tse phenomaster - infrafrontier · expand the time of testing remove the fear/anxiety induced by...
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Copyright © 2016 TSE Systems International Group - All rights reserved.
Automated Multi-Dimensional Phenotyping
of Mouse and Rat Models of Human Diseases
TSE PhenoMaster Metabolic Disease – Metabolic Phenotypes
Harry Knot
TSE Systems [email protected]
Copyright © 2016 TSE Systems International Group - All rights reserved.
Introduction
TSE Vision behind home-cage testing bring the social component back to the laboratory expand the time of testing remove the fear/anxiety induced by handling remove humans bias save manpower, costs and space
Copyright © 2016 TSE Systems International Group - All rights reserved.
Home Cage Environment
Drinking & Feeding & Body Weight Monitoring
Operant Wall
Running Wheel Indirect Calorimetry
Activity Sensor Frames
PhenoMaster – Modular Concept
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PhenoMaster
Metabolism – Indirect Calorimetry CaloSys
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Feeding & Drinking & Body Weight Monitoring
Universal Weighing Technology (consistent accuracy)
Open Cages or CaloCages
Feeding & Drinking & Body Weight Measuring
Choice Experiments
Access Controls (interactive , pairing )
Interchangeable: F / D / BW
F F
D
D D
F
F D
BW
Possible Combinations
Copyright © 2016 TSE Systems International Group - All rights reserved.
PhenoMaster – Calorimetry Module
Indirect Calorimetry
Home Cages / High Animal Welfare
Minimize habituation times
Long-term monitoring
With bedding / Easy Cleaning
Variable Add ons
Small volumes - suited for mice/rats
BUT:
Reasonable dimensions for activity
monitoring (animal welfare !)
Indirect calorimetry
O2, CO2, ( CH4, H2, 13C), H2O
RER (Respiratory Exchange Ratio)
Add-in: Modular Running Wheel
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PhenoMaster – Calorimetry Module CaloSys
Multi Plexing
Air Drying Unit
Sample Out Flow In
Air Out
Air Out Air In
Air In
Air Out Air In
Gas Sensors highly sensitive
Weak spots:
Pumps
Alarms
Flow
Sensors
Water vapor
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Indirect gas calorimetry:
Individual electronic mass flow controllers
MFC for each cage (universal mice / rats)
CO2 sensor:
Infrared spectroscopic differential
O2 sensor:
Paramagnetic differential (high-speed)
Zinkoxide (fuel cell), sensitive but SLOW
High-speed Peltier air drying unit or Dessicant
Choice: PULL (Suction principle) or PUSH
Fully Automated Gas Calibration: O2 & CO2
Choice: PULL or PUSH
Choice: MULTIPLEX or CONTINUOUS
PhenoMaster – Calorimetry Module CaloSys
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Home-Cage Environment Drinking & Feeding BW Monitoring
Indirect Calorimetry
Activity Sensor Frames
PhenoMaster – Modular Concept
Telemetry
Climate Chambers
Copyright © 2016 TSE Systems International Group - All rights reserved.
PhenoMaster – Calorimetry Module CaloSys
Choice: PULL or PUSH Principle
PULL
Advantages: best for small animals (chambers), defined air source, other gases (hypoxia), short lag time between cage and analyzer (continuous mode), enables the use of IsoCages (gnotobiotic / germ-free animals > microbiome, and Immunocompromised / infectious disease models) Disadvantages: potential leak problems
Advantages: “open cage” leak minor problem Disadvantages: sensitive to environmental changes, expired air can leave system, temperature control of the chamber
PUSH
Pressure Air Flow
H2O, CO2 Gas
Analyzer
Suction H2O, CO2 Gas
Analyzer
Environment monitoring system (light,humidity, Human acitvity, O2, CO2
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PhenoMaster – Calorimetry CaloSys Multiplex
O2 / CO2 Sensor Pair
Cage 1 Cage 2 Cage 3 Cage 4 Cage 5 Cage 6 Cage 7 Cage 8
e.g. 1 sample / 9 (27) min cycle
CaloSys
Ref. Cage
Scalable technology, consistent precision, easy expandable, budget friendly
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PhenoMaster – Calorimetry CaloSys Accelerated Multiplex
O2 / CO2 Sensor Pair
O2 / CO2 Sensor Pair
1 sample / 5 (15) min
Cage 1 Cage 2 Cage 3 Cage 4 Cage 5 Cage 6 Cage 7 Cage 8
Ref. Cage
CaloSys CaloSys
Copyright © 2016 TSE Systems International Group - All rights reserved.
