Anna Salvetti

Inserm

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David Durantel

Université Claude Bernard Lyon 1

Philippe Moullier

Gene Therapy Laboratory

Julie Lucifora

Université Claude Bernard Lyon 1

Michel Rivoire

Centre Léon Bérard

Gilliane Chadeuf

Institut du Thorax

Fabien Zoulim

Centre de Recherche en Cancérologie de Lyon

Maud Michelet

Centre de Recherche en Cancérologie de Lyon

Alberto L. Epstein

Université de Versailles Saint-Quentin-en-Yvelines

Hildegard Büning

Medizinische Hochschule Hannover

François‐Loïc Cosset

Inserm

Cooperative Institutions

Inserm

195

Centre National de la Recherche Scientifique

159

Université Claude Bernard Lyon 1

120

École Normale Supérieure de Lyon

European Foundation for the Study of Chronic Liver Failure

Centre de Recherche en Cancérologie de Lyon

Centre International de Recherche en Infectiologie

Université Paris Cité

Sorbonne Université

Centre Léon Bérard

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Delivery of therapeutic proteins from genetically-modified cells

Restorative Neurology and Neuroscience (1995)

Nadia Naffakh Philippe Moullier Anna Salvetti Delphine Bohl Olivier Danos

10.3233/rnn-1995-81216

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Impact of the MRN Complex on Adeno-Associated Virus Integration and Replication during Coinfection with Herpes Simplex Virus 1

Journal of Virology (2015)

Rachel Millet Nelly Jolinon Xuan-Nhi Nguyen Grégory Berger Andrea Cimarelli

Adeno-associated virus (AAV) is a helper-dependent parvovirus that requires coinfection with adenovirus (AdV) or herpes simplex virus 1 (HSV-1) to replicate. In the absence of the helper virus, AAV can persist in an episomal or integrated form. Previous studies have analyzed the DNA damage response (DDR) induced upon AAV replication to understand how it controls AAV replication. In particular, it was shown that the Mre11-Rad50-Nbs1 (MRN) complex, a major player of the DDR induced by double-stranded DNA breaks and stalled replication forks, could negatively regulate AdV and AAV replication during coinfection. In contrast, MRN favors HSV-1 replication and is recruited to AAV replication compartments that are induced in the presence of HSV-1. In this study, we examined the role of MRN during AAV replication induced by HSV-1. Our results indicated that knockdown of MRN significantly reduced AAV DNA replication after coinfection with wild-type (wt) HSV-1 or HSV-1 with the polymerase deleted. This effect was specific to wt AAV, since it did not occur with recombinant AAV vectors. Positive regulation of AAV replication by MRN was dependent on its DNA tethering activity but did not require its nuclease activities. Importantly, knockdown of MRN also negatively regulated AAV integration within the human AAVS1 site, both in the presence and in the absence of HSV-1. Altogether, this work identifies a new function of MRN during integration of the AAV genome and demonstrates that this DNA repair complex positively regulates AAV replication in the presence of HSV-1.Viral DNA genomes trigger a DNA damage response (DDR), which can be either detrimental or beneficial for virus replication. Adeno-associated virus (AAV) is a defective parvovirus that requires the help of an unrelated virus such as adenovirus (AdV) or herpes simplex virus 1 (HSV-1) for productive replication. Previous studies have demonstrated that the cellular Mre11-Rad50-Nbs1 (MRN) complex, a sensor and regulator of the DDR, negatively regulates AAV replication during coinfection with AdV, which counteracts this effect by inactivating the complex. Here, we demonstrate that MRN positively regulates AAV replication during coinfection with HSV-1. Importantly, our study also indicates that MRN also favors integration of AAV genomes within the human AAVS1 site. Altogether, this work indicates that MRN differentially regulates AAV replication depending on the helper virus which is present and identifies a new function of this DNA repair complex during AAV integration.

