My basic research interests are concerned with molecular pathology, diagnosis and therapy of conventional and non-conventional pathogens, including viruses and prions. I am currently investigating the molecular nature of the infectious prion agent associated with transmissible spongiform encephalopathies such as scrapie of sheep, BSE of cattle, CWD of cervids and CJD of humans. I am interested in the interactions of pathogens with the immune system, in particular, peripheral prion pathogenesis and prion neuroinvasion.
My research work has involved the generation of novel anti-PrP monoclonal antibodies to probe the structure of PrPC and PrPSc; the generation of recombinant PrP protein for biophysical and biochemical analysis and resultant computational modelling to predict prion protein structure. In addition, my projects involve the use of novel gene knock-out and transgenic mice, cell culture, flow cytometry, immunohistochemistry, in situ hybridisation, RT-PCR, SDS-PAGE, western blotting, protein misfolding cyclic amplification (PMCA), conformational-dependent immunoassays (CDI) and more recently confocal microscopy and RNASeq analysis. I am actively involved in the development and commercialisation of molecular reagents for prion research and a prion diagnostic test. I also have considerable experience in research into the pathogenesis and chemotherapy of herpes viruses both in vivo and in vitro, using prophylactic and therapeutic strategies.
Analysis of the biochemical pathways associated with ovine prion formation, clearance, and neurotoxicity will be helped by the application of new tractable animal models of this condition. Use of a genetically well-defined host will allow identification of genetic modifiers of ovine prion disease pathogenesis, providing the potential for development of new diagnostic markers and therapeutic targets for prion diseases in general. The fruitfly Drosophila melanogaster has been used to model a number of human neurodegenerative diseases and a major strength of Drosophila model systems is the ability to rapidly generate transgenic flies that express heterologous proteins in a tissue-specific manner. Genetic pathways are extensively conserved between mammalian and Drosophila genomes, which allows fly models of neurodegenerative disease to identify candidate genetic modifiers of the condition in the host species. At present, we are investigating Drosophila as a new animal model for a variety of prion disorders so we can generate a more tractable invertebrate model of transmissible mammalian prion disease.
Research
My basic research interests are concerned with molecular pathology, diagnosis and therapy of conventional and non-conventional pathogens, including viruses and prions. I am currently investigating the molecular nature of the infectious prion agent associated with transmissible spongiform encephalopathies such as scrapie of sheep, BSE of cattle and CJD of humans. I am interested in the interactions of pathogens with the immune system, in particular, peripheral prion pathogenesis and prion neuroinvasion. Analysis of the biochemical pathways associated with ovine prion formation, clearance, and neurotoxicity will be helped by the application of new tractable animal models of this condition. Use of a genetically well-defined host will allow identification of genetic modifiers of ovine prion disease pathogenesis, providing the potential for development of new diagnostic markers and therapeutic targets for prion diseases in general. The fruitfly Drosophila melanogaster has been used to model a number of human neurodegenerative diseases and a major strength of Drosophila model systems is the ability to rapidly generate transgenic flies that express heterologous proteins in a tissue-specific manner. Genetic pathways are extensively conserved between mammalian and Drosophila genomes, which allows fly models of neurodegenerative disease to identify candidate genetic modifiers of the condition in the host species. At present, we are investigating Drosophila as a new animal model of ovine scrapie in order to provide a more tractable invertebrate model of transmissible mammalian prion disease. My research work has involved the generation of novel anti-PrP monoclonal antibodies to probe the structure of PrPC and PrPSc; the generation of recombinant PrP protein for biophysical and biochemical analysis and resultant computational modelling to predict prion protein structure. In addition, my projects involve the use of novel gene knock-out and transgenic mice, cell culture, flow cytometry, immunohistochemistry, in situ hybridisation, RT-PCR, SDS-PAGE, western blotting, protein misfolding cyclic amplification (PMCA), conformational-dependent immunoassays (CDI) and more recently confocal microscopy. I am actively involved in the development and commercialisation of molecular reagents for prion research and a prion diagnostic test. I also have considerable experience in research into the pathogenesis and chemotherapy of herpes viruses both in vivo and in vitro, using prophylactic and therapeutic strategies.
