Rory Hills

All Publications

• Proactive vaccination using multiviral Quartet Nanocages to elicit broad anti-coronavirus responses. Nature nanotechnology Hills, R. A., Tan, T. K., Cohen, A. A., Keeffe, J. R., Keeble, A. H., Gnanapragasam, P. N., Storm, K. N., Rorick, A. V., West, A. P., Hill, M. L., Liu, S., Gilbert-Jaramillo, J., Afzal, M., Napier, A., Admans, G., James, W. S., Bjorkman, P. J., Townsend, A. R., Howarth, M. R. 2024; 19 (8): 1216-1223

  Abstract

  Defending against future pandemics requires vaccine platforms that protect across a range of related pathogens. Nanoscale patterning can be used to address this issue. Here, we produce quartets of linked receptor-binding domains (RBDs) from a panel of SARS-like betacoronaviruses, coupled to a computationally designed nanocage through SpyTag/SpyCatcher links. These Quartet Nanocages, possessing a branched morphology, induce a high level of neutralizing antibodies against several different coronaviruses, including against viruses not represented in the vaccine. Equivalent antibody responses are raised to RBDs close to the nanocage or at the tips of the nanoparticle's branches. In animals primed with SARS-CoV-2 Spike, boost immunizations with Quartet Nanocages increase the strength and breadth of an otherwise narrow immune response. A Quartet Nanocage including the Omicron XBB.1.5 'Kraken' RBD induced antibodies with binding to a broad range of sarbecoviruses, as well as neutralizing activity against this variant of concern. Quartet nanocages are a nanomedicine approach with potential to confer heterotypic protection against emergent zoonotic pathogens and facilitate proactive pandemic protection.

  View details for DOI 10.1038/s41565-024-01655-9

  View details for PubMedID 38710880

  View details for PubMedCentralID PMC11329374

• Computationally designed haemagglutinin with nanocage plug-and-display elicits pan-H5 influenza vaccine responses. Emerging microbes & infections Huang, C. Q., Hills, R. A., Carnell, G. W., Vishwanath, S., Aguinam, E. T., Chan, A. C., Palmer, P., O'Reilly, L., Tonks, P., Temperton, N., Frost, S. D., Tiley, L. S., Howarth, M. R., Heeney, J. L. 2025; 14 (1): 2511132

  Abstract

  The increasing spread of highly pathogenic avian influenza (HPAI) A/H5 viruses poses a pandemic threat. Circulating clade 2.3.4.4b viruses have demonstrated rapid transcontinental dissemination, extensive reassortment, epizootic spread and potential sustained mammal-to-mammal transmission, signifying a heightened risk of becoming a human pathogen of high consequence. A broadly protective, future-proof vaccine against multiple clades of H5 influenza is urgently needed for pandemic preparedness. Here, we combine two novel vaccine technologies to generate a Digitally Immune Optimised and Selected H5 antigen (DIOSvax-H5inter) displayed multivalently on the mi3 nanocage using the SpyTag003/SpyCatcher003 conjugation system. Mice immunized with DIOSvax-H5inter Homotypic Nanocages at low doses demonstrate potent, cross-clade neutralizing antibody and T cell responses against diverse H5 strains. DIOSvax-H5inter Homotypic Nanocages provide a scalable vaccine candidate with the potential for pan-H5 protection against drifted or newly emergent H5 strains. This World Health Organization preferred characteristic is essential for prospective strategic stockpiling in the pre-pandemic phase.