10.1128/jvi.00171-15

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Citations (12)

Aumentata sensibilità cronotropa e ridottqa riserva coronarica si associano a diminuita risposta inotropa allo stimolo beta-adrenergico in paziente con ipertensione lieve-moderata

The Cardiology (1998)

Carlo Palombo M. Kozàkovà G. Bigalli C. Morizzo Alessandra Magagna

Source

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Factors influencing helper-independent adeno-associated virus replication

Virology (2012)

Armel Nicolas Nelly Jolinon Nathalie Alazard-Dany Véronique Barateau Alberto L. Epstein

Helper virus

Adeno-associated virus

Replication

10.1016/j.virol.2012.05.027

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Citations (9)

Zoonoses virales et émergence : la recherche ne fait que commencer

médecine/sciences (2015)

Anna Salvetti Sylvain Baize

> La faune sauvage possede un tres large spectre d’agents infectieux dont certains peuvent occasionner des zoonoses. Dans le cas des virus, ces zoonoses peuvent etre a l’origine d’emergences entrainant des pathologies severes chez l’homme, avec des impacts sanitaires et economiques considerables, comme l’epidemie de maladie a virus Ebola d’une ampleur inedite qui a recemment frappe l’Afrique de l’Ouest [1]. Mais comment definit-on un virus emergent et peut-on anticiper la survenue de ces epidemies ? La definition la plus commune est celle de l’emergence d’un virus precedemment inconnu. Le VIH (virus de l’immunodeficience humaine) en est l’un des exemples les plus notables, a l’origine d’une des plus larges pandemies de l’histoire. Plus recemment, deux nouveaux membres de la famille des Coronavirus1, le SARSCoV (severe advanced respiratory syndrome coronavirus) [2] et le MERS-CoV (middle east respiratory syndrome coronavirus), ont ete identifies comme etant a l’origine des maladies respiratoires severes qui ont touche la Chine en 2003 et l’Arabie saoudite en 2012. Mais un virus deja connu peut aussi donner lieu a une emergence, en touchant une region geographique jusqu’alors indemne, a l’instar du virus West Nile (virus du Nil occidental) [3, 4], un membre de la famille des Flavivirus2, identifie en 1937 en Ouganda et qui reapparait regulierement dans differentes zones geographiques, de la recente epidemie de maladie a virus Ebola en Afrique de l’Ouest, et meme de l’apparition de cas autochtones d’infections par le virus Chikungunya3 [5] ou celui de la dengue4 en France metropolitaine. L’augmentation importante de l’incidence d’une infection virale constitue egalement une emergence, ce qui a ete le cas pour les epidemies de fievres hemorragiques a Filovirus5 frequemment observees en Afrique Centrale au cours des deux dernieres decennies. Enfin, l’apparition de nouvelles souches de virus deja connus presentant une pathogenicite differente

10.1051/medsci/20153112001

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Citations (1)

Soft jamming of viral particles in nanopores

Nature Communications (2024)

Léa Chazot-Franguiadakis Joëlle Eid Gwendoline Delecourt Pauline J. Kolbeck Saskia Brugère

Abstract Viruses have remarkable physical properties and complex interactions with their environment. However, their aggregation in confined spaces remains unexplored, although this phenomenon is of paramount importance for understanding viral infectivity. Using hydrodynamical driving and optical detection, we developed a method to detect the transport of single virus in real time through synthetic nanopores. We unveiled a jamming phenomenon specifically associated with virus confinement under flow. We showed that the interactions of viral particles with themselves and with the pore surface were critical for clog formation. Based on the detailed screening of the physical and chemical determinants, we proposed a simple dynamical model that recapitulated all the experimental observations. Our results pave the way for the study of jamming phenomena in the presence of more complex interactions.

