1. MRC Centre for Neuromuscular Diseases Biobank, London Dr Rahul Phadke Consultant Neuropathologist NHNN, UCL Institute of Neurology & Dubowitz Neuromuscular Centre, GOSH

2. Why a neuromuscular biobank? Rarity of neuromuscular disorders Requirement for access to tissue and cells to - Improve understanding pathophysiology of NMDs - Proof of principle studies regarding novel therapeutic applications - Recruitment of patients in therapeutic trials

3. Why a biobank in the MRC Centre? Three large clinical and pathological services London ION/QSH; London ICH/GOSH; Newcastle ~500 muscle biopsy samples processed every year ~ 100 nerve biopsies Four Nationally Commissioned services, reflecting clinical and research expertise of the groups involved a. ION Channelopathies b. ION + Newcastle Mitochondrial (+ Oxford) c. ICH Congenital muscular dystrophies and myopathies d. Newcastle Limb Girdle Muscular Dystrophies

4. Link basic scientists, clinical investigators and pathologists MRC Biobank

5. Provide an opportunity to train research and clinical scientists from bench to bedside Facilitate links of the MRC centre with other UK institutions Platform for interacting with similar European initiatives, i.e. Eurobiobank/ Treat-NMD MRC Biobank

6. Neuromuscular Biobank – achievements Establishment of biobank at 2 sites (Diana Johnson, Mojgan Reza) Full member of EuroBioBank network and open catalogue More than 2600 NMD samples collected so far (original target: 700) More than 100 different neuromuscular pathologies Samples distributed to more than 65 scientists (centre and external) More than 15 research publications acknowledging biobank Several successful grant proposals with strong biobank element

8. Neuromuscular Biobank – plans Integrate additional sites in the UK in biobank-related activities Additional services: immortalization of primary cell lines or iPS Teaching young researchers in the best use of these biomaterials Increase in research publications and grant opportunities Increased use in biomarker, natural history and therapeutic studies Collaboration with industry, development of fee for service structure Integration in national and international biobanking initiatives Support translational research

9. MRC CNMD Biobank London  One of two Biobanks supported by the MRC Centre for Neuromuscular Diseases  10 years of funding by the MRC (2008 - 2018)  Biobank of tissue (muscle, skin), primary cells (myoblasts, fibroblasts, synovial cells, peripheral blood cells) and biological samples (plasma, serum, DNA, urine) from patients with neuromuscular disease  HTA licensed

10. Access to patient samples  Links with paediatric and adult pathology and clinical services St Mary’s Hospital PI: Mr D Hunt Royal National Orthopaedic Hospital NHS Trust PI: Mr J Lehovsky Guy’s and St Thomas NHS Trust* PAEDIATRIC Muscle Biopsy Service Neuropathology Institute of Neurology ADULT Neuropathology Institute of Neurology MRC CNMD Biobank London Institute of Child Health, London Biobank technician: Diana Johnson * proposed RJAH Orthopaedic Hospital, Oswestry* Great Ormond Street Hospital for Children PI: Prof. F Muntoni National Hospital for Neurology and Neurosurgery PI: Prof M Hanna

11. Institute of Neurology Director Prof Mike Hanna MRC Centre for Neuromuscular Disease Director Prof Mike Hanna Neuropathology Dubowitz Neuromuscular Centre Director Prof Francesco Muntoni Biobank Lead Prof Francesco Muntoni Division of Neuropathology Chair Prof Sebastian Brandner Biobank Technician Ms Diana Johnson Muscle biopsy service Prof Caroline Sewry Dr Rahul Phadke Dr Lucy Feng Nisha Bhardway Darren Chambers Management Structure

12. Inclusion criteria:  Patients with a suspected or confirmed neuromuscular disease  Controls - patients undergoing orthopaedic surgery, usually cerebral palsy or AIS patients

13. How we collect samples  Muscle – diagnostic biopsy or orthopaedic surgery  Skin – diagnostic biopsy, orthopaedic surgery or specifically for research in an outpatient setting  Blood and urine – specifically for research in an outpatient setting

14. Samples stored (to 19/02/2013) Sample typeQuantity stored Myoblasts578 Fibroblasts558 DNA/Plasma/Serum171 PBLs/PBMCs78 Muscle/skin tissue16 Urine16 Synovial cells10 TOTAL1427 Additionally, there is access to thousands of diagnostic muscle tissue biopsies stored with the Paediatric Muscle Biopsy Service at ION, where consent permits

15. Diagnosis

16. Quality assurance  All cells are mycoplasma tested prior to release  All myoblasts are stained for desmin and differentiated to determine purity and ability to fuse into myotubes  All diagnostic muscle specimens have had routine immunohistochemical staining  Diagnosis is crosschecked against genetic, clinical and pathology records

17. Myoblasts Desmin / DAPI 7796 DMD del 42-43 96.5% purity, 85% fusion 7915 Minicore myopathy RYR1 98% purity, 80% fusion 120/258 tested are considered myogenic (≥20% pure, ≥10% fusion) 71 of these are considered very myogenic (≥70% pure, ≥30% fusion)

18. Using patient cells to validate experimental therapies: compounds that upregulate alpha dystroglycan glycosylation The six compounds with a positive outcome in H2K myoblasts are currently being assessed in dystroglycanopathy patients with different genotypes Biobank – achievements and plans

19. Using patient fibroblasts to assess novel genes involved in dystroglycanopathies 2 separate families Mild “MEB-like” phenotype High CK Eye involvement in one (severe myopia) Stirking cerebellar cysts One child is ambulant Control Patient Biobank – achievements and plans

20. Using patient myoblasts to assess novel genes involved in congenital myasthenia Biobank – achievements and plans Limb girdle weakness Decrement on repetitive stimulation No ophthalmoplegia, no ptosis Benefit from esterase inhibitors Tubular aggregates MRC grant awarded 2011

21. Using patient myoblasts to unravel pathomechanism in reversible COX myopathy Biobank – achievements and plans Maternal inheritance Floppy infants Spontaneous recovery Incomplete penetrance MRC grant awarded 2011 (Rita Horvath) 3 month 9 monthMum 14674 tRNA GLU Discriminator base

23. Thank you For all queries please contact: Diana Johnson diana.johnson@ucl.ac.uk