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MRC Prion Unit
From fundamental research to prevention and cure
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Molecular diagnostic strategies in prion disease

The majority of the UK population and a lesser proportion of continental European and the rest of the world were exposed to foodstuffs contaminated with the agent that caused mad cow disease or BSE. This resulted in the new human disease, variant CJD (vCJD) which has fortunately been limited to a small number of cases, the last of which was in 2016. Although the number of patients has remained small, the number of people infected but without any signs of illness could exceed 30,000 in the UK alone. Infected individuals may take over 50 years to develop disease or may never show any signs of illness. However, they themselves may be infectious and there is a risk of spreading the disease through the contamination of medical and dental instruments; the use of contaminated blood for transfusion and the transplantation of infected organs and tissues. All of these dangers pose threats to public health and the protection measures currently in place to limit the spread of prion disease cost 10’s of millions of pounds per annum in the UK alone.

Despite the decline in the number of cases of vCJD the incidence of the classical form of sporadic CJD (sCJD) has been rising consistently for several years (Figure 1). It is likely that improved diagnosis and increased awareness of prion disease following the BSE epidemic can account for much of this, with a further contribution from the UK’s enlarged population and older average age. However, such influences should reach a plateau and the continued rise in the number of cases raises the question of whether BSE-related prion disease is hidden within these statistics.

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GJfig1

Figure 1. The number of people dying as a result of sporadic CJD in the UK per year. (Source : the National CJD Research and Surveillance Unit website [https://www.cjd.ed.ac.uk])

Human PrPc
Human PrPc

Figure 2. The tertiary structure of the human prion protein which contains three alpha-helices and two anti-parallel beta-sheets

Prion disease is characterised by a change that occurs to a normal protein, PrPC, detection of the abnormal protein can be used to show someone has CJD. Unfortunately the amounts of abnormal prion protein are very low in easily obtained samples such as blood and urine, and are difficult to tell apart from the large amounts of the normal protein that are always present in both healthy and infected people. Critical to the utility and benefit of a test for CJD is the specificity of the assay, as any significant number of false results would outweigh the benefits of detecting real cases.

We are particularly interested in the possibility of determining when and if individuals at risk of prion disease may develop disease symptoms. To achieve this we are trying different ways of detecting the rare altered form of the prion protein which accumulates as structured deposits in affected tissues. One approach is to copy the way the prion reproduces itself in the human body during disease. This allows us to ‘amplify’ the very small amounts of prion in patient samples into large amounts that can be easily detected using conventional laboratory tests. In doing so we are also gaining insights into the fundamental processes of prion replication that distinguish this particular protein misfolding event from those involved in other diseases that produce very similar aggregated proteins that do not infect others. Intriguingly, we can induce the replication of prions in a test-tube from proteins made in bacteria which may allow us to develop more automated assays for diagnostic testing.

In addition to monitoring the conversion of normal prion proteins into prions during the course of disease we are identifying other, less specific, marker proteins that indicate the level of active damage occurring in the brain. These markers may vary with the worsening of symptoms during the progression of disease (Figure 3) and conversely with improvement that could occur following the administration of potential new treatments.

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Figure 3. The relationship between the concentration of the serum protein tau to the rate of disease progression in CJD. (Log-tau positively correlates with the rate of disease progression (R2=0.552, p<0.001)).

Our research requires interaction with other Unit programmes and we work closely with Dr Jonathan Wadsworth who is leading research to purify the infectious agent and by combining methods for purification with the ability to replicate prions in a test tube we are aiming to define the molecular mechanisms of prion replication and we are attempting to apply all of our research to generate synthetic prions and determine their molecular organisation and atomic structure.



Peer reviewed articles:
2018
Neurofilament light chain and tau concentrations are markedly increased in the serum of patients with sporadic Creutzfeldt-Jakob disease, and tau correlates with rate of disease progression. 
Thompson AGB, Luk C, Heslegrave AJ, Zetterberg H, Mead SH, Collinge J, Jackson GS J Neurol Neurosurg Psychiatry. 2018 Feb 27.

