Mouse Models of Neurodegenerative Diseases Laboratory (MMON)

Lead Research Organisation: MRC Mammalian Genetics Unit

Abstract

Neurodegeneration is defined by loss of specific types of neurons in the brain, spinal cord, or peripheral nerves, occurring in disorders including motor neuron disease and dementia, which we study in this programme. These diseases afflict millions worldwide, are a leading cause of premature death globally, and have devastating social and personal costs to those affected and those who are close to them. Currently we have at best very limited therapeutic options for most neurodegenerative disorders, and cures for none of them.

We need to better understand the onset and progression of these diseases in order to find effective therapies, but critical early disease mechanisms are difficult to study in human patients due to the inaccessibility of the human central nervous system in living patients, and because the neurodegenerative process starts many years before symptom onset. Therefore, we use mice as a model system, and despite 75 million years of evolution separating mouse and humans, we share much the same biology and genetics. In particular, genes that cause familial forms of neurodegeneration are present in both mice and humans and typically share the same function.

By introducing human disease-causing mutations into mice using the latest genome engineering technologies (including CRISR/Cas9), we can model the pathological changes that these mutations cause, including for example, mutations that cause the encoding protein to aggregate leading to neuronal toxicity. Furthermore, we are now humanising whole genes, not just small mutations, to allow improved relevance for human physiology and improved models for therapeutic testing.

Technical Summary

Neurodegenerative disease encompasses a wide range of disorders afflicting the central and peripheral nervous systems that embody a major unmet biomedical need. There are very limited treatments, and no cures, for most of these diseases, including amyotrophic lateral sclerosis (ALS) and dementia, in which we have a principle interest. The key aims for this programme are (1) to engineer and characterise bespoke models of neurodegeneration and (2) to utilise these and existing models to understand disease pathomechanisms that may lead to new avenues for treatments.

Toward these goals we employ a number of different strategies and genetic tools. Our most straight-forward approach is to make use of the unprecedented range of mouse models available from global collaborations such as the International Knockout Mouse Consortium (IKMC). He we aim to understand the normal function of genes associated with neurodegeneration and neuronal function. We also make use of N-ethyl-N-nitrosourea (ENU) mutant mouse repositories to study point mutations in neurodegenerative disease genes (e.g. ALS genes encoding TDP43 and SOD1).

Our most refined approach involves engineering and studying a new generation of ‘humanised’ knock-in (KI) animals, in which a mouse gene of interest (or specific regions of a mouse gene) is replaced with the orthologous human sequence (e.g. our hFUS and hSOD1 mice). CRISPR/Cas9 technology allows us to additionally introduce any number of human pathogenic mutations to study. The reasons for humanising at the DNA level are so that mouse models express physiological levels of pathogenic genes and proteins analogous to those found in people – with the human splice isoforms and with human protein biochemistry that can be critical for disease pathogenesis. Currently, the most widely used neurodegenerative disease models are transgenic animals that greatly overexpress human pathogenic genes (e.g. SOD1-G93A mice, a widely used ALS model), leading to rapid neurodegeneration (in contrast to humans and KI mice); however, unpicking the effects of overexpression from physiologically relevant pathological changes is difficult in these models. KI models (and similar ‘endogenous’ mutants such as ENU models) develop slowly progressive neurodegenerative disease and thus provide systems for understanding critical early-stage disease mechanisms and biomarkers. Finally, in therapeutic terms, humanised KI models will also allow testing of treatments that target the human transcript or human protein, with potentially better translatability to human patients.

Publications

10 25 50
 
Description Two PhD studentships from internal funding MRC Harwell
Amount £120,000 (GBP)
Organisation Medical Research Council (MRC) 
Sector Public
Country United Kingdom
Start 09/2019 
End 08/2022
 
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 Two new humanised FUS mutant strains P525L and Q519Ifs 
Description Fully humanised FUS gene, wildtype, onto which we have added the P525L mutation and, separately in a second strain of mouse, the Q519Ifs mutation, to try to more faithfully models human FUS motor neuron disease/amyotrophic lateral sclerosis. 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Year Produced 2019 
Provided To Others? Yes  
Impact These mice have been presented at meetings, they are freely available, but we have not yet published on them. 
 
Title new humanised SOD1 mouse 
Description A new Humanised mouse model with the human SOD1 gene; this is a wildtype control for when we go forward to put in human amyotrophic lateral sclerosis mutations into SOD1. 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Year Produced 2019 
Provided To Others? Yes  
Impact This mouse is freely available and has been presented at meetings, but we have not yet published on it. 
 
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 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 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 Tom U of Sheffield for repeat cloning 
Organisation University of Sheffield
Country United Kingdom 
Sector Academic/University 
PI Contribution Tom Cunningham of MRC Harwell developed new cloning method for repeat expansion diseases. In use now in Sheffield.
Collaborator Contribution Cell lines and sequencing
Impact Scientific research collaboration. No publications yet, too early.
Start Year 2019
 
Description Tom with Bill Skarnes, Jax, for repeat cloning 
Organisation The Jackson Laboratory
Country United States 
Sector Charity/Non Profit 
PI Contribution Tom Cunningham for repeat cloning protocol for expanded repeats. To be used by the Jackson laboratory in human cells.
Collaborator Contribution Cell lines, technical expertise, outcome assays.
Impact Laboratory research, publications pending.
Start Year 2020
 
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 with Pietro Fratta for ALS FUS/TDP43 mice 
Organisation University College London
Department Institute of Neurology
Country United Kingdom 
Sector Academic/University 
PI Contribution creation of novel humanised mouse models of ALS
Collaborator Contribution extensive analysis of transcriptomics and other measures.
Impact Potentially novel approach to therapeutics for sporadic ALS.
Start Year 2019
 
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 DS special interest group 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact Wiseman F, LonDownS Consortium (2018) Understanding Down syndrome using Alzheimer's mouse models. Dementia in intellectual disabilities, Special Interest Group Meeting, London, London UK.

Talk to mix of medical and other professionals working with peoplewith Down syndrome.
Year(s) Of Engagement Activity 2018
 
Description Edgar talk to Parkinson's patients at Open Day 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Patients, carers and/or patient groups
Results and Impact Open day at MRC Harwell; PI gave a talk to Parkinson's disease patients and carers.
Year(s) Of Engagement Activity 2017
 
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 New Scientist Live 2018 MRC Harwell 
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 Public/other audiences
Results and Impact New Scientist Live 2018, a public engagement activity for various disciplines in science, on 21 September 2018
Year(s) Of Engagement Activity 2018
 
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 UCL Postgraduate symposium poster 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact De Giorgio A, Devoy A, Milioto C, Zhu F, MacKenzie K, Acevedo Arozena A, Fisher EMC (2018) Humanising the mouse Tardbp gene. UCL Sixth Annual Postgraduate Research Symposium, London, UK

Poster presented to postgraduates outside field.
Year(s) Of Engagement Activity 2018