Normal cellular function of PrP: study of PrP null mice and conditional gene expression studies

Lead Research Organisation: MRC Prion Unit

Abstract

We have established that the normal form of the prion protein is an effective target for treatment in prion diseases using mouse models. We first found that we could cure mice infected with prions by removing the normal form of PrP using a genetically engineered gene switch. We found that the early holes that appear in prion diseased brains disappeared and that the mice survived without symptoms long term. We also found that early memory and motivational problems recovered along with electrical malfunction of the brain. We are using this now to develop a treatment to target PrP as a strategy.||In our group we are using new technologies to deliver this gene treatment to the brain and working to understand the mechanisms underlying the early disease and recovery.

Technical Summary

There is widespread interest in possible therapies for CJD and related diseases. It has been assumed that PrPSc, the disease associated rogue form of one of the bodys normal proteins, prion protein, PrPc is the main neurotoxic molecule in prion disease. However, increasing evidence points to the fact that PrPSc is not directly toxic, but that as yet unidentified transient intermediate forms generated during prion replication and conversion from PrPc to PrPSc are likely to be involved in neuronal dysfunction and death. TO this end we targeted the native form of PrP, PrPc in order to remove the precursor for generation of whatever toxic species is killing cells. We first generated transgenic mice in which genetic deletion of endogenous PrPc resulted in animals being able not only to survive long term after prion infection, but they recovered early prion pathology as well as associated learning, behavioural and neurophysiological defects. We have now used a direct therapeutic approach based on our strategy of removing the native PrPc. We have used lentiviral vectors expressing short hairpin RNAs for RNAinterference directed against PrP. We have stereotactically injected these into the hippocampus of prion infected mice and have shown that this approach protects against neuronal loss, and prion replication in the targeted areas, and leads to a highly significant increased survival of 17% in treated animals after even only a single focal injection. We are now optimising the spatial and temporal requirements for PrP knockdown as an effective therapeutic strategy. We are using an empty-lentivirus for mock treatment and also comparing our results to prion infected mice without viral treatment at all.||We are currently evaluating effects of lentiviral treatment on behavioural outcomes including novel object recognition and burrowing behaviours. Early results that RNAi protects against the loss of burrowing behaviour typical of early prion disease.||We are also interested in the mechanisms of neuroprotection resulting from PrP knockout and are looking both at differential gene expression from microarray data and developing an ex-vivo system of organotypic slice culture to allow a systematic functional analysis of the effects of prion infection and PrP knockdown.

Publications

10 25 50
 
Description MRC Neuroscience strategy board
Geographic Reach National 
Policy Influence Type Participation in advisory committee
 
Description Member of MRC Neuroscience and Mental Health Board
Geographic Reach National 
Policy Influence Type Membership of a guideline committee
 
Description RSM clinical neuroscience council
Geographic Reach Local/Municipal/Regional 
Policy Influence Type Membership of a guidance committee
 
Description leicester neuroscience steering committee
Geographic Reach Local/Municipal/Regional 
Policy Influence Type Membership of a guidance committee
 
Description NEUROPRION Use of RNAi for treatment of inherited prion disease (14.4m euros across 41 participants)
Amount £90,000 (GBP)
Funding ID 506579 
Organisation European Commission 
Sector Public
Country European Union (EU)
Start 09/2007 
End 08/2009
 
Title NFH-Cre/MloxP mice 
Description cre deleter mice in neurons floxed prp mice crossed led to adult onset knockout of prp in neurons 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Year Produced 2006 
Provided To Others? Yes  
Impact reversal prion disease widely used mouse model 
 
Description ISRIB-like substances 
Organisation AstraZeneca
Department MedImmune
Country United Kingdom 
Sector Private 
PI Contribution we will test neuroprotective effects of new compounds in mice
Collaborator Contribution generation of compounds in vitro testing
Impact none yet
Start Year 2016
 
Description ISRIB-like substances 
Organisation University of Cambridge
Department Department of Chemistry
Country United Kingdom 
Sector Academic/University 
PI Contribution we will test neuroprotective effects of new compounds in mice
Collaborator Contribution generation of compounds in vitro testing
Impact none yet
Start Year 2016
 
Description pharmacokinetic studies on PERK inhibitors 
Organisation GlaxoSmithKline (GSK)
Department GSK Oncology
Country United Kingdom 
Sector Private 
PI Contribution mechanism of neurodegeneration in prion disease and use of compounds to modulate disease progression
Collaborator Contribution PK and PD analyses GSK supplied compounds LC/MS-MS Compound production
Impact Oral treatment targeting the unfolded protein response prevents prion neurodegeneration and clinical disease in mice. Moreno JA, Halliday M, Molloy C, Axten JM, Willis AE, Ortori CA, Barrett DA, Fischer PM and Mallucci GR. Science Translational Medicine, 2013, 5,206ra138. Sustained translational repression by eIF2a-P mediates prion neurodegeneration. Moreno JA, Radford H, Peretti D, Steinert JR, Verity N, Guerra Martin M, Halliday M, Morgan J, Dinsdale D, Ortori CA, Barrett DA, Tsaytler P, Bertolotti A, Willis AE, M Bushell and Mallucci GR*. Nature 2012, 485, 507-11
Start Year 2012
 
