Transcriptome Signature of Nipah Virus Infected Endothelial Cells

International audienc

Cloned HTLV-1+CD4+, but not CD8+, T-cells display an oncogenic miRNome

HTLV-1 persistence in vivo relies on the persistent clonal expansion of its host cells. These are CD4+ and CD8+ T cells, yet ATL is regularly CD4+. Accordingly, untransformed HTLV-1+CD4+ but not CD8+ T cells cloned from carriers cumulate the features of preleukemic cells, including multinuclearity, chromatin bridges, increased cell cycling and inappropriate telomerase activity. MicroRNAs (miR) modify the maturation of a plethora of T-cells RNA and their deregulation would therefore constitute an appropriate explanation for the Tax-dependent or -independent pleiotropic changes in the phenotype of HTLV-1+CD4+ T cells. As the miRNome of naturally infected untransformed cells has not been investigated to date, we assessed the miR expression profiling of T cells cloned from carriers. Microarray results, confirmed by quantitative RTPCR, showed that, upon infection, CD4+ and CD8+ clones yielded aberrant expression of 15 distinct miRs including miR-34b and miR-494 that were respectively over- and underexpressed in both compartments. The more prominent effect of the infection consisted in the CD4+-restricted overexpression of the cancer-related miRs miR-21, -27b and -23b associated with the CD4+-restricted downregulation of the proapoptotic miR-15 and -16. Data were extended by the analysis of 40 additional CD4+ clones (20 infected). Crossing the miRNome against the whole transcriptome data identified putative miR-targeted genes. In silico, those targeted by miR-23b and -27b defined 2 hitherto unknown pathways involving the cell cycle and genetic disorders. Therefore HTLV-1 triggers a phenotype-specific miR signature consistent with the preleukemic HTLV-1+CD4+ phenoty

Increasing importance of Bunyaviridae in public and veterinary health illustrated by hantaviruses, and the Schmallenberg and Rift Valley fever viruses

The virus family of Bunyaviridae is very important in terms of public health and veterinary medicine. With over 350 viruses identified to date, it includes viruses mainly transmitted by arthropods (arboviruses) or rodents (roboviruses), infecting mammals and plants for the genus Tospovirus. Humans can be infected by around 60 bunyaviruses sometime with very serious or even fatal consequences. The examples of Schmallenberg and Rift Valley fever viruses and hantavirus genus illustrate perfectly the many questions surrounding the Bunyaviridae family’s capacity to emerge, widely variable pathogenicity for different hosts, and capacity to persist in different vectors such as arthropods or rodents and more recently the soricomorph species (insectivores)La famille des Bunyaviridae est très importante en santé publique et vétérinaire. Avec plus de 350 virus identifiés à ce jour, elle regroupe des virus transmis principalement par des arthropodes (arbovirus) ou des rongeurs (robovirus), responsables d’infections chez les mammifères et chez les plantes pour le genre Tospovirus. L’homme peut être infecté par une soixantaine de ces Bunyaviridae, parfois avec des conséquences très graves, voire fatales. Les exemples du virus de Schmallenberg, du virus de la fièvre de la Vallée du Rift et du genre viral hantavirus illustrent parfaitement les nombreuses incertitudes concernant cette famille virale quant à leur potentiel d’émergence, leur pouvoir pathogène très varié pour des hôtes divers, et leur capacité à persister chez différents vecteurs appartenant aux arthropodes ou aux rongeurs et, plus récemment, aux soricomorphes (insectivores

Lethal Nipah Virus Infection Induces Rapid Overexpression of CXCL10

Nipah virus (NiV) is a recently emerged zoonotic Paramyxovirus that causes regular outbreaks in East Asia with mortality rate exceeding 75%. Major cellular targets of NiV infection are endothelial cells and neurons. To better understand virus-host interaction, we analyzed the transcriptome profile of NiV infection in primary human umbilical vein endothelial cells. We further assessed some of the obtained results by in vitro and in vivo methods in a hamster model and in brain samples from NiV-infected patients. We found that NiV infection strongly induces genes involved in interferon response in endothelial cells. Among the top ten upregulated genes, we identified the chemokine CXCL10 (interferon-induced protein 10, IP-10), an important chemoattractant involved in the generation of inflammatory immune response and neurotoxicity. In NiV-infected hamsters, which develop pathology similar to what is seen in humans, expression of CXCL10 mRNA was induced in different organs with kinetics that followed NiV replication. Finally, we showed intense staining for CXCL10 in the brain of patients who succumbed to lethal NiV infection during the outbreak in Malaysia, confirming induction of this chemokine in fatal human infections. This study sheds new light on NiV pathogenesis, indicating the role of CXCL10 during the course of infection and suggests that this chemokine may serve as a potential new marker for lethal NiV encephalitis

Repurposing of Drugs as Novel Influenza Inhibitors From Clinical Gene Expression Infection Signatures

