don't miss a beat: understanding continuous, real time physiologic monitoring
DESCRIPTION
In vivo, preclinical research encompasses numerous study designs with various species and endpoints being monitored. Having access to all available study data allows the scientist to comprehensively understand the study paradigm and make informed research decisions. During Session 3 of our webseries "Biotelemetry For The Life Sciences", presenters discussed the importance of continuous, real-time monitoring in preclinical research. Case studies included using EEG as a biomarker for CNS activity and drug discovery and using telemetry for disease characterizations and and evaluation of vaccines in Biodefense research. During this exclusive webinar sponsored by Data Sciences International, Steve Fox shares his experience from pharmaceutical development; discussing the importance of continuous EEG monitoring for sleep studies. Anna Honko explains the importance of having access to real-time, continuous data when studying infectious diseases in non-human primates in a Biodefense setting. In addition, Dusty Sarazan reviews how and why continuous, real-time monitoring has become a preferred and essential method for acquiring and studying physiology in today's preclinical research setting. Key Topics: EEG as a biomarker for CNS activity and a platform for pre-clincal drug discovery Sleep/wake patterns and rhythms, and how qEEG signatures allow for accurate clinical predictions of efficacy and CNS adverse event screening Considering the FDA Animal Rule Basic disease characterizations and evaluation of vaccines and therapeutics Non-human primate models of viral biodefense and emerging pathogens Translating pre-clinical study findings to human, clinical populations Guest Speakers: Steve Fox, BS Associate Principal Scientist, Merck & Co., Inc. Anna Honko, PhD Staff Scientist, NIH/NIAID Integrated Research Facility R. Dustan Sarazan, DVM, PhD Vice President & Chief Scientific Officer, Data Sciences InternationalTRANSCRIPT
InsideScientific is an online educational environment designed for life science researchers. Our goal is to aid in the sharing and
distribution of scientific information regarding innovative technologies, protocols, research tools and laboratory services.
Today’s Presenters:
R. Dustan Sarazan, DVM, PhD
Vice President & CSO, Data Sciences International
Steve Fox, BS
Associate Principal Scientist, Merck & Co., Inc.
Anna Honko, PhD
Staff Scientist, NIH/NIAID Integrated
Research Facility
years – 1984 to 2014
Integrated Physiologic Monitoring for Life Science Research
DSI is a pioneering research company focused on systems physiology and pharmacology. We are celebrating 30 years of advancing the field of physiologic monitoring this year. Over the past 30 years, we’ve delivered proven solutions that
have been used to monitor more than 500,000 research subjects and study findings have been published in over 1000 peer-reviewed journal articles. DSI’s 200 scientists, engineers, and personnel are based out of St. Paul, MN, but we also
have offices in Europe and Asia where sales and technical support staff are located to help our customers abroad.
Benefits of Telemetry
1. Freely moving, conscious animals
2. Eliminates exit site infections
3. Chronically instrumented animals can be used sequentially as their own controls, or in several studies, thus improving animal welfare
4. Decreases costs
5. Enhances safety
Additional Considerations
• Number of animals
• Physiologic endpoints of interest
• Study duration
• Interventions
• Data collection schedule
• Data analysis
Utilizing Continuous EEG in Drug Discovery: From Mouse to Man
Steve Fox Associate Principal Scientist
Pharmacology - EEG Merck Research Laboratories
Copyright S. Fox, Data Sciences International, and InsideScientific. All Rights Reserved.
