Curriculum Vitae s328

WIMM PI Curriculum Vitae

Personal Data Name David Beeson Nationality UK Email [email protected]

Present Position Senior Research Fellow, Head of the Neurosciences Group, Weatherall Institute of Molecular Medicine, Nuffield Department of Clinical Neuroscience,

2004 - present Professor in Neuroscience (personal chair) Nuffield Department of Clinical Neuroscience, University of Oxford

1995 - present Department of Clinical Neurology, University of Oxford Honorary University Research Lecturer

Previous Positions

2003 - 2008 MRC Senior Non-Clinical Fellow (Renewal by MRC) Neurosciences Group, Weatherall Institute of Molecular Medicine, Department of Clinical Neurology, University of Oxford

1998 - 2003 MRC Senior Non-Clinical Fellow (MRC) Neurosciences Group, Weatherall Institute of Molecular Medicine, Department of Clinical Neurology, University of Oxford

1988 – 1998 Senior Research Fellow (Muscular Dystrophy Group) Neurosciences Group, Institute of Molecular Medicine, Department of Clinical Neurology, University of Oxford

1985 – 1988 Research Fellow (Muscular Dystrophy Group) Department of Neurology, Royal Free Hospital, University of London.

1984 - 1985 Research Assistant (G.D. Searle) G.D.Searle Research Laboratories, High Wycombe Research Achievements

Over the past 25 years my work has focused on disorders of neuromuscular transmission. We have used molecular genetic techniques combined with electrophysiology to uncover the underlying mechanisms of disease in a series of related genetic syndromes and translated this understanding to the clinic to provide, in many cases, remarkable and dramatic improvements in health.

In the early adoption of molecular genetics we were the first to isolate cDNA and genomic clones for all five human AChR subunits. Additional key molecules at the neuromuscular junction were also cloned and characterised. The cloning work provided the data for mutational screening of candidate genes and the materials for functional studies to demonstrate mutation pathogenicity and investigate underlying disease mechanisms. We have focussed on inherited (congenital) myasthenic syndromes (CMS). Numerous mutations were identified in the AChR subunit genes, while functional analysis showed both the direct correlation of mutations with disease and a diverse series of pathogenic molecular mechanisms. The work also provided proof-of-principle of allele-specific-silencing using RNAi.

We have gone on to define and characterise pathogenic molecular mechanisms for mutations in a series of genes that govern the formation of the neuromuscular synapse. These include showing mutations in Dok-7 underlie a major form of CMS. Recently we identified mutations in DPAGT1, GFPT1, ALG2 and ALG14 as causes of CMS. These genes all code for proteins that are involved in the early stages of N-linked glycosylation and thus, we have identified a new pathway where mutations cause disorders of synaptic transmission.

The success of this program of research on CMS led the National Commissioning Group of the NHS to commission the team in Oxford to provide a national specialist service for CMS. The combination of clinical service and research laboratory provide a prime example of translational research of bedside to bed and back, with the research generating data that can be used directly to make a striking difference to patient well-being.

What are the Future Aims of Your Current Group?

We are able to locate mutations in about 80% of patients with clinically definite CMS (19 genes). We would like to identify the new genes where mutations are located for the remaining 20%. The newly available next generation sequencing techniques are proving invaluable in this work – we have already identified 3 new CMS-associated genes and expect to find a number more. An ongoing aim will be to determine the molecular mechanism of how mutations in these newly identified genes impair neuromuscular transmission, which in turn provides a rational basis for therapy. For example, we now wish to determine how mutations in ubiquitously expressed proteins in the N-linked glycosylation pathway causes a disorder with symptoms restricted to impaired neuromuscular transmission.

At present we subdivide patients into those where the signal transmission across the synapse is directly affected and those where the synaptic structure and stability are affected. In the latter category, treatment with 2-adrenergic receptor agonists (that we have pioneered), has a remarkable beneficial effect, but we do not understand why. We aim to find out the mechanism for this action, with the potential for further optimization of treatment. Part of this work will be to look at the stability of various synaptic membrane proteins and we are keen to apply (with Christian Eggeling) high resolution microscopy techniques in order to visualize synaptic components. For example, we wish to visualize AChR mobility when its anchoring protein, RAPSN, harbours mutations. Finally, we have built a series of animal models that reflect the human disorders. These include models in which the AChR is tagged with EGFP. We would aim to use these models to: i) study beneficial effects of potential new treatments, both new drugs and gene therapies; and ii) to visualize in vivo the changes to the neuromuscular junction that occur during disease progression.

