Institut National De La Santé Et De La Recherche Médicale Research Articles

Neutralizing IFN-γ autoantibodies are rare and pathogenic in HLA-DRB1*15:02 or 16:02 individuals.

BACKGROUNDWeakly virulent environmental mycobacteria (EM) can cause severe disease in HLA-DRB1*15:02 or 16:02 adults harboring neutralizing anti-IFN-γ autoantibodies (nAIGAs). The overall prevalence of nAIGAs in the general population is unknown, as are the penetrance of nAIGAs in HLA-DRB1*15:02 or 16:02 individuals and the proportion of patients with unexplained, adult-onset EM infections carrying nAIGAs.METHODSThis study analyzed the detection and neutralization of anti-IFN-γ autoantibodies (auto-Abs) from 8,430 healthy individuals of the general population, 257 HLA-DRB1*15:02 or 16:02 carriers, 1,063 patients with autoimmune disease, and 497 patients with unexplained severe disease due to EM.RESULTSWe found that anti-IFN-γ auto-Abs detected in 4,148 of 8,430 healthy individuals (49.2%) from the general population of an unknown HLA-DRB1 genotype were not neutralizing. Moreover, we did not find nAIGAs in 257 individuals carrying HLA-DRB1* 15:02 or 16:02. Additionally, nAIGAs were absent in 1,063 patients with an autoimmune disease. Finally, 7 of 497 patients (1.4%) with unexplained severe disease due to EM harbored nAIGAs.CONCLUSIONThese findings suggest that nAIGAs are isolated and that their penetrance in HLA-DRB1*15:02 or 16:02 individuals is low, implying that they may be triggered by rare germline or somatic variants. In contrast, the risk of mycobacterial disease in patients with nAIGAs is high, confirming that these nAIGAs are the cause of EM disease.FUNDINGThe Laboratory of Human Genetics of Infectious Diseases is supported by the Howard Hughes Medical Institute, the Rockefeller University, the St. Giles Foundation, the National Institutes of Health (NIH) (R01AI095983 and U19AIN1625568), the National Center for Advancing Translational Sciences (NCATS), the NIH Clinical and Translational Science Award (CTSA) program (UL1 TR001866), the French National Research Agency (ANR) under the "Investments for the Future" program (ANR-10-IAHU-01), the Integrative Biology of Emerging Infectious Diseases Laboratory of Excellence (ANR-10-LABX-62-IBEID), ANR-GENMSMD (ANR-16-CE17-0005-01), ANR-MAFMACRO (ANR-22-CE92-0008), ANRSECTZ170784, the French Foundation for Medical Research (FRM) (EQU201903007798), the ANRS-COV05, ANR GENVIR (ANR-20-CE93-003), and ANR AI2D (ANR-22-CE15-0046) projects, the ANR-RHU program (ANR-21-RHUS-08-COVIFERON), the European Union's Horizon 2020 research and innovation program under grant agreement no. 824110 (EASI-genomics), the Square Foundation, Grandir - Fonds de solidarité pour l'enfance, the Fondation du Souffle, the SCOR Corporate Foundation for Science, the Battersea & Bowery Advisory Group, William E. Ford, General Atlantic's Chairman and Chief Executive Officer, Gabriel Caillaux, General Atlantic's Co-President, Managing Director, and Head of business in EMEA, and the General Atlantic Foundation, Institut National de la Santé et de la Recherche Médicale (INSERM) and of Paris Cité University. JR was supported by the INSERM PhD program for doctors of pharmacy (poste d'accueil INSERM). JR and TLV were supported by the Bettencourt-Schueller Foundation and the MD-PhD program of the Imagine Institute. MO was supported by the David Rockefeller Graduate Program, the Funai Foundation for Information Technology (FFIT), the Honjo International Scholarship Foundation (HISF), and the New York Hideyo Noguchi Memorial Society (HNMS).

Drug Repurposing Patent Applications April–June 2021

Drug Repurposing Patent Applications April–June 2021

A monocyte/dendritic cell molecular signature of SARS-CoV-2-related multisystem inflammatory syndrome in children with severe myocarditis

BackgroundSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in children is generally milder than in adults, but a proportion of cases result in hyperinflammatory conditions often including myocarditis.MethodsTo better understand these cases, we applied a multiparametric approach to the study of blood cells of 56 children hospitalized with suspicion of SARS-CoV-2 infection. Plasma cytokine and chemokine levels and blood cellular composition were measured, alongside gene expression at the bulk and single-cell levels.FindingsThe most severe forms of multisystem inflammatory syndrome in children (MIS-C) related to SARS-CoV-2 that resulted in myocarditis were characterized by elevated levels of pro-angiogenesis cytokines and several chemokines. Single-cell transcriptomics analyses identified a unique monocyte/dendritic cell gene signature that correlated with the occurrence of severe myocarditis characterized by sustained nuclear factor κB (NF-κB) activity and tumor necrosis factor alpha (TNF-α) signaling and associated with decreased gene expression of NF-κB inhibitors. We also found a weak response to type I and type II interferons, hyperinflammation, and response to oxidative stress related to increased HIF-1α and Vascular endothelial growth factor (VEGF) signaling.ConclusionsThese results provide potential for a better understanding of disease pathophysiology.FundingAgence National de la Recherche (Institut Hospitalo-Universitaire Imagine, grant ANR-10-IAHU-01; Recherche Hospitalo-Universitaire, grant ANR-18-RHUS-0010; Laboratoire d’Excellence ‘‘Milieu Intérieur,” grant ANR-10-LABX-69-01; ANR-flash Covid19 “AIROCovid” and “CoVarImm”), Institut National de la Santé et de la Recherche Médicale (INSERM), and the “URGENCE COVID-19” fundraising campaign of Institut Pasteur.

