Controlling Influenza A Virus Liquid Organelles or LOFlu
This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No 101001521).
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Presenting the project
Viral outbreaks necessitate a front line of antiviral strategies since the on-demand development of vaccines or antibody treatment is not always feasible. The EU-funded LOFlu project will investigate the host-pathogen interactions in influenza A virus (IAV) infection. Particular emphasis will be given to viral inclusions with liquid properties that form during viral assembly and constitute a key step in the virus life cycle. By studying how the host and the virus work in concert to coordinate the formation and material properties of IAV liquid inclusions and their interactions, researchers will shed light on the organisation of viral reactions in space and in time, as well as on the principles governing the assembly of IAV epidemic and pandemic genomes.
building on our preliminary data
LOFlu is concentrating on the fundamental problem of compartmentalizing viral reactions. This is critical in all viral infections, as viruses need to perform several steps in their viral lifecycle using selected material in a particular cellular location at a specific time of infection.
We build on our finding that influenza A virus forms biomolecular condensates also called viral inclusions that display liquid properties (1). We hypothesise that these sites are functional and that the properties are essential for function. Hence, we are learning how to manipulate them may retrieve novel antiviral strategies.
Given that many viruses shown in (2) also build biomolecular condensates with liquid properties, our findings may be of relevance to several viral infections.
(1) Alenquer M, Vale-Costa S, Sousa AL, Etibor TA, Ferreira F and Amorim MJ, Influenza A virus ribonucleoproteins form liquid organelles at endoplasmic reticulum exit sites, 2019, Nature Communications, 10, Article number:1629. https://doi.org/10.1038/s41467-019-09549-4. BioRxiv: https://doi.org/10.1101/410373.
(2) Etibor, T. A., Yamauchi, Y., & Amorim, M. J. (2021, Feb 25). Liquid Biomolecular Condensates and Viral Lifecycles: Review and Perspectives. Viruses, 13(3). https://doi.org/10.3390/v13030366
We build on our finding that influenza A virus forms biomolecular condensates also called viral inclusions that display liquid properties (1). We hypothesise that these sites are functional and that the properties are essential for function. Hence, we are learning how to manipulate them may retrieve novel antiviral strategies.
Given that many viruses shown in (2) also build biomolecular condensates with liquid properties, our findings may be of relevance to several viral infections.
(1) Alenquer M, Vale-Costa S, Sousa AL, Etibor TA, Ferreira F and Amorim MJ, Influenza A virus ribonucleoproteins form liquid organelles at endoplasmic reticulum exit sites, 2019, Nature Communications, 10, Article number:1629. https://doi.org/10.1038/s41467-019-09549-4. BioRxiv: https://doi.org/10.1101/410373.
(2) Etibor, T. A., Yamauchi, Y., & Amorim, M. J. (2021, Feb 25). Liquid Biomolecular Condensates and Viral Lifecycles: Review and Perspectives. Viruses, 13(3). https://doi.org/10.3390/v13030366
outputs
Manuscripts:
Etibor, T. A., Vale-Costa, S., Sridharan, S., Brás, D., Becher, I., Mello, V., Ferreira, F., Alenquer, M., Savitski, M. M., & Amorim, M. J. (2022). Rules for hardening influenza A virus liquid condensates. bioRxiv, 2022.2008.2003.502602. https://doi.org/10.1101/2022.08.03.502602 (this manuscript is being evaluated by Cell Reports).
Jani, R. A., Di Cicco, A., Keren-Kaplan, T., Vale-Costa, S., Hamaoui, D., Hurbain, I., Tsai, F. C., Di Marco, M., Mace, A. S., Zhu, Y., Amorim, M. J., Bassereau, P., Bonifacino, J. S., Subtil, A., Marks, M. S., Levy, D., Raposo, G., & Delevoye, C. (2022, Nov 7). PI4P and BLOC-1 remodel endosomal membranes into tubules. J Cell Biol, 221(11). https://doi.org/10.1083/jcb.202110132
Training:
(3) Registration "Tissue Cleaning Online Workshop", online course organized by the microscopy imaging centrer, CNC (Daniela Brás, 18th-20th May 2021); this course is ideally designed to optimise the staining of animal model tissue sections for microscopy imaging.
