New humanised mouse models for dissecting the pathobiology of disease, using FUS-ALS as a paradigm

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


Humans and mice are descended from a common ancestor and so we share genes and have similar diseases caused by similar mutations in our equivalent genes (known as orthologs). Therefore to understand more about human disease we work with 'mouse models' that have the same mutations and manifest similar symptoms.

Many neurodegenerative diseases run in families as 'dominant' diseases, so if your parent has the disorder there is a 50% chance you will inherit the mutant gene, and also succumb to the disease. These dominant disorders are modelled by mice called transgenics, which carry human mutant genes and so get the disease in a similar way to humans.

Transgenic animals are made by incorporating the human mutant 'transgene' into the mouse DNA. Unfortunately when this happens the human mutant gene is usually incorporated in lots of copies, somewhere fairly random, not just a single copy as it would be normally in humans. One result is the mouse produces considerably more than the usual 'physiological' amount of mutant protein, and more often than not the mouse develops symptoms that simply come from having too much protein, rather than the actual effects of the mutation. This means the mouse is a far less accurate model than it could be.

Also, sometimes the human and mouse biochemistry of the mutant protein can be different, so simply making the same change in the mouse gene does not produce an accurate disease model.

We have developed a new type of mouse model in which we replace the mouse gene with the complete human gene, both the normal human gene and the mutated human gene in different mouse strains. Therefore we can look at what happens to the mouse when the human genetic material is expressed at normal levels, and this will give us new insight into disease processes. Our new mouse models may be a Refinement of existing models.

As an example, we have chosen to work with a gene called FUS, which is mutated in an untreatable neurodegenerative disease called amyotrophic lateral sclerosis (ALS, also known as motor neuron disease). ALS is an incurable and relentless devastating neurodegenerative disorder which causes progressive loss of muscle function and paralysis. ALS leads to death, usually caused by the inability to breathe, on average only 3 years after diagnosis, with a lifetime risk of ~1 in 300 by 85 years old, in UK. ALS is generally a mid-life disease affecting people in their 40s and 50s although there is a wide age-range of symptom onset and aggressive FUS-caused ALS has been described in children as young as 11 years old. FUS is also occasionally mutated in other incurable neurological diseases.

Now we are applying for a three year grant to:

(1) extend our method to make a slightly different type of mouse called a conditional mutant, which allows us to turn the mutant protein on in different tissues at different times. This will help us work out what goes wrong in ALS and other disorders. And,
(2) to fully characterise our new FUS mouse mutants, so we can learn more about the way that mutant FUS causes nerve cells to die. If we know this, we are step further along to therapies for ALS and other neurodegenerative diseases.

Our technology can be used for any gene, not just FUS and for example, we are well along with using this method for SOD1, which is another 'ALS gene'. So we have developed a new technology that we believe will be widely used by other laboratories, and we want to characterise the mice we have already created to understand more of how neurodegeneration happens, so that we can better understand human disease and how to treat it.

Technical Summary

Many genes, including the well-known genes FUS, TDP43, SOD1 that are causative for the neurodegenerative disease amyotrophic lateral sclerosis (ALS) are dosage-sensitive.

ALS is an incurable and essentially untreatable disease that leads to paralysis and death through the progressive loss of motor neurons. ALS is generally a mid-life disorder but patients as young as 11 years of age (with FUS mutations) have been described. Most familial forms of ALS arise from dominant mutations and are modelled in mice by overexpression of the human mutant transgene. However, phenotypes arise in transgenic mice from overexpression per se, rather than from the effects of the mutation.

To create more sophisticated models that overcome issues of dosage-sensitivity and express the biochemically correct human protein at physiological levels, we have developed a new technology that is straight-forward and should be useable by standard-gene targeting laboratories.

We have created 'genomically humanised' animals in which the mouse gene is entirely replaced by the human genomic locus. These mice express the mutant protein at physiological levels. We are now applying for funding to achieve two goals,

(1) to develop our genomic humanisation technology to create conditional mutants and,
(2) to characterise our new mouse lines and shed light on the biology and pathology of our paradigm, FUS, in health and in neurodegeneration.

By the end of this program of research we will have informative new FUS models for the community, and a better understanding of how mutant FUS causes neurons to die. The normal biology of FUS is not well understood and FUS mutations can be particularly aggressive, often causing ALS in young teenagers and adults, for as yet entirely unknown reasons. FUS is also involved in other neurological diseases, and is an important target for our attention.

This technology is applicable to any organism for which ES cells exist, not just mice.

Planned Impact

The impact of this application lies in the fields of neurodegenerative/neurological disease, and in mouse modelling, and the beneficiaries are potentially commercial Pharma and Biotech, patient and carer groups, academics. The work described here could contribute to health and wealth outcomes, and have an impact on biomedicine contributing to quality of life outcomes.

With respect to the mouse modelling aspects of this application, development of this technology will enable all laboratories that carry out gene targeting to humanise mouse models, as the basis of our approach is classical gene-targetting and therefore this project may have significant impact both commercially and academically, in providing more tailored models and so more accurate models for human disease. This has both health and wealth, and quality of life impact.