A Monocyte/Dendritic Cell Molecular Signature of SARS-CoV-2 Related Multisystem Inflammatory Syndrome in Children (MIS-C) with Severe Myocarditis

SARS-CoV-2 infection in children is generally milder than in adults, yet a proportion of cases result in hyperinflammatory conditions often including myocarditis. To better understand these cases, we applied a multi-parametric approach to the study of blood cells of 56 children hospitalized with suspicion of SARS-CoV-2 infection. The most severe forms of MIS-C (multisystem inflammatory syndrome in children related to SARS-CoV-2), that resulted in myocarditis, were characterized by elevated levels of pro-angiogenesis cytokines and several chemokines. Single-cell transcriptomic analyses identified a unique monocyte/dendritic cell gene signature that correlated with the occurrence of severe myocarditis, characterized by sustained NF-κ B activity, TNF-α signaling, associated with decreased gene expression of NF-κ B inhibitors. We also found a weak response to type-I and type-II interferons, hyperinflammation and response to oxidative stress related to increased HIF-1α and VEGF signaling. These results provide potential for a better understanding of disease pathophysiology.Funding: The study was supported by the Institut National de la Santé et de la Recherche Médicale (INSERM), by the “URGENCE COVID-19” fundraising campaign of Institut Pasteur, by the Atip-Avenir, Emergence ville de Paris program and fond de dotation Janssen Horizon and by government grants managed by the Agence National de la Recherche as part of the “Investment for the Future” program (Institut Hospitalo-Universitaire Imagine, grant ANR-10-IAHU-01, Recherche Hospitalo-Universitaire, grant ANR-18-RHUS-0010, Laboratoire d’Excellence ‘‘Milieu Intérieur”, grant ANR-10-LABX-69-01), the Centre de Référence Déficits Immunitaires Héréditaires (CEREDIH), the Agence National de la Recherche (ANR-flash Covid19 “AIROCovid” to FRL and “CoVarImm” to DD and JDS), and by the FASTFoundation (French Friends of Sheba Tel Hashomer Hospital). The LabTech Single-Cell@Imagine is supported by the Paris Region and the “Investissements d’avenir” program through the 2019 ATF funding – Sésame Filières PIA (Grant N°3877871).CdC is the recipient of a CIFRE-PhD (Sanofi). L.B. was a recipient of an Imagine institute PhD international program supported by the Fondation Bettencourt Schueller. L.B. was also supported by the EUR G.E.N.E. (reference #ANR-17-EURE-0013) and is part of the Université de Paris IdEx #ANR-18-IDEX-0001 funded by the French Government through its“Investments for the Future” program. S.M. was a recipient of an INSERM and Institut Imagine post-doctorat program supported by the Fondation pour la Recherche Médicale (FRMN°SPF20170938825). NS was a recipient of the Pasteur-Roux-Cantarini Fellowship. VGP obtained an Imagine international PhD fellowship program supported by the Fondation Bettencourt Schueller. BPP is the recipient of an ANRS post-doctoral fellowship.Conflict of Interest: DD, FRL, JT and MMM are listed as inventors on a patent application related to this technology (European Patent Application no. EP21305197, entitled “Methods of predicting multisystem inflammatory syndrome (MIS-C) with severe myocarditis in subjects suffering from a SARS-CoV-2 infection”).Ethical Approval: The study was approved by the Ethics Committee (Comité de Protection des Personnes Ouest IV, n° DC-2017-2987).

The association of living donor source with patient and graft survival among kidney transplant recipients in the ERA-EDTA Registry - a retrospective study.

SummaryIn this study we aimed to compare patient and graft survival of kidney transplant recipients who received a kidney from a living‐related donor (LRD) or living‐unrelated donor (LUD). Adult patients in the ERA‐EDTA Registry who received their first kidney transplant in 1998–2017 were included. Ten‐year patient and graft survival were compared between LRD and LUD transplants using Cox regression analysis. In total, 14 370 patients received a kidney from a living donor. Of those, 9212 (64.1%) grafts were from a LRD, 5063 (35.2%) from a LUD and for 95 (0.7%), the donor type was unknown. Unadjusted five‐year risks of death and graft failure (including death as event) were lower for LRD transplants than for LUD grafts: 4.2% (95% confidence interval [CI]: 3.7–4.6) and 10.8% (95% CI: 10.1–11.5) versus 6.5% (95% CI: 5.7–7.4) and 12.2% (95% CI: 11.2–13.3), respectively. However, after adjusting for potential confounders, associations disappeared with hazard ratios of 0.99 (95% CI: 0.87–1.13) for patient survival and 1.03 (95% CI: 0.94–1.14) for graft survival. Unadjusted risk of death‐censored graft failure was similar, but after adjustment, it was higher for LUD transplants (1.19; 95% CI: 1.04–1.35). In conclusion, patient and graft survival of LRD and LUD kidney transplant recipients was similar, whereas death‐censored graft failure was higher in LUD. These findings confirm the importance of both living kidney donor types.