(4) Travel/Registration EMBL Introduction to metabolomics analysis (Daniela Brás, 23rd to 28th May 2022), EMBL-EBI Hinxton, Cambridge, UK; this course is key to help Daniela design experimental setup and analyse big data resultant from metabolomics.
(5) Visit from Victor Pessoa to the laboratory of Alex Borodavka at the University of Cambridge, from the 13th – 27th February 2022.
Conferences:
(6) Registration to Cell Symposia: Biological Assemblies: Phase Transitions, an online meeting organized by Elsevier (Temitope Etibor, 1st-3rd November 2021). Here he presented his work on the rules of hardening influenza A virus biomolecular condensates, part of the ERC project.
(7) Registration to EMBL Symposium: Cellular mechanisms driven by phase separation (Daniela Brás, 9th - 12th May 2022). Daniela Brás was exposed to a key meeting in the field of phase transitions.
(8) Registration to Microscopy at the Frontiers of Science (MFS2021), an online meeting organized by INL institute (Sílvia Vale-Costa 29th – 01st October 2021). At this meeting Sílvia presented and received feedback on her work on mechanisms regulating the formation of IAV inclusions, part of the LOFlu project.
(9) Registration/travel/accommodation to XVIII International Conference on Negative Strand Viruses, (Sílvia Vale-Costa; Temitope Etibor, Maria João Amorim 12th -17th June 2022, Braga, PT). At this meeting Sílvia and Etibor presented their projects in poster form and I Maria João Amorim presented an overview of the LOFlu project as a selected oral communication.
(10) Registration/travel/organization of LOFlu conference that took place on the 29th-30th September in Amiera, Portugal. This meeting was key to discuss the progress of LOFlu and critically assess and decide the next steps in LOFlu.
Oral Communications:
(11) Maria João Amorim was invited to give a seminar on “New paradigms in influenza A virus infection”, part of LOFlu for the iMM Monday Lecture series, Lisbon, Portugal, 12th July 2021.
(12) Maria João Amorim was invited to give a seminar on “New paradigms in influenza A virus infection”, part of LOFlu for the SymbNet seminar series, EMBL-IGC, 28th September 2021.
(13) Maria João was invited to give a seminar on “Influenza A virus takes advantage of autophagy machinery to form viral inclusions with liquid properties at endoplasmic reticulum exit sites”, part of LOFlu for the Endoplasmatic reticulum conference, 6-8th September 2021, Paris, France.
Courses:
(14) Maria João was invited to give a seminar on “New paradigms in virology”, part of LOFlu for the Course Fighting Infections, 16th March 2022, University of Coimbra, PT [invited by Isaura Simões].
(15) Maria João was invited to give a seminar on “New paradigms in virology”, part of LOFlu for the Course on phase separation and aggregation, linked with the Twinning project (PhasAGE). 20th April 2022, i3S, PT [invited by Sandra Ribeiro].
Etibor, T. A., Vale-Costa, S., Sridharan, S., Brás, D., Becher, I., Mello, V., Ferreira, F., Alenquer, M., Savitski, M. M., & Amorim, M. J. (2022). Rules for hardening influenza A virus liquid condensates. bioRxiv, 2022.2008.2003.502602. https://doi.org/10.1101/2022.08.03.502602 (this manuscript is being evaluated by Cell Reports).
Jani, R. A., Di Cicco, A., Keren-Kaplan, T., Vale-Costa, S., Hamaoui, D., Hurbain, I., Tsai, F. C., Di Marco, M., Mace, A. S., Zhu, Y., Amorim, M. J., Bassereau, P., Bonifacino, J. S., Subtil, A., Marks, M. S., Levy, D., Raposo, G., & Delevoye, C. (2022, Nov 7). PI4P and BLOC-1 remodel endosomal membranes into tubules. J Cell Biol, 221(11). https://doi.org/10.1083/jcb.202110132
Training:
(3) Registration "Tissue Cleaning Online Workshop", online course organized by the microscopy imaging centrer, CNC (Daniela Brás, 18th-20th May 2021); this course is ideally designed to optimise the staining of animal model tissue sections for microscopy imaging.
