To achieve novel science objectives, validation of a rodent habitat on ISS will enable - In-flight analyses during long duration spaceflight- Use of genetically altered animals- Application of modern analytical techniques (e.g. genomics, proteomics, and metabolomics

Beegle, Janet

Globus, Ruth

NASA Technical Reports Server

Rodent Research-1 
Validation of Rodent Hardware 
Ruth Globus, Ph.D. 
Project Scientist 
Janet Beegle  
Project Manager 
https://ntrs.nasa.gov/search.jsp?R=20140010687 2019-08-31T20:07:30+00:00Z
NRC Decadal Survey for NASA (2011) : panel members 
unanimous in recommendation that ‘an animal habitat 
should be incorporated as soon as possible into the ISS’ 
Rodent research focus areas needed per NRC: 
  Bone, Muscle, Cardiovascular, Immune, Endocrine 
  Interacting systems and factors (including Radiation) 
Direction for Rodent Research on ISS 
Importance and Reason for ISS 
To achieve novel science objectives, validation of a rodent 
habitat on ISS will enable:  
–  In-flight analyses during long duration spaceflight 
–  Use of genetically altered animals 
–  Application of modern analytical techniques  
(e.g. genomics, proteomics, and metabolomics) 
Scientific knowledge to be gained from Rodent Habitat 
capabilities on ISS: 
–  Better understanding of how long term habitation in 
space affects mammalian physiology throughout 
lifespan:  180 days is  ~ 1/3 adult rodent lifespan 
–  Determine basic cellular and molecular mechanisms of 
gravity responses. 
–  Better define factors that impact crew health risk and 
develop effective countermeasures 
Overarching Hypothesis & Goals for 
Rodent Research on ISS 
Research conducted using rodents is foundational for 
advancing basic biological and biomedical scientific 
progress in space as on Earth. 
 
Research conducted using the Rodent Habitat on ISS 
provides the capability to: 
–  Determine biological and health effects of long 
duration spaceflight. 
–  Develop mitigation and countermeasure strategies for 
adverse effects 
 
Background and New Capabilities  
•  Heritage Animal Enclosure Modules (AEM) flew rats or 
mice successfully 27 times on the U.S. Space Shuttle.  
    Longest flight:  19 days 
•  Extend for ISS capability:  up to 180 days 
•  New hardware and operations requirements to conduct 
research on ISS: 
-  transit on unmanned vehicle (SpX Dragon) 
-  life support 
-  telemetry for matching Ground Control group 
-  long duration (30 days minimum per unit) 
-  video observation to minimize crew time 
-  on-orbit sample collection, preservation and return to 
Earth for analysis 
RR-1: Validation Mission Objectives 
Validate that: 
#1 the Rodent Habitat can deliver and maintain healthy 
animals deemed suitable for scientific research 
 
#2  on-orbit activities to support hardware operations can 
be performed successfully  
 
#3  a set of generic on-orbit operations can be performed 
which are needed to support science objectives, including 
euthanasia, dissection and sample preservation 
•  demonstrate capability of obtaining high quality 
science return for application of modern analytic 
methods 
 Hardware Overview 
• Long-duration 
housing for 
rodents on the 
ISS 
• Accommodates 
10 mice  (up to 6 
rats) 
TRANSPORTER 
• Short duration 
housing during 
ascent/descent) 
on Dragon  
• Accommodates 
20 mice, (up to 
12 rats) 
HABITAT 
• On-orbit animal 
access and 
transfer 
Additional kits 
and support 
equipment 
ACCESS UNIT 
RR-1 Validation Methods: Animals 
 
•  Mice:  females, adult (4-mo old), C57Bl/6J strain  
-  common background strain for transgenic/knockout 
animals  
  
•  Flight mice loaded into a Transporter 
  (total 20; 10 for Validation, 10 for CASIS) 
   
•  Control mice (cohorts of flight mice) 
-  Ground Controls-housed in Transporter on time delay 
and in environmental simulator to matched Flight 
conditions (Temperature, Carbon Dioxide, Humidity) 
 -   Vivarium controls (standard housing) 
2. Launch 
3. Ascent  
(~ 3 days) 
4.  ISS Dock/Animal 
transfer  
(1 day) 
5. ISS Habitat: up to 1 month 
1.  Late Load 
on Dragon 8. Descent 
7. Sample 
Cold 
Stowage 
6. Euthanasia 
and tissue 
retrieval 
Access Unit 
Transporter 
Habitat 
Methods: Basic Concept of Operations 
Glovebox 
Measurement Approach 
•  Animal health observations:  
–  Direct:  crew visual observations made during animal 
transfer and dissection 
–  Indirect: video record reviewed by experts on ground 
shortly after collection 
•  Body weights:  
–  Advantage:  quantitative indicator of general health 
–  Achieved by measuring body weights post-flight 
(euthanized animals). 
 
Measurement Approach 
Sample Recovery 
•  Rationale for tissues selected: 
–   tissues affected by spaceflight 
–   simple to dissect intact 
•  Liver and Spleen: quantitative assays for RNA quality 
(spleen) and enzyme content and activities (liver) to: 
–  validate crew & handling operations on ISS: 
•  preservation 
•  past freezing 
–  provide samples for future in-depth genomic and 
proteomic analyses  
•   Ground controls (delayed): tissues harvested with similar 
timing as flight mice 
Validate Tissue Preservation 
Capability 
•  Spleen: preserved on-orbit in RNAlater then frozen (<80 ˚C) 
for ~6 months  
 -total RNA purified from the entire spleen after return 
 -quality of tissue preservation evaluated by: 
  -measuring the amount of total RNA recovered per organ  
   (variance/mean compared to ground controls) 
  -analyze the quality of RNA (degradation) 
    
•  If adequate sample preservation, conduct post-flight global 
gene expression analysis (Genelab) and other tissue sharing 
capabilities 
  -science-enabling and informative about animal health/
    spaceflight responses 
Validate Tissue Freezing Capability 
•  Liver:  quick frozen on-orbit and stored for up to 6 
months 
 -Activity levels of enzymes are highly sensitive to 
temperature/storage conditions. 
 
•  Candidate enzymes: selected as either regulated by 
spaceflight or constitutively expressed. 
 
•  Assays performed on tissue after return of all samples:  
  -total protein content of the liver  
  -enzyme amounts and activities measured after 
   return 
Expected Outcome  
• The Transporter and Habitat are expected to 
support the health & welfare of adult mice through 
all phases of the mission (ground-based 
verification tests)  
-Launch simulation (vibration, hypergravity) in 
Transporter 
  - Biocompatibility in Habitat (32 days) 
 
•    Results from tissue samples analyzed for RNA 
and protein (enzyme activities and content) will 
establish science-enabling capability of all on-orbit 
mission operations related to sample fixation and 
freezing 
Earth Benefits/Spin-off Applications  
•  New insight and possible treatments for major human 
diseases, e.g. 
–  Osteoporosis 
–  Muscle wasting 
–  Cardiovascular disease 
–  Immune dysfunction 
•  Customer base includes 
–  NASA funded, peer reviewed research 
–  Commercial: Pharmaceutical and biotech industries 
–  Other government agencies: DoD, NIH, USDA, etc. 
–  International Partners 


Rodent Research-1 Validation of Rodent Hardware

Abstract

Similar works

Full text

Available Versions

NASA Technical Reports Server