Molecular genetics of herpes simplex virus type 1 (HSV-1)

Group composition

Leader : Alberto L. EPSTEIN

Staff members:
Alberto L. EPSTEIN epstein@cgmc.univ-lyon1.fr
Anna GRECO greco@cgmc.univ-lyon1.fr
Corinne POTEL corinne.potel@cgmc.univ-lyon1.fr
Aldo POURCHET aldo.pourchet@cgmc.univ-lyon1.fr
Matias MELENDEZ matias.melendez@cgmc.univ-lyon1.fr
Aleth CALLÉ calle@cgmc.univ-lyon1.fr
Karine THOINET karine.thoinet@cgmc.univ-lyon1.fr
Coline BIOLLAY coline.biollay@cgmc.univ-lyon1.fr

Grants
- 3 european grants: COMPUVAC (IP, 2005-2008 - Coordinator: D Klatzmann); THOVLEN (STREP, 2006-2008 - Coordinator: AL Epstein); HEVAR (STREP, 2007-2009 - Coordinator: AL Epstein)
- 2 AFM grants: Gene therapy of cardiovascular disease; HSV-1 helper system to produce AAV (2007-2008 - Coordinator A Salvetti)

Scientific themes

We are a team in the fields of fundamental and applied virology, and our working model is herpes simplex virus type 1 (HSV-1). HSV-1 is a human enveloped virus, and the productive infection occurs in the epithelial cells of the oro-facial region, provoking a stomatitis, which ends after some days (primary infection). During this period, the virus infects the sensory neurons located close to the site of the primary infection. Then, HSV-1 establishes latency in the nuclei of these neuronal cells, in the sensory ganglions. The latent genome remains in an episomal state (non integrated into the cellular DNA) and the expression of the viral lytic genes is then widely repressed, explaining the survival of these neurons. In response to some stimuli, such as stress, temperature, sun, or hormonal cycles, the latent episomal genome can reactivate and is transported back along axons to re-infect the epithelial cells that are close to the site of the primary infection, giving place to the secondary infections (also called recurrences), and the most frequent clinical manifestation is the cold sore (or labial herpes).

The laboratory develops two main subjects and also displays an activity of valuation (creation of a start-up BIOVIRON):

1-Development of HSV-1 derived viral vectors and their application to different gene therapy or vaccine research programs.

We set up an original system of production of amplicon vectors, based on the use of the Cre/loxP1 site-specific recombination system, that allows producing significant amounts of good-titre amplicon vectors, free of contaminant helper particles (Logvinoff and Epstein, 2001; Zaupa et al., 2003) (two patents Epstein et al., 2003, 2004). The helper-free amplicon vectors produced using this novel procedure are currently being used in several collaborative projects, including:
* Development of HSV-1-based vector vaccines (Tsitoura et al., 2002; Lauterbach et al., 2004; Lauterbach et al., 2005)
* Gene therapy of cardiovascular disease (Ferrera et al., 2005)
* Anticancer therapy (Cuchet et al., 2005)
* Studies on the contribution of glutamatergic (NMDA) receptors of hippocampal neurons, to the generation of short-term memories in mice (Cheli et al., 2002 Adrover et al., 2003; Cheli et al., 2006; Jerusalinsky and Epstein, 2006) (see above figure: rat hippocampal neurons infected with an amplicon expressing the EGFP protein. In red, glial cells stained with an antibody specific to these cells)
* Development of improved HSV-1 helper system to produce adeno-associated vectors (AAV) (Toublanc et al., 2004; Geoffroy et al., 2004)

2-Study of the interactions between HSV-1 and the host cells: role of some cellular proteins in the outcome of viral infection.

Productive infection of HSV-1, which occurs in the host cell nucleus, is accompanied by dramatic modifications of the nuclear architecture, including profound alterations of nucleoli morphology. In addition, several viral proteins localize at least transiently to nucleoli, and some nucleolar proteins are delocalized from nucleoli during infection.
In the studies on the virus and host cells interactions (Greco et al., 2007, 2005, 2004, 2001, 2000; Diaz et al., 2002), the team has recently showed that during the course of HSV-1 infection, nucleolin - the most abundant nucleolar protein - localizes in the viral replication compartments, then in aggresomes (see figure below), and that HSV-1 replication requires high level of nucleolin expression (Callé et al., 2008). This is the first demonstration for a direct role of a nucleolar protein in HSV biology. The role of nucleolar proteins in the outcome of HSV-1 infection is under investigation.

