New mouse models for tackling motor neuron disease and other neurodegenerative disorders.

Lead Research Organisation: University College London
Department Name: Institute of Neurology

Abstract

This application is for a 4-year research project to create four new valuable mouse models in which to study neurodegeneration, with a long term view to finding therapeutics. These mouse strains complement and are different from existing models. During this award we will make the mice plus control strains, carry out initial ?clinical? analysis, and will make all the mice freely available to all other interested researchers.

Motor neuron diseases (MNDs) are relatively common (in UK 1 in 400 death certificates is issued for some form of MND) and can strike at any age ? spinal muscular atrophy is the biggest single genetic killer of children world-wide, whereas amyotrophic lateral sclerosis (ALS) is a disease of mid-life that has a lifetime risk of up to 1 in 600. These diseases progress inexorably to paralysis and death and we have no treatments that improve quality of life or lifespan of sufferers. MNDs have a genetic component but for ALS only one major effect causative gene, SOD1 has been found (in 1993) and we still have no idea why the mutant form of the protein kills neurons. Recently a protein called TDP43 was identified which aggregates abnormally in the nerve cells of people suffering from ALS and in 2008 rare families have been identified who succumb to ALS with death in mid-life, because they have mutations in the TDP43 gene. TDP43 protein abnormalities have also been found in frontotemporal dementia (the most common form of dementia after Alzheimer?s disease), and Parkinson?s disease, Pick?s disease and other forms of neurodegeneration.

Once a human mutant gene such as SOD1 or TDP43 has been identified, the next step is to genetically alter mice so that they recapitulate the human disease and we can study the pathological processes leading to illness and death. SOD1 transgenic mice have been tremendously helpful in understanding ALS, but have disadvantages. Currently no TDP43 mouse models exist. We want to create two new strains of mice, using newer technologies, which more closely model ALS in humans. These mice will be better for understanding human disease, for finding early ?biomarkers? of disease, and for trialling new therapies because they more closely replicate what is happening in humans. We will make the mice freely available to all researchers, without ties, by distributing them in the major mutant mouse repositories in Europe and USA for sending out to all research labs.

Technical Summary

Amyotrophic lateral sclerosis (ALS) is mostly (~80%) sporadic but familial forms (FALS, ~20%) are known. The SOD1 gene is the most common causative gene for FALS, and recently rare mutations have been identified in the TDP43 gene. This is of interest because TDP43 is the major ubiquitinated protein in inclusions in both sporadic and familial ALS ? although not SOD1 related ALS.

We are requesting funding for one postdoc and one technician (minimum staffing to make this application realistic) to create two novel mouse strains modelling amyotrophic lateral sclerosis (ALS) with the SOD1 human genomic coding region knocked in, and two novel strains with a human TDP43 cDNA knocked in. These mouse strains are essential for understanding ALS because: if the knock in mice have a mid-life disease, this would arise from mutation, not from overexpression of the transgene array, which is known to have an effect in existing SOD1 transgenics; the model is physiologically more relevant than existing transgenics, giving us a better understanding of pathology and a better model for conventional and gene therapeutics ? including siRNA and lentiviruses; this model also gives us a better knowledge of the protein interactions that occur in ALS because it is biochemically more relevant to the human disease; this model helps address the biophysics/biochemistry of mutant:wildtype dimers and enzyme function, in a variety of primary cell lines, including embryonic stem cells and motor neurons; a mouse with late onset disease will direct us to biomarkers, desperately needed by ALS clinicians. Critically, we will make Cre-conditional knock in mice in which the mutation can be turned off in any tissues ? thus informing us, for example, about motor neuron ? glia interactions as we already know SOD1 induced motor neuron death is not cell autonomous and therefore glia and other cells may be targets for therapy. We wish to make knock in mice for both mutant SOD1 and TDP43, because although mutations in these genes may feed into a final common pathway, the lack of TDP43 pathology in SOD1 mutant ALS clearly shows the genes have different initial effects.

A critical feature of this award is that we will make the mice freely available immediately, via the European Mutant Mouse Archive and the Jackson Laboratory, for distribution to all interested laboratories.

