Therapeutic and Research Potential of Human Stem Cells

1. Therapeutic and Research Potential of Human Stem Cells Dr Stephen L Minger Director, Stem Cell Biology Laboratory Wolfson Centre for Age-Related Diseases King’s College London Senior Editor – Regenerative Medicine Stem Cell Expert – UK Gene Therapy Advisory Committee (GTAC)

2. Parkinson’s Disease and the Brain

3. Progressive and sustained improvement in transplant function over ten years in human graft recipient Piccini et al, 1999, Nature Neurosci, 2

4. Stem Cells – Types, Flavours and Sources Pluripotent Multipotent Multipotent Multipotent

5. Adult Neurogenesis Human adult brain contains proliferative neural stem cells in dentate gyrus and lateral ventricle wall (SVZ) that generate new neurons throughout life Eriksson et al, 1998, Nature Med, 4, 1313

6. Positive Enriched Environment Physical Activity (e.g. Running) Antidepressants Caloric Restriction Learning Modulation of Excitation Ischaemia (experimental) Pregnancy (Prolactin) Inflammatory Blockade (NSAIDS) VEGF, G-CSF, NO, SCF Statins (Anti-hypertensives) Lithium Negative Opiates Methamphetamine Seizures Aging Stress (Glucocorticoids) 5-HT Depletion Nitric Oxide Inflammation (Microglial Activation) Interleukin-6 (from activated microglia) Irradiation Nicotine Regulators of Adult Neurogenesis

7. 84-year old woman with progressive vascular disease Stoke 2 weeks prior to death

8. Minger et al, 2007

10. Adult Human SVZ Neural Progenitor Cells

11. Austin Smith, 2003

12. Human Embryo Research in UK • Human Fertilisation and Embryology Authority Created in 1990 • All Reproductive Medicine & Human Embryo Research must be licensed by HFEA • HFEA-Licensed Embryo Research Areas • Treatment of Infertility • Causes of Congenital Diseases • Causes of Miscarriage • Development of more Effective Conception • Improvements in Preimplantation Genetic Diagnosis • Provide Understanding of Human Development • Provide Understanding of Human Disease • Enable Development of Therapies of Human Disease

13. License for Derivation of Human ES cells from UK Human Fertilisation and Embryology Authority Awarded to Drs Susan Pickering, Stephen Minger & Professor Peter Braude in May 2002 – renewed in 2005 for three additional years Derivation from donated embryos with informed consent No financial inducement for donation Cannot create embryos for stem cell derivation All cell lines must be deposited in UK Stem Cell Bank and made freely available to other research groups Cell Nuclear Replacement (Therapeutic) permitted with license Cell Nuclear Replacement (Reproductive) banned with criminal penalties

14. Day 1 Late Day 1 2- cells Day 2 4-cells Day 3 8-cells Early human development in vitro Day 4 Morula Day 4 ICM Blastocyst Day 5 ICM Hatching Day 6 Hatched Late Day 6

15. The inner cell mass - contains pluripotent stem cells

16. A B With Good Karma, Embryos, and Culture Conditions, Human ES Cells Can Be Established At Frequency of ~10% A, B, Cystic Fibrosis Line (CF-1) C HES Cell Line (WT-4) C C

17. Oct-4 b-Tubulin(ectoderm) Albumin (endoderm) Smooth Muscle Actin (mesoderm)

18. Stem Cell Biology Laboratory Therapeutic Targets • CNS Disorders • Spinal Cord (McMahon, Bradbury) - traumatic injury, Multiple Sclerosis • Parkinson’s disease (Duty, Jenner) • Stroke/Vascular Disease (Ballard) – adult neurogenesis • Cardiac Regeneration • Heart Failure/Heart Attack (Mathur/Hirschi & Cardiac Cell Therapy Consortium) • Retinal Regeneration • Macular Degeneration, Retinitis Pigmentosa (Ali, Institute of Ophthalmology) • Endocrine Disorders • Type I Diabetes (Jones, Persuad & King’s Islet Transplant Programme) • Hepatic Regeneration • Hepatic Insufficiency (King’s Liver Transplant Programme) • Joint and Bone Destruction • Cartilage Repair ( De Bari, Dell’Accio) • Bone Replacement (Grigoriadis) • Epithelial Cells • Lung Differentiation (Novartis) • Skin Cells for Genetic Skin Disorders (McGrath) • Stem Cell Bioprocessing • Bioengineering/Scale up (Mason, UCL)