PhenoMaster – Calorimetry CaloSys Continuous
O2 / CO2 Sensor
Pair
O2 / CO2 Sensor
Pair
O2 / CO2 Sensor
Pair
O2 / CO2 Sensor
Pair
O2 / CO2 Sensor
Pair
O2 / CO2 Sensor
Pair
O2 / CO2 Sensor
Pair
O2 / CO2 Sensor
Pair
Cage 1 Cage 2 Cage 3 Cage 4 Cage 5 Cage 6 Cage 7 Cage 8
1 sample / (1 sec)
CaloSys
Ref. Cage
Copyright © 2016 TSE Systems International Group - All rights reserved.
The organs that under standard conditions mainly contribute to BMR (Basal Metabolic Rate):
The muscles (25%) Exercise, XYZ Activity EMG Telemetry The liver (17-20%) Temp The gastrointestinal tract (5-10%) Telemetry, smEMG The kidney (6-7%) GFR The lung (1-4%) Telemetry, RR,RV,TV The brain (3-20%) Cognition, Learning, Memory, EEG The heart (3-11%) Telemetry, BP, ECG, PWA
Note: In small rodents and hibernators, a considerable amount of oxygen can be used by the brown adipose tissue during non-shivering thermogenesis! > Dual Temp Telemetry
Copyright © 2016 TSE Systems International Group - All rights reserved.
PhenoMaster – OPEN System Structure (Hardware & Software – many Home Cages in parallel)
PhenoMaster Control System
HW & SW
Calorimetry
Feeding Drinking
Bodyweight Activity Running
Wheel Operant
Wall
Home Cage
Optogen Infusion
WiredEEG
Stellar
Telemetry
Copyright © 2016 TSE Systems International Group - All rights reserved.
Feeding & Drinking & Body Weight Monitoring
Universal Weighing Technology (consistent accuracy)
Open Cages or CaloCages
Feeding & Drinking & Body Weight Measuring
Choice Experiments
Access Controls (interactive , pairing )
Interchangeable: F / D / BW
F F
D
D D
F
F D
BW
Possible Combinations
Copyright © 2016 TSE Systems International Group - All rights reserved.
CaloCage with integrated Voluntary Running Wheel: Enabled / Disabled (Time & Distance Control on Running)
Workload Control
Paired Running
Operant Control
Running for Food
Running for Liquid
PhenoMaster – Metabolism: Exercise Calorimetry
Copyright © 2016 TSE Systems International Group - All rights reserved.
PhenoMaster – Modular Running Wheel for Exercise Calorimetry
Voluntary Wheel (software controlled):
New application within CaloCage:
Defined work loads by workload control
Pre-programmed workload profiles
Time control by Enabled / Disabled
Analysis of voluntary running
Refined software: automated analysis
for exercise calorimetry
2017: Endurance & Weight training!
Copyright © 2016 TSE Systems International Group - All rights reserved.
PhenoMaster – Metabolism & Cognition
0
20
40
60
80
100
120
1 2 3 4 5 6 7 8 9 10 11 12 13
Nos
e Po
kes
Test Day
Copyright © 2016 TSE Systems International Group - All rights reserved.
CaloCage Mouse:
Indirect Calorimetry
Operant Wall (Liquid Reward)
Automated Liquid Restriction
IntelliMaze integration
PhenoMaster – Metabolism & Cognition
Copyright © 2016 TSE Systems International Group - All rights reserved.