How do These Aims Contribute to the Understanding and/or Management of Human Disease

Our work is closely integrated with the clinic and forms part of the service for the National Referral Centre for the Congenital Myasthenic Syndromes that is jointly based at the West Wing at The John Radcliffe, the genetics laboratories at the Churchill, and the Weatherall Institute. We offer a service from bedside to bench and back, translating the work in the laboratory directly back to patient care.

Over the past 20 years we have defined molecular mechanisms of disease for over 400 different mutations present in 19 different genes. Within a gene it is possible to have mutations that cause disease in at least 5 different ways, each of which requires a different treatment regime. Understanding of the disease mechanism is fed back to the clinicians who can then institute appropriate therapy. It is not yet possible to predict the functional effect of many of the missense mutations we detect, and therefore analysis of the mutations is crucial for the clinicians. Therapy is very much tailored to the individual with six different drugs given in different combinations depending upon which of 19 genes is mutated and underlying molecular mechanism. Optimisation of therapy can lead to dramatic and sustained improvement in patient quality of life, with many children who first come to clinic in a wheel chair able to run or walk about within six months, and many are able to go on to lead a near normal life.

In addition to tailored therapy, the work provides the basis for the standard advice for mendelian disorders of genetic counseling, prenatal diagnosis, and disease prognosis. On a larger scale, what is learnt about synaptic dysfunction and synaptic stability is likely to have implications for a number of disorders in the central nervous system, such as for certain autistic spectrum disorders.

Lay Summary of Research

Muscles contract in response to signals from the brain. Crucial sites for this process are the points where the nerves meet the muscles. These specialised sites, known as neuromuscular junctions, have evolved to allow rapid transfer of signals from nerves to muscles. A group of muscle diseases termed ‘myasthenic disorders’ are caused by defective transmission at the neuromuscular junction. The myasthenic disorders have muscle weakness that can be life-threatening in severe cases, and have a characteristic ‘fatiguability’ whereby the more a muscle is exercised the weaker it becomes. They are caused either through inheriting faulty genes or through a defective immune system where the patient’s own antibodies attack the neuromuscular junction. Our work focuses on inherited defects of nerve-muscle signalling. We have been able to identify mutations in at least 15 different genes that cause myasthenic disorders and believe that there are several more that have not yet been located. Once a mutation is identified we aim to find out how it affects signal transmission; it could generate too much signal, too little signal, or cause structures at the junction to become unstable. Once we identify what is defective, we can feed the information back to clinicians, who can tailor treatment for the individual patient and prescribe appropriate drug therapies. In many cases with the right therapies we can achieve a dramatic improvement in patient quality of life. However, in some cases treatment leads to only modest improvement, and for these cases we aim to investigate new therapies in model systems in the laboratory. We are particularly interested in drug treatments that might stabilise the structures of the neuromuscular junction since these may have implications for treating brain disorders where there are learning difficulties or memory loss. All Publications Over the Past 5 Years

Rodríguez Cruz PM, Sewry C, Beeson D, Jayawant S, Squier W, McWilliam R, Palace J. Congenital myopathies with secondary neuromuscular transmission defects; A case report and review of the literature. Neuromuscul Disord. 2014 Jul 30. pii: S0960-8966(14)00613-0. doi: 10.1016/j.nmd.2014.07.005. [Epub ahead of print]

Illingworth MA, Main M, Pitt M, Feng L, Sewry CA, Gunny R, Vorstman E, Beeson D, Manzur A, Muntoni F, Robb SA. RYR1-related congenital myopathy with fatigable weakness, responding to pyridostigimine. Neuromuscul Disord. 2014 Aug;24(8):707-12. doi: 10.1016/j.nmd.2014.05.003. Epub 2014 May 23

Parr JR, Andrew MJ, Finnis M, Beeson D, Vincent A, Jayawant S. How common is childhood myasthenia? The UK incidence and prevalence of autoimmune and congenital myasthenia. Arch Dis Child. 2014 Jun;99(6):539-42. doi: 10.1136/archdischild-2013-304788. Epub 2014 Feb 5.