Main collaborators
- Professor Candace Mathiason: Colorado State University, USA
- Professor Hermann Schátzl: Calgary Prion Research Unit, Canada
- Professor Matthias Landgraf: University of Cambridge, UK
- Professor Michael Klein: University of Würzburg, Germany
- Dr Olivier Andréoletti: Ecole Nationale Vétérinaire de Toulouse, Toulouse, France
- Dr Walker Jackson: Linköping University, Sweden
- Dr John Spiropoulos: APHA, Weybridge, UK
Publications
Thackray AM, Andréoletti O, Spiropoulos J & Bujdoso R. (Aug 2021). A new model for sensitive detection of zoonotic prions by PrP transgenic Drosophila. Journal of Biological Chemistry 297(2):100878. doi: 10.1016/j.jbc.2021.100878.
Thackray AM, Lam B, Shahira Binti Ab Razak A, Yeo G & Bujdoso R. (Feb 2020). Transcriptional signature of prion-induced neurotoxicity in a Drosophila model of transmissible mammalian prion disease. Biochemical Journal 477(4): 833–852. doi: 10.1042/BCJ20190872 PMCID: PMC7054746.
Huor A, Espinosa JC, Vidal E, Cassard H, Douet JY, Lugan S, Aron N, Marin-Moreno A, Lorenzo P, Aguilar-Calvo P, Badiola J, Bolea R, Pumarola M, Benestad SL, Orge L, Thackray AM, Bujdoso R, Torres JM & Andréoletti O. (Dec 2019). The emergence of classical BSE from atypical/Nor98 scrapie. Proceedings of the National Academy of Sciences USA pii: 201915737. doi: 10.1073/pnas.1915737116 Epub ahead of print. PMID: 31843908.
Sang JC, Meisl G, Thackray AM, Hong L, Ponjavic A, Knowles TPJ, Bujdoso R & Klenerman D. (2018). Direct observation of murine prion protein replication in vitro. Journal of the American Chemical Society. 140 (44): 14789 – 14798 doi: 10.1021/jacs.8b08311.
Thackray AM, Andréoletti O & Bujdoso R. (July 2018). Mammalian prion propagation in PrP transgenic Drosophila. Brain 141(9): 2700-2710. doi: 10.1093/brain/awy183. PMID: 29985975.
Sang JC, Lee JE, Dear AJ, De S, Meisl G, Thackray AM, Bujdoso R, Knowles TPJ & Klenerman D. (2019). Direct observation of prion protein oligomer formation reveals an aggregate mechanism with multiple conformationally distinct species. Chemical Science 10(17): 4588–4597. doi:10.1039/c8sc05627g. PMCID: PMC6492631.
Thackray AM, Andréoletti O & Bujdoso R. (2018). The use of PrP transgenic Drosophila to replace and reduce vertebrate hosts in the bioassay of mammalian prion infectivity. F1000Research 7:595–607. doi: 10.12688/f1000research.14753.1 eCollection 2018 PMID: 29946445.
Thackray AM, Cardova A, Wolf H, Pradl L, Vorberg I, Jackson WS & Bujdoso R. (2017). Genetic human prion disease modelled in PrP transgenic Drosophila. Biochemical Journal 474(19): 3253–3267. doi: 10.1042/BCJ20170462 PMID: 28814578.
Thackray AM, Andréoletti O & Bujdoso R. (2016). Bioassay of prion-infected blood plasma in PrP transgenic Drosophila. Biochemical Journal. 473(23):4399-4412.
Bujdoso R, Landgraf M, Jackson WS & Thackray AM. (2015). Prion-induced neurotoxicity: possible role for cell cycle activity and DNA damage response. World Journal of Virology. 4:188-197.