Influenza virus infections remain a major and recurrent public health burden. The intrinsic ever-evolving nature of this virus, the suboptimal efficacy of current influenza inactivated vaccines, as well as the emergence of resistance against a limited antiviral arsenal, highlight the critical need for novel therapeutic approaches. In this context, the aim of this study was to develop and validate an innovative strategy for drug repurposing as host-targeted inhibitors of influenza viruses and the rapid evaluation of the most promising candidates in Phase II clinical trials. We exploited in vivo global transcriptomic signatures of infection directly obtained from a patient cohort to determine a shortlist of already marketed drugs with newly identified, host-targeted inhibitory properties against influenza virus. The antiviral potential of selected repurposing candidates was further evaluated in vitro, in vivo, and ex vivo. Our strategy allowed the selection of a shortlist of 35 high potential candidates out of a rationalized computational screening of 1,309 FDA-approved bioactive molecules, 31 of which were validated for their significant in vitro antiviral activity. Our in vivo and ex vivo results highlight diltiazem, a calcium channel blocker currently used in the treatment of hypertension, as a promising option for the treatment of influenza infections. Additionally, transcriptomic signature analysis further revealed the so far undescribed capacity of diltiazem to modulate the expression of specific genes related to the host antiviral response and cholesterol metabolism. Finally, combination treatment with diltiazem and virus-targeted oseltamivir neuraminidase inhibitor further increased antiviral efficacy, prompting rapid authorization for the initiation of a Phase II clinical trial. This original, host-targeted, drug repurposing strategy constitutes an effective and highly reactive process for the rapid identification of novel anti-infectious drugs, with potential major implications for the management of antimicrobial resistance and the rapid response to future epidemic or pandemic (re)emerging diseases for which we are still disarmed

Puces à ADN

National audienc

Capturing a Single Cell

International audienceA major problem encountered in genomic and proteomic studies arises from the heterogeneous nature of different tissue. Analysis of a pure cell population is essential for correlating relevant molecular signatures in diseased and disease-free cells. During the last 30 years this challenge has led to the development of different technologies able to isolate cells of interest. Laser capture microdissection (LCM) is the last available technology using the precision of a laser beam to isolate single cells from complex tissue. In this chapter we will review the different technologies available and some applications

Gene expression profiling reveals an inflammatory process in the anx/anx mutant mice

International audienceAnorexia (anx) is a recessive mutation that causes lethal starvation in homozygous mice. Studies of anx/anx mice hypothalamus have shown abnormalities in the orexigenic (NPY/AGRP neurons) and the anorexigenic (POMC/CART neurons) pathways. By gene expression profiling using cDNA and oligonucleotide rnicroarrays, we have shown that a surexpression of genes involved in inflammatory process occurred in anx mice hypothalamus. This inflammatory process could be the cause of the anorexia phenotype observed in these mice

Deregulation of miR-183 and KIAA0101 in Aggressive and Malignant Pituitary Tumors

Changes in microRNAs (miRNAs) expression in many types of cancer suggest that they may be involved in crucial steps during tumor progression. Indeed, miRNAs deregulation has been described in pituitary tumorigenesis, but few studies have described their role in pituitary tumor progression toward aggressiveness and malignancy. To assess the role of miRNAs within the hierarchical cascade of events in prolactin (PRL) tumors during progression, we used an integrative genomic approach to associate clinical-pathological features, global miRNA expression, and transcriptomic profiles of the same human tumors. We describe the specific down-regulation of one principal miRNA, miR-183, in the 8 aggressive (A, grade 2b) compared to the 18 non-aggressive (NA, grades 1a, 2a) PRL tumors. We demonstrate that it acts as an anti-proliferative gene by directly targeting KIAA0101, which is involved in cell cycle activation and inhibition of p53-p21-mediated cell cycle arrest. Moreover, we show that miR-183 and KIAA0101 expression significantly correlate with the main markers of pituitary tumors aggressiveness, Ki-67 and p53. These results confirm the activation of proliferation in aggressive and malignant PRL tumors compared to non-aggressive ones. Importantly, these data also demonstrate the ability of such an integrative genomic strategy, applied in the same human tumors, to identify the molecular mechanisms responsible for tumoral progression even from a small cohort of patients

INCREASING IMPORTANCE OF BUNYAVIRIDAE IN PUBLIC AND VETERINARY HEALTH ILLUSTRATED BY HANTAVIRUSES, AND THE SCHMALLENBERG AND RIFT VALLEY FEVER VIRUSES

National audienceThe virus family of Bunyaviridae is very important in terms of public health and veterinary medicine. With over 350 viruses identified to date, it includes viruses mainly transmitted by arthropods (arboviruses) or rodents (roboviruses), infecting mammals and plants for the genus Tospovirus. Humans can be infected by around 60 bunyaviruses sometime with very serious or even fatal consequences. The examples of Schmallenberg and Rift Valley fever viruses and hantavirus genus illustrate perfectly the many questions surrounding the Bunyaviridae family's capacity to emerge, widely variable pathogenicity for different hosts, and capacity to persist in different vectors such as arthropods or rodents and more recently the soricomorph species (insectivores)