New fellowship explores how C9ORF72 causes MND

Dr Johnathan Cooper-Knock, MRC/MND Association Lady Edith Wolfson Clinical Research Fellow

Dr Johnathan Cooper-Knock from the Sheffield Institute for Translational Neuroscience (SITraN) has been awarded with the fifth Medical Research Council (MRC)/MND Association Lady Edith Wolfson Clinical Research Fellowship.

Through his three-year fellowship, Dr Cooper-Knock will use the MND Association’s DNA bank to study how recently discovered mistakes (known as mutations) in a gene called C9ORF72 can cause the disease.

Dr Johnathan Cooper Knock explains, “I believe that the genetics of MND are a key to understanding both the cause of the disease and how to treat it. The discovery of mutations in C9ORF72 are a great opportunity to get a hold on mechanisms of disease which has so far been elusive. I am excited by the opportunity my fellowship will give me to pursue this important discovery.

“By the end of my fellowship I aim to have contributed significantly to the understanding of disease mechanisms related to C9ORF72 dysfunction in MND. As a result I hope to have identified a number of therapeutic targets for development into new treatments by myself and others.”

C9ORF72: the facts so far

We know that a repeated six-letter code within a gene called C9ORF72 can cause MND and a related condition called fronto-temporal dementia (FTD) for approximately 40% of cases with a positive family history of MND and/or FTD.

Most genetic mistakes found in MND to date have been swaps of genetic letters, which can change the meaning of that part of the gene. The C9ORF72 genetic mistake on the other hand, is a repeat expansion. This means that six letters within the genetic code (CCCCGG) are repeated hundreds of times for people with C9ORF72 MND. In healthy individuals, this repeat is found about 30 times. We already know that the exact size of the repeat varies substantially between people with this genetic mistake. How this repeat causes MND and how the size of the repeat may affect disease progression is currently unknown but this is something that Dr Cooper-Knock wants to find out.

We also don’t know what role C9ORF72 normally has in the body. Even its name, which stands for ‘chromosome 9 open reading frame 72’ refers to where it is in the genetic code and not what it does. This isn’t unusual as it’s currently estimated that we have over 20,000 genes, and understandably, researchers haven’t yet found out what every one of these does – including C9ORF72.

So far, 96 journal articles have been published about C9ORF72 (by searching on Pubmed for C9ORF72). The oldest of these was published in 2011, and describes the original MND/FTD C9ORF72 finding. All subsequent articles on C9ORF72 have been of a direct consequence from this pivotal genetic discovery in the past year.

These 96 studies were focused on finding out how many people have the C9ORF72 genetic repeat and finding out what this mistake ‘looks like’ both clinically in terms of progression rates, age of onset and symptoms; and in terms of post-mortem findings to compare with other forms of MND. Coincidently, the most recent post-mortem and clinical C9ORF72 finding was authored by Dr Cooper-Knock (when searching for C9ORF72 and post mortem on PubMed).

It’s reassuring to know that researchers aren’t resting on their laurels with this genetic finding. There’s a huge international research effort in place to push forward our understanding of C9ORF72, with a number of our own newly funded projects, starting later this year, dedicated to creating new laboratory models of this genetic mistake to better understand how it can cause the disease.

How do we currently think C9ORF72 causes MND?

Due to the sheer size of the repeat expansion in C9ORF72, it’s thought that it causes MND by disruption of the editing process of genetic information.

I’ll explain: In real life terms, our DNA can be thought of as being held within a library, which is the control centre of the cell (the nucleus). Each book (gene) is stored on a particular shelf (chromosome). Gene ‘books’ aren’t allowed to be taken out of the nucleus, but they can be photocopied. These copies (RNA) are edited and transported out of the nucleus to be used as instructions to create proteins that perform specific roles in and sometimes out of the cell.

Unlike real life books, genes are fraught with errors, variations and nonsense from one person to the next. It looks messy, but it’s normal. Genetic editors are needed to edit and chop the RNA into a readable format so that it can be understood by the parts of the cell that use RNA as instructions.

As normal, healthy copies of C9ORF72 hold approximately 30 repeats to be chopped out as RNA, the effect of having much larger repeats may be having a knock-on effect on the efficiency of the editing process. This could then lead to a much higher risk of developing MND.

Finding out exactly how C9ORF72 can cause MND, and whether this theory is right, will provide us with a deeper insight into MND and potentially provide therapeutic targets that can be further investigated.

Dr Cooper-Knock’s fellowship

Dr Cooper-Knock will be using a cutting-edge genetic technique called ‘gene expression profiling’ to study the various levels of RNA in samples provided by people with the C9ORF72 genetic mistake. From this, he’ll find out which genes are switched on and off because of the C9ORF72 repeat expansion.

He will also study whether the size of the C9ORF72 repeat expansion has an effect on symptoms or progression rates to identify factors that may modify disease progression and may therefore be targets for future therapies.

Technology specialised for identifying misassembled RNA will also be applied to skin cells donated by people with the C9ORF72 repeat expansion who have MND/FTD and healthy controls. This will help to elucidate what the C9ORF72 protein does.