Thackray AM, Di Y, Zhang C, Wolf H, Pradl L, Vorberg I, Andréoletti O & Bujdoso R. (2014). Prion-induced and spontaneous formation of transmissible toxicity in PrP transgenic Drosophila. Biochemical Journal. 463: 31-40.
Yang S, Thackray AM, Hopkins L, Monie TP, Burke D & Bujdoso R. (2014). Polymorphisms at amino acid residues 141 and 154 influence conformational variation in ovine PrP. Biomed Research International. Article ID 372491, doi: 10.1155/2014/372491.
Thackray AM, Zhang C, Arndt T & Bujdoso R. (2014). Cytosolic PrP can participate in prion-mediated toxicity. Journal of Virology. 88(14): 8129-8138.
Bujdoso R & Thackray AM. (2014) The emergence of a Drosophila model to measure ovine prion infectivity. CAB Reviews: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources, 8(58): 1-17. DOI 10.1079/PAVSNNR20130058. www.cabi.org/cabreviews.
Li W, Bell NAW, Hernández-Ainsa S, Thacker VV, Thackray AM, Bujdoso R & Keyser UF. (2013). Single protein molecule detection by glass nanopores. ACS Nano. 7(5): 4129-4134.
Thackray AM, Lockey R, Beck KE, Spiropoulos J & Bujdoso R. (2012). Evidence for co-infection of ovine prion strains in classical scrapie isolates. Journal of Comparative Pathology. 147(2-3): 316-329.
Thackray AM, Muhammad F, Zhang C, Di Y, Jahn TR, Landgraf M, Crowther DC, Evers JF & Bujdoso R. (2012). Ovine PrP transgenic Drosophila show reduced locomotor activity and decreased survival. Biochemical Journal. 444(3): 487-495.
Thackray AM, Muhammad F, Zhang C, Denyer M, Spiropoulos J, Crowther DC & Bujdoso R. (2012). Prion-induced toxicity in PrP transgenic Drosophila. Experimental and Molecular Pathology. 92(2): 194-201.
Thackray AM, Hopkins L, Lockey R, Spiropoulos J & Bujdoso R. (2012). Propagation of ovine prions from poor transmitter scrapie isolates in ovine PrP transgenic mice. Experimental & Molecular Pathology. 92(1): 167-174.
Thackray AM, Hopkins L, Lockey R, Spiropoulos J & Bujdoso R. (2011). Emergence of multiple prion strains from single isolates of ovine scrapie. Journal of General Virology. 92(6): 1482-1491.
Zabel M, Greenwood C, Thackray AM, Pulford B, Rens W & Bujdoso R. (2009). Perturbation of T-cell development by insertional mutation of a PrP transgene. Immunology. 127(2): 226-236.
Thackray AM, Hopkins L, Spiropoulos J & Bujdoso R. (2008). Molecular and transmission characteristics of primary-passaged ovine scrapie isolates in conventional and ovine PrP transgenic mice. Journal of Virology. 82(22): 11197-11207.
Yang S, Thackray AM, Fitzmaurice TJ & Bujdoso R. (2008). Copper-induced structural changes in the ovine prion protein are influenced by a polymorphism at codon 112. Biochimica et Biophysica Acta. 1784(4): 683-692.
Fitzmaurice TJ, Burke DF, Hopkins L, Yang S, Yu S, Sy MS, Thackray AM & Bujdoso R. (2008). The stability and aggregation of ovine prion protein associated with classical and atypical scrapie correlates with the ease of unwinding of helix-2. Biochemical Journal. 409(2): 367-375.
Thackray AM, Hopkins L, Klein MA & Bujdoso R. (2007). Mouse-adapted ovine scrapie prion strains are characterized by different conformers of PrPSc. Journal of Virology. 81(22): 12119-12127.