Challenges of Drug Discovery for CNS Indications
a Nature Reviews Drug Discovery 11, 191-200 (March 2012)
Increasing POS of Drug Candidates:
1. Expose at the target site of action over a desired period of time
2. Binding to the pharmacological target as expected for its mode of action
3. Expression of pharmacological activity commensurate with the demonstrated target exposure and target binding
For a development candidate to have potential to elicit the desired effect over the necessary period of time, three fundamental elements need to be demonstrated:
CNS Biomarkers
Body
Brain
Functional Brain Area
Cortex
Mid - Brain Nuclei
ms Second Minute Hour Day Week Months Years
PET
CSF PK
MRI
Genetic Traits
Leve
l of A
nato
mic
Acc
urac
y
Cognition/Mood Space
EEG
EEG Spans a Large Temporal Space
Measure Pharmaco-Dynamic? Pharmaco-Kinetic? Comment:
PET
No
Yes
- Expensive, patient location locked to scanner
MRI
Yes
No
- Expensive, patient location locked to scanner
Genetics No No - Possibly response predictive
EEG
Yes
Yes
- Patient is location and time independent - Spans large temporal and anatomic space - Inexpensive, portable, scaffold for other clinical measures
Symphony in the Brain • Tuba section • Trombone section • French horn section • Trumpet section
EEG • Delta activity (0.5 - 4 Hz) • Theta activity (4 - 8 Hz) • Alpha activity (8 -13 Hz) • Beta activity (14 - 26 Hz)
Beta
Active Cognition
Alpha
Relaxed Wake
Delta
Deep Sleep
Theta
REM Sleep
In Humans: Recording Brain Activity is Done Readily Using External Measurements
Sanna et al. 2004, Int J Sports Med; 25: 457-460
Circadian Physiology
Moore-Ede et al., 1983 NEJM
Sleep is Assessed Using Broadly Useful Measures (EEG, EMG, EOG)
Including: • ECG • cognition • mood • respiration • activity • body temperature • vocalization
& more…
Polysomnography
Sleep is Assessed Using Broadly Useful Measures (EEG, EMG, EOG)
Including: • ECG • cognition • mood • respiration • activity • body temperature • vocalization
& more… NEJM 1974, 290(9) 487
Sleep is Translational by Nature…
Pharmacological Confidence in EEG
1. MRL Sleep / EEG Lab Translation • Rely on natural translation of sleep & EEG
• Employ multiple species (mice, rats, dogs, monkeys & man)
• Add therapeutic area specific measures
2. Sleep & qEEG as Biomarkers • EEG is time scalable
• Dynamic frequencies and transients
• Topography
3. Combine EEG, Cognition & PK • 3-D drug signature
• PK/PD and PD/PD estimates
4. Automation to Drive Probability of Success • Data collection is easy – data analysis is hard
• Alignment of pre-clinical & clinical end points
EEG patterns are conserved among mice, rats, dogs, NHP and humans
- this approach is readily translatable among species…
Ideal Profile for Insomnia Drug:
- reduce sleep latency (sleep onset) and sleep fragmentation (maintenance), improve next-day performance, non-addictive
EEG Translational Strategy for Drug Discovery
• Similar study designs across species
• Control for environmental conditions
• Standardize and optimize analytic approaches
CNS Drug Discovery Improved POS Higher POS Optimal POS Minimal POS
Patients
Human PhI
Rat
Effic
acy,
PK
Patients
Human PhI
Canine
Rat
Mouse (TG)
NHP
Slee
p / q
EEG
/ Ph
arm
aco-
EEG
Effic
acy,
PK
, Cog
nitio
n, T
emp,
EC
G, A
ctiv
ity
Patients
Human PhI
Rat
Mouse (TG)
Slee
p / q
EEG
/ Ph
arm
aco-
EEG
Effic
acy,
PK
Patients
Human PhI
Rat
Mouse (TG)
Slee
p / q
EEG
/ Ph
arm
aco-
EEG
Effic
acy,
PK
, Cog
nitio
n, T
emp,
EC
G, A
ctiv
ity
1. Minimize stress, surgeries, possibilities for infection
2. Maximize predictive ability, animal health, and model durability
3. Measure continuously over multiple circadian cycles/ drug administrations
4. Support discovery and development programs using automated workflows and algorithms
Pre-clinical EEG methods are similar to human but are optimized to:
Measuring Sleep in Pre-Clinical Species
DSI Implant
F20/F40 EET
Quad-ET
D70-EEE
Standard Analytic Approaches for Pre-Clinical Studies
EEG Recording V/t
∆ φ α β γ
Arousal State Average Across Time
Spectral Distribution Spectral Band Average Across Time
Sleep Scoring