2016

Diagnosing Sporadic Creutzfeldt-Jakob Disease by the Detection of Abnormal Prion Protein in Patient Urine. Luk C, Jones S, Thomas C, Fox NC, Mok TH, Mead S, Collinge J, Jackson GS. JAMA Neurol. 2016 Oct 3. doi: 10.1001/jamaneurol.2016.3733. PubMed PMID: 27699415.

Physical, chemical and kinetic factors affecting prion infectivity.  Properzi F, Badhan A, Klier S, Schmidt C, Klöhn PC, Wadsworth JDF, Clarke AR, Jackson GS, Collinge J.  Prion 2016; doi: 10.1080/19336896.2016.1181250

2015

Preclinical detection of infectivity and disease-specific PrP in blood throughout the incubation period of prion disease.
Sawyer EB, Edgeworth JA, Thomas C, Collinge J, Jackson GS. Sci Rep. 2015 Dec 3;5:17742. doi: 10.1038/srep17742.

A systematic investigation of production of synthetic prions from recombinant prion protein.
Schmidt C, Fizet J, Properzi F, Batchelor M, Sandberg MK, Edgeworth JA, Afran L, Ho S, Badhan A, Klier S, Linehan JM, Brandner S, Hosszu LL, Tattum MH, Jat P, Clarke AR, Klöhn PC, Wadsworth JD, Jackson GS, Collinge J. Open Biol. 2015 Dec; 5(12). pii: 150165. doi: 10.1098/rsob.150165.

2014 

Blood test for variant Creutzfeldt-Jakob Disease – reply
Jackson GS, Burk-Rafel J, Mead S, Collinge J. JAMA Neurol 2014: 71(8):1054-5.

Population Screening for Variant Creutzfeldt-Jakob Disease Using a Novel Blood Test: Diagnostic Accuracy and Feasibility Study
Jackson G, Burk-Rafel J, Edgeworth J, Sicilia A, Abdilahi S, Korteweg J, Mackey J, Thomas C, Wang G, Schott J, Mummery C, Chinnery P, Mead S, Collinge J. JAMA Neurol 2014; 71: 421-8.

Variant Creutzfeldt-Jakob disease with extremely low lymphoreticular deposition of prion protein
Mead S, Wadsworth JDF, Porter MC, Linehan J, Pietkiewicz W, Jackson GS, Brandner S, Collinge J.  JAMA Neurol 2014; 71: 340-3.

A highly specific blood test for vCJD
Jackson GS, Burk-Rafel J, Edgeworth JA, Sicilia A, Abdilahi S, Korteweg J, Mackey J, Thomas C, Wang G, Mead S, Collinge J. Blood 2014; 123: 452-3.

2011

Detection of prion infection in variant Creutzfeldt-Jakob disease: a blood-based assay.
Edgeworth JA, Farmer M, Sicilia A, Tavares P, Beck J, Campbell T, Lowe J, Mead S, Rudge P, Collinge J, Jackson GS. Lancet 2011; 377: 487-93.

A Standardised comparison of commercially available prion decontamination reagents using the standard Steel Binding Assay
Edgeworth JA, Sicilia A, Linehan J, Brandner S, Jackson GS, Collinge J.  J Gen Virol 2011; 92: 718-26.

2010
Pharmacological chaperone for the structured domain of human prion protein
Nicoll A, Trevitt C, Risse E, Quaterman E, Ibarra AA, Wright C, Jackson GS, Sessions R, Farrow M, Waltho J, Clarke A, Collinge J.  Proc Natl Acad Sci USA 2010; 107; 17610-5.

The H187R mutation of the human prion protein induces conversion of recombinant prion protein to PrPSc-like form
Hosszu L, Tattum H, Jones S, Trevitt CR, Wells MA, Waltho JP, Collinge J, Jackson GS, Clarke AR.
Biochemistry 2010; 49: 8729-38.