Granulosa cells provide elimination of apoptotic oocytes through unconventional autophagy-assisted phagocytosis.

Do human granulosa cells (GCs) ingest and destroy apoptotic oocytes? Somatic GCs ingest and destroy apoptotic oocytes and other apoptotic substrates through unconventional autophagy-assisted phagocytosis. Most (99%) ovarian germ cells undergo apoptosis through follicular atresia. The mode of cleaning of atretic follicles from the ovary is unclear. Ovarian GCs share striking similarities with testicular Sertoli cells with respect to their origin and function. Somatic Sertoli cells are responsible for the elimination of apoptotic spermatogenic cells through unconventional autophagy-assisted phagocytosis. Human GCs were tested for the ability to ingest and destroy the apoptotic oocytes and other apoptotic substrates. A systemic study of the main phagocytosis steps has been performed at different time points after loading of apoptotic substrates into the GC. Primary cultures of GC retrieved following controlled ovarian stimulation of five women for IVF/ICSI and a human granulosa KGN cell line were incubated with different apoptotic substrates: oocytes which underwent spontaneous apoptosis during the cultivation of immature germ cells for IVF/ICSI; apoptotic KGN cells; and apoptotic membranes from rat retinas. Cultured GC were analyzed for the presence of specific molecular markers characteristic of different steps of phagocytic and autophagy machineries by immunocytochemistry, confocal microscopy, transmission electron microscopy and western blotting, before and after loading with apoptotic substrates. Incubation of human GC with apoptotic substrates resulted in their translocation in cell cytoplasm, concomitant with activation of the phagocytosis receptor c-mer proto-oncogene tyrosine kinase MERTK (P < 0.001), clumping of motor molecule myosin II, recruitment of autophagy proteins: autophagy-related protein 5 (ATG5), autophagy-related protein 6 (Beclin1) and the rise of a membrane form of microtubule-associated protein 1 light chain 3 (LC3-II) protein. Ingestion of apoptotic substrates was accompanied by increased expression of the lysosomal protease Cathepsin D (P < 0.001), and a rise of lysosomes in the GCs, as assessed by different techniques. The level of autophagy adaptor, sequestosome 1/p62 (p62) protein remained unchanged. N/A. The number of patients described here is limited. Also the dependence of phagocytosis on reproductive hormone status of patients should be analyzed. Removal of apoptotic oocytes by surrounding GC seems likely to be a physiological mechanism involved in follicular atresia. Proper functioning of this mechanism may be a new strategy for the treatment of ovarian dysfunctions associated with an imbalance in content of germ cells in the ovaries, such as premature ovarian failure and polycystic ovary syndrome. The study was funded by Rennes Metropole (AIS 2015) and Agence de BioMédecine. This work was supported by funding from Université de Rennes1, Institut National de la Santé et de la Recherche Médicale (INSERM) and CHU de Rennes. A.B. is funded in part by the program Actions Concertées Interpasteuriennes (ACIP) and a research grant from the European Society of Pediatric Endocrinology. This work is supported by the Agence Nationale de la Recherche Grants ANR-17-CE14-0038 and ANR-10-LABX-73. The authors declare no competing interests.

Cardiovascular Toxicities Associated With Hydroxychloroquine and Azithromycin

Keywords: azithromycin;cardiotoxicity;hydroxychloroquine;phamacovigilance EN azithromycin cardiotoxicity hydroxychloroquine phamacovigilance 303 305 3 07/23/20 20200721 NES 200721 Although hydroxychloroquine and azithromycin, alone or in combination, have been proposed for treatment of patients with coronavirus disease 2019 (COVID-19),[1] their potential cardiovascular toxicities had limited consideration in this new clinical environment The proportion that resulted in death for TdP/VT cases was 8 4% (7/83) with hydroxychloroquine and 20 2% (52/257) with azithromycin versus 0% (0/53) and 5 4% (12/223) for LQT without TdP/VT with hydroxychloroquine and azithromycin, respectively ( I P i 0 001 for both) [Extracted from the article] Copyright of Circulation is the property of Lippincott Williams & Wilkins and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission However, users may print, download, or email articles for individual use This abstract may be abridged No warranty is given about the accuracy of the copy Users should refer to the original published version of the material for the full abstract (Copyright applies to all Abstracts )