FFT Analysis
Compound L
• EEG + EMG + Activity
• Polysomnography
• Quantitative EEG (qEEG) – Fast Fourier Transform (FFT)
• Group frequencies by bands (Delta, Theta, Alpha,…)
• Time course of drug effect
Rat Sleep as High-Throughput PK/PD Screen
• Sleep/EEG signatures of CNS compounds
• Focuses target selection, lead optimization, and biomarker assessment
• Wireless implants allow for additional measures in home cages
Paroxetine (Paxil©)
Buproprion (Wellbutrin©)
Buspirone (Buspar©)
High-Throughput EEG Screen for Seizure Liability
• WAG-Rij rats genetically susceptible to absence-like seizures
• EEG evaluates brain penetrance and thalamic calcium channel target engagement
• Automated scoring reduces cycle time while maintaining accuracy 0%
20%
40%
60%
80%
100%
0 2 4 6 8 10 12 14 16 18 20 22
Perc
ent C
umul
ativ
e Se
izur
e Ti
me
(%)
Untreated (n=32)
200mpk (8)400mpk (8)
Dosed
50mpk (8)
ACTIVE IN-ACTIVE ACTIVE
Hours After Dosing
0%
20%
40%
60%
80%
100%
0 2 4 6 8 10 12 14 16 18 20 22
Perc
ent C
umul
ativ
e Se
izur
e Ti
me
(%)
Untreated (n=32)
0%
20%
40%
60%
80%
100%
0 2 4 6 8 10 12 14 16 18 20 22
Perc
ent C
umul
ativ
e Se
izur
e Ti
me
(%)
Untreated (n=32)
200mpk (8)200mpk (8)400mpk (8)400mpk (8)
Dosed
50mpk (8)50mpk (8)50mpk (8)
ACTIVE IN-ACTIVE ACTIVE
Hours After Dosing
5 sec
seizure
Automatically scored seizure
Ethosuximide
Summary
= statistically significant effects 0.05, 0.01, 0.001
0 2 4 6 8 10 12 14 16
OlanzapineVehicle
Active Wake
Light Sleep
Delta Sleep
REM Sleep
Hours
Dur
atio
n (M
in)
Olanzapine• 10 mpk, PO• Vehicle- 0.5% Methylcellulose• Analysis of all nights • Dose Time=3:30• Time 0 = 3:00pm• n=8 rats & nights: 56
Study details
-30369
1215
3
6
9
12
15
5
10
15
20
25
0
2
4
6
8
1. 8 hour decrease in active wake starting immediately post dose
2. Increase in light sleep for 2 hours
3. 10.5 hour increase in delta sleep for starting 3 hours post dose
4. 1 hour increase in REM sleep immediately post dose; 5 hour decrease in REM starting 2 hours post dose
Summary
= statistically significant effects 0.05, 0.01, 0.001
0 2 4 6 8 10 12 14 160 2 4 6 8 10 12 14 16
OlanzapineVehicleOlanzapineVehicle
Active Wake
Light Sleep
Delta Sleep
REM Sleep
Hours
Dur
atio
n (M
in)
Olanzapine• 10 mpk, PO• Vehicle- 0.5% Methylcellulose• Analysis of all nights • Dose Time=3:30• Time 0 = 3:00pm• n=8 rats & nights: 56
Study details
-30369
1215
3
6
9
12
15
5
10
15
20
25
0
2
4
6
8
1. 8 hour decrease in active wake starting immediately post dose
2. Increase in light sleep for 2 hours
3. 10.5 hour increase in delta sleep for starting 3 hours post dose
4. 1 hour increase in REM sleep immediately post dose; 5 hour decrease in REM starting 2 hours post dose
Rat Sleep as Adverse Event Screen
• Sleep, EEG, & cognition are also adverse event detection tools
• Seizures also detected automatically if needed
• Zyprexa causes “excessive sedation” when given PO to rats
Rat Sleep Study: Olanzapine
Ancillary Measures Using Quad-ET in Rats
• Acute and chronic changes in SC body temperature and heart rate
• Indicative of adverse effects, though insomnia drugs may reduce body temperature and heart rate to sleep-phase levels
• Activity data not shown, but is also analyzed
Heart Rate SC Body Temperature
Innovative Research
• Flexibility to design and run interesting studies
• Sleep Disruption
• Lights-on, Sounds
• Develop new translatable analysis
• Combine EEG + Cognition for residual sleep effects / arousability
0
2
4
6
0
5
10
15
0
5
10
15
-0.10
0.10.20.30.4
0
10
20
30
Dur
atio
n (M
in)
REM Sleep
Delta Sleep II
Delta Sleep I
Active Wake
Quiet Wake
REM Sleep
Delta Sleep II
Delta Sleep I
Active Wake
Quiet Wake
0 2 4 6 8 10 12 14 16 18 20 220 2 4 6 8 10 12 14 16 18 20 22
Sleep Disruption
Human “Home Cage” EEG
Recursive Vertical Integration
• PD/PK across species using common PD/PK endpoints
• Improve next-generation drug quality
• Decrease clinical discovery time
Cellular Assays (Specificity, Potency)
Rodent Assays (Safety, EEG)
Monkey Assays (EEG, Cognition)
Identify Specific Signals
Identify Efficacy (Success per Year)
2MM
3000
200
3-5
Human Assays (EEG, Cognition)
