactions of biomarkers. Epigenetic mechanisms are unquestionably tied to the tumorigenic process and should be considered as a grand new source of information not only for identification of prognostic and predictive biomarkers, but also for the development of new, possibly tumor- and therefore patient-specific, anti-cancer therapies.UBL - phd migration 201

Environmental behaviour of inorganic pollutants present in raw and desalinated French marine sediments

International audienceIn the frame of long-term management of contaminated dredged sediments, this paper is centered on determinating the mobility of inorganic contaminants. A methodology derived from waste characterization has been developed and applied to marine sediments from Lazaret bay (Toulon, southern France) to determine the potential mobilization of inorganic pollutants in specific conditions. It consists of mineral and textural analysis combined with leaching tests. This methodology was applied to untreated, 5.8 % organic matter, light sandy silt harbor sediment and to the same sediment after a desalinization treatment. In both untreated and desalinated sediments, the contaminant content was around 26.1, 0.18, 42.5, 34, 31, 35 and 99 mg kg-1 for As, Cd, Cr, Cu, Ni, Pb and Zn, respectively. After 24 hours of time contact between deionized water and sediments, contaminant release of metals was very low (ca. <0.7 total mass %, for all studied elements) due to low solubility of the bearing solid phases (organic matter, carbonates and sulfides), while Mo and B were widely released. After 48 hours, Cd, As, Mo and B release was higher while more significant but no clear differences for other metal appeared

Evolution of iron speciation during hydration of C4 AF

International audienceIt is now well accepted and demonstrated that calcium silicate, calcium aluminate and calcium sulfo aluminate (ettringite, AFm) phases exhibit a good capability to fix metals and metalloids. Unfortunately the role of minor phases and especially calcium-ferric aluminate phase, shorthand C(4)AF is not well defined. In other systems like in soils or sediments iron phases play a key role in the fixation of pollutant. In cement sorption isotherms, indicated that various metals can be retained by the C(4)AF hydrated products. Therefore the capabilities of those phase to retain heavy metal should not be neglected. Previous investigations have shown that the minerals formed during the hydration of C(4)AF are similar to those formed from C3A (pure tri-calcium aluminate) under comparable conditions. Nevertheless no investigation was conducted at the molecular level and there is still a controversy whether Fe substitutes for Al in the hydrated minerals in whole or in part, or if it forms FeOOH clusters scattered throughout the matrix. In this context we have conducted XAS experiments using synchrotron radiation. It was found that the hydration of C(4)AF forms C(3)AH(6) (hydrogarnet) in which Fe randomly substitutes for Al as well as an amorphous FeOOH phase. Intermediate products like AFm (i.e., an ill organized lamellar phase) are also formed but rapidly evolve to C(3)AH(6); iron does not seem to be incorporated in the AFm structure

Design and Synthesis of Thiazolo[5,4-f]quinazolines as DYRK1A Inhibitors, Part I

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PO-033 Identification and functional evaluation of monoclonal antibodies specifically targeting human carbonic anhydrase IX

Introduction Poor vascularisation of solid tumours leads to inadequate nutrient and oxygen supplies which forces tumour cells to reprogram their metabolism. As a consequence the tumour cell's environment becomes acidic and hypoxic. This, in turn, triggers signalling cascades involving for example heterodimeric hypoxia-inducible factor (HIF). Activation of this hypoxia-induced transcriptional program is crucial for the survival of tumour cells in their hostile microenvironment but also their ability to metastasize. One of the genes upregulated through the HIF pathway is carbonic anhydrase (CA)-IX (CAIX, gene G250/MN-encoded transmembrane protein). CA-IX catalyses carbon dioxide (CO2) thereby generating a proton (H+) and bicarbonate (HCO3-), the latter of which is transported back into the cell and utilised to help safeguard intracellular pH (pHi) stability. Except for the stomach and the gallbladder, CA-IX expression is negligible in normal tissues. In contrast, a broad range of tumours express high levels of CA-IX, where the protein can serve as a biomarker for the early stages of tumour development but also as tumour marker of hypoxia associated with resistance to chemotherapy and radiotherapy. Material and methods Preclinical and clinical studies have shown that CA-IX is a promising therapeutic target for detection and therapy for several cancer types. To date only a limited number of ant-CAIX monoclonal antibodies (mAbs) have been available for clinical testing as therapeutic and imaging agents. In the current study, we generated and functionally categorised a panel of 51 mouse mAbs that specifically bind to human CA-IX. Results and discussions Characterisation of the mAbs revealed that of the mAbs with the best biophysical characteristics, three3 mAbs are suitable as an antibody-drug conjugate (ADC), two2 mAbs inhibit the CA-IX enzyme activity, and one1 mAb that is suitable for CA-IX imaging purposes. Conclusion These preliminary data presented here could thus form the basis for the development of novel CA-IX targeted immunotherapies and diagnostic tools for the treatment of cancer

