Transcript James Battey, Director NIDCD, NIH.

Presentation Overview
1.
2.
3.
4.
Use of mice in biomedical research
KOMP: Activity and progress to date
Rationale and planning for phenotyping
Request for continued support
Mice in Biomedical Research
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•
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The laboratory mouse has been considered the premiere experimental model of human biology and
disease since 1902 when it was first used to demonstrate how genetic traits could be transferred from
parents to offspring via classical or “Mendelian” inheritance in mammals.
In just over a century, an impressive array of genetic tools, reagents and processes has been developed in
the mouse, including:
– Homozygous inbred strains
– Recombinant inbred and consomic strains
– Transgenic and knockout methods
– Monoclonal antibodies
– iPS cells
Evidence that mice have played a key role in biomedical research is provided by the fact that at least 18
Nobel prizes have been awarded for work done using the mouse. Most recently, the 2007 Nobel Prize was
awarded to Mario Capecchi, Martin Evans, and Oliver Smithies for their discovery of the "principles for
introducing specific gene modifications in mice by the use of embryonic stem (ES) cells.”
The Premiere Model Organism
Janan Eppig, Jax
KOMP Vision
R01s
KOMP2 (2011-21)
ARRA (2010-11)
KOMP (2006-11)
Austin et al., Nature Genetics, 2004
Funded KOMP Projects
The Completion of a Comprehensive Mouse Knockout Resource
($47 M - 5 years - funded 9/06)
CHORI-Sanger-UC Davis
Regeneron Pharmaceuticals, Inc.
Development and Improvement of Inbred ES Cell Lines for Use in Generation of Mouse Mutants
($1.9 M - 2 years - funded 9/06)
U Pennsylvania
Samuel Lunenfeld Research Institute
A Data Coordination Center for the Knockout Mouse Project (KOMP)
($2.5M - 5 years - funded 9/06)
The Jackson Lab
The KOMP Repository
($4.8M - 4 years - funded 8/07)
UC Davis – CHORI
Goals and Progress
CSD
Regeneron
EuCOMM
NorCOMM
TIGM
Total
Goal (ES)
5,000
3,500
8,000
500
Vectors
4991
3116
4271
596
Targeted ES
3064
2038
2883
236
8,221
Mice
189
160
351
12
700
17,000
The KOMP
Website
Target List
www.knockoutmouse.org
The KOMP
Website
Details
Complete
information on
allele design,
vector,
genotyping and
sequencing
primers are
provided to the
end-user.
www.knockoutmouse.org
The KOMP Repository
The
repository
offers
vectors, KO
ES cells,
parental ES
lines, and
support
services.
Orders are
processed
using a
standard
“shopping
cart.”
www.komp.org
KOMP Repository Activities
Orders
120
WT-ordered
mice-ordered
cryo-recovery-ordered
germplasm-ordered
GLT-ordered
microinj-ordered
premium-ESC-pckg-ordered
ESC-ordered
100
80
60
40
20
Jan-10
Oct-09
Jul-09
Apr-09
Jan-09
Oct-08
Jul-08
Apr-08
Jan-08
Oct-07
Jul-07
Apr-07
Jan-07
0
Orders have recently increased to
about 100/month.
Over 3,000 users have created
accounts and registered interest in
~2,500 genes.
Kent Lloyd, UCDavis
Rationale for KOMP2
• Supporting a broad phenotyping effort would provide the
following advantages:
– A single cohort of mice would go through multiple phenotyping assays, so the
cost of producing multiple cohorts in different laboratories for phenotyping
would be eliminated.
– Each mutant mouse strain would be characterized for a broad set of
phenotypes in a way that will allow direct comparisons and result in a more
thorough description of gene function.
– Quality standards will be established and maintained, so the data will be of
the highest reliability.
– The risk of not finding a phenotype will be greatly reduced.
– Important, but unpublishable, negative results will be captured.
Planning for next phase: Phenotyping
• 3 workshops: Rome in 2007, Bar Harbor and Toronto in
2008 to establish vision for an IMPC & discuss
international, coordinated phenotyping efforts – agreed
that the way forward is to develop a business plan
• Medical Research Council/Wellcome Trust workshops in
Nov 2008 and Oct 2009 to engage UK scientific community
• NIH Phenotyping meeting, Bethesda October 2009
(survey)
• EC–funded EUMODIC (Helmholtz, Munich; ICS, Strasbourg,
MRC Harwell, WTSI) project is now doing broad-based
phenotyping of 500 mutant lines – completion 2011
The International Mouse Phenotyping Consortium
Members:
The Wellcome Trust
The Medical Research Council
The Wellcome Trust Sanger Center
MRC Harwell
European Commission
The National Institutes of Health
German Mouse Clinic
Toronto Center for Phenogenomics
Pending members:
Australia Phenomics Network
Czech Republic
Institut Clinique de la Souris
Nanjing University
Universitat Autonoma de Barcelona
Current funding:
EuMODIC - $7M/yr + matching funds, 150 strains
Sanger Mouse Genetics Program - $9M/yr, 200 strains
Australia Phenomics Network - $10.5M/yr
EMPReSSslim Primary Phenotyping Pipelines
Steve Brown, Harwell, MRC
Phenotyping of KOMP CSD mice at Sanger
Metabolism
• Weight gain curve
• Food intake and energy expenditure)
• Fat/lean tissue assessment by DEXA
• Glucose tolerance test
• Serum Insulin
• Blood Clinical Chemistry
Cardiovascular
• Blood clinical Chemistry
• Heart weight (hypertrophy)
• Heart histology
• Eye fundus (retinal vessels)
General
• Visible malformations
• Adult organ lacZ expression
• Necropsy
• Tissue banking for histology
Immune system
• Complete blood count
• White blood cell profile
• Immunoglobulin subclass profile
Infectious disease susceptibility
• Salmonella (bacterial)
Neurology
• SHIRPA assessment
• Hot plate (pain)
• Open field (anxiety)
• Stress-induced hyperthermia
• Brain anatomy
Musculo-skeletal
• Grip strength
• X-rays (skeleton)
• Bone mineral density
• Bone mineral content
Developmental Biology
• Citrobacter (bacterial)
• Plasmodium (malaria)
• Flu (viral)
Cancer
Reproduction
• Micronuclei analysis
• Fertility
http://www.sanger.ac.uk/mouseportal/
• Viability
• Embryonic lacZ pattern
• Viability at E14.5
• MRI at E14.5
Hair and skin
Senses
• Slit lamp (front of the eye)
• Eye fundus (retina)
• Auditory brain response (hearing
circuits)
• Hair growth rate
• Hair follicle stem cell
Ramiro Ramirez-Solis, WTSI
Genentech/Lexicon Mouse Phenotype Project
472 Mouse knockouts were broadly phenotyped
130 (27%) strains had 1 phenotype
245 (52%) strains had 2-5 phenotypes
Andy Peterson, Genentech
KOMP Phenotyping Scheme
Database
Web server
Mouse Production
KOMP Repository
Mice/Embryos
Tracking
Analysis
Display
Microinjection
Germline Transmission
LacZ staining
Lethality/Fertility
Cryopreservation
KOMP ES cells
EuCOMM ES cells
Mice/Embryos
Data
ES cells
Data
Mice/Embryos
Cohort breeding
Phenotyping
Data Upload
Cohort breeding
Phenotyping
Data Upload
Cohort breeding
Phenotyping
Data Upload
Mouse Phenotyping Center(s)