+ ID
+ ID
+ ID
30
1 FDA Approved!
Can the flow of medicines be improved? Fundamental pharmacokinetic and pharmacological principles toward improving Phase II survival
Sleep Lab Translational Strategy…
Paul Morgan, Piet H. Van Der Graaf, John Arrowsmith, Doug E. Feltner, Kira S. Drummond, Craig D. Wegner and Steve D.A. Street
Drug Discover Today – Volume 17, Number 9/10 – May 2012
EEG as a Model for CNS Drug Discovery
1. Sleep & qEEG as biomarkers for CNS penetration and vertical translation to patients
2. EEG is high throughput & highly translational 3. Multi-species for optimal human predictions
• Sleep • qEEG • Cognition • Aging
4. Ambulatory PSG/EEG • FIM • CNS Penetration • Dose selection
5. Database building • CNS target specific EEG signals • PK/PD & PD/PD across species
Patients
Human
Non-Human Primate
Canine
Rat
Mouse
Telemetry for Biodefense Research Applications
Anna Honko, PhD Staff Scientist
NIH/NIAID Integrated Research Facility
Frederick, MD
Copyright A. Honko, Data Sciences International, and InsideScientific. All Rights Reserved.
What will we cover today…
1. Introduce what makes biodefense research unique from other pharm/tox or drug discovery applications in the context of disease models
2. Discuss some of the different study designs where telemetry can be utilized
Biosafety Level 3 (BSL-3)
• Agents associated with serious or lethal human disease for which preventive or therapeutic interventions may be available
• Personnel may have to change into scrubs before entering the laboratory and take a complete shower before exiting
• Other personal protective equipment may be required as well, depending on the tasks to be performed
CDC Biosafety in Microbiological and Biomedical Laboratories (BMBL) 5th Edition
Biosafety Level 4 (BSL-4)
• BSL-4 is required for work with dangerous and exotic agents that pose a high risk of aerosol-transmitted laboratory infections and life-threatening disease that is frequently fatal, for which there are no approved vaccines or treatments
CDC Biosafety in Microbiological and Biomedical Laboratories (BMBL) 5th Edition
BSL-4 viral agents:
Ebola virus Marburg virus
Smallpox (Variola virus)
Lassa Fever Virus Junin Virus
Crimean-Congo hemorrhagic
fever virus
BSL-3 bacterial agents:
Yersinia pestis Bacillus
anthracis Francisella tularensis
Mycobacterium tuberculosis
BSL-3 viral agents:
Venezuelan equine encephalitis virus
Eastern equine encephalitis virus Western equine
encephalitis virus SARS coronavirus Yellow Fever Virus
West Nile Virus Chikungunya
Rift Valley Fever Virus
Due to engineering design requirements of laboratory, telemetry system reliability is critical… 1. Only persons whose presence in the facility or individual laboratory
rooms is required for scientific or support purposes are authorized to enter
2. Equipment must be decontaminated using an effective and validated method before repair, maintenance, or removal from the laboratory
This means… • Only able to install or make major repairs when laboratory is decontaminated, or • Personnel must be able to install or make minor adjustments if necessary • Telemetry device sterilization/decontamination
[Either one-time use or decontaminate and replace/exchange (factor into costs)]
CDC Biosafety in Microbiological and Biomedical Laboratories (BMBL) 5th Edition
Sequence of Entry Rooms
• Since there is frequently a physical separation between the telemetry base station and the animals being monitored, communication is critical…
Biodefense research and the FDA Animal Rule
1. Non-traditional regulatory pathway for new drug approval when human efficacy studies are not ethical or feasible
2. The Animal Rule states that FDA will rely on evidence from animal studies to provide substantial evidence of effectiveness only when the following four criteria are met: 1. There is a reasonably well-understood pathophysiological mechanism of the
toxicity of the substance and its prevention or substantial reduction by the product;
2. The effect is demonstrated in more than one animal species expected to react with a response predictive for humans, unless the effect is demonstrated in a single animal species that represents a sufficiently well-characterized animal model for predicting the response in humans;
3. The animal study endpoint is clearly related to the desired benefit in humans, generally the enhancement of survival or prevention of major morbidity; and
4. The data or information on the kinetics and pharmacodynamics of the product or other relevant data or information, in animals and humans, allows selection of an effective dose in humans. Paraphrased from “Guidance for Industry, Product
Development Under the Animal Rule, May 2014”
3. Adequate and well-controlled animal efficacy studies are required for approval under the Animal Rule = well-documented and GLP if possible
Purpose of natural history studies for rare diseases
From a presentation by Anne R. Pariser, M.D. , former Associate Director for Rare Diseases, CDER, FDA:
• “The natural course of a disease from the time immediately prior to its inception, progressing through its presymptomatic phase and different clinical stages to the point where it has ended and the patient is either cured, chronically disabled or dead without external intervention”1
There is limited natural history data in humans for high containment diseases for a number of reasons:
• Sporadic nature of the outbreaks and the remote locations of the cases, availability of resources at sites
• Variability in dose, route of exposure and time to presentation for individual cases
1Posada de la Paz M; Groft SC. 2010. Rare diseases epidemiology. Vol. 686
Benefits of natural disease progression studies in non-human primates using telemetry
1. Studies are observational and longitudinal, allowing for
the characterization of the pathophysiology of the disease in a more controlled system
• Does the disease in nonhuman primates model the disease in humans?