HIV infection drives IgM and IgG3 subclass bias in Plasmodium falciparum-specific and total immunoglobulin concentration in Western Kenya

BACKGROUND: HIV infection is associated with more frequent and severe episodes of malaria and may be the result of altered malaria-specific B cell responses. However, it is poorly understood how HIV and the associated lymphopenia and immune activation affect malaria-specific antibody responses. METHODS: HIV infected and uninfected adults were recruited from Bondo subcounty hospital in Western Kenya at the time of HIV testing (antiretroviral and co-trimoxazole prophylaxis naïve). Total and Plasmodium falciparum apical membrane antigen-1 (AMA1) and glutamate rich protein-R0 (GLURP-R0) specific IgM, IgG and IgG subclass concentrations was measured in 129 and 52 of recruited HIV-infected and uninfected individuals, respectively. In addition, HIV-1 viral load (VL), CD4+ T cell count, and C-reactive protein (CRP) concentration was quantified in study participants. Antibody levels were compared based on HIV status and the associations of antibody concentration with HIV-1 VL, CD4+ count, and CRP levels was measured using Spearman correlation testing. RESULTS: Among study participants, concentrations of IgM, IgG1 and IgG3 antibodies to AMA1 and GLURP-R0 were higher in HIV infected individuals compared to uninfected individuals (all p < 0.001). The IgG3 to IgG1 ratio to both AMA1 and GLURP-R0 was also significantly higher in HIV-infected individuals (p = 0.02). In HIV-infected participants, HIV-1 VL and CRP were weakly correlated with AMA1 and GLURP-R0 specific IgM and IgG1 concentrations and total (not antigen specific) IgM, IgG, IgG1, and IgG3 concentrations (all p < 0.05), suggesting that these changes are related in part to viral load and inflammation. CONCLUSIONS: Overall, HIV infection leads to a total and malaria antigen-specific immunoglobulin production bias towards higher levels of IgM, IgG1, and IgG3, and HIV-1 viraemia and systemic inflammation are weakly correlated with these changes. Further assessments of antibody affinity and function and correlation with risk of clinical malaria, will help to better define the effects of HIV infection on clinical and biological immunity to malaria

Risk of first ischaemic stroke and use of antidopaminergic antiemetics: nationwide case-time-control study

OBJECTIVE: To estimate the risk of ischaemic stroke associated with antidopaminergic antiemetic (ADA) use. DESIGN: Case-time-control study. SETTING: Data from the nationwide French reimbursement healthcare system database Système National des Données de Santé (SNDS). PARTICIPANTS: Eligible participants were ≥18 years with a first ischaemic stroke between 2012 and 2016 and at least one reimbursement for any ADA in the 70 days before stroke. Frequencies of ADA reimbursements were compared for a risk period (days -14 to -1 before stroke) and three matched reference periods (days -70 to -57, -56 to -43, and -42 to -29) for each patient. Time trend of ADA use was controlled by using a control group of 21 859 randomly selected people free of the event who were individually matched to patients with stroke according to age, sex, and risk factors of ischaemic stroke. MAIN OUTCOME MEASURES: Association between ADA use and risk of ischaemic stroke was assessed by estimating the ratio of the odds ratios of exposure evaluated in patients with stroke and in controls. Analyses were adjusted for time varying confounders (anticoagulants, antiplatelets, and prothrombotic or vasoconstrictive drugs). RESULTS: Among the 2612 patients identified with incident stroke, 1250 received an ADA in the risk period and 1060 in the reference periods. The comparison with the 5128 and 13 165 controls who received an ADA in the same periods yielded a ratio of adjusted odds ratios of 3.12 (95% confidence interval 2.85 to 3.42). Analyses stratified by age, sex, and history of dementia showed similar results. Ratio of adjusted odds ratios for analyses stratified by ADA was 2.51 (2.18 to 2.88) for domperidone, 3.62 (3.11 to 4.23) for metopimazine, and 3.53 (2.62 to 4.76) for metoclopramide. Sensitivity analyses suggested the risk would be higher in the first days of use. CONCLUSIONS: Using French nationwide exhaustive reimbursement data, this self-controlled study reported an increased risk of ischaemic stroke with recent ADA use. The highest increase was found for metopimazine and metoclopramide

