MouseAge slideshare
- 1. Know Your Models! (Know your Disease) Multiple Sclerosis Models & Experience with Clinical Translation MouseAGE 2015 Prof. David Baker david.baker@qmul.ac.uk Slides Available on www.ms-res.org (Slideshare)
- 2. MOUSEAGE 2015 • KNOW YOUR DISEASE-DESCRIPTION OF MULTIPLE SCLEROSIS • KNOW YOUR MODEL- DESCRIPTION OF MS MODELS • KNOW THE LIMITATIONS: FAILURE TO TRANSLATE • LIMIT YOUR LIMITATIONS: ARRIVE REPORTING GUIDELINES • DRUG DEVELOPMENT: BENCH TO BEDSIDE Slides Available on www.ms-res.org (Slideshare)
- 3. Multiple sclerosis (MS) is a chronic (auto)immune-mediated CNS- confined demyelinating disease affecting 2,500,000 people worldwide (Restricted Distribution: Northern European Dissent) Disease onset is usually between 20 and 40 years of age (F:M ratio, 2:1. Polygenic-150 genes identified so far) (30% concordance in Identical Twins, 100% Infected with EBV) MS is clinically characterized by a relapsing-remitting course usually followed by a progressive phase MS is pathologically heterogeneous – inflammation, demyelination and neurodegeneration (axonal loss and neuronal damage) Spontaneous repair occurs but MS invariably progresses (ambulatory problems in 70-80% patients at 25 years from onset) KNOW YOUR DISEASE-BIOLOGY OF MS
- 4. NERVE LOSS PROGRESSIVE MSRELAPSING-REMITTING MS DISABILITY Frequent inflammation, demyelination, axonal transections, plasticity and remyelination Inflammation, Persistent Demyelination & Gliosis Infrequent inflammation, Gliosis, Chronic Neurodegeneration CLINICAL THRESHOLD INFLAMMATION Symptoms Clinical Effects are Due to Altered Nerve Conduction CLINICAL COURSE Immune-Mediated Beta-interferons, Alemtuzumab (CD52), Cladribine, Bone Marrow Transplantation, Fingolimod (S1P1-modulator) , Rituximab (CD20) are ACTIVE Neurodegeneration Beta-interferon, Alemtuzumab, Cladribine, Rituximab Bone Marrow Transplantation, Fingolimod are INACTIVE KNOW YOUR DISEASE-BIOLOGY OF MS
- 5. Eyes,Brain & Spinal Cord You are told you have Eyes,Brain & Spinal Cord You are told you have White Blood Cells No Repair • INFLAMMATORY, DEMYELINATING, NEURODEGENERATIVE DISEASE OF CNS •MULTIFOCAL OLIGODENDROCYTE LOSS MS plaques KNOW YOUR DISEASE-BIOLOGY OF MS Myelin (blue) stain
- 6. NORMAL AGING ATROPHY RATE 0.1% p.a. MULTIPLE SCLEROSIS ATROPHY RATE 0.4-1.0 % MS is a Neurodegenerative Disease KNOW YOUR DISEASE-BIOLOGY OF MS
- 7. Brain atrophy occurs across all stages of the disease De Stefano, et al. Neurology 2010 n= 963 pwMS KNOW YOUR DISEASE-BIOLOGY OF MS
- 8. ATROPHY KNOW YOUR DISEASE-BIOLOGY OF MS
- 9. Onset of Progressive MS is Age-Sensitive and Independent of Pre-Progression Disease Course Confavreaux et al. 2006, Tutuncu et al. 2012 0 10 20 30 40 50 60 70 80 100 80 60 40 20 0 PPMS SPMS 0 10 20 30 40 50 60 70 80 100 80 60 40 20 0 PPMS SPMS Mean Age at MS Onset (Years) Mean Age at PMS Onset (Years) Percentage Percentage KNOW YOUR DISEASE-BIOLOGY OF MS
- 10. in vitro monkeys rodents clinic Mechanisms of Disease Design of Therapies Safety & Ethics monkeys clinic discovery/screening Validation/safety safety/ preclinical clinical trials KNOW YOUR DISEASE-BIOLOGY OF MSMODEL-WHY USE ANIMALS?