  View details for DOI 10.1080/22221751.2025.2511132

  View details for PubMedID 40476519

  View details for PubMedCentralID PMC12258178

• mRNA delivery of genetically encoded mosaic-8 pan-sarbecovirus RBD vaccines. bioRxiv : the preprint server for biology Cohen, A. A., Keeffe, J. R., Manasyan, L., Madireddy, I., Fils, A. I., Dam, K. A., Stober, H. E., Hills, R. A., Moon, W. J., Lin, P. J., Howarth, M. R., Hoffmann, M. A., Bjorkman, P. J. 2025

  Abstract

  Global health remains threatened by spillovers of zoonotic SARS-like betacoronaviruses (sarbecoviruses) that could be mitigated by a pan-sarbecovirus vaccine1. We described elicitation of potently neutralizing and cross-reactive anti-sarbecovirus antibodies by mosaic-8 nanoparticles (NPs) displaying eight different sarbecovirus spike receptor-binding domains (RBDs) as 60 copies of eight individual RBDs2-6 (mosaic-8 RBD-NPs) or 30 copies of two "quartets," each presenting four tandemly-arranged RBDs7 (dual quartet RBD-NPs). To facilitate manufacture of a broadly protective mosaic-8 vaccine, we generated membrane-bound RBD quartets that can be genetically encoded and delivered via mRNA: dual quartet RBD-mRNA and dual quartet RBD-EABR-mRNA, which utilizes ESCRT- and ALIX-binding region (EABR) technology that promotes immunogen presentation on cell surfaces and circulating enveloped virus-like particles (eVLPs)8. Immunization with mRNA immunogens elicited equivalent or improved binding breadths, neutralization potencies, T cell responses, and targeting of conserved RBD epitopes across sarbecoviruses, demonstrating successful conversion of protein-based mosaic-8 RBD vaccines to mRNA formats. Systems serology9 showed that the mRNA vaccines elicited balanced IgG subclass responses with increased Fcγ receptor-binding IgGs, consistent with potentially superior Fc effector functions. A new technique, Systems Serology-Polyclonal Epitope Mapping (SySPEM), revealed distinct IgG-subclass-specific epitope targeting signatures across mRNA and protein-based vaccine modalities. These results demonstrate successful conversion of mosaic-8 RBD-NPs to mRNA or EABR-mRNA vaccines that provide easy manufacturing and enhanced protection from future pandemic sarbecovirus outbreaks.

  View details for DOI 10.1101/2025.10.21.683747

  View details for PubMedID 41280075

  View details for PubMedCentralID PMC12633426

• Mpox multiprotein virus-like nanoparticle vaccine induces neutralizing and protective antibodies in mice and non-human primates. Nature communications Belghith, A. A., Cotter, C. A., Ignacio, M. A., Earl, P. L., Hills, R. A., Howarth, M. R., Yee, D. S., Brenchley, J. M., Moss, B. 2025; 16 (1): 4726

  Abstract

  The upsurge of mpox in Africa and the recent global outbreak have stimulated the development of new vaccines and therapeutics. We describe the construction of virus-like particle (VLP) vaccines in which modified M1, A35 and B6 proteins from monkeypox virus (MPXV) clade Ia are conjugated individually or together to a scaffold that accommodates up to 60 ligands using the SpyTag/SpyCatcher nanoparticle system. Immunisation of female mice with VLPs induces higher anti-MPXV and anti-vaccinia virus (VACV) neutralizing antibodies than their soluble protein (SP) counterparts or modified VACV Ankara (MVA). Vaccination with individual single protein VLPs provides partial protection against lethal respiratory infections with VACV or MPXV clade IIa, whereas combinations or a chimeric VLP with all three antigens provide complete protection that is superior to SPs. Additionally, the VLP vaccine reduces the replication and spread of the virus at intranasal and intrarectal sites of inoculation. VLPs induce higher neutralizing activity than the Jynneos vaccine in rhesus macaques, and the VLP-induced antiserum provides better protection against MPXV and VACV than the Jynneos-induced antiserum when passively transferred to female mice. These data demonstrate that an mpox VLP vaccine derived from three MPXV clade Ia proteins protects against clade IIa MPXV and VACV, indicating cross-reactivity for orthopoxviruses.

