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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
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
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)