1425P - Towards a screening test for cancer by circulating DNA analysis

BackgroundCirculating DNA (cfDNA) has emerged as a potential biomarker in cancer, and is the subject of extensive studies in translational and clinical research. Our group has been interested in its implication and clinical significance in the field of oncology for many years, and is now focused on evaluating its potential for early cancer detection. MethodsWe recently developed a screening test (MNR: Multi Normalized Ratio), based on various cfDNA parameters determined by a specific q-PCR based method, targeting both nuclear and mitochondrial sequences. ResultsWhen applied to the supernatant of cell culture, the MNR had a discriminative potential of 100% between normal and cancer cell lines. An extensive evaluation of this test was carried out in plasma samples of 289 healthy subjects and 987 cancer patients (CRC, breast, liver, pancreatic, ovarian) of all stages. Preliminary results revealed a high potential with an AUC of 0.81 (0.78-0.84, 95% CI), a 70% sensitivity (Se) and 77% specificity (Sp). In breast cancer (N=169), an AUC of 0.82 (0.78-0.86, 95% CI) with 72% Se and 80% Sp were observed. In all stages CRC patients (N=795), the results showed an AUC of 0.80 (0.78-0.84, 95%, CI), 75% Se and 70% Sp; for CRC stages 0/I/II (N=426), an AUC of 0.79 (0.75-0.82, 95% CI), 70% Se and 72% Sp; and for CRC stage IV (N=186), a 0.86 AUC (0.82-0.89, 95% CI) with 75% Se and 80% Sp. When combining the MNR to a total cfDNA concentration threshold value (AUC = 0.81 (0.79-0.83, 95% CI), 72% Se and 76% Sp for all stage cancers (N=987)), in a test cohort of 173 stages 0/I/II CRC patients and 132 healthy individuals, we increased the sensitivity and specificity to 74% and 95% respectively. Furthermore we recently discovered that cfDNA fragmentation, as determined by Whole Genome Sequencing using either double or single strand library, is also a parameter enabling discrimination between healthy and cancer individuals. ConclusionsThe implementation of a multi-parametric test combining total cfDNA quantification, MNR and fragmentation biomarkers, with help of a decision tree in machine learning, is currently on-going. Our data suggest that our strategy in targeting cfDNA structural features might be powerful for early cancer detection, and appears as an alternative or a synergistic combination to the detection of mutations. Legal entity responsible for the studyAlain R. Thierry - INSERM (Institut national de la santé et de la recherche médicale). FundingMSDAvenir - Mitest / Alain R. Thierry is supported by INSERM (Institut national de la santé et de la recherche médicale). DisclosureAll authors have declared no conflicts of interest.

3O - Systemic gut microbial metabolites limit the anti-tumour effect of CTLA-4 blockade in hosts with cancer

BackgroundGut microbiota composition might influence the clinical benefit of immune checkpoints in patients with advanced cancer but mechanisms underlying this relationship remained unclear. Molecular mechanism whereby the gut microbiota influences immune responses is mainly assigned to gut microbial metabolites. The main metabolites of the gut microbiota are short-chain fatty acids (SCFAs), which are produced in large amounts in the colon through bacterial fermentation of dietary fiber. We evaluated in mice and in patients treated with anti-CTLA-4 if SCFAs levels could be related to the clinical outcome. MethodsThirty-nine patients with metastatic melanoma were treated with ipilimumab were prospectively enrolled. Fecal microbiota composition was assessed using 16S metagenomic analysis and quantitative PCR (Q-PCR) analyses for Faecalibacterium prausnitzii (F. prausnitzii) at baseline. Stool and serum concentrations of acetate, propionate and butyrate were also evaluated in two independent cohorts in accordance with clinical outcomes. Peripheral blood lymphocytes immunophenotypes were studied in parallel. The anti-CTLA-4 anti-tumor effect was also evaluated in mice models in with or without butyrate supplementation. ResultsSystemic butyrate was linked to Faecalibacterium prausnitzii enrichment in the stool of patients. High blood SCFAs levels (mainly propionate or butyrate) were associated with resistance to CTLA-4 blockade and were associated with a rise of Treg cells. During the course of anti-CTLA-4 treatment, butyrate limited the up-regulation of CD80/CD86 expression on dendritic cells, the induction of tumor-specific T cells and the accumulation of memory T cells in mice with cancer. In patients, restricted accumulation of memory T cells and IL-2 impregnation after ipilimumab treatment was observed. ConclusionsAltogether, these results suggested that gut microbial metabolites might favor an immune tolerance profile that limits anti-CTLA-4 activity. Legal entity responsible for the studyInstitut Gustave Roussy. FundingThis study was funded by Gustave Roussy Cancer Campus, Fondation Gustave Roussy, the Institut national de la santé et de la recherche médicale (INSERM), the Centre national de la recherche scientifique (CNRS), SIRIC SOCRATE (INCa DGOS INSERM 6043), SIRIC SOCRATE 2.0 (INCa-DGOS-INSERM_12551), MMO program: ANR-10IBHU-0001); Direction General de l’Offre de Soins (DGOS; TRANSLA 12-174); Institut National du Cancer (INCa; 2012-062 N_ Canceropole: 2012-1-RT-14-IGR-01). Dr. Clélia Coutzac was supported by fellowships from Fondation pour la Recherche Medicale (FRM) from 2015 to 2016. DisclosureC. Coutzac: Honoraria (self): AMGEN; Honoraria (self): Servier. L. Lacroix: Advisory/Consultancy, Speaker Bureau/Expert testimony: Roche; Advisory/Consultancy, Speaker Bureau/Expert testimony: AstraZeneca; Advisory/Consultancy: BMS; Advisory/Consultancy: Genomic Health; Advisory/Consultancy: Illumina; Advisory/Consultancy: Qiagen; Advisory/Consultancy: Novartis Thermofisher; Speaker Bureau/Expert testimony: Amgen; Speaker Bureau/Expert testimony: Dyn; Speaker Bureau/Expert testimony: Vela diagnostics; Speaker Bureau/Expert testimony: Luye Pharma. F. Carbonnel: Advisory/Consultancy: enterome; Advisory/Consultancy: Amgen; Advisory/Consultancy: Astra; Advisory/Consultancy: MSD; Advisory/Consultancy: BMS; Advisory/Consultancy: Janssen; Advisory/Consultancy: Pfizer; Advisory/Consultancy: Abbvie; Advisory/Consultancy: Mayoly Spindler; Advisory/Consultancy: Takeda; Advisory/Consultancy: Pileje; Advisory/Consultancy: Roche. P.A. Ascierto: Advisory/Consultancy: Bristol Myers-Squibb; Advisory/Consultancy, Research grant/Funding (self): Roche-Genentech; Advisory/Consultancy: Merck Sharp & Dohme; Advisory/Consultancy, Research grant/Funding (self): Array; Advisory/Consultancy: Novartis; Advisory/Consultancy: Amgen; Advisory/Consultancy: Merck Serono; Advisory/Consultancy: Pierre Fabre; Advisory/Consultancy: Incyte; Advisory/Consultancy: NewLink Genetics; Advisory/Consultancy: Genmab; Advisory/Consultancy: Medimmune; Advisory/Consultancy: AstraZeneca; Advisory/Consultancy: Syndax; Advisory/Consultancy: SunPharma; Advisory/Consultancy: Sanofi; Advisory/Consultancy: Idera. C. Robert: Advisory/Consultancy: Roche; Advisory/Consultancy: GSK; Advisory/Consultancy: Merck Sharp & Dohme; Advisory/Consultancy: Novartis; Advisory/Consultancy: Amgen; Advisory/Consultancy: BMS. N. Chaput: Research grant/Funding (self): Cytune Pharma; Research grant/Funding (self): GSK; Speaker Bureau/Expert testimony, Research grant/Funding (self): Sanofi; Advisory/Consultancy, Speaker Bureau/Expert testimony: AstraZeneca. All other authors have declared no conflicts of interest.