Webster R, Liu WW, Chaouch A, Lochmüller H, Beeson D. Fast-channel congenital myasthenic syndrome with a novel acetylcholine receptor mutation at the α-ε subunit interface. Neuromuscul Disord. 2014 Feb;24(2):143-7. doi: 10.1016/j.nmd.2013.10.009. Epub 2013 Nov 6.

Koneczny I, Cossins J, Waters P, Beeson D, Vincent A. MuSK myasthenia gravis IgG4 disrupts the interaction of LRP4 with MuSK but both IgG4 and IgG1-3 can disperse preformed agrin-independent AChR clusters. PLoS One. 2013 Nov 7;8(11):e80695. doi: 10.1371/journal.pone.0080695. eCollection 201

Klein A, Pitt MC, McHugh JC, Niks EH, Sewry CA, Phadke R, Feng L, Manzur AY, Tirupathi S, Devile C, Jayawant S, Finlayson S, Palace J, Muntoni F, Beeson D, Robb SA. DOK7 congenital myasthenic syndrome in childhood: Early diagnostic clues in 23 children. Neuromuscul Disord. 2013 Jul 3. doi:pii: S0960-8966(13)00171-5. 10.1016/j.nmd.2013.06.002. [Epub ahead of print] Webster RG, Cossins J, Lashley D, Maxwell S, Liu WW, Wickens JR, Martinez-Martinez P, de Baets M, Beeson D. A mouse model of the slow channel myasthenic syndrome: Neuromuscular physiology and effects of ephedrine treatment. J. Exp Neurol. 2013 Jun 21. doi:pii: S0014-4886(13) 00185-4.10.1016/j.expneurol.2013.06.012. [Epub ahead of print] Basiri K, Belaya K, Liu WW, Maxwell S, Sedghi M, Beeson D. Clinical features in a large Iranian family with a limb-girdle congenital myasthenic syndrome due to a mutation in DPAGT1. Neuromuscul Disord. 2013; 23:469-72. Zoltowska K, Webster R, Muller J, Maxwell S, Cossins J, Lochmuller H, Beeson D. Mutations in GFPT1 that underlie limb-girdle congenital myasthenic syndrome result in reduced cell-surface expression of muscle AChR. Hum Mol Genet. 2013; 22(14):2905- 13. Finlayson S, Palace J, Belaya K, Walls T, Burke G, Holton J, Pascual Pascual S, Cossins J Beeson D. Clinical features of congenital myasthenic syndrome due to mutations in DPAGT1 J Neurol Neurosurg Psychiatry 2013 Feb 27. [Epub ahead of print] Cossins J, Belaya K, Hicks D, Salih M, Finlayson S,Carboni N, Liu WW, Maxwell S, Zoltowska K, Golara Farsani G, Laval S, Seidhamed M, "WGS500 consortium", Donnelly P, Bentley D, McGowan S, Müller J, Palace J, Lochmüller H, Beeson D. Congenital myasthenic syndromes due to mutations in ALG2 and ALG14. Brain 2013 Mar;136(Pt 3):944-56. Burke G, Hiscock A, Klein A, Niks E, Mansur A, Ng J, de Vile C, Beeson D, Muntoni F and Robb, S. (2012). Salbutamol benefits children with congenital myasthenic syndrome due to DOK7 mutations. Neuromuscul. Disord. 2013 Feb;23(2):170-5. Finlayson S, Beeson D, Palace J. Congenital myasthenic syndromes: An update. Pract Neurol 2013 Apr;13(2):80-91. Finlayson S, Spillane J, Kullmann DM, Howard R, Webster R, Palace J, Beeson D. Slow channel congenital myasthenic syndrome responsive to a combination of fluoxetine and salbutamol. Muscle Nerve. 