As well as skin cells from people with MND/FTD, this study will use post-mortem brain and spinal cord tissue from people with the C9ORF72 repeat expansion and healthy controls within the Sheffield tissue bank; as well as cells from the blood of C9ORF72 patients and healthy controls obtained from the MND Association’s DNA Bank.

Talking about the importance of people with MND having provided these numerous samples Dr Cooper-Knock said “Without the participation of patients and their families MND research will get nowhere; and equally with their participation, doors are opened towards new and exciting treatments. At this time, with discoveries like the mutations in C9ORF72 to build from, we can do even more with the participation of those who have been affected by this disease, who like us are passionate to see it cured.”

More information:
Read our official news release on our MND Association website.
Find out more about our DNA bank.

MND Association funded researcher Dr Martin Turner wins ENCALS Young Investigator Award

We’re pleased to announce that Dr Martin Turner has been awarded with the European Network for the Cure of ALS (ENCALS) Young Investigators Award 2012.

Dr Martin Turner, MRC/MND Association Lady Edith Wolfson Clinical Research Fellow

Dr Turner was awarded with the MRC/ MND Association Lady Edith Wolfson Clinical Research Fellowship in 2008 for his study to identify biomarkers in MND (called BioMOx). Since then, Dr Turner has already published two findings from his five-year disease marker study in the prestigious journals Neurology and Brain. Using advanced brain scanning technology, his study has identified a common pattern of nerve damage in the brains of MND patients. This holds the promise of a much-needed disease marker.

Talking about why he thinks the ENCALS award is so important, Dr Turner said:

“The ENCALS award marks a major highlight in my career.”

“I am passionate about MND, and feel privileged to help care for those living with the most challenging of diseases. To be recognised as having made a useful contribution to research as well, by international leaders in the field, means an enormous amount.

“It is 13 years since I began as a PhD student under Professor Nigel Leigh, whose ground-breaking ideas about brain changes in MND first sparked my interest. I was fortunate to meet Professor Kevin Talbot in 2003, and through his support and partnership I have been able to develop these ideas alongside leading brain imaging neuroscientists at Oxford University.

“I have never felt more sure that progress is accelerating in MND research, and I am pleased to be adding something to the wider global effort.”

Funding promising researchers

One of our research aims, is to develop the research workforce. Dr Turner talks more about how our funding has helped to develop his career:

“The Lady Edith Wolfson Clinical Research Fellowship scheme, uniquely linked to the Government-funded Medical Research Council through the MND Association, has been critical to my development as an MND researcher.

“These highly competitive 5-year Fellowships don’t simply provide the funding for the experimental studies, but crucially allow me to devote most of my time as a consultant neurologist solely to the care and research of MND patients. There is no simple way to specialise like this within the standard NHS framework, and such schemes are a vital way to help develop a strong UK academic neurology workforce in MND.”

Commenting on this story, our Director of Research Development, Dr Brian Dickie said “We’re delighted that one of our Lady Edith Wolfson Fellows has won this prestigious international award. The Fellowships were created to attract and retain the brightest and the best young clinicians to MND research and it is a fitting tribute to the knowledge, expertise and dedication that Dr Turner brings to this important field of MND research.”

More information:

Our official news release

Go to the BioMOx website to find out more about this project

Find out more about ENCALS

Our research aims

BioMOx findings:

• Common signature of nerve damage has been identified in the brains of people living with MND using an advanced MRI technique.
• Increase in functional connectivity and activity in other parts of the brain, associated indirectly with movement.

May The Fourth Be With You – PhD Studentship Applications

Spring has finally sprung and so it’s now time to open our online summary application form for our next round of research grant applications.

This round is for PhD studentship applications, for projects starting in October 2013. The deadline for summary applications is Friday 4 May 2012.

Attracting promising researchers
Through our successful PhD studentship programme we have a track record of attracting and funding promising young scientists to develop their careers in MND research. Since 1998 we have funded 30 studentships, 12 of these are currently ongoing.

We need to continue to develop the UK basic research capacity by attracting more young scientists to develop careers in MND research. We can do this by funding PhD studentships.

Funding the best of the best
As with all of the research projects funded by the MND Association, our rigorous application process allows us to ensure we only fund studentships of the highest quality and of direct relevance to MND.

The way that we fund research starts with a summary application, which is a concise outline of the proposed project. After the deadline date has passed a decision is made as to whether the summary is relevant to ‘classical’ MND and the project aims fit with our Research Strategy. If the summary does not fit, it’s rejected. If all criteria are met, the summary is reviewed by our Biomedical Research Advisory Panel (BRAP).

The reviewer’s comments and scores are then assessed using a two thirds majority rule. Each reviewer scores the summary application. A score under 50 is classed as unsuitable for funding, if it’s over 50 then the applicant is invited to submit a full application.

We hope this year holds an exciting round of PhD studentship applications!

More information:

For further information please see the Prize PhD Studentship Flyer or visit our website www.mndassociation.org

Sharing and networking in Liverpool

From Sunday morning to Tuesday evening last week, there was a lot of talk of MND research going on in Liverpool. The reason for this ‘hotspot’ of discussions was due to the annual meeting of an international consortium of MND researchers taking place at the University of Liverpool. The 10th International Consortium on SOD1 and ALS (ICOSA) meeting took place last weekend (4 – 5 March).

