James Briscoe & Christopher Shaw

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Neurodevelopmental characterisation of a novel cytoplasmic RNA binding protein mutated in smyotrophic lateral sclerosis

Joint Crick/King's College London position

Amyotrophic Lateral Sclerosis (ALS) causes motor neurons in the spinal cord, brainstem, and motor cortex to degenerate leading to progressive muscular paralysis. Disabilities accumulate relentlessly until patients are unable to move their limbs, eat or speak and death from respiratory failure usually occurs within 3-5 years of symptom onset. The tragedy of this illness sparked a viral fundraising effort 'The Ice Bucket Challenge'.

We have identified 10 different novel mutations in an RNA binding protein in 24 familial and sporadic ALS cases. One mutation appears in 12 individuals and another in four individuals providing powerful proof that these mutations are pathogenic. The few publications that exist on this gene report that it has putative RNA binding domains. The position and nature of the mutations have strong parallels with two other RNA binding proteins that we previously demonstrated to be pathogenic in ALS, TDP-43 (Sreedharan, 2008) and FUS (Vance, 2009). The discovery of mutations in these genes placed abnormal RNA processing centre stage as a disease mechanism in Neurodegeneration. Both of these gene discoveries were published in the journal Sciencewith past PhD students as first authors and each have >1,000 citations.

Very little is known about this new ALS gene but there are important differences from other ALS-linked genes. While TDP-43 and FUS are ubiquitously expressed the new gene is expressed within neurons in very specific sites within the nervous system. It also has a very interesting subcellular localisation. While TDP-43 and FUS are predominantly nuclear and are involved in gene splicing the new gene is largely cytoplasmic and is likely to have a different role in RNA processing. We now know that vital aspects of neuronal RNA processing occur within axons and dendrites but the normal function of this protein and the mechanisms by which mutations cause motor neuron degeneration are unknown.

The candidate will have an opportunity to join two world's leading laboratories; studying ALS-genetics (Professor Shaw) and motor neuron development (Dr Briscoe). This discovery is unpublished at this stage and the successful candidate will be in a unique position to map the expression of this protein during central nervous system development and define its normal function(s) in neurons. They will define the regulatory elements of this gene in order to understand the specificity of its expression. They will also explore the pathological effects of mutations in a range of patient-derived induced pluripotent stem cell lines and transgenic mouse models. The student will learn cutting-edge tools of cellular and molecular biology including gene cloning, mutagenesis and genome editing with CRISPR/Cas9 to generate and analyse mouse and human stem cells, their differentiation and the effect of gene knockout, functional deletions and ALS-disease specific mutations.

1. Sreedharan, J., Blair, I. P., Tripathi, V. B., Hu, X., Vance, C., Rogelj, B., Ackerley, S., Durnall, J. C., Williams, K. L., Buratti, E., Baralle, F., de Belleroche, J., Mitchell, J. D., Leigh, P. N., Al-Chalabi, A., Miller, C. C., Nicholson, G. and Shaw, C. E. (2008)
TDP-43 mutations in familial and sporadic amyotrophic lateral sclerosis.
Science 319: 1668-1672. PubMed abstract

2. Vance, C., Rogelj, B., Hortobágyi, T., De Vos, K. J., Nishimura, A. L., Sreedharan, J., Hu, X., Smith, B., Ruddy, D., Wright, P., Ganesalingam, J., Williams, K. L., Tripathi, V., Al-Saraj, S., Al-Chalabi, A., Leigh, P. N., Blair, I. P., Nicholson, G., de Belleroche, J., Gallo, J.-M., Miller, C. C. and Shaw, C. E. (2009)
Mutations in FUS, an RNA processing protein, cause familial amyotrophic lateral sclerosis type 6.
Science 323: 1208-1211. PubMed abstract

3. Smith, B. N., Topp, S. D., Fallini, C., Shibata, H., Chen, H.-J., Troakes, C., King, A., Ticozzi, N., Kenna, K. P., Soragia-Gkazi, A., Miller, J. W., Sato, A., Dias, D. M., Jeon, M., Vance, C., Wong, C. H., de Majo, M., Kattuah, W., Mitchell, J. C., Scotter, E. L., Parkin, N. W., Sapp, P. C., Nolan, M., Nestor, P. J., Simpson, M., Weale, M., Lek, M., Baas, F., Vianney de Jong, J. M., Ten Asbroek, A. L. M. A., Redondo, A. G., Esteban-Pérez, J., Tiloca, C., Verde, F., Duga, S., Leigh, N., Pall, H., Morrison, K. E., Al-Chalabi, A., Shaw, P. J., Kirby, J., Turner, M. R., Talbot, K., Hardiman, O., Glass, J. D., de Belleroche, J., Maki, M., Moss, S. E., Miller, C., Gellera, C., Ratti, A., Al-Sarraj, S., Brown, R. H., Jr., Silani, V., Landers, J. E. and Shaw, C. E. (2017)
Mutations in the vesicular trafficking protein annexin A11 are associated with amyotrophic lateral sclerosis.
Science Translational Medicine 9: eaad9157. PubMed abstract

4. Gouti, M., Tsakiridis, A., Wymeersch, F. J., Huang, Y., Kleinjung, J., Wilson, V. and Briscoe, J. (2014)
In vitro generation of neuromesodermal progenitors reveals distinct roles for Wnt signalling in the specification of spinal cord and paraxial mesoderm identity.
PLOS Biology 12: e1001937. PubMed abstract

5. Kicheva, A., Bollenbach, T., Ribeiro, A., Valle, H. P., Lovell-Badge, R., Episkopou, V. and Briscoe, J. (2014)
Coordination of progenitor specification and growth in mouse and chick spinal cord.
Science 345: 1254927. PubMed abstract