- 11. MS was called an Autoimmune, Demyelinating Disease of the White Matter MYELIN = BROWN DEMYELINATION MONONUCLEAR CELL INFILTRATE BLOOD VESSEL (VENULE) KNOW YOUR DISEASE-BIOLOGY OF MS/MODEL-BIOLOGY OF MS
- 12. MS was called an Autoimmune, Demyelinating Disease of the White Matter CHEMICAL-INDUCED DEMYELINATION VIRAL-INDUCED DEMYELINATION AUTOIMMUNE-INDUCED DEMYELINATION EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS (EAE) IN MAMMALS (MYELIN-REACTIVE AUTOIMMUNITY) SEMLIKI FOREST VIRUS MOUSE HEPATITIS VIRUS (NEUROTROPHIC VIRUS) CUPRIZONE FEEDING LYSOLECITHIN INJECTION (OLIGODENDROCYTE TOXIN) MONONUCLEAR CELL INFILTRATE TRANSGENE-INDUCED DEMYELINATION NO ANIMAL GETS SPONTANEOUS MS-LIKE DISEASE KNOW YOUR DISEASE-BIOLOGY OF MS/MODEL-BIOLOGY OF MS
- 13. Myelin Neurofilament Demyelination Axonal Transection Axonal Transections are Associated with Inflammation Control White Matter <1transection/mm2 Core of Chronic Lesion 875transection/mm2 Edge of Chronic Active Lesion ~3000transection/mm2 Active Lesion ~11000transection/mm2 MS was called an Autoimmune, Demyelinating Disease of the White Matter KNOW YOUR DISEASE-BIOLOGY OF MS/MODEL-BIOLOGY OF MS AUTOIMMUNE-INDUCED DEMYELINATION EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS (EAE) IN MAMMALS (MYELIN-REACTIVE AUTOIMMUNITY) MONONUCLEAR CELL INFILTRATE Grey Matter Demyelination Grey Matter Lesions have few T cells & macrophages
- 14. Experimental Autoimmune Encephalomyelitis 1. Active EAE - induced with myelin proteins or peptides in Complete Freund’s Adjuvant. Myelin proteins in CFA 2. Adoptive EAE - induced by T cell transfer from mice immunised for active EAE. T cells to myelin proteins Myelin proteins in CFA 3. Myelin TCR Transgenic - Develop Spontaneous EAE. KNOW YOUR MODEL-BIOLOGY OF EAE
- 15. No Clinical Disease-No Spinal Cord Infiltration- No Cytokine X, Y or Z Limp tail Impaired righting reflex hindlimb paralysis Moribund partial paralysis Normal Remission 0 1 2 3 4 5 (1) Clinical Score Day 7 Spinal cord homogenate in Freund’s complete adjuvant Day 0 Spasticity & Tremors Develop KNOW YOUR MODEL-BIOLOGY OF EAE Video is Coming Next 43
- 16. No Clinical Disease-No Spinal Cord Infiltration- No Cytokine X, Y or Z Limp tail Impaired righting reflex hindlimb paralysis Moribund partial paralysis Normal Remission 0 1 2 3 4 5 (1) Clinical Score Day 7 Spinal cord homogenate in Freund’s complete adjuvant Day 0 Spasticity & Tremors Develop KNOW YOUR MODEL-BIOLOGY OF EAE Video is Coming Next 43
- 17. MULTIPLE SCLEROSIS MODELS KNOW YOUR MODEL-BIOLOGY OF EAE
- 18. Time Post-Induction (Days) 10 15 20 25 30 35 40 MeanNeurologicalScore 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 Acute (ABH, SJL) EAE Nerve Loss Chronic (C57BL/6) EAE Tail Paralysis HindLimb Paralysis Video is Coming Next KNOW YOUR MODEL-BIOLOGY OF EAE Conduction Block Strain Antigen C57BL/6 MOG35-55 ABH PLP56-70 ABH MOG8-22 ABH MOG35-55 PL/J MBP1-9 SJL MBP89-101 SJL PLP139-151 SJL PLP178-191 MOG Myelin oligodendrocyte glycoprotein, MBP Myelin Basic Protein, PLP Proteolipid Protein
- 19. Normal Remission 1 Remission 2 Remission 4 NfH(µg/mgtotalprotein) Number of Attacks N RM1 RM2 RM3 50 100 150 200 250 300 350 400 Axonal Content Assessed By Neurofilament ELISA www.msbrainhealth.org No-evidence of Disease Activity (NEDA) KNOW YOUR MODEL-BIOLOGY OF EAE Stop Relapses Save Brain
