Pablo Gastaminza Landart
Group Leader
Research summary
Our laboratory is interested in deciphering the molecular events underlying the functional dependence of HCV infection on lipid and lipoprotein metabolism and how to exploit these pathways to inhibit HCV infection. We use a cell culture infection system based on the infectious molecular clone JFH-1 and the Huh-7 hepatoma cell line and derived subclones to evaluate the impact that genetic and pharmacological manipulation of cellular lipid metabolism has on HCV infection and pathogenesis.
Publications
Sepúlveda-Crespo D, Jiménez JL, Gómez R, De La Mata FJ, Majano PL, Muñoz-Fernández MÁ, Gastaminza P. Polyanionic carbosilane dendrimers prevent hepatitis C virus infection in cell culture. Nanomedicine 2017; 13: 49-58
Pérez-Berná AJ, Rodríguez MJ, Chichón FJ, Friesland MF, Sorrentino A, Carrascosa JL, Pereiro E, Gastaminza P. Structural Changes In Cells Imaged by Soft X-ray Cryo-Tomography During Hepatitis C Virus Infection. ACS Nano 2016; 10: 6597-611
Carnero E, Barriocanal M, Prior C, Pablo Unfried J, Segura V, Guruceaga E, Enguita M, Smerdou C, Gastaminza P, Fortes P. Long noncoding RNA EGOT negatively affects the antiviral response and favors HCV replication. EMBO Rep 2016; 17: 1013-28
Benedicto I, Gondar V, Molina-Jiménez F, García-Buey L, López-Cabrera M, Gastaminza P, Majano PL. Clathrin mediates infectious hepatitis C virus particle egress. J Virol 2015; 89: 4180-90
Vasallo C, Gastaminza P. Cellular stress responses in hepatitis C virus infection: Mastering a two-edged sword. Virus Res 2015; 209: 100-17
Hepatitis C virus (HCV) is a pathogen that infects 3% of the human population worldwide. Despite great efforts to control this pandemic, 3 to 4 million people become infected and about 350,000 individuals die of HCV-related diseases every year. Our laboratory is interested in the cellular and molecular processes that underlie different aspects of HCV biology and pathogenesis, to discover new targets for antiviral therapy.
Using a cell culture model of HCV infection, we identified a host factor, the sigma-1 receptor (S1R), with a specific role at the onset of the HCV lifecycle. This cell factor is an important component of mitochondria-associated endoplasmic reticulum (ER) membranes (MAM) and regulates bidirectional interorganellar transport of lipids and Ca2+ ions between mitochondria and the ER. Silencing of this factor resulted in a proportional decrease in susceptibility to HCV infection. Mechanistic studies indicated that early steps in viral RNA replication downstream of translation of the incoming viral genomes are rate-limited by cellular S1R levels. These findings raise the possibility that HCV uses MAM as a gateway to the cell machinery necessary for efficient viral replication. We are currently determining the molecular mechanisms by which HCV uses S1R and the potential pathological consequences, using broad approaches that involve determination of alterations in the proteomic composition of S1R-containing macromolecular complexes during HCV infection.
In addition to these basic studies, we sought new molecules with antiviral potential against HCV. Using a screening system designed in-house, we interrogated a chemical library of 478 compounds approved for use in clinical practice for non-HCV applications. The rationale is that prior knowledge of the molecular mechanisms of their pharmacological action as well as their cell targets, toxicity, bioavailability and pharmacokinetics could expedite translation to the clinic. In this study, we identified a set of 12 compounds, of which two (hydroxyzine and benztropine) were selected for further characterisation. At micromolar concentrations, these compounds selectively blocked HCV entry; hydroxyzine was antiviral at clinically achievable doses, preferentially for genotype 2 viruses.