Description pharmacokinetic studies on PERK inhibitors 
Organisation University of Cambridge
Department Cambridge Institute for Medical Research (CIMR)
Country United Kingdom 
Sector Academic/University 
PI Contribution mechanism of neurodegeneration in prion disease and use of compounds to modulate disease progression
Collaborator Contribution PK and PD analyses GSK supplied compounds LC/MS-MS Compound production
Impact Oral treatment targeting the unfolded protein response prevents prion neurodegeneration and clinical disease in mice. Moreno JA, Halliday M, Molloy C, Axten JM, Willis AE, Ortori CA, Barrett DA, Fischer PM and Mallucci GR. Science Translational Medicine, 2013, 5,206ra138. Sustained translational repression by eIF2a-P mediates prion neurodegeneration. Moreno JA, Radford H, Peretti D, Steinert JR, Verity N, Guerra Martin M, Halliday M, Morgan J, Dinsdale D, Ortori CA, Barrett DA, Tsaytler P, Bertolotti A, Willis AE, M Bushell and Mallucci GR*. Nature 2012, 485, 507-11
Start Year 2012
 
Description pharmacokinetic studies on PERK inhibitors 
Organisation University of Nottingham
Department Medicinal and Biological Chemistry
Country United Kingdom 
Sector Academic/University 
PI Contribution mechanism of neurodegeneration in prion disease and use of compounds to modulate disease progression
Collaborator Contribution PK and PD analyses GSK supplied compounds LC/MS-MS Compound production
Impact Oral treatment targeting the unfolded protein response prevents prion neurodegeneration and clinical disease in mice. Moreno JA, Halliday M, Molloy C, Axten JM, Willis AE, Ortori CA, Barrett DA, Fischer PM and Mallucci GR. Science Translational Medicine, 2013, 5,206ra138. Sustained translational repression by eIF2a-P mediates prion neurodegeneration. Moreno JA, Radford H, Peretti D, Steinert JR, Verity N, Guerra Martin M, Halliday M, Morgan J, Dinsdale D, Ortori CA, Barrett DA, Tsaytler P, Bertolotti A, Willis AE, M Bushell and Mallucci GR*. Nature 2012, 485, 507-11
Start Year 2012
 
Description pharmacokinetic studies on PERK inhibitors 
Organisation University of Nottingham
Country United Kingdom 
Sector Academic/University 
PI Contribution mechanism of neurodegeneration in prion disease and use of compounds to modulate disease progression
Collaborator Contribution PK and PD analyses GSK supplied compounds LC/MS-MS Compound production
Impact Oral treatment targeting the unfolded protein response prevents prion neurodegeneration and clinical disease in mice. Moreno JA, Halliday M, Molloy C, Axten JM, Willis AE, Ortori CA, Barrett DA, Fischer PM and Mallucci GR. Science Translational Medicine, 2013, 5,206ra138. Sustained translational repression by eIF2a-P mediates prion neurodegeneration. Moreno JA, Radford H, Peretti D, Steinert JR, Verity N, Guerra Martin M, Halliday M, Morgan J, Dinsdale D, Ortori CA, Barrett DA, Tsaytler P, Bertolotti A, Willis AE, M Bushell and Mallucci GR*. Nature 2012, 485, 507-11
Start Year 2012
 
Description Anne Silk Lecturer, Royal Society of Medicine, London, UK 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact public lecture on progress in treatment of neurodegeneration
Year(s) Of Engagement Activity 2014
URL https://www.rsm.ac.uk/events/events-listing/2013-2014/groups/public-engagement-programme/pee07-anne-...
 