- 20. Leg moved to full flexion for assessment Spastic Leg RESISTANCEFORCETOHINDLIMBFLEXION(N) 0.00 0.05 0.10 0.15 0.20 0.25 0.30 0.35 0.40 0.45 Left leg Left legLeft legRight Leg Right Leg Right Leg Non-spastic Remission Paralysed Relapse Spastic Remission 0.08 ± 0.01# 0.03 ± 0.01* 0.17 ± 0.10 *,# SYMPTOMATIC MODELS OF MS Chronic EAE RM4(6 months ) Normal Post Relapse-Remission Spastic Limbs Bladder Problems KNOW YOUR MODEL-BIOLOGY OF EAE
- 21. PERIPHERAL IMMUNITY Slow RELAPSE RATE NEURODEGENERATION Slow PROGRESSION SYMPTOM CONTROL Improve QUALITY OF LIFE REPAIR Reverse Deficits OUTCOME OF CONTROLDISEASE PROCESS Increasing Number of Effective Drugs Effective Drugs with Unpleasant Side-Effects No Treatments No Treatments Adaptive Immune –Dependent Inflammation Adaptive Immune–Independent, Innate Dependent Recently Demyelinated Chronic Demyelinated/Gliotic Lesions KNOW YOUR MODEL-BIOLOGY OF EAE
- 22. • PRE-CLINICAL FAILURE • Model does not reflect human disease biology • Drug does not target biology relevant to human application • Lack of appreciation of human disease • Dogma & overstating effect • Model used in a way that does not reflect human indication • Drug doses are not used in at physiological doses • Drugs are not delivered in a way appropriate to how used in humans • Studies are not transparent & not reproducible (Ineffective Study Design) • CLINICAL FAILURE • Lack of clear understanding of human pathology • Drug is seldom investigated by scientists developing the Idea. • Over-interpretation of significance of pre-clinical studies • Drug is not used at a dose relevant to the pre-clinical studies • Population does not respond as predicted. (Ineffective Trial Design) • Dose-limiting side-effects • Study Underpowered, too short or unrealistic expectations • Measurement Instruments Inadequate Clinical Outcomes and Surrogate Markers • Wrong Group of pwMS studied (IneffectiveTrial Design) • Commercial Interests Mechanism is all Important. Relevance of Slight Delay of a Few Days, Slight Diminution Prophylactic/Therapy “Toxicity leading to Stress” “Route & Timing” Reporting Issues KNOW THE LIMITATIONS-FAILURE TO TRANSLATE Two thousand drugs tested in EAE, only nine-ten classes of drugs approved Time Post Induction (Days) 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 MeanClinicalScore 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 Vehicle n= 10/10 Test Drug n = 10/10 Test Drug 2 n=0/10 or n = 7/10
- 23. ARRIVE Guidelines (UK) Kilkenny C, Browne WJ, Cuthill IC, Emerson M & Altman DG. Improving bioscience research reporting: the ARRIVE guidelines for reporting animal research. PLoS Biol. 2010; 8(6):e1000412 (2010) National Research Council (USA). Institute for Laboratory Animal Research. Guidance for the Description of Animal Research in Scientific Publications. National Academies Press, Washington (DC) (2011). Guidelines on EAE Studies Baker D, Amor S. Publication guidelines for refereeing and reporting on animal use in experimental autoimmune encephalomyelitis. J Neuroimmunol. 2012 242:78-83 LIMIT YOUR LIMITATIONS-EXPERIMENTAL DESIGN RANDOMISATION BLINDING SAMPLE-SIZE DATA HANDLING REPLICATION NINDS GUIDELINE Nature 2012; 490:187 >10% >20% >1%