In 2001, five laboratories came together to form ICOSA, where the aim was to share knowledge to design better-informed experiments to understand the rare, inherited SOD1 form of MND. MND Association grantee, Prof Samar Hasnain was one of its founding members. Success of this philosophy of sharing knowledge prior to publication has resulted in several leading groups joining the effort, looking at other causes of inherited MND too.

A tradition of ICOSA meetings is to hold an open meeting for sharing latest results with a wider audience, following their closed meeting. Thus, on Tuesday 6 March, an open meeting was held to allow the exchange of the latest results and ideas between ICOSA members and the UK MND research community.

I attended this one day meeting in Liverpool and I’ve written a mini report on the meeting below, including a couple of highlights.

The first few presenters demonstrated the truly international nature of this collaboration – they had travelled from the snowy landscape of northern Sweden, the sweltering heat (at least in August!) of mid-state Florida and from RIKEN, the large natural sciences research centre, in Japan .

The researchers represented were a mixture of physicists, biochemists and neurologists – an unusually broad spectrum of knowledge and speciality for an MND research meeting. Essentially, their core, joint interest was in understanding how the structure of a protein has such a marked change leading to MND developing or the disease progressing.

The structure of a protein is essentially about folding. The correct folding will mean that the protein can do its job. Folded incorrectly the protein won’t be able to work. An example of incorrectly folded protein is the protein clumps or ‘aggregates’ seen within motor neurones in MND. There is a whole chain of events that lead the appearance of these clumps of protein – and researchers at the meeting discussed every step along the way.

How do proteins fold and why is it important?

When the instructions for making a protein (ie genes) are read and edited by DNA and RNA respectively, they are reading or editing instructions to arrange a set of building blocks in a particular order – there are 20 different types of building block – our amino acids. ALL of our proteins within our bodies are made from specific arrangements of this core set of 20 building blocks. The arrangement of the building blocks determines where the protein folds, in which direction and the shape it makes. There are many possible folding arrangements a protein could make, but it will always try and fold itself into the lowest energy shape (a good way to think about this is the shape where the protein is ‘most comfortable’).

Geneticists know a lot about the beginning of the process (what the sequence of building blocks will be) and biochemists and pathologists know a lot about the end of this process (what the protein does and a what it looks like in the cell when it clumps together) – but the physicists of the MND research world are working on the bit in the middle (precisely where which building block is, in the folded protein).

A change to the sequence of the building blocks, as seen in the proteins made from mutated genes that cause MND, will lead to unusual folding, and damage to the cell – due to the loss of normal function or a trigger for toxicity. So having a complete picture of a protein ‘lifespan’ is really important in understanding what goes wrong in MND and how to fix it.

Unravelling questions about SOD1

People with the SOD1 form of the rare, inherited type of MND have a mistake in the assembly of one building block in the instruction to make the SOD1 protein. Over 160 different, single building block mistakes have been found in this form of MND so far. All of them lead to the development of MND. So that means 160 damaging variations in the folding of the SOD1 protein.

Over 70 other delegates and I heard the latest on how mimicking the effects of these mutations (by changing building blocks of the protein) in SOD1 mouse models tells us more about this cause of MND. It’s even possible to study the different effects of the toxic protein on different cell types essential for motor neurone function. (Although motor neurones carry the messages, they are supported by groups of ‘glia’ cells around them).

Where (the) ‘FUS’ is

Prof Larry Hayward presented his research on a protein called ‘FUS’; mutations in this gene causes another form of the rare inherited MND. The damaged ‘FUS’ protein is found in a completely different place in motor neurones than usual. Images of motor neurones where the FUS is in the centre of motor neurones, as usual, looked a bit like fried eggs; but the location of the damaged FUS in the outside of the cell reminded me of ring donuts! By stressing motor neurones, he showed a video of the proteins moving from the centre to the outside of the cell; and back to the centre when the stress was removed. This all happens very quickly, in a matter of minutes!

C9orf72 – a hot topic

Another highlight of the meeting was the presentation by MND Association grantee Prof Huw Morris on both how the C9orf72 gene mistake was found last year, and also on what’s happened since the results of this finding were announced. In the five and a half months since the 21 September announcement, another 26 reports have been published in this area of MND research. That’s slightly more than one report a week! (To put this in context there are roughly 36 MND reports published a week, total, across a broad range of topics). He commented that one factor that kept him focussed in the long search for this gene defect was the people with MND in his care.

Drug scaffolding to correct damaged folding

Above I mentioned that the physicists work out the precise folding of proteins, knowing where each of the building blocks is within its final shape. They do this by isolating the protein they want to study and placing it in increasingly high concentrations of salt solution to remove literally every molecule of water, until the protein itself comes out of solution and forms crystals. These crystals are then analysed by x-ray crystallography and other analytical chemistry techniques.

For a protein made from a mutated SOD1 gene, x-ray crystallography studies found a hole in the protein folding that may explain why it forms clumps within motor neurones. MND Association funded researcher Dr Neil Kershaw from the University of Liverpool presented the latest results from his research in designing a drug that will ‘prop up’ incorrectly folded SOD1, in the hope that this will remove its damaging effects.

I hope that this report demonstrates that in between the ‘big news’ stories about MND research, steady progress continues to be made in understanding MND and searching for treatments for it.