Publications

2008
Callé A, Ugrinova I, Epstein AL, Diaz JJ, Bouvet P and Greco A (2008). Nucleolin is required for efficient herpes simplex virus type 1 infection. Journal of Virology (Epub ahead of print).
Foka P, Epstein AL and Mavromara P (2008). Cancer virotherapy using recombinant oncolytic herpes simplex virus type 1 (HSV-1)-derived vectors. Med Reviews (in press)
Marconi P. Argnani R, Epstein AL and Manservigi R (2008). HSV as a vector in vaccine development and gene therapy. In Pharmaceutical Biotechnoogy, edited by Carlos A. Guzman and Giora Feuerstein. Landes Bioscience (in press).
Morfin F, Frobert E, Callé A, Thouvenot D, Diaz J-J et Greco A (2008). Nouvelles cibles pour le développement de molécules anti-herpétiques. Virologie (in press)

2007
Cuchet D, Potel C., Thomas J and Epstein AL (2007). HSV-1 amplicon vectors: a promising and versatile tool for gene delivery. Expert Opinion in Biological Therapy 7: 975-995.
Greco A, Diaz JJ, Thouvenot D, et Morfin F (2007). Novel targets for the development of anti-herpes compounds. Infect Disord Drug Targets 7: 11-8.
Manservigi R, Argnani R, Marconi P and Epstein AL (2007). Herpesvirus-based vectors for gene transfer, gene therapy, and the development of novel vaccines. In Virus Expression Vectors, pp205-246. Edited by Kathleen L. Hefferon. Transworld Research Network.
Thomas J, Cuchet D, Potel C et Epstein AL (2007). Les vecteurs amplicon derives du virus HSV-1: un système souple et puissant pour le transfert de genes. Virologie 11: 339-350.

2006
Cheli V, Adrover M, Blanco C, Ferrari C, Cornea A, Pitossi F, Epstein AL, and Jerusalinsky D (2006). Knocking-down the NMDAR1 subunit in a limited amount of neurons in the rat hippocampus impairs learning. Journal of Neurochemistry 97 (S1) 68-73.
Couté Y., Burgess J.A., Diaz J-J., Chichester C., Lisacek F., Greco A. Sanchez J-C (2006). Deciphering the human nucleolar proteome. Mass Spec Rev 25: 215-34.
Epstein AL (2006). Gene transfer to the nerve system using HSV-1-derived amplicon vectors. In Gene Therapy for Neurological Disorders pp17-31. Edited by MG Castro and PR Lowenstein. Taylor and Francis group.
Epstein AL (2006). Editorial. 25 years of amplicons ! In Herpes simplex virus type 1-based amplicon vectors. Current Gene Therapy 6: 275-276.
Jerusalinsky D and Epstein AL (2006). Amplicon vectors as outstanding tools to study and modify cognitive functions. In Herpes simplex virus type 1 amplicons. Current Gene Therapy 6: 351-360.

2005
Cuchet D, Ferrera R, Lomonte P and Epstein AL (2005). Characterisation of the antiproliferative and cytotoxic properties of the HSV-1 immediate-early ICP0 protein. Journal of Gene Medicine 7: 1187-1199.
Epstein AL (2005). Present and future of herpes simplex virus type 1 (HSV-1)-based amplicon vectors. Pathologie/Biologie 53: 1-3.
Epstein AL, Marconi P, Argnani R, and Manservigi R (2005). HSV-1-derived recombinant and amplicon vectors for gene transfer and gene therapy. Current Gene Therapy 5: 445-458.
Epstein AL (2005). HSV-1-derived recombinant and amplicon vectors for preventive or therapeutic gene transfer. An overview. Gene Therapy 12, S153.
Epstein AL (2005). HSV-1-based amplicon vectors. Design and applications. Gene Therapy 12, S154-S158.
Ferrera R, Cuchet D, Zaupa C, Michel P, Revol-Guyot V, Ovize M and Epstein AL (2005). Efficient and non-toxic gene transfer to cardiomyocytes using novel generation amplicon vectors derived from HSV-1. Journal of Molecular and Cellular Cardiology 38: 219-223.
Greco A, Callé A, Morfin F, Thouvenot D, Cayre M, Kindbeiter K, Martin L, Levillain O, Diaz J-J (2005) S-adenosyl methionine decarboxylase activity is required for the outcome of herpes simplex virus type 1 infection and represents a new potential therapeutic target. FASEB J. 19: 1128
Lauterbach H, Ried C, Epstein AL, Marconi P and Brocker T (2005). Reduced immune responses after vaccination with a recombinant Herpes Simplex Virus Type 1 vector in the presence of antiviral immunity. Journal of General Virology 86: 2401-2410.
Thiry E, Meurens F, Muylkens B, McVoy M, Gogev S, Thiry J, Vanderplasschen A, Epstein AL, Keil G and Schynts F (2005). Recombination in alphaherpesviruses. Reviews in Medical Virology 15: 89-103.

