Transcript EAE Model - Neuro ACF
Stanford University Department of Comparative Medicine Developing Best Care Practices and Endpoint Guidelines for Rodent Models of Neurologic Disease Lisa Heath, DVM, DACLAM
Rodent Models
• Experimental Allergic Encephalomyelitis
(EAE)
• Amyotrophic Lateral Sclerosis (ALS) • Huntington’s Disease
Guidelines and Endpoints Developing Guidelines
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Resources
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Working with Researchers
Multiple Sclerosis in People
Fatigue
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Bowel/bladder dysfunction
Numbness
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Cognitive dysfunction
Gait, balance, coordination problems
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Pain
Multiple Sclerosis
Health.howstuffworks.com
Multiple Sclerosis
•http://www.elements4health.com/images/stories/conditions/myelin-sheath.jpg
EAE Models
Active EAE: immunized with myelin antigen or peptide emulsified in CFA
Monophasic or relapsing-remitting disease
Disease process commences 10-15 days post immunization
75-80% incidence of disease
EAE Models
Passive or adoptively transferred EAE: Recipient mice injected with activated, myelin-specific T cells (harvested from immunized donor mice)
Relapsing-remitting disease
EAE Guidelines Benchmarking
We benchmarked with other institutions to see if their IACUC had specific guidelines to cover EAE rodent models
64 institutions were surveyed regarding whether or not they had rodent EAE models
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19 institutions responded
Survey Results Data and graphs courtesy of Sonja Wallace
EAE Benchmarking Survey
Breakdown of Respondents 5% 5% 11% 26% 53% University Biotech/Pharma Private Institution Government CRO
EAE Benchmarking Survey
IACUC Guidelines 6% 26% 68% Specific EAE Guideline Humane Endpoint Guideline No Guideline Specified
EAE Benchmarking Survey
% Weight Loss Endpoint
11% 5% 5% 5% 5% 69%
10% Loss 15% Loss 20% Loss 25% Loss 30% Loss Not Specified
EAE Benchmarking Survey
Separate Debilitated Animals
16% 21% 63%
Separate Don't Separate Not Specified
Summary
Majority of respondents had an IACUC guideline specifically for EAE models
Some respondents used their humane endpoint guideline for EAE models
Majority of respondents used 20% percent weight loss as an endpoint
Majority of respondents separated symptomatic from asymptomatic animals
Refining Guidelines
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Intervention/Monitoring: When and how often
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Nutrition and Hydration: What, when and how often
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Include the Researchers
EAE Scoring
Score 0 = No overt signs of disease
Requirements: Label Special Request Card with “EAE Model” on Experimental Day 1
Baseline Body Weight recorded on Day 1
Assess and record scoring observations at least 3 times per week.
EAE Scoring
Score 1 = Limp tail, flaccidity of tail Requirements (same as for Score 0)
EAE Scoring
Score 2 = Paraparesis; limp tail and hindlimb weakness, waddling gait.
Provide gel pack and/or moistened food on cage floor, or other appropriate supplements.
Weigh and/or assess body condition score at least 3 times per week.
Tail Suspension Test A.
B.
Normal Plantar Reflex Hindlimb Clasping
Animal Welfare Information Center Bulletin, Winter 1999/2000, Vol. 10 No. 3-4
EAE Scoring Score 3 = Hindlimb paralysis. Total loss of moment in hindlimbs. Mouse moves itself with forelimbs.
Daily Scoring and monitoring.
Weigh and/or assess BCS at least once daily.
Assess hydration status (body weight and skin turgor).
Assess bladder for evidence of urinary retention. Express bladder as needed.
Provide fluid supplementation, SC or oral gavage.
EAE Scoring
Score 4: Subdivided into 4A and 4B (for guidelines only*)
4a = Hindlimb paralysis and mild forelimb weakness
4b. Hindlimb paralysis and moderate to severe forelimb paralysis; lateral recumbency.
EAE Scoring
Score 4b. Hindlimb paralysis and moderate to severe forelimb paresis; lateral recumbency.
Mice must be monitored at least twice daily for no more than 24 hours.
Any animal that does not remit to score 3 within the time frame must be euthanized.
EAE Scoring
Score 5 : Moribund, or complete hindlimb paralysis with moderate to severe forelimb paresis with no remittance after 24 hrs and intensive supportive care.
Euthanize immediately.
EAE Scoring
Any animal which loses >20% body weight or has a body condition score <2 must also be euthanized, regardless of EAE score.
A Sidenote on Body Weight Recording
Each mouse’s body weight data is unique
No averages!
Data recording: pre-disease weight and pre-calculated 20% loss cut-off
EAE Scoring: Mouse Posture
0 : Tail has tone.