623P - Plasma clearance of RAS mutation under therapeutic pressure is a rare event in metastatic colorectal cancer

BackgroundIn metastatic colorectal cancer (mCRC), circulating tumour DNA (ctDNA) monitoring can be used to genotype tumors and track clonal evolution. We investigated the clearance of RAS mutated clones under chemotherapy pressure by ctDNA analysis in patients with a RAS mutated mCRC. MethodsPatients with a RAS mutated tumour included in the prospective PLACOL study, were monitored for ctDNA. Analyzes were based on optimized targeted next generation sequencing and/or droplet-based digital PCR (ddPCR). For plasma samples without detectable mutations, we tested the methylation status of WIF1 and NPY genes using methylation-ddPCR (met-ddPCR) to validate the presence of ctDNA. ResultsAmong the 36 patients with positive plasma samples for RAS mutations at inclusion, 28 (77.8%) remained RAS positive at disease progression, and 8 (22.2%) became negative. Subsequent met-ddPCR for methylated markers showed that only 2 out of the 8 patients with RAS negative plasma had detectable ctDNA at progression. Therefore, only two samples among 36 were confirmed for clearance of RAS mutation in our series. ConclusionsThis study suggests that the clearance of RAS mutations in patients treated by chemotherapy for a RAS mutated mCRC is a rare event. Monitoring tumor mutations in plasma samples should be combined with a strict control of the presence of ctDNA. The therapeutic impacts of RAS clearance need to be further explored. Legal entity responsible for the studyThe authors. FundingThe Ministère de l’Enseignement Supérieur et de la Recherche, the Université Paris-Descartes, the Centre National de la Recherche Scientifique (CNRS), the Institut National de la Santé et de la Recherche Médicale (INSERM), the Institut National du Cancer (INCA, n° 2009-1-RT-03-US-1 and 2009-RT-03-UP5-1), the Association pour la recherche contre le cancer (ARC, no. SL220100601375), the Agence Nationale de la Recherche (ANR Nanobiotechnologies; no. ANR-10-NANO-0002-09), the SIRIC CARPEM, the ligue nationale contre le cancer (LNCC, Program “Equipe labelisée LIGUE”; no. EL2016.LNCC/VaT) and Advanced Merieux Research Grant and canceropole funding (no. 2011-1-LABEL-UP5-2). DisclosureH. Blons: Advisory / Consultancy: AstraZeneca; Advisory / Consultancy: Merck; Advisory / Consultancy: Amgen; Advisory / Consultancy: Boehringer Ingelheim; Advisory / Consultancy: Pfizer. V. Taly: Advisory / Consultancy: Servier; Advisory / Consultancy: Boehringer Ingelheim; Advisory / Consultancy: Raindance Technologies. W. Shu-Fang: Research grant / Funding (institution): Servier. S. Pernot: Advisory / Consultancy: Amgen; Advisory / Consultancy: Sanofi. J. Taieb: Advisory / Consultancy, Speaker Bureau / Expert testimony: Merck; Advisory / Consultancy, Speaker Bureau / Expert testimony: Roche; Advisory / Consultancy, Speaker Bureau / Expert testimony: Amgen; Advisory / Consultancy, Speaker Bureau / Expert testimony: Eli Lilly; Advisory / Consultancy, Speaker Bureau / Expert testimony: Sanofi; Advisory / Consultancy, Speaker Bureau / Expert testimony: Celgene; Advisory / Consultancy, Speaker Bureau / Expert testimony: Sirtex; Advisory / Consultancy, Speaker Bureau / Expert testimony: Shire; Advisory / Consultancy, Speaker Bureau / Expert testimony: MSD; Advisory / Consultancy, Speaker Bureau / Expert testimony: Servier; Advisory / Consultancy, Speaker Bureau / Expert testimony: Pierre Fabre. P. Laurent-Puig: Advisory / Consultancy: Amgen; Advisory / Consultancy: Merck-Serono; Advisory / Consultancy: Sanofi; Advisory / Consultancy: Boehringer Ingelheim; Advisory / Consultancy: Roche; Advisory / Consultancy: Lilly. A. Zaanan: Advisory / Consultancy: Amgen; Advisory / Consultancy: Baxter; Advisory / Consultancy: Lilly; Advisory / Consultancy: Merck-Serono; Advisory / Consultancy: MSD; Advisory / Consultancy: Sanofi; Advisory / Consultancy: Roche; Advisory / Consultancy: Servier. All other authors have declared no conflicts of interest.