Disappointing results from UK based lithium clinical trial

Yesterday, we announced on our website the disappointing news that the UK-based lithium clinical trial showed that lithium carbonate is ineffective at treating MND.

Commenting on the lithium clinical trial, Dr Brian Dickie, our Director of Research Development said:

“As many people will know, when lithium was first proposed as having benefit in MND, a couple of small, short-term trials were performed to establish whether the drug had a large and rapid effect on physical changes in disease progression. This trial, by contrast, was developed to ask whether the drug had a more subtle benefit over a longer time course, as is the case with riluzole, using survival times as the primary measure. The only way to answer this question was by performing larger, lengthier and more comprehensive studies.

“While the result is deeply disappointing, we now have a clear answer.

“Lithium can be described as a messy drug. It can act in multiple ways in the body, producing potentially beneficial effects as well as possible unwanted side effects. An overall beneficial effect, even modest, would have refocused scientific interest in the drug to try and separate ‘the good from the bad’ with the longer-term goal of developing more effective compounds. This is a strategy that is presently being pursued with regard to riluzole, in a project co-funded by the ALS Association, the University of Reading and ourselves.

“This trial was the first of its type in the UK, devised and run by clinicians without the need for drug company funding. A number of MND clinics that previously had little or no experience in clinical drug trials for MND have developed vital expertise and confidence in delivering trials to the highest standards. This can only help make the UK a more attractive place in the future for drug companies looking to push potential treatments from lab to clinic.”    

Two hundred and fourteen people with MND took part in this trial, each giving up their time to help find us the answers. We’d like to thank those that have taken part in this trial.

One person who took part in the UK lithium clinical trial was Colin Knight. We spoke to him a few years ago about his views on taking part. Please be aware that in the film clip, Colin speaks frankly about his diagnosis.

 

Read our official press release.

Cogane produces encouraging results in MND Association-funded study

Prof Linda Greensmith

Thanks to funding and some strategic ‘match-making’ by the MND Association, a new drug may have taken one step closer to beginning clinical trials in MND after producing promising results in an animal model of the disease.

The drug, known as Cogane, was developed by the biotechnology company Phytopharm. It had already demonstrated in laboratory tests that it could help to protect neurones by promoting the production of natural, nerve nourishing substances called neurotrophic factors and early animal testing had hinted at its potential beneficial effects in MND. However, its journey towards clinical testing in MND had hit a road block because it hadn’t been extensively put through its paces in large numbers of the most widely used animal model of the disease, the SOD1 mouse. Without robust data from this model, there would have been little to encourage further investment in Cogane’s development.

So up stepped the Association to introduce Phytopharm to Professor Linda Greensmith at University College London, a leading MND researcher with considerable expertise in SOD1 mouse testing. With funding from the Association, Prof Greensmith and her team were able to conduct a rigorous study of the effects of Cogane, administered to the mice after they had developed MND-like symptoms.

The drug produced some significant improvements in muscle strength and motor neurone survival and managed to produce positive effects even in mice that had reached the later stages of the disease. To give more substance to these preliminary but very encouraging results, the research team will now go on to the painstaking work of examining more closely Cogane’s effects on the motor neurones and other key cells that play a critical role in the progression of MND. 

After the disappointment of the Trophos trial results, it’s great to be able to share some positive news on the drug development front. We know from long experience that it’s wise to limit our excitement over positive results from the mouse model – after all, plenty of drugs have shown promise at this stage and have then gone on to fail in clinical trials. However, Prof Greensmith’s experience and expertise mean that Cogane will have been tested with the utmost rigor. As she herself commented, the results indicate that “Cogane has significant potential as a therapy for ALS and merits further evaluation”.  We don’t yet know what Phytopharm’s next steps will be – these may become clearer once the more detailed data from Prof Greensmith’s work have been published, which could take the best part of a year. Let’s hope that we have a given Cogane enough of a boost to push it out of the drug development ‘doldrums’.

Read the Phytopharm press release.

Happy New Year – Quiz answers and round up of 2011!

And the answers to our Christmas Quiz are:

1. How many neurones does a human have? Billions
2. Which animal has the largest brain? Bottlenose dolphin
3. How much does a human brain weigh in comparison with our total average body weight (in percent)? 2
4. How many DNA samples does the MND Association’s DNA bank hold? 3,400
5. How many research projects do we currently fund? 44
6. How much does our research project portfolio currently come to? £7.6m
7. How many PhD studentships do we currently fund? 12
8. How many times a year do we have research grant funding rounds? 2
9. How many unproven MND treatments have ALSUntangled investigated so far? 13
10. How many stem cell research projects do we fund? 2

At the beginning of a new year, it’s always encouraging to look back on how far we’ve come. The list of MND research achievements continues to grow exponentially every year, and I’m pleased to say that last year was no exception, demonstrating that we really are living in exciting times.

2011 had some important discoveries in the world of MND research to find the answers to what causes MND. A number of MND causing gene mistakes were discovered including C9ORF72, Ubiquilin2 and SQSTM1. With these findings, we now know the cause of approximately 70% of cases of inherited MND – a massive leap from approximately 25-30% of known genetic mistakes the previous year.