Thackray AM, Hopkins L & Bujdoso R. (2007). Proteinase K-sensitive disease-associated ovine prion protein revealed by conformation-dependent immunoassay. Biochemical Journal. 401(2): 475-483.
Thackray AM, Fitzmaurice TJ, Hopkins L & Bujdoso R. (2006). Ovine plasma prion protein levels show genotypic variation detected by C-terminal epitopes not exposed in cell-surface PrPC. Biochemical Journal. 400(2): 349-358.
Thackray AM & Bujdoso R. (2006). Elevated PrPC expression predisposes to increased HSV-1 pathogenicity. Antiviral Chemistry & Chemotherapy. 17(1): 41-52.
Bujdoso R, Burke DF & Thackray AM. (2005). Structural differences between allelic variants of the ovine prion protein revealed by molecular dynamics simulations. Proteins. 61(4): 840-849.
Thackray AM, Ryder SJ & Bujdoso R. (2005). Modification of blood cell PrP epitope exposure during prion disease. Biochemical Journal. 390(2): 563-571.
Yang S, Thackray AM & Bujdoso R. (2005). Generation of ovine recombinant prion protein (25-232): Characterisation via anti-PrP monoclonal antibodies and CD spectroscopy. Korean Journal of Veterinary Service. 28(4): 393-405.
Thackray AM & Bujdoso R. (2004). A role for pro- and anti-inflammatory cytokines in the regulation of prion disease pathogenesis. Immunology. 113: (S1) 98-137 (OP159).
Thackray AM, McKenzie AN, Klein MA, Lauder A & Bujdoso R. (2004). Accelerated prion disease in the absence of Interleukin-10. Journal of Virology. 78(24): 13697-13707.
Thackray AM, Yang S, Wong E, Fitzmaurice T, Morgan-Warren R & Bujdoso R. (2004). Conformational variation between allelic variants of cell-surface ovine prion protein. Biochemical Journal. 381(1): 221-229.
Wong E, Thackray AM & Bujdoso R. (2004). Copper induces increased beta-sheet content in the scrapie-susceptible ovine prion protein PrPVRQ compared with the resistant allelic variant PrPARR. Biochemical Journal. 380(1): 273-282.
Thackray AM, Wong E, Yang S, Morgan-Warren R & Bujdoso R. (2003). Conformational mapping of cell surface ovine prion protein. Immunology. 110: (S1) 97.
Wong E, Thackray AM & Bujdoso R. (2003). Copper induction of beta-sheet conformation in the ovine prion protein. Immunology. 110: (S1) 97.
Thackray AM, Klein MA & Bujdoso R. (2003). Subclinical prion disease induced by oral inoculation. Journal of Virology. 77(14): 7991-7998.
Thackray AM, Madec J-Y, Wong E, Morgan-Warren R, Brown D, Baron T & Bujdoso R. (2003). Detection of bovine spongiform encephalopathy, ovine scrapie prion-related protein (PrPSc) and normal PrPC by monoclonal antibodies raised to copper-refolded prion protein. Biochemical Journal. 370(1): 81-90.
Shaw MM, Gürr WK, Thackray AM, Watts PA, Littler E & Field HJ. (2002). Temporal pattern of herpes simplex virus type 1 infection and cell death in the mouse brain stem: influence of guanosine nucleoside analogues. Journal of Virological Methods. 102(1-2): 93-102.
Thackray AM & Bujdoso R. (2002). PrPc expression influences the establishment of herpes simplex virus type1 latency. Journal of Virology. 76(5): 2498-2509.
Thackray AM, Klein MA, Aguzzi A & Bujdoso R. (2002). Chronic subclinical prion disease induced by low dose inoculum. Journal of Virology. 76(5): 2510-2517.
Thackray AM, Knight R, Haswell SJ, Bujdoso R & Brown DR. (2002). Metal imbalance and compromised antioxidant function are early changes in prion disease. Biochemical Journal. 362(1): 253-258.