  View details for DOI 10.1038/s41467-025-59826-8

  View details for PubMedID 40399314

  View details for PubMedCentralID PMC12095655

• Structural basis for a conserved neutralization epitope on the receptor-binding domain of SARS-CoV-2 NATURE COMMUNICATIONS Huang, K. A., Chen, X., Mohapatra, A., Nguyen, H., Schimanski, L., Tan, T., Rijal, P., Vester, S. K. K., Hills, R. A. A., Howarth, M., Keeffe, J. R. R., Cohen, A. A. A., Kakutani, L. M. M., Wu, Y., Shahed-Al-Mahmud, M., Chou, Y., Bjorkman, P. J. J., Townsend, A. R. R., Ma, C. 2023; 14 (1): 311

  Abstract

  Antibody-mediated immunity plays a crucial role in protection against SARS-CoV-2 infection. We isolated a panel of neutralizing anti-receptor-binding domain (RBD) antibodies elicited upon natural infection and vaccination and showed that they recognize an immunogenic patch on the internal surface of the core RBD, which faces inwards and is hidden in the "down" state. These antibodies broadly neutralize wild type (Wuhan-Hu-1) SARS-CoV-2, Beta and Delta variants and some are effective against other sarbecoviruses. We observed a continuum of partially overlapping antibody epitopes from lower to upper part of the inner face of the RBD and some antibodies extend towards the receptor-binding motif. The majority of antibodies are substantially compromised by three mutational hotspots (S371L/F, S373P and S375F) in the lower part of the Omicron BA.1, BA.2 and BA.4/5 RBD. By contrast, antibody IY-2A induces a partial unfolding of this variable region and interacts with a conserved conformational epitope to tolerate all antigenic variations and neutralize diverse sarbecoviruses as well. This finding establishes that antibody recognition is not limited to the normal surface structures on the RBD. In conclusion, the delineation of functionally and structurally conserved RBD epitopes highlights potential vaccine and therapeutic candidates for COVID-19.

  View details for DOI 10.1038/s41467-023-35949-8

  View details for Web of Science ID 000982581400002

  View details for PubMedID 36658148

  View details for PubMedCentralID PMC9852238

• SpySwitch enables pH- or heat-responsive capture and release for plug-and-display nanoassembly. Nature communications Vester, S. K., Rahikainen, R., Khairil Anuar, I. N., Hills, R. A., Tan, T. K., Howarth, M. 2022; 13 (1): 3714

  Abstract

  Proteins can be empowered via SpyTag for anchoring and nanoassembly, through covalent bonding to SpyCatcher partners. Here we generate a switchable version of SpyCatcher, allowing gentle purification of SpyTagged proteins. We introduce numerous histidines adjacent to SpyTag's binding site, giving moderate pH-dependent release. After phage-based selection, our final SpySwitch allows purification of SpyTag- and SpyTag003-fusions from bacterial or mammalian culture by capture at neutral pH and release at pH 5, with purity far beyond His-tag methods. SpySwitch is also thermosensitive, capturing at 4 °C and releasing at 37 °C. With flexible choice of eluent, SpySwitch-purified proteins can directly assemble onto multimeric scaffolds. 60-mer multimerization enhances immunogenicity and we use SpySwitch to purify receptor-binding domains from SARS-CoV-2 and 11 other sarbecoviruses. For these receptor-binding domains we determine thermal resilience (for mosaic vaccine development) and cross-recognition by antibodies. Antibody EY6A reacts across all tested sarbecoviruses, towards potential application against new coronavirus pandemic threats.