Within the team, we’ve also made some promising headway toward our aims set out in our research strategy, by funding and promoting cutting edge research both within the UK and around the world. For example, our groundbreaking biomarker project led by Dr Martin Turner at Oxford yielded its second set of promising results, just three years into the five-year project. Dr Martin Turner also gave an enthralling talk at last year’s International Symposium on ALS/MND on neuroimaging (brain scanning) and he’s regarded as ‘the man’ to speak to in terms of MND neuroimaging on an international level.

As well as the research projects that we fund yielding positive results, and following progress on an international level, we’re also a major player in promoting research. The key to defeating MND lies in fostering strong collaboration between leading researchers around the world  and sharing new understanding of the disease as rapidly as possible. In 2011, we made two huge steps in this:

In January 2011, in conjunction with two leading members of the International Consortium of Stem Cell Networks (the Canadian Stem Cell Network and the UK Stem Cell Network), The New York Stem Cell Foundation and the ALS Association of the USA, we organised an MND stem cell conference. Our workshop brought together 60 of the world’s leading stem cell research experts to shape the development of future international MND stem cell research and to form new research collaborations. We were privileged to organise this event and the research community now have a solid foundation of understanding of where we are in terms of MND stem cell research. Dr Brian Dickie, our Director of Research now also has the honour of being a co-author on the scientific paper from the conference – published in the journal ALS.

In July 2011, we made a further step forward in sharing new understanding rapidly by joining a group of research-funding organisations to fund UK PubMed Central, an online research database containing over two million research articles. This is the first step in the Association’s aim to establish a comprehensive resource for the global MND research community.

We also had a fantastic year for improving the way we fund research and maintaining our high standards.

For our first grants round of the year, a record-breaking 19 full applications were considered for funding by our Biomedical Research Advisory Panel. Only one in five research applications is considered of a high enough standard for funding, but through our rigorous process we can provide our donors with the assurance that they are supporting the ‘very best of the best’ MND research.

Before our second grants round, we announced the successful launch of our online summary application form for researchers applying for grants and PhD studentships. By evolving our summary application process to use an online system, we are able to ensure that our high standards are maintained and that we are using our time efficiently and effectively to fund high-quality research.

We also proudly received our certificate for best practice for our rigorous procedures for funding research from the Association of Medical Research Charities (AMRC) in the UK with a comment saying that we are “considered as setting the standard within the audit”.

You can find out more information on the research projects we currently fund on our research we fund information sheet.

One of our highlights from last year, and the result of over a year’s work in preparation from the research team and our conference team, was the International Symposium on ALS/MND held in Sydney, Australia. We are proud to organise this vital worldwide event every year, and are pleased that last year was successful. Holding the event in different countries around the world enables us to draw new people into the international research community, bringing new ideas and expertise to the field and creating new alliances in the fight against MND.

We took you behind the scenes of last year’s symposium by writing daily blog articles on a multitude of topics. If you’ve not already read these, you can find an introduction to these with links on our blog. Please remember to complete our survey on what you thought of our reporting, as it really helps us to determine whether we should continue to report from the symposium, and whether we should change anything.

We’ve definitely set the bar in 2011 and have a lot to live up to in 2012. We’re really looking forward to see what 2012 holds for MND research, and we hope that you’ll continue to follow our progress on our blog throughout the year.

We wish you a very Happy New Year from all of us in the Research Development Team at the MND Association.

Fighting a faulty recycling machine in MND, with Prof John Mayer

A recent gene finding suggests that recycling within our cells is key to all forms of MND. This story captivated many people affected by MND and our blog broke its previous record for the number of hits in one week at over 4,000. It was also linked to, as a reliable and informative piece, from a number of worldwide MND/ALS Associations and forums.

Prof John Mayer, University of Nottingham

Due to the popularity of this story, Prof John Mayer from University of Nottingham will be taking you on a whirlwind tour of the recycling process within our cells. Prof Mayer is currently the chair of our Biomedical Research Advisory Panel, which ensures that we fund the most promising laboratory based research projects to investigate the causes, develop treatments and find markers of disease progression.

He’s also been pioneering the investigation of the recycling process within our cells to learn more about neurodegenerative diseases such as MND for the past 24 years. Below, Prof Mayer explains more about how he’s been involved with this story, and where this could lead us in the future:

The beginning…
Twenty four years ago, Prof Jim Lowe and I discovered that pile-ups of proteins in neurones in MND, and other chronic neurodegenerative diseases eg Parkinson’s disease and Alzheimer’s disease, contained ‘tagged’ proteins. We’ve been trying to understand why ever since!

We did it by detecting those tags, and ‘staining’ tissue sections from the brains and spines of patients who had died of MND in Derby and Nottingham to see if we could ‘see’ those piles of proteins in surviving neurones. We did!

From that day on, we knew that the protein recycling system must be deeply involved in neurological disease and that the system must fail or be overwhelmed in the neurones of people with MND.

How it works
Proteins can be thought of as the building blocks of our cells and all proteins are made and broken down continuously –this is called protein turnover. It is essential because faulty proteins can be made or proteins can be damaged in each cell, including neurones. Protein turnover in neurones is vital because the vast majority do not divide – once these cells are laid down we are stuck with them. Any problem protein in a neurone must be removed or it may die.

