In an era of increasing human longevity, it is becoming ever more critical to prioritize no only improved treatment of disease but also approaches for maintaining wellness to preserve the quality of life for aging adults. Spaceflight-induced physiological changes represent deviations from this state of wellness, with some changes serving as accelerated models of Earth-based conditions, including musculoskeletal disease, immune dysfunction, and delayed wound healing. This session will highlight research onboard the ISS National Lab that seeks to exploit these spaceflight-induced physiological effects to improve human health on Earth through studies that advance the prevention, detection, and treatment of disease. In parallel, spaceflight research and development efforts also seek to improve the human condition by employing the space environment to advance diagnostic tools and drug discovery, development and delivery systems.
Seismic Method Estimate velocity from seismic data.pptx
ISS Research to Advance Health on Earth
1. ISS Research to Advance Health on Earth
• DR. BESS DAWSON-HUGHES, SENIOR SCIENTIST AND
DIRECTOR, BONE METABOLISM LABORATORY, TUFTS
UNIVERSITY
• DR. MILLIE HUGHES-FULFORD, SCIENTIFIC ADVISOR TO
THE UNDER SECRETARY OF THE DEPARTMENT OF
VETERANS AFFAIRS, SAN FRANCISCO VA HEALTH CARE
SYSTEM
• ROSAMUND SMITH, RESEARCH FELLOW,
BIOTECHNOLOGY DISCOVERY RESEARCH, ELI LILLY AND
COMPANY
• DR. EUGENE BOLAND, CHIEF SCIENTIST, TECHSHOT, INC.
SPEAKERS:
2. Using ISS to Study T Cell Activation
Dr. Millie Hughes-Fulford
milliehf@gmail.com
14. Are T cells in other mammals activated normally
in spaceflight?
• Test the effects of microgravity in vivo in spaceflight in the mouse
model on STS-131.
22. 2016 Hughes-Fulford
TRANSCRIPTION
TRANSLATION
TNFa
INFg
GMCSF
CCL3
• The messenger (mRNA) is
converted to protein through a
process called translation.
• It has long been known that the
translation step is regulated by
transcription factors in the
promoter regions, however new
data has proven that there is a
second level of regulation by
miRNA.
40. Eli Lilly and Company
We make medicines that help people live longer, healthier, more active lives
♦ Company founded May 10, 1876 by U.S. Civil War
veteran Colonel Eli Lilly
♦ Headquarters located in Indianapolis, Indiana, U.S.A.
♦ Approximately 40,000 employees worldwide
♦ Products marketed in 125 countries
♦ More than 8,000 employees engaged in research and
development, or 20% of total workforce
♦ Lilly products treat cancer, depression,
schizophrenia, diabetes, osteoporosis, psoriasis and
many other conditions
♦ Expertise in musculoskeletal conditions
• Osteoporosis, Muscle atrophy
• Preclinical and clinical measures of bone and
muscle
• Myostatin inhibition
41. Skeletal Muscle Atrophy occurs with Many
Diseases
• Over 124 conditions/diseases have been associated with skeletal muscle
wasting/atrophy
• Includes certain forms of cancer, ALS, muscular dystrophy, sarcopenia, COPD, critical illness
myopathy
• And of course astronauts in space
• Its not the size but the decrease in muscle function that is most critical to the patient
• Decreased muscle power, strength, endurance
• Decreased muscle function leads to diminished functional performance
• Decreased ability to climb stairs, withstand medical treatments, perform activities of daily
living
• Increased chance of falls, injuries from falls, fractures, hospitalizations, death
• Often leads to lack of independence
• Associated with poor prognosis and outcomes
42. Muscle Atrophy Represents a Large Unmet
Medical Need
• The only treatment currently for muscle atrophy is physical
therapy/exercise
• May not be sufficient
• Poor compliance with the ill/elderly
• Is it possible to develop a drug to combat muscle atrophy?
• Some drugs are working their way through clinical trials, but nothing
near FDA approval
43. Mouse Models of Muscle Atrophy
• Mechanisms of muscle wasting highly conserved between mice and man
– Similar cell signaling pathways and key genes are involved
• In drug discovery it is important to test compounds in mouse disease
models before progressing into the clinic
• Mouse models of muscle atrophy are not ideal
– Unloading models, eg hindlimb suspension, limb casting
– Tumor models
– Genetic models eg SOD1 transgenic mice, mdx mice
• In microgravity, muscles are ‘unloaded’ and evidence to date suggests they
undergo atrophy
44. Evidence for Muscle Wasting in Mice under
Conditions of Microgravity
Species
Number
/group
Age at
onset
Duration
Days
Included Measures Transporter Ref
Rat
Fisher 344
12 17 -5% BW; -24% gastroc Space shuttle
Lalini et al
2000
Mouse
C57Bl/6J
12 9 week 11
Decreased muscle CSA; -
5%BW from baseline; atrophy
soleus> gastroc
Space shuttle
(Amgen)
Harrison et al
2003
Mouse
C57Bl/6J
12 9 week 13
-8.5% BW; decreased CSA
particularly type II fibers (IIb,
IIx). Soleus -30%; gastroc -15%
wt.; masticater muscles not
affected
Space Shuttle
(Amgen)
Ferguson et al
2012; Sung et
al 2013;
Philippou et al
2015
RR-3
Mouse
Balb/c
10 12 week 42
Body composition,
grip strength
ISS Lilly
45. Rodent Research 3
• Goal: To determine if a novel investigational compound
is able to prevent the muscle wasting that is expected to
occur on the ISS
• Primary goal:
– Use the results to help in discovery research to find ways to
develop new medicines for patients in need with muscle
wasting conditions
• Secondary goals:
– Continue to learn how animal cellular functions are affected
by longer term microgravity
46. Unique Features of RR-3
• Novel investigational compound (myostatin inhibitor)
• New strain of mice, Balb/c
• 6 week experiment