Publications

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Bunton-Stasyshyn RK (2015) SOD1 Function and Its Implications for Amyotrophic Lateral Sclerosis Pathology: New and Renascent Themes. in The Neuroscientist : a review journal bringing neurobiology, neurology and psychiatry

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Devoy A (2011) Genomically humanized mice: technologies and promises. in Nature reviews. Genetics

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Fratta P (2014) Profilin1 E117G is a moderate risk factor for amyotrophic lateral sclerosis. in Journal of neurology, neurosurgery, and psychiatry

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Joyce PI (2011) SOD1 and TDP-43 animal models of amyotrophic lateral sclerosis: recent advances in understanding disease toward the development of clinical treatments. in Mammalian genome : official journal of the International Mammalian Genome Society

 
Description Acad Med Sci humanisation of animals - giving evidence
Geographic Reach National 
Policy Influence Type Citation in other policy documents
 
Description ALSA project grant
Amount $160,000 (USD)
Funding ID Fisher 15-IIP-198 
Organisation ALS Association 
Sector Charity/Non Profit
Country United States
Start 08/2014 
End 07/2016
 
Description MRC Research Grant (Fus humanised)
Amount £994,155 (GBP)
Funding ID MR/L021056/1 
Organisation Medical Research Council (MRC) 
Sector Public
Country United Kingdom
Start 08/2014 
End 07/2017
 
Description Motor Neurone Disease Association PhD studentship (Fus)
Amount £112,690 (GBP)
Organisation Motor Neurone Disease Association (MND) 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2014 
End 09/2017
 
Description PhD studentship MNDA Humanizing the Tardbp (TDP43) locus in the mouse
Amount £88,690 (GBP)
Funding ID Fisher (10/442) 
Organisation Motor Neurone Disease Association (MND) 
Sector Charity/Non Profit
Country United Kingdom
Start 12/2015 
End 11/2018
 
Description Rosetrees award
Amount £9,601 (GBP)
Organisation Rosetrees Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 03/2015 
End 02/2017
 
Title A new antibody - mouse system for analysing FUS ALS, the Delta14 FUS mouse 
Description Novel antibody that detects mutant FUS only, including in our genome engineered mouse. 
Type Of Material Antibody 
Year Produced 2016 
Provided To Others? Yes  
Impact Mice/antibody used currently by collaborators e.g. Fratta/Ule/Schiavo. Will make freely available after our first publication. 
 
Title Mouse humanised wildtype FUS model 
Description Humanised genomic wildtype FUS gene in mouse 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Year Produced 2015 
Provided To Others? Yes  
Impact Pending 
 
Title Mouse model Sod1 D83G 
Description Mouse with endogenous mutation in ALS gene (Sod1 D83G) 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Year Produced 2011 
Provided To Others? Yes  
Impact Paper submitted currently, after which the mouse will be made freely available. Second paper being written. We anticipate this mouse will be of interest to the ALS research community. 
 
Title mouse cells with humanised FUS wt and mutant 
Description Engineered mouse embryonic stem cell with knocked in human FUS gene - wildtype and mutant 
Type Of Material Cell line 
Provided To Others? No  
Impact Research on these cell lines only just beginning. 
 
Title mouse embryonic stem cells for Down syndrome 
Description Manipulated mouse embryonic stem cells to model aspects of Down syndrome 
Type Of Material Cell line 
Year Produced 2006 
Provided To Others? Yes  
Impact academic papers 
 
Title mouse models for fus mnd delta 14 only 
Description Mouse models of motor neuron degeneration, with a specific mutation in the gene FUS (delta 14) 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Year Produced 2014 
Provided To Others? Yes  
Impact Papers pending, and model is basis of two new grant applications (both successful) 
 
Title Engineered mouse FUS humanised ES cells. 
Description Mouse cell line that we may analyse rather than working with whole animals, thus helps with aims of NC3Rs. 
Type Of Material Data analysis technique 
Provided To Others? No  
Impact None yet. 
 