Patent Highlights December 2018–January 2019

Patent Highlights December 2018–January 2019

Highlight on Endothelial Activation and Beyond.

Highlight on Endothelial Activation and Beyond.

Drops in Blood Pressure, BMI Common Prior to Dementia Diagnosis

Back to table of contents Previous article Next article Clinical and Research NewsFull AccessDrops in Blood Pressure, BMI Common Prior to Dementia DiagnosisNick ZagorskiNick ZagorskiSearch for more papers by this authorPublished Online:23 Oct 2018https://doi.org/10.1176/appi.pn.2018.9a16AbstractA study that tracked seniors over 14 years is one of the first to explore how temporal changes in metabolic risk factors affect dementia risk.Metabolic problems are a known risk factor for dementia, but studies examining the association between metabolic status at certain ages and dementia risk have found varied results. High blood pressure and low blood pressure at certain ages have both been linked with dementia risk, as have obesity and being underweight.iStock/Sasha_SuziIn a study published in JAMA Psychiatry, researchers at the University of Bordeaux in France described how rather than examining the relationship between metabolic status at a specific age and dementia risk, they tracked the trajectories of these risk factors. They found evidence to suggest that exaggerated drops in body mass index (BMI) and blood pressure could signify oncoming dementia. They also reaffirmed that high blood glucose in older age is a significant dementia risk factor.The findings were based on analysis of health data from nearly 4,000 individuals who were part of the Three-City (3C) study. The study followed adults aged 65 and older in three French cities (Bordeaux, Dijon, and Montpelier) for 14 years to ascertain connections between heart disease and dementia.The research team identified 841 people in the 3C cohort who developed dementia at some point during the study (the average length of time to diagnosis was about eight years). The researchers then tried to match each person with dementia with four control individuals with similar demographics at the time of diagnosis; they successfully matched 785 dementia cases with 3,140 controls for the final analysis.The researchers then plotted out all prior assessments of BMI; systolic and diastolic blood pressure; and high-density and low-density lipoprotein cholesterol (HDL-C and LDL-C), triglyceride, and blood glucose levels for these 3,925 people.The metabolic trajectories revealed that people who developed dementia showed much steeper declines in BMI in the years preceding their diagnosis, with the most rapid weight loss starting at about seven years before diagnosis. Over the 14-year period, the average BMI of dementia patients dropped from 26.1 to 24.8, whereas BMI in controls only dropped from 25.7 to 25.3. People who developed dementia also experienced a greater drop in blood pressure over time compared with controls. Although both groups of patients experienced rising blood glucose level with age, those with dementia had consistently higher glucose levels over the 14-year period. Lead study author Maude Wagner, M.P.H., of the University of Bordeaux said that the changes seen in dementia patients likely represent reverse causation—that is, subtle cognitive decline may be accelerating normal metabolic changes. In the case of BMI, early dementia may cause someone to eat less, which would increase age-related weight loss. Likewise, some research has suggested that early-stage dementia disrupts brain processes involved in blood pressure regulation.Because low blood pressure reduces the delivery of oxygen to the brain, Wagner and colleagues noted that it is also possible that lower blood pressure contributes to dementia.“This study reinforces that the relationship between metabolism and dementia is complex and highly dependent on timing,” Gary Small, M.D., director of geriatric psychiatry at the David Geffen School of Medicine at the University of California, Los Angeles, told Psychiatric News. Even so, he cautioned that it’s too soon for clinicians to make predictions using these measures.“We need more longitudinal studies to try to tease out this gray area of what changes are cause versus effect,” he said. The 3C is conducted under a partnership agreement between the Institut National de la Santé et de la Recherche Médicale (INSERM), the University of Bordeaux, and Sanofi. ■“Evaluation of the Concurrent Trajectories of Cardiometabolic Risk Factors in the 14 Years Before Dementia” can be acccessed here. ISSUES NewArchived

Meet the First Authors.

Meet the First Authors.

