Identify CHIKV receptor and identify cellular proteins link to alphavirus host susceptibility. Understanding the virus cell entry mechanism at the molecular level is essential for the development of new medicines of the therapeutic class of entry inhibitors. However, the entry process of alphavirus is not understood enough at the molecular level. Our objective is to clone the alphavirus receptors involve in interspecies infection and identify cellular proteins link to host susceptibility to alphaviruses. Our laboratory is using different molecular tools (alphavirus pseudoparticles (pp), infectious clones, replicon) and cutting edge technologies. We are currently using proteomics and CRISPR/Cas9 genome wide screen to identify any factors involved in the entry and infectious cell cycle of Chikungunya virus and Semliki Forest Virus which differ in term of tropism. Different candidates are being validated and we will characterize the function of these proteins in the virus cell cycle of different alphaviruses. In parallel, to complement these in vitro studies in mammals and insect cells, we are developing in vivo studies. We developed the use of mosquitoes and the use of Drosophila and its genetic engineering as a powerful “tool” to address the role of different molecules in alphavirus replication in insects.

Characterize the consequences of Alphavirus/Flavivirus co-infection. Pathology associated to infection is often considered as resulting from the reciprocal interaction of a given pathogen with a given host. This paradigm looks however over-simplistic since the pathology, as well as the epidemiology of a pathogen, relies also on the interactions between several infectious agents present within an organism. It is therefore crucial to consider to which extent a host infected by a first microorganism is modified and whether its reaction to the infection by a second is consequently altered. This consideration is especially relevant for arboviruses. Indeed, one consequence of the rapid spreading of arboviruses worldwide is an increased overlapping distribution. Overlapping in the distribution of the arboviral diseases leads to a higher detection of co-infections in humans. This co-infection is also possible in mosquitoes as DENV, ZIKV and CHIKV can share Aedesaegypti and albopictus vectors. At the cellular level, DENV, ZIKV and CHIKV share interactions with common cellular pathways. These possible similar host-virus interactions probably impact the outcome of one virus during co-infection. Our purpose is to define more precisely the molecular and cellular mechanisms of interference between Alphaviruses and Flaviviruses infection in human and mosquito target cells.

As a first approach, we are studying successive co infection of different mosquito cell lines derived from Aedesalbopictus or Aedesaegypti. We are using different molecular virology technic to quantify infection, production, assembly and spreading of the viruses. In order to provide more relevant analysis, we are also doing in vivo experiments by infecting sequentially Aedesaegypyi mosquito.

Zikavirus cell host interactions. Zika virus (ZIKV) is a mosquito borne flavivirus, which was a neglected tropical pathogen until it emerged and spread across the Pacific Area and the Americas, causing large human outbreaks associated with fetal abnormalities and neurological disease in adults. The factors that contributed to the emergence, spread and change in pathogenesis of ZIKV are not well understood. Using sequence alignments of different ZIKV strains of a range of spatiotemporal history of isolation and representing African and Asian lineages, we identified possible differences in the post translational modification (PTM) of different proteins. We are analysis the effect of some PTM in the function of ZIKV proteins and on the pathogenesis.

Collaboration

-        PrGang LONG and Pr Pei HAO, IPS-CAS, onZika virus cell host interaction

-        PrJin ZHONG and PrZhong HUANG, IPS-CAS, on HCV inhibitors development

-        Dr Carine Maisse-Paradissi, UMR754 INRA-UCBL-EPHE "Virus infection and comparative Pathogenesis", Lyon, France

-        Dr Valerie CHOUMET, ERI/CIBU, Institut Pasteur Paris, France

-        Dr Nathalie Pardigon, ERI/CIBU,, Institut Pasteur Paris, France

-        Dr Nathalie DEJUCQ-RAINSFORD, IRSET-Inserm U1085, University os Rennes

-        Dr Alexander PLOSS and DrFlorian DOUAM, Princeton University, U.S.A.

-        DrOusmane FAYE and Gamou FALL, “Arbovirus and Hemorrhagic fever Unit”, Institut Pasteur de Dakar, Senegal

Funding

2017. International grant “Chinese Ministry of Science and Technology for EU-China Inter-governmental S&T Cooperation grant” (MOST). (grant number 2016YFE0133500). 06/2017-05/2019. Coordinator.

2016. ZIKAlliance consortium. A global alliance for Zika virus control and prevention. Horizon 2020 European Commission ？Research and Innovation Action？ (H2020-SC1-2016-RTD-Zika) in WP3 ？ Virology and antivirals ？ Role as Coordinator-Legal Entity Appointed Representative (LEAR) for IPS. 7/2016-6/2019.

2016. ？ Flavizome ？ Role of PDZ-binding motif from Flavivirus non-structural proteins in flaviviral life cycle. Programmes Transversaux de Recherche PTR Pasteur Paris with Nathalie PARDIGON (Unité de recherche et d'expertise Environnement et risques infectieux CIBU), Yves JACOB and Nicolas Wolff. 2017-2019. Participant.

2016. Cai Yuan Pei program; French ministry of foreign affairs (international exchanges) with Carine Maisse-Paradissi coordinator France (Virus Infection and Comparative Pathology, INRA UMR754, Lyon). Coordinator for China.