2004
Epstein AL and Manservigi R (2004). Herpesvirus/retrovirus chimeric vectors. Current Gene Therapy 4: 409-416.
Epstein AL, Zaupa C, Logvinoff C, Lomonte P, and Thomas J (2004). Demande PCT N° FR2004051205. Improved method for producing non-pathogenic helper virus-free preparations of herpes virus amplicon vectors, the helper virus and the cells used in this method, the corresponding genetic tools, as well as the applications of these non-pathogenic amplicon vectors.
Epstein AL (2004). De cómo tratar una enfermedad grave utilizando vectores virales, sin necesariamente matar a los pacientes en el intento. In : "ADN: 50 años no es nada". Comps. A. Diaz y D. Golombek. Siglo XXI editores, Buenos Aires, Argentine, p120-133.
Geoffroy M-C, Epstein AL, Toublanc E, Moullier P and Salvetti A (2004). The herpes simplex virus type 1 ICP0 protein mediates the activation of adeno-associated virus type 2 rep gene expression from a latent integrated form. Journal of Virology 78: 10977-10986.
Goutagny N, Vieux C, Decullier E, Ligeoix B, Epstein A, Trepo C, Couzigou P, Inchauspe G, Bain C. (2004). Quantification and functional analysis of plasmacytoid dendritic cells in patients with chronic hepatitis C virus infection. Journal of Infectious Diseases 189: 1646-1655
Greco A, Couté Y, Giraud S and Diaz J-J (2004). Modifications of host cell proteomes induced by herpes simplex virus type 1. In Biomedical Applications of Proteomics (Sanchez, J.C., Corthals, G., Hochstrasser, D.F. eds) p263-83, Wiley-VCH Verlag GmbH & Co. KgaA, Weinheim
Lauterbach H, Kerksiek K, Busch D, Berto E, Bozac A, Mavromara P, Manservigi R, Epstein AL, Marconi P and Brocker T (2004). Protection from bacterial infection by a single vaccination with replication-deficient mutant herpes simplex virus-1. Journal of Virology 78: 4020-4028.
Lomonte P, Thomas J, Texier P, Caron C, Khochbin S and Epstein AL (2004). Functional interaction between class II HDACs and ICP0 of herpes simplex virus type 1. Journal of Virology 78: 6744-6757.
Toublanc E, Benraiss A, Bonnin D, Blouin V, Brument N, Cartier N, Epstein A L, Moullier P and Salvetti A (2004). Identification of a replication defective Herpes Simplex virus for recombinant adeno-associated virus type 2 (rAAV2) particle assembly using stable producer cell lines. Journal of Gene Medicine 6: 555-564.

2003
Adrover M, Revol-Guyot V, Cheli V, Blanco C, Vidal R, Alché L, Kornisiuk E, Epstein AL and Jerusalinsky D (2003). Hippocampal infection with HSV-1-derived vectors expressing a NMDAR1 antisense modifies behavior. Genes, Brain and Behavior 2, 103-113.
Couté Y, Scherl A, Dieckmann R, Déon C, Callé A, Hoogland C, Kindbeiter K, Sanchez J-C, Greco A, Diaz J-J. and Hochstrasser D (2003). Subcellular proteomes : nucleolar proteomics. Nature Encyclopedia of the human genome p 405-8, Macmillan Publishers Ltd, Nature Publishing Group.
Epstein A, Zaupa C and Logvinoff C (2003). Improved method for producing non-pathogenic helper virus-free preparations of herpes virus amplicon vectors, the helper virus and the cells used in this method, the corresponding genetic tools, as well as the applications of these non-pathogenic amplicon vectors. Brevet américain N°10/438.019.
Levillain O, Greco A, Diaz J-J, Augier R, Didier A, Kindbeiter K, Catez F and Cayre M (2003). Influence of testosterone on regulation of ODC, antizyme and N1-SSAT gene expression in mouse kidney. Am J Physiol Renal Physiol. 285:F498-506
Schynts F, McVoy MA, Meurens F, Detry B, Epstein, AL and Thiry E (2003). The structures of bovine herpesvirus 1 virion and concatemeric DNA: implications for cleavage and packaging of herpesvirus genomes. Virology 314, 326-335.
Tang Q, Li L, Ishov AM, Revol V, Epstein AL and Maul GG (2003). Determination of minimum HSV-1 components to localize transcriptionally active DNA to ND10. J. Virol. 77, 5821-5828.
Zaupa C, Revol-Guyot V and Epstein AL (2003). Improved packaging system for generation of high levels non-cytotoxic HSV-1 amplicon vectors using Cre-loxP site-specific recombination to delete the packaging signals of defective helper genomes. Human Gene Therapy 14, 1049-1063.