Further, this technology is applicable to all organisms for which ES cells capable of homologous recombination exist, the number of such organisms is increasing dramatically and includes both rat and human. In human we can see uses in gene therapy in correcting relatively large regions of the genome, for example in iPS cells.

With respect to neurodegenerative disease/neurological disease, we are specifically interested in analysing FUS humanised mouse mutants, and this project came about because of the lack of FUS models due to the dosage sensitivity of FUS in such animals -- and similar difficulties in other models (e.g. with the TDP43 gene), being created to model human neurodegenerative disease. Thus we have invested in developing a new technology to create animals with the correct human protein biochemistry arising from the mutant human gene, and in expressing the gene at endogenous levels. We see this as essential for understanding not only how mutant FUS causes disease, but how other dosage-sensitive genes, in ALS and all other areas of biomedicine, give rise to pathology.

Thus the impact of our specific biological investigation lies in dissection mechanisms of FUS-ALS and neurodegeneration, and in a better understanding of the role of disruption of RNA metabolism in human disease. The beneficiaries are commercial organisations investigating such diseases - and we note our excellent ties to Pharma and Biotech, through current collaborations and through the extensive opportunites for networking and development provided by UCL and UCLBusiness. The beneficiaries are also patient organisations ultimately as treatments and cures are eventually found for ALS (and other forms of neurodegeneration). We also note that new - commercial and academic -- approaches to treating ALS entail reduction of levels of transcripts from mutant alleles and our mouse models offer the only genomically accurate models for developing such therapies.

We also note we are excellently placed to follow up findings with translational neuroscience, as the adjacent National Hospital for Neurology and Neurosurgery hosts one of the largest ALS clinics in Europe and clinicians at that clinic are joint appointed to our Departments as clinician-scientists working on projects related to this application.

With respect to training, we have a range of students at all levels working in our labs including undergraduate neuroscientists and geneticists from UCL, UK and abroad, Master's students from UCL and elsewhere in UK, PhD students, MD students, and a range of clinician scientists. Thus we provide an unusual and important training in the creation, validation and characterisation of novel engineered mouse models of neurodegeneration, for new generations of scientists and clinicians.

We hope that the new models we are developing will Refine the use of mice as disease models, in accordance with the 3Rs.


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Fisher EMC (2019) Mouse models of neurodegeneration: Know your question, know your mouse. in Science translational medicine

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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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Nair RR (2019) Uses for humanised mouse models in precision medicine for neurodegenerative disease. in Mammalian genome : official journal of the International Mammalian Genome Society

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Sivakumar P (2018) TDP-43 mutations increase HNRNP A1-7B through gain of splicing function. in Brain : a journal of neurology

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
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 A new mouse model of motor neuron degeneration (FUS ALS) 
Description A new genetically engineered mouse model of FUS ALS 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Year Produced 2017 
Provided To Others? Yes  
Impact Several labs now working with this model 
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. 
Title FUS homozygotes MEFs 
Description Working with a mouse model, an in vivo model, to produce IMMORTILISED cell lines so that we can drop our animal useage. 
Type Of Material Database/Collection of data 
Year Produced 2017 
Provided To Others? Yes  
Impact Reduced mouse numbers 
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 FUS mouse transcriptome 
Organisation University College London
Department Institute of Neurology
Country United Kingdom 
Sector Academic/University 
PI Contribution Provided new mouse models for analysis by the two separate groups (Sobell and Molecular Neuroscience)
Collaborator Contribution Specialist analysis in transcriptomics
Impact Pending, molecular biology of different types
Start Year 2015
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 Labs at UCL for bespoke mouse models of neurodegeneration 
Organisation University College London
Department Department of Neuroscience, Physiology and Pharmacology
Country United Kingdom 
Sector Academic/University 
PI Contribution Genome engineering expertise
Collaborator Contribution In depth knowledge of specific forms of neurodegeneration Fratta, Isaacs, Greensmith, Schiavo, Wiseman.
Impact None yet.
Start Year 2017
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 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
Title Antibody for FUS Delta 14 disease epitope. 
Description A disease specific antibody for our FUS Delta14 mouse model of motor neuron disease/amyotrophic lateral sclerosis. 
IP Reference  
Protection Protection not required
Year Protection Granted 2017
Licensed Yes
Impact Antibody for FUS Delta 14 sent out for academic use.
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 Research seminar given at Inaugural ALS Symposium, Queen Square 2019, and in Umea, Sweden, and in Yeditepe University, Turkey 2019 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Three seminars given in UK, Turkey, Sweden, to specialist audiences of researchers, on the disease ALS.
One outcome was collaboration and grant writing with the group of expert biochemists in Sweden.
Year(s) Of Engagement Activity 2019
Description Set up new ALS Seminar Series 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact Set up an academic seminar series on Amyotrophic Lateral Sclerosis and similar topics, but as this is open to the lay public we have had a surprising interest and attendance from sixth form pupils and undergraduates.
Year(s) Of Engagement Activity 2014
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