Proteins are ‘tagged’ for removal by chemically attaching a small protein to them called ubiquitin – actually chains of ubiquitins all linked to one another to create a long ‘tag’ which will be easily ‘seen’ by the machine that will destroy the tagged protein.

The machine is an enormous entity in the cell called the 26S proteasome. The tagged proteins have to be fed into large caverns in the middle of the machine for destruction, with the tags removed first to be used again. The mechanism is called the Ubiqutin Proteosome System of protein destruction in the cell, the UPS for short (and not to be confused with the ‘logistics’ company!).

It just would not do if proteins could be destroyed anywhere in the cell, like by those proteases in biological washing powders, the cell proteins would always be at risk of degradation. So, the destructive sites are hidden inside the proteasome machine, the proteins are tagged and they’re fed inside!

There are also a group of cousins of ubiquitin that transfer the tagged proteins to the proteaseome machine. These proteins have a ‘docking site’ for tags at one end and a different tag at the other end which docks to special sites on the proteasome machine. The transfer proteins seek and find tagged proteins and take them to the proteasome machine where they dock and the tagged proteins are fed in for destruction after removal of the tags.

Ubiquilin 2 is one of the carrier proteins which, when made with errors, has been found to cause MND.

Medical science is most comfortable when there is genetic proof of the importance of a process – the discovery of mistakes in ubiquilin 2 has now done this for us!

Mimicking MND by deleting ‘machine’ genes
We have used modern gene targeting in mice to demonstrate that if we deliberately deleted a gene for one of those proteins in the 26S proteasome machine conditionally in neurones in the brain, so we would not cause problems anywhere else in the body, we could ‘mimic’ different neurological diseases.

The way we did it was to target the neurones that die in Parkinson’s disease and the neurones that die in the second most common cause of dementia after Alzheimer’s disease – dementia with Lewy bodies. This was published in 2008 and our genetic approach worked!

By depleting one of the 26S proteasomes machine parts, in the section of the brain which dies in Parkinson’s disease or dementia with Lewy bodies we caused neuronal death –and there were pile ups of tagged proteins in surviving neurones – a key hallmark of disease.

Implications for the future of our MND research
The MND Association has given a pilot grant of £10,000 to Dr Lynn Bedford, who carried out this work, to see if it is possible to delete the gene in motor neurones and innervated muscles to cause MND. She is still working on this (only one pair of hands!) but the tissue sections are now ready to see if MND can be caused this way. We expect that this will be the case and we should know soon!

Keeping open minded
Research into complex disease needs open-minds and different areas of research. Genetics provide clues to familial disease, like for ubiquilin 2, but families are just a small number of people with the disease. The finding of ubiquilin 2 in pile ups with TDP-43, FUS etc shows the generality of the UPS response in MND and ubiquilin 2 will probably be in pile ups of proteins in the other disease too.

The discovery of mistakes in one gene, ubiquilin 2, whose protein product is involved in protein degradation, is fantastic to try to understand MND and other chronic neurodegenerative diseases but there is much more…

Rare mistakes in the genes for three other proteins involved in protein degradation that cause neurodegeneration have recently come to light. Mistakes in a gene called VCP cause MND and a related disease called frontotemporal dementia, mistakes in a gene called optineurin cause MND and mistakes in the p62 protein gene cause MND.

Lightning generally does not strike in the same place twice, yet alone four times! So, to have mistakes in at least four genes causing MND whose protein products are involved in protein degradation dramatically increases the likelihood that problems with this system are central to neurodegenerative disease.

For general effects in disease, researchers must have a pathway that when misbehaving or overwhelmed causes disease (not to mention to provide a therapeutic target). It is one thing to have the capability to find these genetic errors, but it is another to map out the steps in a pathway(s) that cause disease. If a pathway is identified, like through ubiquilin 2 (and the other three genes plus our other ubiquitin-related work), and in general the UPS in other neurological diseases, then I believe that this pathway should be the focus of investment to try to find a cure.

Putting my money where my mouth is
I published a review in the journal Nature Reviews Drug Discovery on the ‘druggability’ of the UPS for many unrelated diseases. Towards the end I had a ‘dream’: if the UPS could be stimulated then neurodegenerative disease could be controlled.

I could not believe it, but some time later, at the end of 2010, my friend Dan Finley and colleagues answered my dream, at least conceptually. They published in the prestigious journal Nature, work on a drug that activated 26S proteasomes to degrade proteins including some involved in neurodegeneration.

So, what are we waiting for? Answer, all the work that goes into converting an initial discovery into a novel therapeutic approach…Watch this space!

Our final thoughts

The story of the recycling process and ubiquilin 2 is indeed an exciting one that is constantly evolving to provide us with more answers as to what causes MND and how we can fight it in the future. As described by Prof Mayer, ‘the machine’,  the proteasome, is normally part of a well oiled process and it is clear that if spanners are thrown into the works that the system can go terribly wrong and cause a number of neurodegenerative diseases, including MND. It will definitely be interesting to watch this research story unravel its secrets in the future.

One thing is certain though – that keeping on top of recycling is very important!

Chromosome 9 finally reveals its secrets

It’s taken a huge international collaboration, including 3 MND Association-funded scientists, to discover a genetic mistake that appears to cause almost 40% of cases of familial (inherited) MND – that’s nearly twice as many as are caused by mutations in the SOD1 gene and more than three times as many as are caused by TDP-43 and FUS combined. Yet despite the fact that it’s relatively common, the rogue gene proved especially difficult to find.