2002
Bienvenut WV, Hoogland C, Greco A, Heller M, Gasteiger E, Appel RD, Diaz JJ, Sanchez JC, and Hochstrasser DF (2002). Hydrogen/deuterium exchange for higher specificity of protein identification by peptide mass fingerprinting. Rapid Commun Mass Spectrom. 16:616-626.
Cheli VT, Adrover MF, Blanco C, Rial Verde E, Guyot-Revol V, Vidal R, Martin E, Alché L, Sanchez G, Acerbo M, Epstein AL and Jerusalinsky D (2002). Gene transfer of NMDAR1 subunit sequences to the rat CNS using Herpes Simplex Virus vectors interfered with habituation. Cell Mol Neurobiol. 3, 303-314.
Diaz J-J, Giraud S and Greco A (2002). Alteration of ribosomal proteins maps in herpes simplex virus type 1 infection. J Chromat B, 771:237-249
Scherl A, Couté Y, Déon C, Callé A, Kindbeiter K, Sanchez J-C, Greco A, Hochstrasser D and Diaz J-J (2002). Functional proteomic analysis of the human nucleolus. Mol Biol Cell, 13:4100-0909.
Schynts F, Meurens F, Muylkens B, Epstein AL, McVoid M et Thiry E (2002). Réplication, clivage-encapsidation et recombinaison de lADN des herpèsvirus. Virologie 6, 343-352.
Tsitoura E, Lucas M, Revol-Guyot V, Epstein AL, Manservigi R and Mavromara P (2002). Expression of hepatitis C virus envelope glycoproteins by herpes simplex virus-1 based amplicon vectors. J. Gen. Virol. 83, 561-566.

2001
Berthommé H, Thomas J et Epstein AL (2001). Les vecteurs herpétiques Dans La Thérapie Génique. Odile Cohén-Haguenauer Coordonnatrice. Editions Tec & Doc-Editions Médicales Internationales, pp. 167-186.
Berthommé H, Thomas J et Epstein AL (2001). Vecteurs neurotropes atténués et défectifs dérivés du virus de lherpès simplex de type 1 (HSV-1). Virologie 5 N°2, 131-145.
Berthommé H, Thomas J, Texier P, Epstein AL and Feldman LT (2001). The enhancer and long term expression functions of herpes simplex virus type 1 latency-associated promoter are both located in the same region. J. Virol. 75, 4386-4393.
Braz J, Beaufour C, Coutaux A, Epstein AL, Cesselin F, Hamon M, Cesselin F and Pohl M (2001). Therapeutic efficacy in experimental polyarthritis of viral-driven enkephalin overproduction in sensory neurons. Journal of Neuroscience 21, 7881-88.
Duisit G, Savard N, Epstein AL, Moullier P et Cosset FL (2001). Production de rétrovirus recombinants par des vecteurs viraux hybrides. Dans La Thérapie Génique. Odile Cohén-Haguenauer Coordonatrice. Editions Tec & Doc-Editions Médicales Internationales, pp. 187-203.
Giraud S, Greco A, Brink M, Diaz J-J and Delafontaine P (2001). Translation initiation of the insulin-like growth factor I receptor mRNA is mediated by an internal ribosome entry site. J Biol Chem. 276:5668-5675
Greco A, Bienvenut W, Sanchez JC, Kindbeiter K, Hochstrasser D, Madjar JJ, and Diaz JJ (2001). Identification of ribosome-associated viral and cellular basic proteins during the course of infection with herpes simplex virus type 1. Proteomics. 1:545-59.
Logvinoff C and Epstein AL (2001). A novel approach for HSV-1 amplicon vector production using Cre-loxP recombination system to remove the helper virus. Human Gene Therapy 12, 161-167.

2000
Greco A, Bausch N, Couté Y and Diaz, J-J (2000). Characterization by two-dimensional gel electrophoresis of host proteins whose synthesis is sustained or stimulated during the course of herpes simplex virus type 1 infection. Electrophoresis, 21:2522-30
Logvinoff C and Epstein AL (2000). Genetic engineering of herpes simplex virus and vector genomes carrying loxP sites in cells expressing Cre recombinase. Virology 267, 102-110.
Logvinoff C and Epstein AL (2000). Intracellular Cre-mediated deletion of the unique packaging signal carried by an HSV-1 recombinant and its relationship with the cleavage-packaging process. J. Virol. 74, 8402-8412.
Zaupa C and Epstein AL (2000). Herpesvirus/Retrovirus Chimeras for Gene Delivery. In Viral Vectors: Basic Science and Gene Therapy. A Cid-Arregui and A Garcia-Carrança, eds. Eaton Publishing, Natick, MA, pp.251-263.

Last modified : 03/17/2009