- 24. ARRIVE Guidelines (UK) Kilkenny C, Browne WJ, Cuthill IC, Emerson M & Altman DG. Improving bioscience research reporting: the ARRIVE guidelines for reporting animal research. PLoS Biol. 2010; 8(6):e1000412 (2010) National Research Council (USA). Institute for Laboratory Animal Research. Guidance for the Description of Animal Research in Scientific Publications. National Academies Press, Washington (DC) (2011). Guidelines on EAE Studies Baker D, Amor S. Publication guidelines for refereeing and reporting on animal use in experimental autoimmune encephalomyelitis. J Neuroimmunol. 2012 242:78-83 LIMIT YOUR LIMITATIONS-EXPERIMENTAL DESIGN RANDOMISATION BLINDING SAMPLE-SIZE DATA HANDLING REPLICATION NINDS GUIDELINE Nature 2012; 490:187 >10% >20% >1%
- 25. Parametric (47%) t- test (46%) Non Parametric 39% Not Reported (14%) Parametric (65%) t test (67%) Non-Parametric (4%) Not Reported (31%) All EAE Publications 6 months 1:12:2011-31:5:2012 n=175 All EAE Publications 2 Years 1:1:2010-11:9:2012 n=26 All PUBMED PUBLICATIONS BIG 6 PUBLICATIONS Nature, Nat Med, Nat Immunol, Nat. Neurosci, Science, Cell T-test Criteria Continuous Normally Distributed Equal Variances LIMIT YOUR LIMITATIONS-EXPERIMENTAL DESIGN
- 26. Figure legend. Clinical scores of two independent EAE experiments at d23 post disease induction. Individual scores as well as the mean score of two independent experiments are shown. Control: n=10, vehicle: n=13, xxxxx-345: n=11. Control versus vehicle: P=0.620, control versus xxxxx-345: P=0.017, vehicle versus xxxxx-345 P=0.029. * indicate P values <0.05 and ** indicate P values <0.005 based on a non-paired Student’s t test. Error bars are s.e.m Nature Paper 2015 Drug-treated animals the scores appear to be: 0, 0 , 0, 0.5, 0.5, 2, 2.5, 3, 3.5, 3.5, 3.5 n=11 Vehicle scores appear to be: 0.5, 2.5, 2.5, 2.5, 2,75, 2.75, 3, 3.5, 3.5, 3.5, 3.5, 3.5, 3.5 n=13. Do a t test drug verses vehicle p=0.029 The assumptions of a t test (a) Data is normally distributed. You test this and it passes the test p=0.152, (b) data groups have equal variances Test for that and it fails P<0.05. (c) Data is Parametric. Fail. It is non-parametric. t test is not a valid test Mann Whitney test. P=0.082.........Ooooops. LIMIT YOUR LIMITATIONS-EXPERIMENTAL DESIGN
- 27. • PRE-CLINICAL FAILURE • Model does not reflect human disease biology • Drug does not target biology relevant to human application • Lack of appreciation of human disease • Dogma & overstating effect • Model used in a way that does not reflect human indication • Drug doses are not used in at physiological doses • Drugs are not delivered in a way appropriate to how used in humans • Studies are not transparent & not reproducible (Ineffective Study Design) • CLINICAL FAILURE • Lack of clear understanding of human pathology • Drug is seldom investigated by scientists developing the Idea. • Over-interpretation of significance of pre-clinical studies • Drug is not used at a dose relevant to the pre-clinical studies • Population does not respond as predicted. (Ineffective Trial Design) • Dose-limiting side-effects • Study Underpowered, too short or unrealistic expectations • Measurement Instruments Inadequate Clinical Outcomes and Surrogate Markers • Wrong Group of pwMS studied (IneffectiveTrial Design) • Commercial Interests Mechanism is all Important. Relevance of Slight Delay of a Few Days, Slight Diminution Prophylactic/Therapy “Building Site Effect” “Route & Timing” “Placebo Effect” Less Circuitry so Less Compensation Capacity Non-Responders Immune (T/B cell) or Neurodegeneration Professional Trialists Reporting Issues Two thousand drugs tested only nine-ten classes of drugs approved KNOW THE LIMITATIONS-FAILURE TO TRANSLATE