Digging for genes

Our genetic code is arranged into 23 pairs of subunits called chromosomes. Earlier work had homed in on an area on chromosome 9 that appeared to be significantly associated with both MND and the related neurodegenerative disease frontotemporal dementia (FTD), but nobody could drill down as far as the problem gene itself. As a result, chromosome 9 became something of an ‘archaeological dig site’ for MND researchers, with several groups using cutting edge techniques to try and excavate the elusive causative gene that they knew was lurking somewhere in the short arm of this chromosome. The successful international team, which included almost 60 scientists at 37 institutes, finally discovered the exact location and nature of the aberrant genetic code by looking in the most unlikely of places – in the stretches of DNA that do not actually provide any instructions for building proteins, otherwise known as non-coding DNA.

What did the researchers unearth?

The research team studied DNA samples from a Welsh family affected by inherited MND and FTD that was already known to be associated with chromosome 9, as well as samples from a similar Dutch family and a large number of Finnish inherited and non-inherited MND cases. In among the non-coding DNA in a chromosome 9 gene called C9ORF72, the researchers found a 6-letter genetic ‘word’ which, in healthy individuals, is consecutively repeated up to about 20 times. However, in the Welsh and Dutch families and a large proportion of the Finnish familial cases, the 6-letter word was repeated as many as 250 times. This phenomenon is known as a ‘repeat expansion’. The researchers went on to check for this repeat expansion in familial MND cases from North America, Germany and Italy, and found it cropped up in 38% of them. They even found it in a much smaller proportion of sporadic cases from Finland, suggesting that it could be an important risk factor in at least some people with the  non-inherited form of the disease.

What does the discovery mean for MND research?

Despite the fact that the repeat expansion does not directly affect the instructions for building a protein, there is good reason to believe that it can still lead to significant neuronal damage. At the moment it is not fully understood how this happens, but one possibility is that it leads to the production of excessive and consequently toxic quantities of RNA, the molecule that provides the cell with a more usable copy of DNA. Disruption to RNA processing has already been implicated as a disease mechanism in MND – this is the pathway through which faulty TDP-43 and FUS are thought to exert their effects – so C9ORF72 may provide scientists with another piece of the RNA jigsaw.

The effect of the repeat expansion is clearly open to influence. Among those people with the repeat expansion, some experienced only FTD, others showed only muscle weakness, and some had both MND and FTD.  The reasons for this variation in symptoms will be just one area that scientists will now want to look into. This overlap between MND and FTD is something that researchers are very keen to understand, and the C9ORF72 discovery may be the key to solving this puzzle. They will also want to better understand how the repeat expansion causes damage, and that will include trying to find out what C9ORF72 actually does – at the moment this is unknown. (Maybe it’ll get a more interesting name along the way!) Building on the new finding in this way could help move us closer to an effective treatment.

For now, a more tangible consequence of the discovery could be a genetic test for people already diagnosed with familial MND who want to understand more about the basis of their disease. Such a test will take a little time to develop but should become available in the UK in the next few months. When it does, it will be accessible to genetics labs across the country. Anyone interested should speak to their doctor or specialist nurse.  

Dead heat

Just as archaeologists might question whether a newly discovered artefact is the real thing, so scientists need double-checking when they claim to have made a new discovery. Fortunately, a second team hit upon C9ORF72 at exactly the same time, and their results will be published alongside the work described here, in the journal ‘Neuron’. The race to the ‘Lost Ark’ of chromosome 9 ended in a tie, but has provided the research community with a major piece of the MND puzzle on which to build future discoveries.

Article: Renton A, Majounie E, Waite A et al. A hexanucleotide repeat expansion in C9ORF72 is the cause of chromosome 9p21-linked amyotrophic lateral sclerosis-frontotemporal dementia. Neuron (2011).

Read our press release on the C9ORF72 story.

Prof Siddharthan Chandran talks about the recipe for stem cell success at our Annual Conference

Neural Progenitor cells, courtesy of Prof Chandran lab, University of Edinburgh

We invited Prof Siddharthan Chandran to be our keynote speaker at our Annual Conference and below we’ve provided a brief overview of his presentation which we hope you’ll find useful as either a recap if you attended or as an insight into MND and stem cells if you couldn’t make it on the day.

About Prof Chandran

Prof Chandran is Professor of Neurology at the University of Edinburgh and Director of the Euan MacDonald Centre for MND Research which is based at the university. He is leading the Association’s largest stem cell research programme which pulls together world-class researchers from leading institutes in Edinburgh, London and New York. Working together, the international research teams are manipulating stem cells to provide a unique tool for studying MND and developing new drugs. It’s research programmes like this, that really demonstrate our role as a leader in funding and promoting cutting-edge MND research. Naturally we were only too pleased to introduce Prof Chandran to our conference delegates.