PhenoMaster – Swivel Applications with Operant Control
Automated Infusion
by Syringe Pump
Optogenetics
Blood Sampling
Microdialysis
Electrophysiology
Electrostimulation
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PhenoMaster – Multiple Fields of Applications
PhenoCage Configurations
PhysioCage CircadianCage
CogniCage ChronoCage
ObesityCage AnxietyCage
AddictionCage SocialCage > IntelliCage
Metabolism: Obesity&Diabetes, Cognition Learning & Memory, Exercise & Sports, Immunology & Cancer
Copyright © 2016 TSE Systems International Group - All rights reserved.
PhenoMaster: IDO Helmholtz Munich
32 Mice System Shown:
Climate Chamber
with 8 Places
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MPI Psychiatry
Munich
PhenoMaster Software Acquisition – Analysis - Modeling
> Aquisition CaloPM: Integration with IntelliMaze
> OriginLab, Advanced analysis and Presentation
with Statistics plugin (SPSS, R-scripts)
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PhenoMaster – Calorimetry Data Analysis
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PhenoMaster 1.0 > 2.0 – Software
Analysis & Batch Processing Easier to customize Report Tables in graphs New Statistics such as 3-way ANOVA, Partial Correlation, etc. Word Template for Report Creation in Batch Processing
Programming R Console and support for Rserve to exchange data between Origin and R
Graphing Smart Plotting with Clonable Graph Templates
Copyright © 2016 TSE Systems International Group - All rights reserved.
MPI Psychiatry
Munich
Metabolic Modeling in PhenoMaster Systems
Clinical strength PKPD Software
Features predictive modeling
Experimental design & planning
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PhenoMaster – UTSW 4/2016
Consistent reproducible, robust high quality data after 10+ years continuous operation
Torpor !
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Energy expenditure increases with decreasing environmental temperature – Climate Control
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Typical diurnal changes in energy expenditure and locomotor activity of laboratory mice
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PhenoMaster - Applications
• Pharmacology of Diabetes / Obesity
• Space Physiology – Metabolism
• Microbiota & Energy metabolism
• Phenotyping – Huntington Disease - Activity
• Phenotyping – Running - Open Field
Copyright © 2016 TSE Systems International Group - All rights reserved.
A rationally designed monomeric peptide triagonist corrects obesity and diabetes in rodents Nature Medicine 21, 27–36 (2015)
We report the discovery of a new monomeric peptide that reduces body weight and diabetic complications in rodent models of obesity by acting as an agonist at three key metabolically-related peptide hormone receptors: glucagon-like peptide-1 (GLP-1), glucose-dependent insulinotropic polypeptide (GIP) and glucagon receptors.
Copyright © 2016 TSE Systems International Group - All rights reserved.
(a–j) Effects on body weight change (a), cumulative food intake (b), fat mass change (c), lean mass change (d), body weight–correlated average energy expenditure (e), cumulative energy expenditure (f), real-time respiratory quotient (g)
Figure 4: Addition of glucagon activity contributes thermogenic character to the triagonist
http://www.nature.com/nm
Copyright © 2016 TSE Systems International Group - All rights reserved.
Dual melanocortin‐4 receptor and GLP‐1 receptor agonism amplifies metabolic benefits in diet‐induced obese mice.
EMBO Molecular Medicine (2015) 7, 288-298
“The superior metabolic efficacy of this combination therapy is attributed to the anorectic and glycemic actions of both drugs, along with the ability of RM‐493 to increase energy expenditure”
Copyright © 2016 TSE Systems International Group - All rights reserved.
Effect of liraglutide and RM‐493 co‐treatment on body weight and body composition in DIO mice A–CFive‐day treatment of DIO male mice with vehicle (white), liraglutide (10 nmol/kg)
(gray), RM‐493 (3.6 μmol/kg) (black), or liraglutide (10 nmol/kg) and RM‐4...
