BENG0011-Lecture 7.pdf

BENG0011- Manufacturing Regenerative Medicines: from Lab Bench to Industry Lecture 7:Neural Regenerative Medicine Dr Rana Khalife Overview Brain and CNS anatomy Basal ganglia disorders and cell replacement therapy Parkinson’s disease Huntington’s disease Current stage Spinal co...

BENG0011- Manufacturing Regenerative Medicines: from Lab Bench to Industry Lecture 7:Neural Regenerative Medicine Dr Rana Khalife Overview Brain and CNS anatomy Basal ganglia disorders and cell replacement therapy Parkinson’s disease Huntington’s disease Current stage Spinal cord Aims and objectives Understand the importance of Brain and CNS anatomy Define the cause of Parkinson’s disease ,Huntington’s disease, Spinal cord injuries as the cause of injury, type of neurone affected , pathway affected and how to cure it/potential drug and cell therapy (cell source ) Be able to explain the Current pre-clinical and clinical trials for these diseases  CNS Anatomy  CNS Anatomy Nuclei of the basal ganglia Striatum Caudate nucleus, putamen and nucleus accumbens Inhibitory GABAergic neurons Globus pallidus - internal (GPi) and external (GPe) Inhibitory GABAergic neurons Substantia nigra - pars reticulata (SNr) and pars compacta (SNc) Dopaminergic nigrostriatal neurons Subthalamic nucleus (STN) Excitatory glutamatergic neurons  Parkinson’s Disease 1 in 500 people in the UK affected Loss of dopaminergic neurons in the substantia nigra results in loss of neurotransmitter dopamine in the striatum.  Dopamine Signaling in Parkinson's Disease Interactive Pathway Lewy body pathology in the substantia nigra of a PD patient Dopamine signalling in Parkinson’s Disease https://www.cellsignal.co.uk/contents/science-cst-pathways-neuroscience/dopamine-signaling-in-parkinson-s-disease-interactive- pathway/pathways-park Treatment Levodopa Dopamine signalling in Parkinson’s Disease  Current pharmacological treatments – Symptomatic relief – Levodopa – primary treatment for PD Rapid fluctuations in clinical state, chronic side effects, difficult to optimize dosage On-off – switch between mobility (on) and immobility (off) Deep brain stimulation – Great in some patients but serious complications can arise in others Depression, hallucinations, compulsive gambling, hypersexuality  Neural transplantation in Parkinson’s disease Offers alternative therapy aiming to replace dopamine in the striatum to: – Stop disease progression – Repair damage – Overcome issues with current drug treatments What cells? How can they be implanted? From Möllers et al (2011) in “Towards New Therapies for Parkinson’s Disease” (http://bit.ly/2i9ezhM) How can neural transplantation in PD be a scalable treatment option? Neural transplantation in Parkinson’s disease Foetal cells Committed dopaminergic neuron precursors from developing ventral mesencephalon (VM) Advantages Disadvantages Neuronal differentiation Availability Regional specification Limited potential for expansion Safety Viability No tumorigenesis Variability Quality control Ethical Sensitivity  Graft preparation Cell suspension grafts Collect Dissect Tissue is dissected from the developing brain Dissociated cells contain Enzyme digestion precursor population of neurons Transplantation to an Dissociate animal model, eg. Lesioned rat brain Wash Transplant Allows differentiation and integration of cells to be nigra graft in studied Can be transplanted as striatum 6- OHD A dissociated cells or whole lesion pieces of tissue Diagrams - SB Dunnett  Pre-clinical assessment of graft survival VM transplants rVM hV Survive in the adult rodent M brain and differentiate into dopaminergic neurons Form synapses within the host brain Release dopamine in appropriately regulated manner Restore function in animal models of PD Lelos et al (2016) Heuer et al (2013)  Amphetamine-induced rotation Immunohistological analysis of hVM tissue at 20 weeks post-graft Stimulus response learning The “Carli” task (Carli, Evenden & Robbins, Nature 1985; 313: 679-682) Can nigral grafts alleviate the 6-OHDA graft contralateral 6-OHDA lesion 3 months train t e s t re-test deficit? stimu lus lights photocel well blanks l detectors food well (Dowd & Dunnett, Eur J Neurosci 2004; 20: 1953- 59) See also: Lelos et al (2016) Exp Neurol 278:54- 61  Barker, R. A. et al. (2015), Nat. Rev. Neurol. doi:10.1038/nrneurol.2015.123 [18F]fluorodopa PET imaging of dopaminergic activity in PD basal ganglia pre- and post-transplantation with human foetal ventral mesencephalon By Piccini and Brooks, from Dunnett & Björklund (1999), Nature 399, A32-A39 Neural transplantation in Parkinson’s disease: Clinical data Foetal transplants have shown proof-of-principle for cell therapy in PD Safety and clinical efficacy developed BUT Variable outcomes between trials Some outcomes very positive, two double-blind placebo-controlled trials (Denver, Florida) showed negative results – Poor survival – Poor functional response, – Side effects – dyskinesias Outcomes depend on trial design – Non-standard techniques for transplantation – Issues with immunosuppression/rejection – Patient selection  Aims To improve the consistency and efficacy of dopaminergic cell replacement in Parkinson’s disease To eliminate any off-state dyskinesias after dopaminergic cell https://www.youtube.com/watch?v= transplants PQMWs6UMUL8&feature=youtu.be https://youtu.be/JoiVrXofzgQ?list=P To develop a protocol that can L1ixQpbHed3bMzACwNWL4DX3s6 serve as a template for future clinical QsjN-pT trials in cell therapy https://scienceofparkinsons.com/lectures/the-2016-gretschen-amphlet-memorial-lecture/  Making dopaminergic neurons for transplantation Arenas et al (2015), Development 142:11, 1918-1936 ESC derived dopaminergic neurons for transplantation Grealish et al (2014), Cell Stem Cell 15, 65:i rom human ventral mesencephalon (VM)  iPSC derived dopaminergic neurons for transplantation Transplantation of iPSC derived DA neurons in a primate model of PD Graft integration up to 2 years, no overgrowth Improvements in spontaneous movement recorded Kikuchi et al (2017), Nature 548, 592–596  Neural transplantation in Parkinson’s disease Current stage Ongoing work aims to optimize – Differentiation protocols – Scale-up – CTP preparation and delivery – Clinical outcome measures – Screening criteria predictive of a good CTP European stem cell consortium for neural cell replacement, reprogramming and functional brain repair Better Cells - Better Trials http://www.neurostemcellrepair.org/ A NEW GLOBAL INITIATIVE AROUND STEM CELL BASED THERAPIES FOR PARKINSON’S DISEASE Barker et al (2015), npj Parkinson's Disease 1 Neural transplantation in Parkinson’s disease Current stage Kirkeby et al (2017), Prog. In Brain Res. Vol 230, Ch17