19. Developments Required for Therapeutic Application of Human ES Cells To Parkinson’s Disease – Preclinical • Generation of A9-Mesencephalic Dopaminergic Neural Stem Cells by Genetic Engineering – not A10 • Characterisation of Dopaminergic Progenitor Cells • Testing of A9 Stem Cells in Rat PD Models • Identification of Cell-Surface Proteins Unique to A9 Dopa Stem Cells by Mass Spectroscopy (if required) • Selection of A9 Neural Stem Cells by Sorting • Long-term Safety and Efficacy Testing of A9 Cells in Rodent -> Primate Models of PD

20. Developments Required for Therapeutic Application of Human ES Cells To Parkinson’s Disease Therapy – Clinical Issues • Delivery of Cells for Therapeutic Applications – Good Experience with Foetal Cell Transplantation, Efficacy Proportional to Number of DA Neurons Implanted • Need for Standardised Neurosurgical Protocol • Requirement for Strict Inclusion/Exclusion Criteria for Patient Selection – Reduce Variability between Patients • Generation of Animal-Free, Chemically Defined Culture Conditions -> No Xenoproducts • Generation of Therapeutic-Grade Lines – Requires Specialised GMP-Level Facilities – King’s 2008 • Immune Response – Short term (2-3 yrs) minimal immunosuppression supportive/required for graft survival

21. Somatic Cell Nuclear Replacement (Cloning) Can it work in Humans? Should it be allowed? How can we use this technology for therapy and/or research?

24. Strengths of UK Stem Cell Research • World-class academic research centres • Tight regulation on Assisted Reproduction & Human Embryo Research by HFEA • Government Commitment to Stem Cell Research (~£45 million – 2003-2004); £350-800 million 10-year strategy from Chancellor’s Office announced Dec 2005 • Government Funded UK Stem Cell Bank for foetal, adult & embryonic human stem cell lines • London Regenerative Medicine Network – drive towards clinical applications • UK Stem Cell Network – integration of all UK regional network

25. Stem Cell Biology Laboratory Wolfson Centre for Age-Related Diseases Sarah Pringle Siobhán Connor Zhenling Luo Katerina Varanou Woojin Lee Julie Ghosh Daniel Webber Jessica Cooke Hannah Taylor Antigoni Ekonomou SH Cedar Chetan Shatapathy Yue Wu

26. Stem Cell Biology Laboratory Collaborators Human ES Cells Peter Braude Sara Hall Glenda Cornwall Emma Stephenson Pancreatic Islet Cells Peter Jones Shanta Persaud Cartilage Generation Cosimo De Bari Francesco Dell’Accio Blood Stem Cells Paul Fairchild (Oxford) Karl Karlsson (Oxford) Hepatic Stem Cells Anill Dhawan Robin Hughes Ragai Mitry Huntington’s Disease Gillian Bates Eva Sirinathsinghji Retinal Regeneration Robin Ali (UCL/Moorfields) Spinal Cord Repair Steve McMahon Liz Bradbury Paul Felts Merion Davies Lung Differentiation Carol Jones (Novartis) Phil Kemp (Novartis) Bioethics & Politics Claire Williams Steve Wainright Sarah Franklin (LSE) Brian Salter (Norwich) Herbert Gottweis (Vienna) Neural Differentiation Jonathan Corcoran Bia Gonclaves Bioprocessing Chris Mason (UCL) Tooth Replacement Paul Sharpe Parkinson’s Disease Peter Jenner Susan Duty Vascular Development Karen Hirschi (Baylor) Qingbo Xu Adult Neurogenesis Clive Ballard Omar Pathmanaban Chirstos Tolias Elaine Perry (Newcastle) Robert Perry (Newcastle) Aaron Cheung (GSK) Proteomics Tony Ng Simon Ameer-Beg Sukhi Bansal Giampietro Schiavo (CRUK) Jonas Bergquist (Uppsala) Electrophysiology Reginald Docherty Bone Development Agi Grigoriadis Cardiac Repair Anthony Mathur (Barts) Matthew Lovell (Barts) Skin Differentiation John McGrath Medical Research Council, Edmond J Safra Foundation, BBSRC, ESRC, EPSRC, European Commission, Oliver Bird Foundation, UK DTI, Francesca Patrizi, High Q Foundation, Wellcome Trust, Huntington’s Disease Society, John &Helen Robertson, Alzheimer’s Research Trust, Liz & John Hancock, Guy’s & St Thomas’ Charitable Trust