• Are we able to identify potential triggers for therapeutic intervention, biomarkers or correlates of disease?
Benefits of natural disease progression studies in non-human primates using telemetry
2. Some disease manifestations that are common in human cases, such as shortness of breath or fever, are difficult to objectively measure in nonhuman primates cageside • Telemetry is considered a “refined technique” for collecting this data
3. Additionally, if mortality is used as a primary endpoint, considerations must be made for the effects of euthanasia criteria • Real-time monitoring using telemetry has provided an additional means
of objective euthanasia criteria, reducing observational study bias
• Increasing sample integrity since terminal sample collection can occur
Study designs that benefit from telemetry data acquisition
1. Longitudinal natural history studies and basic pathophysiology modeling studies
2. Comparison of nonhuman primate species
3. Agent dose-seeking studies
4. Identification / application of therapeutic interventions, establishment of euthanasia criteria
5. Efficacy trials of potential vaccine or therapeutic
Longitudinal natural history studies and basic pathophysiology modeling studies
Characterization of disease progression differences between fatal and non-fatal viral infection
Non-survivor Survivor
1
Longitudinal natural history studies and basic pathophysiology modeling studies 1
Ability to match physiological parameters by telemetry with commonly collected clinical parameters
Plasma RNA levels Clinical serum chemistry
Febrile response of macaques following EEE virus exposure
Longitudinal natural history studies and basic pathophysiology modeling studies 1
Comparisons of viral disease in different NHP models
Comparison of Non-human Primate Species 2
Comparison of febrile responses between dose groups of aerosol exposed macaques (WEE)
Low Dose High Dose
Agent Dose-Seeking Studies 3
Trigger to initiate therapeutic intervention • Results of natural disease modeling studies can be used to determine triggers for treatment
• In combination with RT-PCR assessment, fever as calculated by ≥1.5°C above pre-challenge baseline using remote telemetry monitoring was used to determine starting point for mAb treatment for Ebola
Identification / application of therapeutic interventions, establishment of euthanasia criteria 4
Comparison of vaccine efficacy: Sham-immunized controls challenged against EEEV
Efficacy trials of potential vaccine or therapeutic 5
Comparison of vaccine efficacy: SIN-SAEEEV vaccinated (nonprotective – 17% survival)
Efficacy trials of potential vaccine or therapeutic 5
Comparison of vaccine efficacy: SIN/NAEEEV vaccinated (significant protection – 82% survival)
Efficacy trials of potential vaccine or therapeutic 5
Summary
Real-time monitoring of physiological parameters using less-invasive implanted telemetry devices will provide critical data for:
1. Development of animal models for biodefense or emerging diseases
2. Evaluation of potential vaccines and therapeutics
3. Establishment of objective criteria for therapeutic interventions or endpoints
Thank You! For additional information on telemetry applications and various DSI solutions for continuous monitoring of physiology in pre-clinical research studies please visit:
www.datasci.com
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InsideScientific is an online educational environment designed for life science researchers. Our goal is to aid in the sharing and
distribution of scientific information regarding innovative technologies, protocols, research tools and laboratory services.