Environmental assessment of the behavior of a BOF steel slag used in road construction : the PRECODD-ECLAIR research program

International audienceSteel production generate great amounts of by-products as steel slags. The use of Basic Oxygen Furnace slags (BOF slags) has been restrained due to insufficient volume stability, and due to the lack of environmental regulations. The purpose of the PRECODD-ECLAIR research program is to develop a behavior model based on a multi-scale physico-chemical, mechanical, hydrodynamic and ecotoxicological characterizations of a BOF slag used in a public works scenario. This paper aims at presenting the overall ECLAIR research program, the equipped experimental platform constructed using a BOF steel slag, and the first results of the slag characterization

Digital Multiplex Ligation-Dependent Probe Amplification for Detection of Key Copy Number Alterations in T- and B-Cell Lymphoblastic Leukemia

Recurrent and clonal genetic alterations are characteristic of different subtypes of T- and B-cell lymphoblastic leukemia (ALL), and several subtypes are strong independent predictors of clinical outcome. A next-generation sequencing based multiplex ligation-dependent probe amplification variant (digitalMLPA) has been developed enabling simultaneous detection of copy number alterations (CNAs) of up to 1000 target sequences. This novel digitalMLPA assay was designed and optimized to detect CNAs of 56 key target genes and regions in ALL. A set of digital karyotyping probes has been included for the detection of gross ploidy changes, to determine the extent of CNAs, while also serving as reference probes for data normalization. Sixty-seven ALL patient samples (including B- and T-cell ALL), previously characterized for genetic aberrations by standard MLPA, array comparative genomic hybridization, and/or single-nucleotide polymorphism array, were analyzed single blinded using digitalMLPA. The digitalMLPA assay reliably identified whole chromosome losses and gains (including high hyperdiploidy), whole gene deletions or gains, intrachromosomal amplification of chromosome 21, fusion genes, and intragenic deletions, which were confirmed by other methods. Furthermore, subclonal alterations were reliably detected if present in at least 20% to 30% of neoplastic cells. The diagnostic sensitivity of the digitaLMLPA assay was 98.9%, and the specificity was 97.8%. These results merit further consideration of digitalMLPA as a valuable alternative for genetic work-up of newly diagnosed ALL patients.Peer reviewe

Real-life management of neovascular age-related macular degeneration (nAMD) in France: a nationwide observational study using retrospective claims data

AIMS: Intravitreal anti-vascular endothelial growth factor (anti-VEGF) therapy is standard care for neovascular age-related macular degeneration (nAMD), but the recommended monthly injection regimen is burdensome. Evidence suggests low injection/monitoring frequencies in clinical practice and suboptimal vision outcomes. This observational cohort study uses administrative claims data from the French national healthcare system to assess anti-VEGF treatment patterns and nAMD-specific healthcare resource demands and costs. PATIENTS AND METHODS: nAMD patients ≥50 years initiating intravitreal ranibizumab, aflibercept or bevacizumab treatment (2014‒2015), and propensity score-matched non-nAMD patients (controls), were identified from the Echantillon Généraliste de Bénéficiaires database. Outcomes of interest included anti-VEGF treatment patterns, and healthcare resource utilization and associated costs of patients vis-à-vis controls over 24 months. RESULTS: Study patients (n = 355) received (mean) 5.2 and 2.4 anti-VEGF injections over 0‒12 and 12‒24 months, respectively. Most patients (79.0%) remained on their initial anti-VEGF agent; among treatment switchers the most common transition was from ranibizumab to aflibercept. During follow-up, nAMD patients were more likely than controls to require ophthalmology visits (99.7% vs 44.8%), ocular procedures (optical coherence tomography/angiography/fundoscopy) (96.9% vs 27.2%), cataract surgery (13.0% vs 6.7%), and medical transports (38.0% vs 31.9%). Mean numbers of ophthalmology visits (25.1 vs 1.2) and medical transports (6.0 vs 3.5) were higher (p<.01) among nAMD patients. Total reimbursed costs were two-fold higher for nAMD patients than controls (mean €16,799 vs €8255) due to higher treatment costs (€6847 vs €1156), medical fees (€1858 vs €295), hospital fees (€6396 vs €5235), and transport costs (€358 vs €259). Excess total healthcare cost was (mean) €5279 and €7918 over the first 12 and 24 months of treatment, respectively. CONCLUSIONS: Current intravitreal anti-VEGF treatment and monitoring requirements place considerable economic burden on the French healthcare system. New intravitreal therapies with extended dosing intervals and predictable efficacy might reduce demand on ophthalmology services