  View details for DOI 10.1038/s41467-022-31193-8

  View details for PubMedID 35764623

  View details for PubMedCentralID PMC9240080

• Metabolism of a hybrid algal galactan by members of the human gut microbiome. Nature chemical biology Robb, C. S., Hobbs, J. K., Pluvinage, B., Reintjes, G., Klassen, L., Monteith, S., Giljan, G., Amundsen, C., Vickers, C., Hettle, A. G., Hills, R., , Xing, X., Montina, T., Zandberg, W. F., Abbott, D. W., Boraston, A. B. 2022; 18 (5): 501-510

  Abstract

  Native porphyran is a hybrid of porphryan and agarose. As a common element of edible seaweed, this algal galactan is a frequent component of the human diet. Bacterial members of the human gut microbiota have acquired polysaccharide utilization loci (PULs) that enable the metabolism of porphyran or agarose. However, the molecular mechanisms that underlie the deconstruction and use of native porphyran remains incompletely defined. Here, we have studied two human gut bacteria, porphyranolytic Bacteroides plebeius and agarolytic Bacteroides uniformis, that target native porphyran. This reveals an exo-based cycle of porphyran depolymerization that incorporates a keystone sulfatase. In both PULs this cycle also works together with a PUL-encoded agarose depolymerizing machinery to synergistically reduce native porphyran to monosaccharides. This provides a framework for understanding the deconstruction of a hybrid algal galactan, and insight into the competitive and/or syntrophic relationship of gut microbiota members that target rare nutrients.

  View details for DOI 10.1038/s41589-022-00983-y

  View details for PubMedID 35289327

  View details for PubMedCentralID 5388186

• Virus-like particles against infectious disease and cancer: guidance for the nano-architect. Current opinion in biotechnology Hills, R. A., Howarth, M. 2022; 73: 346-354

  Abstract

  Virus-like particles (VLPs) can play important roles in prevention and therapy for infectious diseases and cancer. Here we describe recent advances in rational construction of VLP assemblies, as well as new approaches to enhance long-lasting antibody and CD8+ T cell responses. DNA origami and computational protein design identified optimal spacing of antigens. Chemical biology advances enabled simple and irreversible VLP decoration with protein or polysaccharide antigens. Mosaic VLPs co-displayed antigens to generate cross-reactive antibodies against different influenza strains and coronaviruses. The mode of action of adjuvants inside VLPs was established through knock-outs and repackaging of innate immune stimuli. VLPs themselves showed their power as adjuvants in cancer models. Finally, landmark clinical results were obtained against malaria and the SARS-CoV-2 pandemic.

  View details for DOI 10.1016/j.copbio.2021.09.012

  View details for PubMedID 34735984

  View details for PubMedCentralID PMC8555979

• Implementing an integrated multi-technology platform for drug checking: Social, scientific, and technological considerations. Drug testing and analysis Wallace, B., Hills, R., Rothwell, J., Kumar, D., Garber, I., van Roode, T., Larnder, A., Pagan, F., Aasen, J., Weatherston, J., Gozdzialski, L., Ramsay, M., Burek, P., Azam, M. S., Pauly, B., Storey, M. A., Hore, D. 2021; 13 (4): 734-746

  Abstract

  The illicit drug overdose crisis in North America continues to devastate communities with fentanyl detected in the majority of illicit drug overdose deaths. The COVID-19 pandemic has heightened concerns of even greater unpredictability in the drug supplies and unprecedented rates of overdoses. Portable drug-checking technologies are increasingly being integrated within overdose prevention strategies. These emerging responses are raising new questions about which technologies to pursue and what service models can respond to the current risks and contexts. In what has been referred to as the epicenter of the overdose crisis in Canada, a multi-technology platform for drug checking is being piloted in community settings using a suite of chemical analytical methods to provide real-time harm reduction. These include infrared absorption, Raman scattering, gas chromatography with mass spectrometry, and antibody-based test strips. In this Perspective, we illustrate some advantages and challenges of using multiple techniques for the analysis of the same sample, and provide an example of a data analysis and visualization platform that can unify the presentation of the results and enable deeper analysis of the results. We also highlight the implementation of a various service models that co-exist in a research setting, with particular emphasis on the way that drug checking technicians and harm reduction workers interact with service users. Finally, we provide a description of the challenges associated with data interpretation and the communication of results to a diverse audience.

  View details for DOI 10.1002/dta.3022

  View details for PubMedID 33646611