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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]

mailto:[email protected]




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


Home Cage Environment

Drinking & Feeding & Body Weight Monitoring

Operant Wall

Running Wheel Indirect Calorimetry

Activity Sensor Frames

PhenoMaster – Modular Concept


PhenoMaster

Metabolism – Indirect Calorimetry CaloSys


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


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


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


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



Home-Cage Environment Drinking & Feeding BW Monitoring


Activity Sensor Frames

PhenoMaster – Modular Concept

Telemetry

Climate Chambers



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


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

Copyright © 2016 TSE Systems International Group - All rights reserved. 12

PhenoMaster – Calorimetry CaloSys Accelerated Multiplex



1 sample / 5 (15) min


Ref. Cage

CaloSys CaloSys


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


1 sample / (1 sec)

CaloSys

Ref. Cage


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


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


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


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


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!


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


CaloCage Mouse:


Operant Wall (Liquid Reward)

Automated Liquid Restriction

IntelliMaze integration

PhenoMaster – Metabolism & Cognition


PhenoMaster – Swivel Applications with Operant Control

Automated Infusion

by Syringe Pump

Optogenetics

Blood Sampling

Microdialysis

Electrophysiology

Electrostimulation


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


PhenoMaster: IDO Helmholtz Munich

32 Mice System Shown:

Climate Chamber

with 8 Places


MPI Psychiatry

Munich

PhenoMaster Software Acquisition – Analysis - Modeling

> Aquisition CaloPM: Integration with IntelliMaze

> OriginLab, Advanced analysis and Presentation

with Statistics plugin (SPSS, R-scripts)


PhenoMaster – Calorimetry Data Analysis


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


MPI Psychiatry

Munich

Metabolic Modeling in PhenoMaster Systems

Clinical strength PKPD Software

Features predictive modeling

Experimental design & planning


PhenoMaster – UTSW 4/2016

Consistent reproducible, robust high quality data after 10+ years continuous operation

Torpor !


Energy expenditure increases with decreasing environmental temperature – Climate Control


Typical diurnal changes in energy expenditure and locomotor activity of laboratory mice


PhenoMaster - Applications

• Pharmacology of Diabetes / Obesity

• Space Physiology – Metabolism

• Microbiota & Energy metabolism

• Phenotyping – Huntington Disease - Activity

• Phenotyping – Running - Open Field


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.


(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


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”


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


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


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


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


PhenoMaster – New developments

• Climate Chambers

• BAT/WAT Telemetry

• New sensors (13C, CH4, H2, VOCs)

• New applications (IsoCage / Push mode)


PhenoMaster – Calorimetry Module


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


Mouse Cage Systems:

Tecniplast Greenline IVC / Allentown NexGEN Easy IVC


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


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!

tse phenomaster - infrafrontier · expand the time of testing remove the fear/anxiety induced by...

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