2013 Feb;47(2):279-82. Cossins J, Belaya K, Zoltowska K, Koneczny I, Maxwell S, Jacobson L, Leite MI, Waters P, Vincent A, Beeson D. The search for new antigenic targets in myasthenia gravis. Ann N Y Acad Sci. 2012 Dec;1275(1):123-8. Beeson D. Synaptic dysfunction in congenital myasthenic syndromes. Ann N Y Acad Sci. 2012 Dec;1275(1):63-9. Belaya K, Finlayson S, Cossins J, Liu WW, Maxwell S, Palace J, Beeson D. Identification of DPAGT1 as a new gene in which mutations cause a congenital myasthenic syndrome. Ann N Y Acad Sci. 2012 Dec;1275(1):29-35. Vincent A, Waters P, Leite MI, Jacobson L, Koneczny I, Cossins J, Beeson D. Antibodies identified by cell-based assays in myasthenia gravis and associated diseases. Ann N Y Acad Sci. 2012 Dec;1274(1):92-8. Belaya K, Finlayson S, Slater CR, Cossins J, Liu WW, Maxwell S, McGowan SJ, Maslau S, Twigg SR, Walls TJ, Pascual Pascual SI, Palace J, Beeson D.Mutations in DPAGT1 cause a limb-girdle congenital myasthenic syndrome with tubular aggregates. Am J Hum Genet. 2012; 91:193-201. Webster R, Maxwell S, Spearman H, Tai K, Beckstein O, Sansom M, Beeson D. A novel congenital myasthenic syndrome due to decreased acetylcholine receptor ion-channel conductance. Brain. 2012; 135:1070-1080. Chaouch A, Beeson D, Hantaï D, Lochmüller H. 186th ENMC International Workshop: Congenital myasthenic syndromes 24-26 June 2011, Naarden, The Netherlands. Neuromuscul Disord. 2012; 22:566-576 Maselli RA, Fernandez JM, Arredondo J, Navarro C, Ngo M, Beeson D, Cagney O, Williams DC, Wollmann RL, Yarov-Yarovoy V, Ferns MJ. LG2 agrin mutation causing severe congenital myasthenic syndrome mimics functional characteristics of non-neural (z-) agrin. Hum Genet. 2012; 131:1123-1135. Guergueltcheva V, Müller JS, Dusl M, Senderek J, Oldfors A, Lindbergh C, Maxwell S, Colomer J, Mallebrera CJ, Nascimento A, Vilchez JJ, Muelas N, Kirschner J, Nafissi S, Kariminejad A, Nilipour Y, Bozorgmehr B, Najmabadi H, Rodolico C, Sieb JP, Schlotter B, Schoser B, Herrmann R, Voit T, Steinlein OK, Najafi A, Urtizberea A, Soler DM, Muntoni F, Hanna MG, Chaouch A, Straub V, Bushby K, Palace J, Beeson D, Abicht A, Lochmüller H. Congenital myasthenic syndrome with tubular aggregates caused by GFPT1 mutations. J Neurol. 2012; 259:838-850. Palace J, Lashley D, Bailey S, Jayawant S, Carr A, McConville J, Robb S, Beeson D. Clinical features in a series of fast channel congenital myasthenia syndrome. Neuromuscul Disord. 2011; 22:112-117. Senderek J, Müller JS, Dusl M, Strom TM, Guergueltcheva V, Diepolder I, Laval SH, Maxwell S, Cossins J, Krause S, Muelas N, Vilchez JJ, Colomer J, Mallebrera CJ, Nascimento A, Nafissi S, Kariminejad A, Nilipour Y, Bozorgmehr B, Najmabadi H, Rodolico C, Sieb JP, Steinlein OK, Schlotter B, Schoser B, Kirschner J, Herrmann R, Voit T, Oldfors A, Lindbergh C, Urtizberea A, von der Hagen M, Hübner A, Palace J, Bushby K, Straub V, Beeson D, Abicht A, Lochmüller H. Hexosamine biosynthetic pathway mutations cause neuromuscular transmission defect. Am J Hum Genet. 2011;88:162- 172 Robb SA, Sewry CA, Dowling JJ, Feng L, Cullup T, Lillis S, Abbs S, Lees MM, Laporte J, Manzur AY, Knight RK, Mills KR, Pike MG, Kress W, Beeson D, Jungbluth H, Pitt MC, Muntoni F. Impaired neuromuscular transmission and response to acetylcholinesterase inhibitors in centronuclear myopathies. Neuromuscul Disord. 2011; 21:379-386. Salih MA, Oystreck DT, Al-Faky YH, Kabiraj M, Omer MI, Subahi EM, Beeson D, Abu- Amero KK, Bosley TM. Congenital myasthenic syndrome due to homozygous CHRNE mutations: report of patients in Arabia. J Neuroophthalmol. 