- 28. KNOW THE LIMITATIONS-FAILURE TO TRANSLATE Once Drugs Become Available, Clinicians Voice their Opinion that Animal Experiments are not worth while, because of the failures “It is now possible to conduct a Phase II trial for anti-inflammatory MS drug candidates within a few months…..it is not useful to pre-screen potentially effective drugs using the EAE model (Ransohoff 2006)”_______________________________________________________________________________________ EAE MS ____________________________________________________________________________________________________ MS DRUGS Copaxone Inhibition Inhibition Beta Interferon Inhibition Inhibition CD49d-specific mAb Inhibition Inhibition Gilenya/Fingolimod Inhibition Inhibition Aubagio/teriflunomide Inhibition Inhibition Tecfidera/Dimethy Fumarate Inhibition Inhibition CD52-specific mAb Inhibition Inhibition (CD20-specific mAb) Inhibition Inhibition /Context Dependent /Context Dependent /Worsening /No effect/Worsening FAILURES CD25-specific mAb Worsening Inhibition Gamma Interferon Inhibition Worsening TNF-specific mAb Inhibition Worsening PDE4 inhibitor (rolipram) Inhibition No Effect/Worsening Antigen-Specific Treatments Inhibition No Effect/Worsening Th2 Cytokines (IL-4/IL-10) Inhibition No Effect CD4-specific mAb Inhibition No Effect IL-12/23-specific mAb Inhibition No Effect ____________________________________________________________________________________________________ Treg cells important Th1/Th17 cells important
- 29. If 65% T cell Depletion does not Stop EAE, Why would it be expected to stop MS? The Trial in MS was doomed before it even started? ACUTE EAE IL-12/23 KO C57BL/6 EAE non-susceptible (IL12p40 + IL12p35 = IL-12) anti-IL23 (p40) C57BL/6 EAE inhibited (IL-12p40 + IL23p19 = IL-23) anti-IL23 (p40) ABH EAE inhibited RELAPSING EAE anti-IL-23 (p40) ABH No inhibitory effect (Heremanns et al. 1999) RELAPSING MS anti-IL23(p40) Human MS No inhibitory effect Th17 IL-23 IL-6/TGFb Th1 IL-12 Th2 IL-4 B IL-4, IL10, IL-13 IFNg IL-4 IL-10 IL-13 IL-17 IL-21 IL-22 IFN-g Ustekinumab (Segal et al 2008) KNOW THE LIMITATIONS-FAILURE TO TRANSLATE
- 30. Time Post-inoculation (Days) 0 7 14 21 28 35 42 49 56 63 70 77 DevelopmentofClinicalEAE(%) 0 10 20 30 40 50 60 70 80 90 100 Untreated CD4b mAb 3 weeks CD4d&CD8d mAb D12 + CD4b mAb 3 Wks “Anti-CD4 antibody Cures EAE” KNOW THE LIMITATIONS-FAILURE TO TRANSLATE
- 31. Time Post-inoculation (Days) 0 7 14 21 28 35 42 49 56 63 70 77 DevelopmentofClinicalEAE(%) 0 10 20 30 40 50 60 70 80 90 100 Untreated CD4b mAb 3 weeks CD4d&CD8d mAb D12 + CD4b mAb 3 Wks Anti-CD4 antibody Causes Transient Immunosuppression in EAE KNOW THE LIMITATIONS-FAILURE TO TRANSLATE
- 32. • CD4 T cell depletion inhibits EAE • CD4 T cell deletion inhibits virtually every T and B Cell mediated autoimmunity • CD4 T cell deletion does not inhibit multiple sclerosis (Lindsey et al. 1994 Neurol 44:413 : 810; van Oosten et al. 1998 Mut Scler 1:339) . • CD4 T cell deletion does not inhibit other human autoimmune diseases Problem with the Study Planned <60% Depletion Naïve CD45RA preferentially affected Primed CD45RO T cells relatively unaffected (Llewellyn-Smith et al. 1997 Neurol 48:810; 48) MS IS NOT CONTROLLED BY T CELL DEPLETION No effect on Gadolinium enhancing lesions If 60% T cell Depletion does not Stop EAE, Why would it be expected to stop MS? The CD4 trial in MS was Probably doomed before it even started KNOW THE LIMITATIONS-FAILURE TO TRANSLATE Degree of CD4 depletion was important with regard to treatment efficacy. There was a significant 41% reduction in relapses.
