Alejandro Matia

All Publications

• Spatio-temporal analysis of Vaccinia virus infection and host response dynamics using single-cell transcriptomics and proteomics bioRxiv Matia, A., McCarthy, F., Woosley, H., Turon-Lagot, V., Platzer, S. W., Liu, J., Lorenzo, M. M., Borja, M., Shetty, K., Winkler, J., Elias, J. E., Blasco, R., Arias, C., Hein, M. Y. 2024

  View details for DOI 10.1101/2024.01.13.575413v1

• Identification of β2 microglobulin, the product of B2M gene, as a Host Factor for Vaccinia Virus Infection by Genome-Wide CRISPR genetic screens. PLoS pathogens Matía, A., Lorenzo, M. M., Romero-Estremera, Y. C., Sánchez-Puig, J. M., Zaballos, A., Blasco, R. 2022; 18 (12): e1010800

  Abstract

  Genome-wide genetic screens are powerful tools to identify genes that act as host factors of viruses. We have applied this technique to analyze the infection of HeLa cells by Vaccinia virus, in an attempt to find genes necessary for infection. Infection of cell populations harboring single gene inactivations resulted in no surviving cells, suggesting that no single gene knock-out was able to provide complete resistance to Vaccinia virus and thus allow cells to survive infection. In the absence of an absolute infection blockage, we explored if some gene inactivations could provide partial protection leading to a reduced probability of infection. Multiple experiments using modified screening procedures involving replication restricted viruses led to the identification of multiple genes whose inactivation potentially increase resistance to infection and therefore cell survival. As expected, significant gene hits were related to proteins known to act in virus entry, such as ITGB1 and AXL as well as genes belonging to their downstream related pathways. Additionally, we consistently found β2-microglobulin, encoded by the B2M gene, among the screening top hits, a novel finding that was further explored. Inactivation of B2M resulted in 54% and 91% reduced VV infection efficiency in HeLa and HAP1 cell lines respectively. In the absence of B2M, while virus binding to the cells was unaffected, virus internalization and early gene expression were significantly diminished. These results point to β2-microglobulin as a relevant factor in the Vaccinia virus entry process.

  View details for DOI 10.1371/journal.ppat.1010800

  View details for PubMedID 36574441

  View details for PubMedCentralID PMC9829182

• Tools for the targeted genetic modification of poxvirus genomes. Current opinion in virology Matía, A., Lorenzo, M. M., Blasco, R. 2020; 44: 183-190

  Abstract

  The potential of viruses as biotechnology platforms is becoming more appealing due to technological advances in synthetic biology techniques and to the increasing accessibility of means to manipulate virus genomes. Among viral systems, poxviruses, and their prototype member Vaccinia Virus, are one of the outstanding choices for different biotechnological and medical applications based on heterologous gene expression, recombinant vaccines or oncolytic viruses. The refinement of genetic engineering methods on Vaccinia Virus over the last decades have contributed to facilitate the manipulation of the genomes of poxviruses, and may aid in the improvement of virus variants designed for different goals through reverse genetic approaches. Targeted genetic changes are usually performed by homologous recombination with the viral genome. In addition to the classic approach, recent methodological advances that may assist new strategies for the mutation or edition of poxvirus genomes are reviewed.

  View details for DOI 10.1016/j.coviro.2020.10.006

  View details for PubMedID 33242829

• Vaccinia Virus Strain MVA Expressing a Prefusion-Stabilized SARS-CoV-2 Spike Glycoprotein Induces Robust Protection and Prevents Brain Infection in Mouse and Hamster Models. Vaccines Lorenzo, M. M., Marín-López, A., Chiem, K., Jimenez-Cabello, L., Ullah, I., Utrilla-Trigo, S., Calvo-Pinilla, E., Lorenzo, G., Moreno, S., Ye, C., Park, J. G., Matía, A., Brun, A., Sánchez-Puig, J. M., Nogales, A., Mothes, W., Uchil, P. D., Kumar, P., Ortego, J., Fikrig, E., Martinez-Sobrido, L., Blasco, R. 2023; 11 (5)

  Abstract

  The COVID-19 pandemic has underscored the importance of swift responses and the necessity of dependable technologies for vaccine development. Our team previously developed a fast cloning system for the modified vaccinia virus Ankara (MVA) vaccine platform. In this study, we reported on the construction and preclinical testing of a recombinant MVA vaccine obtained using this system. We obtained recombinant MVA expressing the unmodified full-length SARS-CoV-2 spike (S) protein containing the D614G amino-acid substitution (MVA-Sdg) and a version expressing a modified S protein containing amino-acid substitutions designed to stabilize the protein a in a pre-fusion conformation (MVA-Spf). S protein expressed by MVA-Sdg was found to be expressed and was correctly processed and transported to the cell surface, where it efficiently produced cell-cell fusion. Version Spf, however, was not proteolytically processed, and despite being transported to the plasma membrane, it failed to induce cell-cell fusion. We assessed both vaccine candidates in prime-boost regimens in the susceptible transgenic K18-human angiotensin-converting enzyme 2 (K18-hACE2) in mice and in golden Syrian hamsters. Robust immunity and protection from disease was induced with either vaccine in both animal models. Remarkably, the MVA-Spf vaccine candidate produced higher levels of antibodies, a stronger T cell response, and a higher degree of protection from challenge. In addition, the level of SARS-CoV-2 in the brain of MVA-Spf inoculated mice was decreased to undetectable levels. Those results add to our current experience and range of vaccine vectors and technologies for developing a safe and effective COVID-19 vaccine.

  View details for DOI 10.3390/vaccines11051006

  View details for PubMedID 37243110

  View details for PubMedCentralID PMC10220993

• MaGplotR: a software for the analysis and visualization of multiple MaGeCK screen datasets through aggregation bioRxiv Matia, A., Lorenzo, M. M., Peng, D. 2023

  View details for DOI 10.1101/2023.01.12.523725