(4) Travel/Registration EMBL Introduction to metabolomics analysis (Daniela Brás, 23rd to 28th May 2022), EMBL-EBI Hinxton, Cambridge, UK; this course is key to help Daniela design experimental setup and analyse big data resultant from metabolomics.
(5) Visit from Victor Pessoa to the laboratory of Alex Borodavka at the University of Cambridge, from the 13th – 27th February 2022.
Conferences:
(6) Registration to Cell Symposia: Biological Assemblies: Phase Transitions, an online meeting organized by Elsevier (Temitope Etibor, 1st-3rd November 2021). Here he presented his work on the rules of hardening influenza A virus biomolecular condensates, part of the ERC project.
(7) Registration to EMBL Symposium: Cellular mechanisms driven by phase separation (Daniela Brás, 9th - 12th May 2022). Daniela Brás was exposed to a key meeting in the field of phase transitions.
(8) Registration to Microscopy at the Frontiers of Science (MFS2021), an online meeting organized by INL institute (Sílvia Vale-Costa 29th – 01st October 2021). At this meeting Sílvia presented and received feedback on her work on mechanisms regulating the formation of IAV inclusions, part of the LOFlu project.
(9) Registration/travel/accommodation to XVIII International Conference on Negative Strand Viruses, (Sílvia Vale-Costa; Temitope Etibor, Maria João Amorim 12th -17th June 2022, Braga, PT). At this meeting Sílvia and Etibor presented their projects in poster form and I Maria João Amorim presented an overview of the LOFlu project as a selected oral communication.
(10) Registration/travel/organization of LOFlu conference that took place on the 29th-30th September in Amiera, Portugal. This meeting was key to discuss the progress of LOFlu and critically assess and decide the next steps in LOFlu.
Oral Communications:
(11) Maria João Amorim was invited to give a seminar on “New paradigms in influenza A virus infection”, part of LOFlu for the iMM Monday Lecture series, Lisbon, Portugal, 12th July 2021.
(12) Maria João Amorim was invited to give a seminar on “New paradigms in influenza A virus infection”, part of LOFlu for the SymbNet seminar series, EMBL-IGC, 28th September 2021.
(13) Maria João was invited to give a seminar on “Influenza A virus takes advantage of autophagy machinery to form viral inclusions with liquid properties at endoplasmic reticulum exit sites”, part of LOFlu for the Endoplasmatic reticulum conference, 6-8th September 2021, Paris, France.
Courses:
(14) Maria João was invited to give a seminar on “New paradigms in virology”, part of LOFlu for the Course Fighting Infections, 16th March 2022, University of Coimbra, PT [invited by Isaura Simões].
(15) Maria João was invited to give a seminar on “New paradigms in virology”, part of LOFlu for the Course on phase separation and aggregation, linked with the Twinning project (PhasAGE). 20th April 2022, i3S, PT [invited by Sandra Ribeiro].
our achievements & future aims
In LOFlu, we have started to elucidate the molecular underpinnings on the formation and influenza A liquid inclusions. We are expanding our efforts to elucidate how the material properties are maintained and regulated in the cell. We have already shown that it is possible to harden viral inclusions and have defined the best strategies. This work supports the development of antivirals targeting the material properties of biomolecular condensates in viral infections (SEE FIGURE ABOVE FOR VIRALS IDENTIFIED TO UTILISE BIOMOLECULAR CONDENSATES TO DATE). It also provides a framework for the selection of compounds with this activity for general application and thus provides an advance in disease therapy. We will now concentrate on mechanisms that may modulate targeting the molecular interactions established within viral inclusions to provide novel antiviral strategies. Finally, we have identified which viral and host components undergo phase transitions during infection and are now understanding how phase transitions regulate host cell function and response to infection whilst facilitating or inhibiting viral replication. Such understanding will shed new light on how phase transitions operate as a new regulatory layer in the cell.