Nanopore

Infectivity

10.1038/s41467-024-50059-9

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Identification of a replication‐defective herpes simplex virus for recombinant adeno‐associated virus type 2 (rAAV2) particle assembly using stable producer cell lines

The Journal of Gene Medicine (2004)

Estelle Toublanc Abdellatif Benraiss Delphine Bonnin Véronique Blouin Nicole Brument

Abstract Background The development of stable producer cell lines for recombinant adeno‐associated virus (rAAV) assembly is a strategy followed by many groups to develop scalable production methods suitable for good manufacturing practice (GMP) requirements. The major drawback of this method lies in the requirement for replicating adenovirus (Ad) for rAAV assembly. In the present study, we analyzed the ability of several replication‐defective herpes simplex type 1 (HSV‐1) helper viruses to induce rAAV2 particle production from stable producer cell lines. Methods Several stable rAAV producer cell clones were infected with wild‐type and replication‐defective HSV strains and analyzed for rep‐cap gene amplification, viral protein synthesis and rAAV titers achieved. In vivo analysis following rAAV injection in the murine brain was also conducted to evaluate the toxicity and biopotency of the rAAV stocks. Results We demonstrated that an HSV strain mutated in the UL30 polymerase gene could efficiently be used in this context, resulting in rAAV titers similar to those measured with wild‐type HSV or Ad. Importantly, with respect to clinical developments, the use of this mutant resulted in rAAV stocks which were consistently devoid of contaminating HSV particles and fully active in vivo in the murine central nervous system with no detectable toxicity. Conclusions This study, together with our previous report describing a rAAV chromatography‐based purification process, contributes to the definition of an entirely scalable process for the generation of rAAV particles. Copyright © 2004 John Wiley & Sons, Ltd.

Adeno-associated virus

10.1002/jgm.542

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Citations (26)

Efficient recombinant adeno‐associated virus production by a stable rep‐cap HeLa cell line correlates with adenovirus‐induced amplification of the integrated rep‐cap genome

The Journal of Gene Medicine (2000)

Gilliane Chadeuf David S. Favre Jacques Tessier Nathalie Provost Pascale Nony

HeLa

Adeno-associated virus

10.1002/1521-2254(200007/08)2:4<260::aid-jgm111>3.3.co;2-

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Citations (1)

Fast Differentiation of HepaRG Cells Allowing Hepatitis B and Delta Virus Infections

Cells (2020)

Julie Lucifora Maud Michelet Anna Salvetti David Durantel

HepaRG cells are liver bipotent progenitors acquiring hepatocytes features when differentiated in the presence of dimethylsulfoxide (DMSO). Differentiated HepaRG (dHepaRG) are considered the best surrogate model to primary human hepatocytes (PHH) and are susceptible to several hepatotropic viruses, including Hepatitis B Virus (HBV) and Hepatitis Delta Virus (HDV) infection. Despite these advantages, HepaRG cells are not widely used for the study of these two viruses because of their long differentiation process and their rather low and variable infection rates. Here, we tested the use of a cocktail of five chemicals (5C) combined or not with DMSO to accelerate the cells' differentiation process. We found that NTCP-mediated HDV entry and replication are similar in HepaRG cells cultivated for only 1 week with 5C and DMSO or differentiated with the regular 4-week protocol. However, even though the NTCP-mediated HBV entry process seemed similar, cccDNA and subsequent HBV replication markers were lower in HepaRG cells cultivated for 1 week with 5C and DMSO compared to the regular differentiation protocol. In conclusion, we set up a new procedure allowing fast differentiation and efficient HDV-infection of HepaRG cells and identified differential culture conditions that may allow to decipher the mechanism behind the establishment of the HBV minichromosome.

cccDNA

10.3390/cells9102288

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Citations (10)

Modification of the intermediate filaments distribution in HeLa cells expressing the tax protein from HTLV-I

Biology of the Cell (1991)

Anna Salvetti Marie‐Madeleine Portier Pierre Gounon Alain Lilienbaum D. Paulin

HeLa

10.1016/0248-4900(91)90130-f

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