2011;31:42-47 Yang L, Maxwell S, Leite MI, Waters P, Clover L, Fan X, Zhang D, Yang C, Beeson D, Vincent A. Non-radioactive serological diagnosis of myasthenia gravis and clinical features of patients from Tianjin, China. J Neurol Sci. 2011;301:71-76. Munot P, Lashley D, Jungbluth H, Feng L, Pitt M, Robb SA, Palace J, Jayawant S, Kennet R, Beeson D, Cullup T, Abbs S, Laing N, Sewry C, Muntoni F Congenital fibre type disproportion associated with mutations in the tropomyosin 3 (TPM3) gene mimicking congenital myasthenia. Neuromuscul Disord. 2010; 20:796-800. Nicholl DJ, Hilton-Jones D, Palace J, Richmond S, Finlayson S, Winer J, Weir A, Maddison P, Fletcher N, Sussman J, Silver N, Nixon J, Kullmann D, Embleton N, Beeson D, Farrugia ME, Hill M, McDermott C, Llewelyn G, Leonard J, Morris M. Open letter to prime minister David Cameron and health secretary Andrew Lansley. BMJ. 2010; 341:c6466. doi: 10.1136/bmj.c6466. Munot P, Lashley D, Jungbluth H, Feng L, Pitt M, Robb SA, Palace J, Jayawant S, Kennet R, Beeson D, Cullup T, Abbs S, Laing N, Sewry C, Muntoni F. Congenital fibre type disproportion associated with mutations in the tropomyosin 3 (TPM3) gene mimicking congenital myasthenia. Neuromuscul Disord. 2010; 20:796-800. Clark RH, McTaggart JS, Webster R, Mannikko R, Iberl M, Sim XL, Rorsman P, Glitsch M, Beeson D, Ashcroft FM. Muscle dysfunction caused by a KATP channel mutation in neonatal diabetes is neuronal in origin. Science. 2010;329:458-461. Jephson CG, Mills NA, Pitt MC, Beeson D, Aloysius A, Muntoni F, Robb SA, Bailey CM.Congenital stridor with feeding difficulty as a presenting symptom of Dok7 congenital myasthenic syndrome. Int J Pediatr Otorhinolaryngol. 2010;74:991-994. Spillane J, Beeson D, Kullmann DM Myasthenia and related disorders of the neuromuscular junction. J Neurol Neurosurg Psychiatry. 2010; 81:850-857. Irani SR, Bera K, Waters P, Zuliani L, Maxwell S, Zandi MS, Friese MA, Galea I, Kullmann DM, Beeson D, Lang B, Bien CG, Vincent A N-methyl-D-aspartate antibody encephalitis: temporal progression of clinical and paraclinical observations in a predominantly non- paraneoplastic disorder of both sexes. Brain. 2010;133:1655-1667 Lashley D, Palace J, Jayawant S, Robb S, Beeson D. Ephedrine treatment in congenital myasthenic syndrome due to mutations in DOK7 . Neurology 2010;74:1517-23. Gattenlöhner S, Jörissen H, Huhn M, Vincent A, Beeson D, Tzartos S, Mamalaki A, Etschmann B, Muller-Hermelink HK, Koscielniak E, Barth S, Marx A. A human recombinant autoantibody-based immunotoxin specific for the fetal acetylcholine receptor inhibits rhabdomyosarcoma growth in vitro and in a murine transplantation model. J Biomed Biotechnol. 2010;2010:187621. Burke G, Allen D, Arunachalam R, Beeson D, Hammans S.A treatable muscle disease. Pract Neurol. 2009;9:233-236. Faber CG, Molenaar PC, Vles JS, Bonifati DM, Verschuuren JJ, van Doorn PA, Kuks JB, Wokke JH, Beeson D, De Baets M. AChR deficiency due to epsilon-subunit mutations: two common mutations in the Netherlands. J Neurol. 2009; 256:1719-23. Vogt J, Morgan V, Marton T, Maxwell S, Harrison B, Beeson D, Maher. ER.Germline mutation in DOK7 associated with Foetal Akinesia Deformation Sequence. J Med Genet. 