Mammary epithelium‐specific conditional expression of human mutant Ki‐Ras in mice generates estrogen receptor alpha (ERα)‐positive mammary adenocarcinomas

BackgroundSeveral genetically modified mouse models (GEMMs) have been generated for mechanistic and therapeutic studies of mammary tumors. Although a number of cancer GEMMs have been developed, an individual model may recapitulate only some aspects of this complex disease. Indeed, breast cancer features include at least five biologically and clinically distinct intrinsic subtypes, including luminal A, luminal B, human epidermal growth factor receptor 2 (HER2)‐enriched, basal‐like, and normal‐like, with luminal A and B subtypes representing ~ 70% of breast cancers, and thus being of major clinical interest. We have engineered a novel transgenic mouse line expressing human Ki‐Ras bearing an activating mutation (Ki‐Ras(G12V)) selectively in the mammary epithelium and characterized them.MethodsTo allow conditional Ki‐Ras(G12V) expression in mammary epithelium, we generated C‐Ki‐Ras(G12V) transgenic mice that express, after Cre‐mediated DNA recombination during lactation, Ki‐Ras(G12V) under the control of the mammary tumor virus long terminal repeat (MMTV‐LTR) promoter. Tumor development was followed in 48 females after the start of lactation. Tumor characterization was performed by histopathological analysis, immunohistochemistry and real time PCR. Two cell lines were isolated from mouse mammary tumors and their phenotypes were evaluated by immunoblotting, ‘in vitro’ cell growth assays and ‘in vivo’ tumor xenograft studies.ResultsAll mice developed homogenous invasive ductal adenocarcinomas with 100% incidence within 13–35 weeks after Ki‐Ras(G12V) expression. Immunohistochemical analysis revealed high levels of estrogen receptor (ER) α in more than 90% of epithelial cells and focal progesterone receptor (PR) staining in neoplastic epithelial cells, while no HER2 staining was observed. Mammary tumor cells exhibited less than 14% nuclei positive for the proliferation marker Ki‐67, a feature that can segregate the luminal A subtype. The cytokeratin profile, as well as quantification of transcript levels of specific markers in the tumor cells, also reflected a luminal cell phenotype. Cell lines isolated from such tumors have similar characteristics as the primary tumors, are estrogen‐responsive and transplantable. Importantly, palpable tumors were detected faster than those derived from the human luminal breast cancer MCF‐7 cells (one week versus one month).ConclusionsMG‐Ki‐Ras(G12V) mice develop within a couple of months at full penetrance mammary tumors that recapitulate the most common human breast cancer subtype, classified as ductal ER‐positive invasive adenocarcinoma. Thus, this animal model together with the derived primary cancer cell line represent valuable tools with high potential of translational relevance to investigate the biology of estrogen‐dependent mammary tumors and to test various therapeutic agents.Support or Funding InformationThis work was supported by: PRIN2015 #2015B7M39T (to SA), Fondazione Italiana per la Ricerca sul Cancro (AIRC) grants: IG no. 11595 (to SA), MFAG no. 16899 (to IB), PROGRAMMA ‘FUTURO IN RICERCA’ Anno 2012 no.RBFR12FI27 (to IB), AIRC Fellowships for abroad no. 19552 (to DR), EMBO ASTF no. 18‐2010 (to RM), European Social Fund operational programme of the Calabria region (to SP), Faculté de Médecine, Université de Strasbourg (to CE) and by the Centre Nationalpour la Recherche Scientifique (CNRS), the Institut National de la Santé et de laRecherche Médicale (INSERM), the Université de Strasbourg and French state funds through the Agence Nationale de la Recherche ANR‐10‐LABX‐0030‐INRT under the frame programme Investissements d'Avenir labelled ANR‐10‐IDEX‐0002‐02.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Interpretation of health news items reported with or without spin: protocol for a prospective meta-analysis of 16 randomised controlled trials

IntroductionWe aim to compare the interpretation of health news items reported with or without spin. ‘Spin’ is defined as a misrepresentation of study results, regardless of motive (intentionally or unintentionally)...

Dietary flavonoid intake and colorectal cancer risk in the European prospective investigation into cancer and nutrition (EPIC) cohort.

Flavonoids have been shown to inhibit colon cancer cell proliferation in vitro and protect against colorectal carcinogenesis in animal models. However, epidemiological evidence on the potential role of flavonoid intake in colorectal cancer (CRC) development remains sparse and inconsistent. We evaluated the association between dietary intakes of total flavonoids and their subclasses and risk of development of CRC, within the European Prospective Investigation into Cancer and Nutrition (EPIC) study. A cohort of 477,312 adult men and women were recruited in 10 European countries. At baseline, dietary intakes of total flavonoids and individual subclasses were estimated using centre-specific validated dietary questionnaires and composition data from the Phenol-Explorer database. During an average of 11 years of follow-up, 4,517 new cases of primary CRC were identified, of which 2,869 were colon (proximal = 1,298 and distal = 1,266) and 1,648 rectal tumours. No association was found between total flavonoid intake and the risk of overall CRC (HR for comparison of extreme quintiles 1.05, 95% CI 0.93-1.18; p-trend = 0.58) or any CRC subtype. No association was also observed with any intake of individual flavonoid subclasses. Similar results were observed for flavonoid intake expressed as glycosides or aglycone equivalents. Intake of total flavonoids and flavonoid subclasses, as estimated from dietary questionnaires, did not show any association with risk of CRC development.

Persistence and clearance of Ebola virus RNA from seminal fluid of Ebola virus disease survivors: a longitudinal analysis and modelling study.