2015. 100 Talent of Chinese Academy of Sciences. Alphaviruses and Flaviviruses interspecies transmission and host cell response.

2015. 1000 Talent of Shanghai Municipality. Alphaviruses and Flaviviruses interspecies transmission.

Publications

_ Co-infection of mosquitoes with Chikungunya and Dengue Viruses reveals modulation of the replication of both viruses in midguts and salivary glands of Aedesaegypti mosquitoes. Le Coupanec A, Tchankouo-Nguetcheu S, Roux P, Khun H, Huerre M, Morales-Vargas R, Enguehard M, Lavillette D, Missé D, Choumet V. Int J Mol Sci. 2017 Aug 4;18(8). pii: E1708. doi: 10.3390/ijms18081708. PMID: 28777313

_ Specialization of Hepatitis C Virus envelope glycoproteins for B-lymphocytes in chronically infected patients. Douam F, Bobay LM, Maurin G, Fresquet J, Calland N, Durand T, Cosset FL, Féray C and Lavillette D. J Virol. January 2016 vol. 90  no. 2  992-1008.

_ Marnata C, Saulnier A, Mompelat D, Krey T, Cohen L, Boukadida C, Warter L, Fresquet J, Vasiliauskaite I, Escriou N, Cosset FL, Rey FA, Lanford RE, Karayiannis P, Rose NJ, Lavillette D, Martin A. Determinants Involved in Hepatitis C Virus and GB Virus B Primate Host Restriction. J Virol. 2015 Dec 1;89(23):12131-44. doi: 10.1128/JVI.01161-15

_ Fusil F, Calattini S, Amirache F, Mancip J, Costa C, Robbins JB, Douam F, Lavillette D, Law M, Defrance T, Verhoeyen E, Cosset FL. A Lentiviral Vector Allowing Physiologically Regulated Membrane-anchored and Secreted Antibody Expression Depending on B-cell Maturation Status.MolTher.2015 Aug 18.doi: 10.1038/mt.2015.148.

_ Maurin G, Halgand B, Bruscella P, Fresquet J, Duclos-Vallée JC, Roque-Afonso AM, Cosset FL, Samuel D, Féray C*, Lavillette D*. Low cross neutralization of Hepatitis C virus correlates with liver disease in immunocompromized patients. AIDS. 2015 Jun 1;29(9):1025-33. doi: 10.1097/QAD.0000000000000651.

_ Douam F, Dao Thi VL, Maurin G., Fresquet J., Mompelat D., Cosset FL and Lavillette D. A critical interaction between E1 and E2 glycoproteins determines binding and fusion properties of hepatitis C virus during cell entry. Hepatology. 2014 Mar;59(3):776-88.

_ Murphy L, Varela M, Desloire S, Ftaich N, Murgia C, Golder M, Neil S, Spencer TE, Wootton SK, Lavillette D, Terzian C, Palmarini M, Arnaud F. The sheep tetherinparalog oBST2B blocks envelope glycoprotein incorporation into nascent retroviral virions. J Virol. 2015 Jan;89(1):535-44. doi: 10.1128/JVI.02751-14. Epub 2014 Oct 22.

_ Girard-Gagnepain A, Amirache F, Costa C, Lévy C, Frecha C, Fusil F, Nègre D, Lavillette D, Cosset FL, Verhoeyen E. Baboon envelope pseudotypedlentiviral vectors outperform VSV-G pseudotypedlentiviral vectors for gene transfer into cytokine-stimulated and resting hematopoietic stem cells. Blood.2014 Jun 20.

_ Lévy C, Aerts L, Hamelin Mè, Granier C, Szécsi J, Lavillette D, Boivin G, Cosset FL. Virus-like particle vaccine induces cross-protection against human metapneumovirus infections in mice. Vaccine. 2013 Jun 7;31(25):2778-85

_ Garrone P, Fluckiger AC, Mangeot PE, Gauthier E, Dupeyrot-Lacas P, Mancip J, Cangialosi A, Du Chéné I, Legrand R, Mangeot I, Lavillette D, Bellier B, Cosset FL, Tangy F, Klatzmann D, Dalba C. A prime-boost strategy using virus-like particles pseudotyped for HCV proteins triggers broadly neutralizing antibodies in macaques. SciTransl Med. 2011 Aug 3;3(94):94

_ Maurin G, Fresquet J, Granio O, Wychowski C, Cosset FL and Lavillette D. Identification of interactions in the E1E2 heterodimer of Hepatitis C virus important for cell entry. J Biol Chem. 2011 Jul 8;286(27):23865-76

_ Lupberger J, Zeisel MB, Xiao F, Thumann C, Fofana I, Zona L, Davis C, Mee CJ, Turek M, Gorke S, Royer C, Fischer B, Zahid MN, Lavillette D,Fresquet J, Cosset FL, Rothenberg SM, Pietschmann T, Patel AH, Pessaux P, Doffo？l M, Raffelsberger W, Poch O, McKeating JA, Brino L, Baumert TF. EGFR and EphA2 are host factors for hepatitis C virus entry and possible targets for antiviral therapy. Nat Med. 2011 May;17(5):589-95.