Project ID CM-HD2025_05

ThemeCM-HD

Co Supervisor 1A Prof Jody Rosenblatt Faculty of Life Sciences & Medicine, School of Basic & Medical Biosciences, Randall Centre for Cell & Molecular BiophysicsEmail

Co Supervisor 1B Dr Rui Pedro Galao Faculty of Life Sciences & Medicine, School of Immunology & Microbial Sciences, Department of Infectious DiseasesEmail

New roles for extrusion in learned innate immune response to SARS-CoV2

Background: While most viral response studies have focused on adaptive immunity, to launch an infection, a virus must breach the epithelial barrier that protects our organs. We found that epithelial monolayers can rapidly eliminate rhinovirus, responsible for most common colds, by extruding cells. Importantly, these extruded virally-infected cells act as viral bombs that can infect fresh epithelia, but do not re-infect previously infected epithelia, suggesting a new type of epithelial-based learned innate immunity. Recent evidence that innate response can effectively protect mice without T- or B-cells, emphasizes the importance of this finding and suggests that we could boost learned innate immunity.

Rationale and aims: Here, we want to learn if epithelia eliminate SARS-CoV2 similarly by extrusion, yielding an infection-resistant monolayer in collaboration with Rui Galao. We have preliminary data suggesting that the original wave 1 SARS-CoV2 virus is eliminated by extrusion. Yet, evidence suggest that SARS-CoV2 variants of concern (VOCs) evolved mechanisms of innate immune evasion. To investigate this hypothesis, the student will be working with the Galao and Rosenblatt labs at KCL and the Crick to:

1) Test if epithelial eliminate various variants of SARS CoV2-infected cells by extrusion

2) Investigate if the remaining monolayer is resistant to further infections

3) If monolayer acquires learned innate immunity, does it apply to all variants?

Approach and objectives:

The student will learn: how to work with viruses safely, how to do extrusion assays in both live and fixed monolayers (Year 1), learn high resolution and live microscopy (all years), and transcriptomic approaches to investigate changes after infection (Year 2). Identify pathways for cells to protect themselves against infections, using knockdowns, inhibitors, and viral bomb assays (Year 3). The student will be expected to be self-motivated in learning techniques, reading literature, and networking to become an engaged and successful scientist.

Representative Publications

1. Bronchoconstriction damages airway epithelia by crowding-induced excess cell extrusion. Bagley DC, Russell T, Ortiz-Zapater E, Stinson S, Fox K, Redd PF, Joseph M, Deering-Rice C, Reilly C, Parsons M, Brightling C, Rosenblatt J. Science. 2024 Apr 5;384(6691):66-73. doi: 10.1126/science.adk2758. Epub 2024 Apr 4. PMID: 38574138. pdf

2. Epithelial cell extrusion: Pathways and pathologies. Gudipaty SA, Rosenblatt J. Semin Cell Dev Biol. 2017 Jul;67:132-140. doi: 10.1016/j.semcdb.2016.05.010. Epub 2016 May 19. Review. PMID: 27212253. pdf

3. Crowding induces live cell extrusion to maintain homeostatic cell numbers in epithelia.Eisenhoffer GT, Loftus PD, Yoshigi M, Otsuna H, Chien CB, Morcos PA, Rosenblatt J. Nature. 2012 Apr 15;484(7395):546-9. doi: 10.1038/nature10999. PMID: 22504183. pdf

Lista MJ, Winstone H, Wilson HD, Dyer A, Pickering S, Galão RP, De Lorenzo G, Cowton VM, Furnon W, Suarez N, Orton R, Palmarini M, Patel AH, Snell L, Nebbia G, Swanson C, Neil SJD. The P681H mutation in the Spike glycoprotein confers Type I interferon resistance in the SARS-CoV-2 alpha variant. J Virol. 2022 Dec 14;96(23):e0125022. doi:10.1128/jvi.01250-22. Pdf

Galão RP#, Wilson H, Schierhorn KL, Debeljak F, Bodmer BS, Goldhill D, Hoenen T, Wilson SJ, Swanson CM, Neil SJD. TRIM25 and ZAP target the Ebola virus ribonucleoprotein complex to mediate interferon-induced restriction. PLoS Pathogens. 2022, 18(5), e1010530. # co-corresponding author. doi:10.1371/journal.ppat.1010530. Pdf

Pickering S, Batra R, Merrick B, Snell LB, Nebbia G, Douthwaite S, Reid F, Patel A, Kia Ik MT, Patel B, Charalampous T, Alcolea-Medina A, Lista MJ, Cliff PR, Cunningham E, Mullen J, Doores KJ, Edgeworth JD, Malim MH, Neil SJD, Galão RP. Comparative performance of SARS-CoV-2 lateral flow antigen tests and association with detection of infectious virus in clinical specimens: a single-centre laboratory evaluation study. Lancet Microbe. 2021 Sep;2(9):e461-e471. doi:10.1016/S2666-5247(21)00143-9. Pdf