Origins of understanding the power of stem cells

Prof Chandran began his talk on a mythological level with the story of Prometheus, who was punished by the Greek God Zeus by being chained to a rock and having his liver eaten daily by an eagle, only to have it grow back the next day to endure the torture again. Not a very nice story, but Prof Chandran went on to explain that through this myth, the Greeks had stumbled onto the origins of understanding the nature of stem cells. The liver is one of the only solid organs that we have that has the power to regenerate itself when damaged. Although this wasn’t the moral of the myth, it’s still an important historical reference that demonstrates that the potential of stem cells as a regenerative tool is not a new concept.

From science fiction to science fact

If Prof Chandran, while at university, had suggested that in the future it would be possible to create stem cells from a skin sample, he said that he would have been ridiculed and the idea would’ve been seen as pure science fiction. Yet here we are, now living in ‘the future’ and this technology is a reality, the newest finding of which was the discovery of stem cell-like cells called ‘induced pleuripotent stem cells’ (or iPS cells for short) in 2008 by a Japanese research group. By delivering a cocktail of chemicals to skin cells donated by a living person, they were able to turn back the clock of the skin cells to turn them into iPS cells. This finding is now the cornerstone of many new stem cell research projects, which has arguably revolutionised the field.

Future treatment potential, but currently regeneration is impractical

There are many ways that stem cells could be used in the future to treat MND, but using them to regenerate motor neurones is not currently a practical solution. But why isn’t this practical? In his talk, Prof Chandran explained…

Crossing wires

The brain is a very complex organ and can be related to a ball of wiring, with each wire being linked to a specific place within the brain and body. If this were to be wired up inaccurately, then it would cause pandemonium in our bodies, with movement instructions meant for our feet to possibly end in our hands, mouth or elbow for example – something we’d definitely not want to happen!

Prof Chandran went onto explain that each neurone has its own ‘postcode’ in the brain, and depending on where it ‘lives’, he explained that its function will vary.

The function of each motor neurone will also intuitively denote what muscle it’s supposed to connect to. The way that our neurones grow toward a muscle is an extremely well orchestrated affair, with chemical messages throughout the body that either attract, or repulse it. However, as our bodies develop in the womb, this system is switched off – meaning that any new motor neurones trying to grow from scratch in the brain will find it near to impossible to know where it’s supposed to go.

It is therefore a very complicated issue to try and regenerate motor neurones in humans to ensure that the motor neurone firstly starts in the right place, and secondly that the neurone has the right instructions in place to guide it toward its target muscle.  However, these aren’t the only issues that researchers face…

Being sure it’s a motor neurone

In our search for using stem cells as a treatment for MND, there is also an issue of making sure that stem cells turn into the cells you want them to be, and Prof Chandran eloquently explained this by using a video of heart cells, generated using stem cell technology and saying that you definitely wouldn’t want these cells beating away in your brain instead of your motor neurones!

But how do researchers turn stem cells into the ‘right’ sort of cell? Prof Chandran explained that this is done quite simply, by giving them the right recipe of chemical ingredients to tell them what to become when they’re older.

Neurones are slow growers

Even if researchers could somehow ensure that ‘new’ motor neurones could be created and would connect to the right ‘postcode’ of the brain, neurones are very slow growing. As some of our motor neurones would have to grow over a metre to reach its target muscle, the amount of time that it would take to regenerate motor neurones would be implausible in terms of using them as a treatment. There just isn’t a way to speed this up at the moment.

For all of the above reasons, this is why stem cells cannot currently be used to regenerate motor neurones as a treatment for MND. However, this is not to say that they don’t have other uses…

Using stem cells to learn more about MND

Stem cells are great tools for recreating diseases in a dish, as they are able to divide to create large numbers of cells and are able to turn (with the right receipe) into any type of cell, such as a motor neurones.

In his laboratory, Prof Chandran’s research group have created living human motor neurones grown in a dish from skin cells donated by people with an inherited form of MND using iPS cell technology. In his presentation, he showed us that within 100 days, his laboratory is able to create a billion (10¹², referred to as a trillion in USA) cells from a stem cell. He has also shown that these motor neurones generated from stem cells connect to muscle cells and are electrically active – which means that to all intents and purposes, they are real motor neurones.

He then explained that his MND Association funded project is creating these motor neurones and support cells from a skin biopsy of somebody with MND with faults in a gene called TDP-43. They can then use these new cells as a tool to investigate the disease process and hopefully in the future to test the effectiveness of therapies in this model.

Realising the potential of stem cells

As well as using stem cells to create new models in the laboratory, to discover new medicines, stem cells could potentially be used in a different way to treat the disease. These treatments would not aim to regenerate the motor neurones, but instead would attempt to slow down, or even stop the disease.

Realistically, researchers could use neurone support cells to provide a protective environment to lasting motor neurones – in fact, there are plans in place to test such a treatment which is estimated to being enrolling in 2014 (see stem cell conference blog article for more information).

Overall, Prof Chandran’s talk was extremely well received with delegates commenting to us that “Prof Chandran was the best speaker I can recall” and Prof Chandran’s talk was: “clear, hopeful, excellent. He inspired confidence and spoke in language I could understand”

We’re pleased that so many people who attended our AGM and Annual Conference enjoyed Prof Chandran’s talk, with 91.2% of delegates saying that it was “excellent” (from our survey of 57 people who attended).

Find out more about stem cells on our website.

Stay up-to-date with news on our next conferences by following our conference team on Twitter @mndconference