- 33. Red = abundant CB1 receptors Black = moderately abundant CB1 receptors PHARMACEUTICAL CANNABIS BENCH TO BESIDE – SYMPTOM CONTROL Time Post-Injection (Min) 0 10 20 30 40 50 60 70 80 90 100 110 120 ChangeinHindlimbStiffness(%)±SEM -50 -40 -30 -20 -10 0 10 20 Sativex Vehicle
- 34. Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION OR NOT Placebos did not progress as predicted Zajicek J et al. 2013 P<0.01 CUPID Cannabinoid Use in Progressive Inflammatory brain Disease. Animal Studies show that Cannabinoids are Neuroprotective Compounds in cannabis THC & CBD cause Neuroprotection Cannabis Receptor Loss CB1 KO exhibit Neurodegeneration Natural Ligand Increase FAAH KO looses Neuroprotection FAAH inhibitors Neuroprotection
- 35. Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION OR NOT Placebos did not progress as predicted Zajicek J et al. 2013 P<0.01 Ambulatory People Starting EDSS less than 5.5 CUPID Cannabinoid Use in Progressive Inflammatory brain Disease. Animal Studies show that Cannabinoids are Neuroprotective Compounds in cannabis THC & CBD cause Neuroprotection Cannabis Receptor Loss CB1 KO exhibit Neurodegeneration Natural Ligand Increase FAAH KO looses Neuroprotection FAAH inhibitors Neuroprotection
- 36. INFLAMMATORY PENUMBA Animal Studies indicate that sodium channel blockers can be neuroprotective by: •Inhibiting metabolic overload in nerves •Blocking microglial activity Lamotrigine trial in secondary progressive MS fails Brain atrophy MRI shows (pseudo) atrophy greater than placebo. Drug was poorly tolerated (Kapoor et al. 2010). 50% of people in trial are not drug compliant. Neurofilament Biomarker is reduced in Drug Compliant individuals (Gnanapavan et al. 2013) Animal studies show that sodium channel blockers are particularly active during the inflammatory penumbra. (Al-Izki et al. 2014) PLP (brown) myelin Stain) Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION
- 37. Optic Neuritis is Often First Sign of MS Most accessible part of Human CNS Courtesy of Roy Weller Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION
- 38. EYE OPTIC NERVE VISUAL CORTEX The eye is the window to the brain… OCCIPITAL LOBE The Visual SystemX Damage in MS Optic Neuritis Common First Sign of MS Occurs in Over 50% of MS X Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION
- 39. EYE OPTIC NERVE VISUAL CORTEX The eye is the window to the brain… OCCIPITAL LOBE The Visual SystemX Damage in MS Optic Neuritis Common First Sign of MS Occurs in Over 50% of MS X Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION
- 40. Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION C57BL/6-Tg(Tcra2D2,Tcrb2D2)1Kuch /J
- 41. Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION C57BL/6-Tg(Thy1-CFP)23Jrs /J
- 42. Develops Spontaneous/Induced Optic Neuritis. Subclinical Spinal Cord Disease Less Severe than Classical EAE Fluorescent RGC Detection of Nerve Loss in Living Eye Repeated Monitoring Not Requiring Histology Disease is Concentrated in the Visual System Human Relevant Outcome Measures Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION C57BL/6-Tg(Tcra2D2,Tcrb2D2)1Kuch.Cg-Tg(Thy1-CFP)23Jrs /J = FLASH GORDON
- 43. Low Contrast Eye Chart Human Test Visual acuity is a measure of clearness of vision and is used as a measure as to how you can see Mouse Test Visual tracking drum Netpositiveheadmovements 0 2 4 6 8 10 12 14 Before Optic Neuritis After Optic Neuritis Loss of vision Head movements are Reduced after Optic Neuritis (Nerve Loss) Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION MEASURING SIGHT-VISUAL ACUITY