Description BBC radio interview 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact naked scientist radio interview about hibernation and Alzheimer's
Year(s) Of Engagement Activity 2016
URL http://www.thenakedscientists.com/HTML/podcasts/naked-scientists/show/20160119/
 
Description Frank May Medal for Excellence in Research and Outstanding Future Promise; University of Leicester, UK 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Other audiences
Results and Impact public lecture discussing medical advances
Year(s) Of Engagement Activity 2012
 
Description MRC PEN event 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact public panel debate between writers and scientists " barriers to creativity"
Royal geographical society
public debate and questions

MRC website
PEN website
Year(s) Of Engagement Activity 2009
 
Description Media interviews for STM paper 2013 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact MRC scientists halt neurodegeneration in mice (BBC 4 Today programme, 07.12) also interviews on BBC, ITV, Channel 5, Sky, CNN, ABC, Al jazeera, local radio-stations
The Independent (front page) with additional online commentary
The Daily Express (front page)
Daily Mirror (front page)
The Sun (page 4) Online version unavailable
Daily Mail (page 5)
The Guardian (page 6)
The Daily Record (page 5)
The Scotsman p13
The Herald p5 Online version unavailable
BBC online
ITV
Sky News
Channel 4
The Daily Telegraph (page 14)

The story was also reported without an MRC mention in The Times (front page, online version unavailable)

Plus...innumerable regional newspapers, radio and TV outlets - particularly in the Leicester area!! - and international outlets from CNN to ABC to The Times of India, Time magazine, Spanish, Italian, Brazilian, Polish, Turkish, French and German outlets.
A pill for neurodegenerative disease? Not yet, but we're getting closer (BBC News Online)
The Guardian
SKY News
BBC World Service
ITV
BBC World Business Report
Radio 5
BBC Breakfast

The Daily Mirror (page 8) highlighted the research and asked what other diseases can we hope to defeat. Online version unavailable
The Sunday Times, News Review page 2, Vivienne Parry
(BBC News Online) The Guardian
SKY News
BBC World Service
ITV
BBC World Business Report
Radio 5

The Daily Mirror (page 8) highlighted the research and asked what other diseases can we hope to defeat. Online version unavailable
The Sunday Times, News Review page 2, Vivienne Parry

Huge interest into the Unit's research
Year(s) Of Engagement Activity 2013,2014
 
Description Nature podcast 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact nature podcast about discovery of hibernation and synaptic regeneration and role in neuroprotection
Year(s) Of Engagement Activity 2015
URL http://www.nature.com/nature/podcast/index-2015-01-15.html
 
Description Science Festival in Genova 2012 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact public interest high

public interest
Year(s) Of Engagement Activity 2012
 
Description Television interviews 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact discussed our research which was hailed as "turning point in search for medicines to control and prevent Alzheimer's"
Year(s) Of Engagement Activity 2013
URL http://www.bbc.co.uk/news/health-24462699.
 
Description UK Conference of Science Journalists 2012 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? Yes
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact dialogue with science writers

interest in science reporting
Year(s) Of Engagement Activity 2012
 
Description Unit Open Day 2013 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? Yes
Type Of Presentation Workshop Facilitator
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Over 500 members of the public visited the Unit

Members of the public were very appreciative.
Students asked to come and carry out work experience in the Unit.
Year(s) Of Engagement Activity 2013
URL http://tox.mrc.ac.uk/
 
Description biology in action 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact Biology in Action is a very popular annual day, which sets out to give students a stimulating day of biology in five wide-ranging sessions, and helps them and their teachers keep up to date with developments and issues. We aim to provide students with inspiration to pursue their studies at a higher level through talks that go beyond the curriculum.
Year(s) Of Engagement Activity 2016
 
Description media interviews 2007 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact numerous interviews to BBC radio, US radio
science press
national press

widespread reporting in science and mass media
radio 4 interview 'science now'
Year(s) Of Engagement Activity 2007
 
Description mrc annual report 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Participants in your research and patient groups
Results and Impact interviewed as one of MRC's 'people behind discovery'

positive feedback, applicants to work with me
Year(s) Of Engagement Activity 2007
 
Description nottingham inspire 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Professional Practitioners
Results and Impact 400 medical students
careers in science and medicine
Year(s) Of Engagement Activity 2014
 
Description nottingham inspire 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Professional Practitioners
Results and Impact 400 medical students
careers in science and medicine
Year(s) Of Engagement Activity 2014
 
Description press interviews 2008 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact press interviews broadsheets science journals BBC Radio 4 science now

widespread dissemination of research
Year(s) Of Engagement Activity 2008
 
Description schools careers in science talks 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Local
Primary Audience Schools
Results and Impact pupils interested in science
work experience in my lab

work experience and science interests
Year(s) Of Engagement Activity 2010,2011,2012
 
Description toxicology open day 2010 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact presentation to graduate students

top performer voted by students!
Year(s) Of Engagement Activity 2010
 
Description work experience 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? Yes
Geographic Reach Local
Primary Audience Schools
Results and Impact work student for 1 week in my lab
work experience undergraduate for 10wks

positive experience further applicants
Year(s) Of Engagement Activity 2010,2012,2013,2014
 
Description work highlighted British Neuroscience Bulletin 2012 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact highlight of groups reseracj

generated much interest adn phd applications
Year(s) Of Engagement Activity 2012