Spontaneous generation of mammalian prions
Edgeworth J, Gros N, Alden J, Joiner S, Wadsworth JDF, Linehan J, Brandner S, Jackson GS, Weissmann C, Collinge J.  Proc Natl Acad Sci USA 2010; 107: 14402-6.

A highly sensitive immunoassay for the detection of prion-infected material in whole human blood without the use of proteinase K.
Tattum MH, Jones S, Pal S, Khalili-Shirazi A, Collinge J, Jackson GS. Transfusion 2010; 50: 2619-27.

PRNP allelic series from 19 years of prion protein gene sequencing at the MRC Prion Unit
Beck JA, Poulter M, Campbell TA, Adamson G, Uphill JB, Guerreiro R, Stephens JC, Jackson GS, Manji H, Collinge J, Mead S.  Hum Mut 2010; 31: E1551- 63.

Superoxide dismutase 1 and tgSOD1 mouse spinal cord seed fibrils, suggesting a propagative cell death mechanism in amyotrophic lateral sclerosis
Chia R, Tattum HM, Jones S, Collinge J, Fisher EMC, Jackson GS.  PlosOne 2010; 5: e10627.

The legs at odd angles (Loa) mutation in cytoplasmic dynein ameliorates mitochondrial function in SOD1G93A mouse model for motor neuron disease
El Kadi AM, Bros-Facer V, Deng W, Philpott A, Stoddart E, Banks G, Jackson GS, Fisher EM, Duchen MR, Greensmith L, Moore AL, Hafezparast M.  J Biol Chem 2010; 285: 18627-39.

Modification of superoxide dismutase 1 (SOD1) properties by a GFP tag--implications for research into amyotrophic lateral sclerosis (ALS)
Stevens JC, Chia R, Hendriks WT, Bros-Facer V, van Minnen J, Martin JE, Jackson GS, Greensmith L, Schiavo G, Fisher EM.  PLoS ONE 2010; 5: e9541.

Discrimination between prioninfected and normal blood samples by protein misfolding cyclic amplification
Tattum MH, Jones S, Pal S, Collinge J, Jackson GS. Transfusion 2010; 50: 996-1002.

2009

The R1441C mutation alters the folding properties of the ROC domain of LRRK2
Li Y, Dunn L, Greggio E, Krumm B, Jackson GS, Cookson MR, Lewis PA, Deng J.
Biochim Biophys Acta 2009; 1792: 1194-7.

Conformational properties of beta -PrP
Hosszu LL, Trevitt CR, Jones S, Batchelor M, Scott DJ, Jackson GS, Collinge J, Waltho JP, Clarke AR.  J Biol Chem 2009; 284: 21981-90.

Crystal structure of human prion protein bound to a therapeutic antibody
Antonyuk SV, Trevitt CR, Strange RW, Jackson GS, Sangar D, Batchelor M, Cooper S, Fraser C, Jones S, Georgiou T, Khalili-Shirazi A, Clarke AR, Hasnain SS, Collinge J. Proc Natl Acad Sci USA 2009; 106: 2554-8.

Highly sensitive, quantitative cell-based assay for prions adsorbed to solid surfaces
Edgeworth JA, Jackson GS, Clarke AR, Weissmann C, Collinge J.  Proc Natl Acad Sci USA 2009; 106: 3479-83.

A role of cellular prion protein in programming T-cell cytokine responses in disease
Ingram RJ, Isaacs JD, Kaur G, Lowther DE, Reynolds CJ, Boyton RJ, Collinge J, Jackson GS, Altmann DM.  FASEB J 2009; 23: 1672-84.

Folding kinetics of the human prion protein probed by temperature jump.
Hart T, Hosszu LL, Trevitt CR, Jackson GS, Waltho JP, Collinge J, Clarke AR.
Proc Natl Acad Sci USA 2009; 106: 5651-6.