2009; 46:338-40. Ten key publications throughout your career Cossins J, Belaya K, Hicks D, Salih M, Finlayson S,Carboni N, Liu WW, Maxwell S, Zoltowska K, Golara Farsani G, Laval S, Seidhamed M, "WGS500 consortium", Donnelly P, Bentley D, McGowan S, Müller J, Palace J, Lochmüller H, Beeson D. Congenital myasthenic syndromes due to mutations in ALG2 and ALG14. Brain 2013 Mar;136(Pt 3):944-56. Belaya K, Finlayson S, Slater CR, Cossins J, Liu WW, Maxwell S, McGowan SJ, Maslau S, Twigg SR, Walls TJ, Pascual Pascual SI, Palace J, Beeson D. Mutations in DPAGT1 cause a limb-girdle congenital myasthenic syndrome with tubular aggregates. Am J Hum Genet. 2012 ;91:193-201. Webster R, Maxwell S, Spearman H, Tai K, Beckstein O, Sansom M, Beeson D. A novel congenital myasthenic syndrome due to decreased acetylcholine receptor ion-channel conductance. Brain. 2012 ;135:1070-80. Clark RH, McTaggart JS, Webster R, Mannikko R, Iberl M, Sim XL, Rorsman P, Glitsch M, Beeson D, Ashcroft FM. Muscle dysfunction caused by a KATP channel mutation in neonatal diabetes is neuronal in origin. Science. 2010;329:458-61. Giraud M, Taubert R, Vandiedonck C, Ke X, Levi-Strauss M, Pagani F, Baralle FE, Eymard B, Tranchant C, Gajdos P, Vincent A, Willcox N, Beeson D, Kyewski B, Garchon HJ. An IRF8-binding promoter variant and AIRE control CHRNA1 promiscuous expression in thymus. Nature. 2007; 448:934-7 Palace J, Lashley D, Newsom-Davis J, Cossins J, Maxwell S, Kennett R, Jayawant S, Yamanashi Y, Beeson D. Clinical features of the DOK7 neuromuscular junction synaptopathy. Brain 2007; 130:1507-1515 Beeson D, Higuchi O, Palace J, Cossins J, Spearman H, Maxwell S, Newsom-Davis J, Burke G, Fawcett P, Motomura M, Muller J, Lochmuller H, Slater C, Vincent A, Yamanashi Y. Dok-7 mutations underlie a neuromuscular junction synaptopathy. Science 2006; 313:1975-1978. Abdelghany A, Wood M, Beeson D. Allele-specific gene silencing of a pathogenic mutant muscle acetylcholine receptor subunit by RNA interference. Hum Mol Genet. 2003; 12:2637-2644. Croxen R, Young C, Slater C, Haslam S, Brydson M, Vincent A and Beeson D. Endplate - and -subunit mRNA levels in AChR deficiency syndrome due to -subunit null mutations. Brain 2001; 124:1362-1372. Beeson D, Morris A, Vincent A and Newsom-Davis J. The human muscle acetylcholinereceptor  subunit exists as two isoforms: A novel exon. EMBO J 1990;9:2101-2106

Current Research Support

Medical Research Council Programme Grant MR/M006824/1 Disease mechanisms and therapies for inherited disorders of the neuromuscular synapse £1,500,000 1.10.14 - 30.09.19

Myasthenia Gravis association/John Moulton Charitable Foundation Genes, mechanisms and models of congenital myasthenic syndromes £80,000 1.10.13-30.9.14

John Moulton Charitable Foundation Defining defective acetylcholine receptor function for definitive diagnosis and appropriate treatments of congenital myasthenic syndromes £90,000 1.10.11 – 30.09.14 Muscular Dystrophy Campaign Prize Studentship Therapy to stabilise synaptic structure for both genetic and autoimmune forms of myasthenia £95,000 1.10.2011-30.09.2014

Other Funding Source Named applicant and member of the management committee for OXION: Ion Channels and Disease Initiative strategic award by the Wellcome Trust ~ £5M 1.10.11 - 30.09.17

Based on the research output from the laboratory the National Specialist Commissioning Service (NSC) of the NHS commissioned and provides funding for a National Service for the treatment and genetic diagnosis of patients with congenital myasthenic syndromes based at the John Radcliffe Hospital, in Oxford. ~ £90,000/year for the laboratory