Description Abraham Acevedo Arozena 
Organisation Hospital Universitario Insular de Gran Canaria
Department La Fundación Canaria Instituto de Investigación Sanitaria de Canarias
Country Spain 
Sector Public 
PI Contribution PhD student time and effort to develop a new mouse model
Collaborator Contribution PhD supervision, DNA analysis, breeding and phenotypic analysis of a cohort of mice.
Impact Posters at meetings
Start Year 2016
 
Description Analysing the metabolome Griffin, Cambridge 
Organisation University of Cambridge
Department Department of Earth Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution Access to novel mice and mouse crosses
Collaborator Contribution Analysis of the metabolome including lipidomics
Impact No outputs yet
Start Year 2016
 
Description Analysis of the FUS mouse translatome, Fratta, UCL 
Organisation University College London
Department Marie Curie Palliative Care Research Department
Country United Kingdom 
Sector Academic/University 
PI Contribution contribution of the unique FUS Delta14 mouse model
Collaborator Contribution RiboTagging and ChatCre breeding to pull down polysomes from the Delta14 mouse
Impact Multidisiplinary output. No outcomes yet as just started.
Start Year 2016
 
Description Looking at gliosis in neurodegeneration 
Organisation University of Queensland
Country Australia 
Sector Academic/University 
PI Contribution Access to unique mouse models and crosses
Collaborator Contribution Analysis of gliosis and potentially the inflammasome
Impact No outputs yet
Start Year 2017
 
Description MMON 
Organisation MRC Harwell
Country United Kingdom 
Sector Academic/University 
PI Contribution Collaboration with the Mouse Models of Neurodegeneration lab at MRC Harwell, analysis of homozygous and heterozygous mice
Collaborator Contribution Breeding, inbreeding onto another background, and phenotypic analysis of homozygous and heterozygous mice.
Impact Inbred mice on different backgrounds. Cohorts of mice of different ages, sex-matched with littermate controls, wildtype, heterozygous, homozygous, for phenotypic analysis. Analysis of different phenotypes ranging from behavioural through to physiological.
Start Year 2017
 
Description NIMR and mouse engineering 
Organisation Medical Research Council (MRC)
Department MRC National Institute for Medical Research (NIMR)
Country United Kingdom 
Sector Academic/University 
PI Contribution a postdoc, grant funding, expertise.
Collaborator Contribution expertise in manipulating ES cells.
Impact grant and papers
 
Description studying ribosomal proteins 
Organisation University of Padova
Department Department of Neurosciences
Country Italy 
Sector Hospitals 
PI Contribution Access to a unique mouse model of FUS ALS (Delta14)
Collaborator Contribution Analysis of ribosomal proteins
Impact No outputs yet
Start Year 2017
 
Description Genetic engineering workshop on humanisation 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Type Of Presentation Workshop Facilitator
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact High level workshop for 30 international researchers to discuss issues on humanisation of the mouse genome.

Collaborations and networking of expertise
Year(s) Of Engagement Activity 2012
 
Description Interviewed on BBC Radio 4 for 'The Life Scientific' 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Interviewed by Professor Jim Al-Khalili for a BBC Radio 4 broadcast on the 'Life Scientific'.
Year(s) Of Engagement Activity 2019
 
Description Open day for patients 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? Yes
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Regional
Primary Audience Participants in your research and patient groups
Results and Impact Talking directly to patients and carers about animal research and how we work with animals to understand motor neuron disease.

Telling patients about the current state of our work.
Year(s) Of Engagement Activity 2012
 
Description Private meeting with major funders 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Supporters
Results and Impact Private meeting to inform major funders of research in motor neuron disease supported by the charity in question.

Maintaining funders enthusiasm and interest and showing value for money.
Year(s) Of Engagement Activity Pre-2006,2012
 
Description Visit by fundraisers from MNDA 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Fundraisers from MNDA visited to learn more of current research. These people are well-informed on issues regarding MND and care for people with MND, but are not scientists and do not necessarily have easy access to researchers.

Better informed fundraisers.
Year(s) Of Engagement Activity 2014