Christoffer Clemmensen et al. EMBO Mol Med. 2015;7:288-298
©2015 by European Molecular Biology Organization
Copyright © 2016 TSE Systems International Group - All rights reserved.
Effect of liraglutide and RM‐493 co‐treatment on energy metabolic
parameters in DIO mice A–GFive‐day treatment of DIO male mice with
vehicle (white), liraglutide (10 nmol/kg) (gray), RM‐493
(3.6 μmol/kg) (black), or liraglutide (10 nmol/kg) and RM‐493 (3...
Christoffer Clemmensen et al. EMBO Mol Med. 2015;7:288-298
©2015 by European Molecular Biology Organization
Copyright © 2016 TSE Systems International Group - All rights reserved.
Mice in Bion-M 1 Space Mission: Training and Selection
PLOS ONE | www.plosone.org 1 August 2014 | Volume 9 | Issue 8 |
This paper presents an overview of the scientific goals, the experimental design and the mouse training/selection program. The aim of mice experiments in the Bion-M 1 project was to elucidate cellular and molecular mechanisms, underlying the adaptation of key physiological systems to long-term exposure in microgravity. The studies with mice combined in vivo measurements, both in flight and post-flight (including continuous blood pressure measurement), with extensive in vitro studies carried out shortly after return of the mice and in the end of recovery study
Cognitive & Metabolic Profiling pre/post flight, telemetry in Bion-M2
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https://www.alnmag.com/news/2016/06/how-changes-microbiome-cause-obesity#disqus_thread
How Changes to the Microbiome Cause Obesity
Acetate mediates a microbiome–brain–β-cell axis to promote metabolic syndrome
http://www.nature.com/nature/journal/v534/n7606/full/nature18309.html
Correlations of Fecal Metabonomic and Microbiomic Changes Induced by High-fat Diet in the Pre-Obesity State
http://www.nature.com/articles/srep21618
Impact of dietary resistant starch type 4 on human gut microbiota and immunometabolic functions http://www.nature.com/articles/srep28797
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PhenoMaster – New developments
• Climate Chambers
• BAT/WAT Telemetry
• New sensors (13C, CH4, H2, VOCs)
• New applications (IsoCage / Push mode)
Copyright © 2016 TSE Systems International Group - All rights reserved.
PhenoMaster – Calorimetry Module
Indirect Calorimetry
Home Cages / High Animal Welfare
Minimize habituation times
Long-term monitoring
With bedding / Easy Cleaning
Variable Add-ons > Applications
Climate Chambers / Telemetry
Indirect Calorimetry Innovations
O2, CO2, H2O gas sensors
RER (Respiratory Exchange Ratio for quantification of substrate utilization) & Water balance.
CH4, H2, VOCs gas sensors
Microbial activity of gut microbes
New Metabolic PhenoCage, urine/feces collection for EE and Microbiome analysis
13C/12C gas sensor
Advanced substrate utilization studies
13C-Glu, 13C-Fru, 13C-Gal, 13C-FAs
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Mouse Cage Systems:
Tecniplast Greenline IVC / Allentown NexGEN Easy IVC
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All-purpose use Bioexclusion Biocontainment
Indirect Calorimetry D/F/BW/Ac Exercise Cognition Optogenetics Telemetry
Indirect Calorimetry D/F/BW/Ac Gnotobiotic Germ-free Animals Microbiome Ext CH4/H2/H2O/VOCs
Indirect Calorimetry D/F/BW/Ac Immunocompromised Animals Infections / Inhalation Tox
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Phenomaster Summary: Most flexible and modular metabolic phenotyping platform on the market Most advanced capabilities and integration towards scientific endpoints Number one choice for mission critical metabolic studies (Pharma) Only system tried and tested in push mode for hypoxia and gnotobiotic research Only system capable of temperature challenges for BAT/WAT research with Telemetry 24/7 support by a team of 12 PhD‘s and a dozen Engineers. Over 350 systems installed worldwide (USA, EU, ASIA, AU) Over 500 publications of which 30% in the top 20 journals in their respective fields!