- 44. Reference electrode Recording electrode Heating pad Visual Evoked Potential (VEP) Z Z Z Electrophysiology can be used to measure the Visual Evoked Potential (VEP). This is a measure of the neurotransmission from the eye to the visual cortex. Human Test Mouse Test Latency Latency of VEP is Increased after Optic Neuritis (Demyelination) Before Optic Neuritis After Optic Neuritis Loss of Nerve Conduction Amplitude Amplitude of VEP is Reduced after Optic Neuritis (Nerve Loss) Amplitude(µV) 0 10 20 30 40 Before Optic Neuritis After Optic Neuritis Loss of vision Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION MEASURING SIGHT-ELECTROPHYSIOLOGY
- 45. Optic Nerve Head Ganglion Cell Layer (GCL) OCT is a non-invasive retinal imaging tool to look at the structure of the retina. Human Test Mouse Test Optic Nerve Head OCT imageHistology image Eye from Living AnimalEye from Dead Animal Outer Nuclear Layer Photo- Receptors Ganglion Cell Layer Inner Nuclear Layer Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION OPTICAL COHERENCE TOMOGRAPHY (OCT)
- 46. Before Induction After Induction Optic Nerve Head Optic Nerve Head OCT scan OCT Can Detect Loss of RGC in Mouse with Optic Neuritis Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION OPTICAL COHERENCE TOMOGRAPHY (OCT)
- 47. cSLO is a high resolution retinal imaging tool to look at the structure of the retina Optic NeuritisPre-Disease After Disease Onset Retinal Ganglion Cell Loss Before Disease Onset Retinal Ganglion Cell Loss cSLO Image Blood Vessel Optic Nerve Head Retinal Ganglion Cell Optic Nerve Head Pre-Disease 2018 cells/mm2 Optic Neuritis 285 cells/mm2 RGC Loss P<0.001 Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION Confocal SCANNING LASER OPHTHALMOSCOPY (cSLO)
- 48. Day 0 2 10 14 21 Demyelinating mAb (250μg Z12 mAb i.p.) Disease Induction (150ng Pertussis Toxin i.p.) T cell-Mediated Optic Neuritis Retinal Flatmount Microscopy Treatment Period 5mg/kg i.p.) Optic Neuritis A D BA D CB Vehicle Sodium Channel Blocker A D CB Low Power Normal Reduced Nerve Damage Normal mouse Meanretinacelldensity(cells/mm2) 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900 OPTIC NEURITIS + Vehicle OPTIC NEURITIS + drug Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION NEUROPROTECTION WITH SODIUM CHANNEL BLOCKERS
- 49. OPTIC NEURITIS Gabilondo I et al. 2015 Change in Retinal Nerve Fibre Layer Thickness after Optic Neuritis Double blind, randomised placebo- controlled, parallel group design Initial dose 15 mg/kg, daily 4 mg/kg (max 300mg) , treatment duration 3 months Blinded assessing and treating physicians; 2 imaging sites (Sheffield, London) Outcomes OCT, VEP, MRI, Vision Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION NEUROPROTECTION WITH SODIUM CHANNEL BLOCKERS
- 50. OPTIC NEURITIS TRIAL Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION NEUROPROTECTION WITH SODIUM CHANNEL BLOCKERS
- 51. 300 pwMS Year 1 Year 2 Year 3 600 pwM 300 pwMS Active tablet Placebo tablet Year -2 Year 4 Month 6 Active tablet Placebo tablet -6month month 18 Month 12 LP1 LP2 LP3 STANDARD TRIAL DESIGN NOVEL TRIAL DESIGN 30 pwMS 30 pwMS 60 pwMS Red = abundant CB1 receptors Black = moderately abundant CB1 receptors BENCH TO BESIDE – NEUROPROTECTION NEUROPROTECTION WITH SODIUM CHANNEL BLOCKERS DMT + Nerve protector DMT
- 52. UCL-INSTITUTE OF NEUROLOGY Queen Square UCL-INSTITUTE OF NEUROLOGY Queen Square UCL-INSTITUTE OF NEUROLOGY Queen Square THANK YOU FOR LISTENING Spasticity Studies David Baker Gareth Pryce Gavin Giovannoni Neuroprotection Studies David Baker Gareth Pryce Sarah Al-Izki Katie Lidster Sam Jackson Gavin Giovannoni Optic Neuritis Trial Raj Kapoor, Rhian Raftopoulos, Simon Hickman, Basil Sharrock Klaus Schmierer, Gavin Giovannoni David H Miller & Others Slides Available on www.ms-res.org (Slideshare)