By January, 2016, all known transmission chains of the Ebola virus disease (EVD) outbreak in west Africa had been stopped. However, there is concern about persistence of Ebola virus in the reproductive tract of men who have survived EVD. We aimed to use biostatistical modelling to describe the dynamics of Ebola virus RNA load in seminal fluid, including clearance parameters. In this longitudinal study, we recruited men who had been discharged from three Ebola treatment units in Guinea between January and July, 2015. Participants provided samples of seminal fluid at follow-up every 3-6 weeks, which we tested for Ebola virus RNA using quantitative real-time RT-PCR. Representative specimens from eight participants were then inoculated into immunodeficient mice to test for infectivity. We used a linear mixed-effect model to analyse the dynamics of virus persistence in seminal fluid over time. We enrolled 26 participants and tested 130 seminal fluid specimens; median follow up was 197 days (IQR 187-209 days) after enrolment, which corresponded to 255 days (228-287) after disease onset. Ebola virus RNA was detected in 86 semen specimens from 19 (73%) participants. Median duration of Ebola virus RNA detection was 158 days after onset (73-181; maximum 407 days at end of follow-up). Mathematical modelling of the quantitative time-series data showed a mean clearance rate of Ebola virus RNA from seminal fluid of -0·58 log units per month, although the clearance kinetic varied greatly between participants. Using our biostatistical model, we predict that 50% and 90% of male survivors clear Ebola virus RNA from seminal fluid at 115 days (90% prediction interval 72-160) and 294 days (212-399) after disease onset, respectively. We also predicted that the number of men positive for Ebola virus RNA in affected countries would decrease from about 50 in January 2016, to fewer than 1 person by July, 2016. Infectious virus was detected in 15 of 26 (58%) specimens tested in mice. Time to clearance of Ebola virus RNA from seminal fluid varies greatly between individuals and could be more than 13 months. Our predictions will assist in decision-making about surveillance and preventive measures in EVD outbreaks. This study was funded by European Union's Horizon 2020 research and innovation programme, Directorate-General for International Cooperation and Development of the European Commission, Institut national de la santé et de la recherche médicale (INSERM), German Research Foundation (DFG), and Innovative Medicines Initiative 2 Joint Undertaking.

Modifiable causes of premature death in middle-age in Western Europe: results from the EPIC cohort study.

BackgroundLife expectancy is increasing in Europe, yet a substantial proportion of adults still die prematurely before the age of 70 years. We sought to estimate the joint and relative contributions of tobacco smoking, hypertension, obesity, physical inactivity, alcohol and poor diet towards risk of premature death.MethodsWe analysed data from 264,906 European adults from the EPIC prospective cohort study, aged between 40 and 70 years at the time of recruitment. Flexible parametric survival models were used to model risk of death conditional on risk factors, and survival functions and attributable fractions (AF) for deaths prior to age 70 years were calculated based on the fitted models.ResultsWe identified 11,930 deaths which occurred before the age of 70. The AF for premature mortality for smoking was 31 % (95 % confidence interval (CI), 31–32 %) and 14 % (95 % CI, 12–16 %) for poor diet. Important contributions were also observed for overweight and obesity measured by waist-hip ratio (10 %; 95 % CI, 8–12 %) and high blood pressure (9 %; 95 % CI, 7–11 %). AFs for physical inactivity and excessive alcohol intake were 7 % and 4 %, respectively. Collectively, the AF for all six risk factors was 57 % (95 % CI, 55–59 %), being 35 % (95 % CI, 32–37 %) among never smokers and 74 % (95 % CI, 73–75 %) among current smokers.ConclusionsWhile smoking remains the predominant risk factor for premature death in Europe, poor diet, overweight and obesity, hypertension, physical inactivity, and excessive alcohol consumption also contribute substantially. Any attempt to minimise premature deaths will ultimately require all six factors to be addressed.Electronic supplementary materialThe online version of this article (doi:10.1186/s12916-016-0630-6) contains supplementary material, which is available to authorized users.

Loss of C9 ORF 72 impairs autophagy and synergizes with polyQ Ataxin‐2 to induce motor neuron dysfunction and cell death

An intronic expansion of GGGGCC repeats within the C9ORF72 gene is the most common genetic cause of amyotrophic lateral sclerosis and frontotemporal dementia (ALS-FTD). Ataxin-2 with intermediate length of polyglutamine expansions (Ataxin-2 Q30x) is a genetic modifier of the disease. Here, we found that C9ORF72 forms a complex with the WDR41 and SMCR8 proteins to act as a GDP/GTP exchange factor for RAB8a and RAB39b and to thereby control autophagic flux. Depletion of C9orf72 in neurons partly impairs autophagy and leads to accumulation of aggregates of TDP-43 and P62 proteins, which are histopathological hallmarks of ALS-FTD SMCR8 is phosphorylated by TBK1 and depletion of TBK1 can be rescued by phosphomimetic mutants of SMCR8 or by constitutively active RAB39b, suggesting that TBK1, SMCR8, C9ORF72, and RAB39b belong to a common pathway regulating autophagy. While depletion of C9ORF72 only has a partial deleterious effect on neuron survival, it synergizes with Ataxin-2 Q30x toxicity to induce motor neuron dysfunction and neuronal cell death. These results indicate that partial loss of function of C9ORF72 is not deleterious by itself but synergizes with Ataxin-2 toxicity, suggesting a double-hit pathological mechanism in ALS-FTD.