1: Tail no longer stands up, loss of tail tone. Walks normally.
2: Limp tail, hindlimb dysfunction.
3: No longer holds onto underside of cagelid with hindlimbs. Moves hindlimbs.
Current Protocols in Neuroscience
4: Hindlimbs drag behind, not used for movement.
EAE Scoring
Models of Multiple Sclerosis. ACNR. Vol 6. No 6. 2007
Amyotrophic Lateral Sclerosis
Fatal neurodegenerative disease
Familial and Sporadic
Lou Gehrig’s Disease Loss of upper and lower motor neurons
Paralysis of muscles
www.alsa.org
ALS Models
SOD1 mice
ALS Mouse Models
Muscle atrophy
Weight loss
Limb weakness and paralysis
Oral and pharyngeal dysphagia
Respiratory weakness and paralysis
ALS Scoring System
Score of 0 Full extension of hind legs away from lateral midline when mouse is suspended by its tail, and mouse can hold this for two seconds, suspended two or three times. Score of 1 Collapse or partial collapse of leg extension towards lateral midline (weakness) or trembling of hind legs during tail suspension. Score of 2 Toes curl under at least twice during walking of 12 inches, or any part of foot is dragging along cage bottom/table.* Score of 3 Rigid paralysis or minimal joint movement, foot not being used for generating forward motion.
Score of 4 Mouse cannot right itself within 30 seconds after being placed on either side. Working with ALS Mice: Guidelines for preclinical testing & colony management. The Jackson Laboratory
ALS Scoring System
*If one hind leg is scored as 2, food pellets are left on bedding.
If both hind legs are scored as 2, Nutra-Gel is provided as food in addition to food pellets on bedding and a long sipper tube is placed on the water bottle.
Working with ALS Mice: Guidelines for preclinical testing & colony management. The Jackson Laboratory
ALS Scoring System:
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8 Point Scale
CS 0 CS 1 CS1.5
No evidence of disease Shaking or splaying of hind legs when suspended by tail Weakness in one hind leg CS2* Change in gait (used as disease onset when attained on two consecutive days *disease onset CS 2.5 Extreme weakness in one hind leg (inability to dorsiflex) CS 3 CS 3.5
Extreme weakness in both hind leg Functional paralysis in one hind leg Solomon, et.al. One Universal Common Endpoint in Mouse Models of Amyotrophic Lateral Sclerosis. June 2011, vol. 6, issue 6.
Endpoint Scoring for ALS: 8 Point Scale
CS4 Both hind legs functionally paralyzed CS4+ CS5 CS4 plus the earliest time mice attained one of the following: a) weight loss > 20% vs. body weight immediately prior to a clinical score of 2* b) weight loss > 20% vs. peak body weight c) body condition score <2 d) righting reflex > 20s (clinical score of 5) CS 4 plus a righting reflex >20s
Huntington’s Disease
Huntington’s Disease Scoring: Part 1
Score of 0 Normal Score of 1 Clasps hind limbs within 30s of being suspended Score of 2 Clasps hind limbs within 5s but recovers quickly after 30s Score of 3 Clasps hind limbs within 5s, difficulty recovering when released after 30s
Auerbach, et.al. Human Molecular Genetics, 2001, Vol. 10, No. 22.
Huntington’s Disease Scoring: Movement Abnormalities
Tail stiffness or paralysis
Hind-limb stiffness or paralysis
Reduced left-right alteration of hindlimbs (“hopping”)
Resting tremors
Difficulties walking after handling
Seizure-like episodes
Huntington’s Disease Scoring: Part 2
Score of 0 Normal Score of 1 One movement abnormality Score of 2 Two movement abnormalities Score of 3 Three movement abnormalities
Huntington’s Disease Scoring: Combining Part 1 and 2
Scores range from 0:0 (normal, no disease) to 3:3 (greatest expression of clinical signs)
Choose score/score combinations for adding care :
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food and water accessibility, supplements
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heat source, nesting materials
Huntington’s Disease
Reference:
Guyenet, et.al. (2010). A Simple Composite Phenotype Scoring System for Evaluating Mouse Models of Cerebellar Ataxia. JoVE. 39. ID 1787. Tests: ledge test, hindlimb clasping, gait, kyphosis
Huntington’s Disease
Noise sensitivity: construction, cage changes, hoods and ventilation
Seizure-like activity and deaths related to noise and other disturbances
Hypothermia in late stage disease: external heat source; nestlets vs. paper nesting material Enviro dri®
(Shepherd Specialty Papers) www.ssponline.com
Summary
Meet with investigators: look at their scoring and weight data
Provide specific solutions and options
Include investigators on guidelines development