• Measure muscle function with grip strength meter
• Anaesthesia followed by recovery on the mice to enable interim
body composition measures
• Body composition with DEXA Instrument (Bone Densitometer,
TechSHOT), 2nd experiment to use this capability
47. Experimental Design
Groups:
1. Baseline, n=10
2. Flight mice, control treated, n=10
3. Flight mice, drug treated, n=10
4. Ground control, control treated, n=10
5. Ground control, drug treated, n=10
Female Balb/c mice, approx 12 weeks of age at launch
Duration of experiment= 6 weeks
Ground controls offset 3 days from flight animals
Animals randomized on body weight to groups
Control and drugs delivered by sub-cutaneous injection at 0,2,4 weeks
Treated and control mice mixed in each habitat (3,2)
• Tail tattoo used for rapid identification of treatment (treated vs. control)
• Mice identified by implanted microchip
48. Pre-flight Adjustments
• Mice acclimatized to drinking apparatus and food bars 4 weeks
prior to launch
• Mice acclimatized to lack of bedding and wire floors 4 weeks prior to
launch
• Mice moved into Dragon habitat (10 mice/habitat) at L-24h
50. Life Onboard ISS
• Dragon docked 1 day 17 hours post-launch
• Mice moved into ISS habitat
51. Live Phase Measures: Grip Strength
Columbus Instruments, OH
Grip strength measured pre-flight, interim and
termination for both flight and ground control mice
• 4 measures taken
per mouse per time
point
• 2 middle values
averaged
52. Live Phase Measures: Body composition
• Body composition measured by DEXA (dual energy x-ray absorptiometry) at
interim (approx 4 weeks) and at termination (approx. 6 weeks)
– Lean mass, fat mass, bone mineral content and density
Bone scan of anesthetized mouse
53. Live Phase Measures:
Anaesthesia Recovery
• DEXA performed on anesthetized mice
• First time to perform anesthesia followed by recovery on
mice in space
• Mixture of ketamine/xylazine used, followed by
atipamezole to speed recovery
• Mice kept separately in newly designed hardware
(Anesthesia/Recovery System or ARS)
54. Termination Sampling
• Serum (cardiac puncture)
• Hindlimb put into formalin for later histology
measures
• Carcass frozen at -80oC
• Samples due back to Earth in late August on
SPX-9
55. Later Measures
• Muscle weights (including gastrocnemius, quadriceps, soleus)
• Brain weights
• Muscle hindlimb histology, myofiber cross-sectional area
• Muscle gene expression
• Compound exposure (serum)
• Bone parameters including biomechanics
• Other measures still to be determined
• Plan is to give unused tissues from control mice to other
spaceflight researchers
56. Myostatin - a regulator of
muscle mass
• Compound tested is a myostatin inhibitor
(LSN2478185, YN41)
– Myostatin is a growth factor that is a negative
regulator of muscle mass
– Has been shown to prevent muscle wasting in
terrestrial mouse models of muscle wasting (Smith et
al 2015; unpublished)
• A less specific inhibitor of myostatin, called ActRIIB-
Fc, was tested by Amgen on a Space Shuttle mission
(13 day flight)
– Was able to reduce or prevent muscle wasting
(Ferguson et al, 2012)
Smith et al 2015
Indicates significance relative to the control p< 0.05.
**
57. Preliminary Results from Live Phase Study
– Muscle Function
♦ Grip strength in ground controls relatively stable
• Increase at week 6 likely age-related increase in size
♦ Grip strength decreased in flight animals at week 4
• Recovered towards baseline by week 6
• Increase at week 6 likely due to age-related increase
in size
• Note different operator and orientation of grip
strength meter of baseline vs weeks 4 and 6
measures
♦ In both ground control and flight animals myostatin
inhibition significantly increased grip strength over
control animals at both weeks 4 and 6
• To baseline values in flight animals
• Above baseline values in ground controls
Effects of myostatin inhibition on
skeletal muscle function can occur
under conditions of microgravity
(*), refers to significance relative to its respective
control at each time point, ($), refers to significance
relative to the flight control group. All p values < 0.05.
*
*
$
*
$
*
58. Preliminary Results from Live Phase Study
– Lean Mass (DEXA)
♦ Increase in lean mass seen with myostatin inhibitor
compared to respective controls at both 4 and 6 weeks
♦ Ground and flight controls show increase in lean mass with
time
• Likely age related, as body weight increases with time for
ground controls
♦ Surprisingly, flight controls did not have decreased lean
mass compared to ground controls
• Previous shuttle missions had shown loss of body weight, lean
mass and individual hind limb muscles but these were earlier
time points
• Discordance with grip strength data
RR-3 Lean Mass (DXA)
Weeks
0 1 2 3 4 5 6 7
%ChangefromBaseline
-5
0
5
10
15
20
25
Flight Control
Flight YN41
Ground Control
Ground YN41
*
*
*
*
(*), refers to significance relative to its respective
control at each time point, All p values < 0.05.
Effects of myostatin inhibition on skeletal muscle
mass can occur under conditions of microgravity
59. Preliminary Results from Live Phase Study
– Lean Mass (DEXA)
♦ Explanations for apparent lack of muscle wasting
• Are the ground controls doing poorly?
– Equivalent ground control experiment performed
independently at Lilly in regular housing and results show
ground controls performed as expected in terms of body
weight gain
• Wasting may have occurred earlier but mice
recovered/overcompensated?
• Fluid distribution in space affects DEXA readings?
• Greater levels of altered types of exercise by flight animals?
♦ Important to get muscle weights to validate DEXA results
♦ Determine if changes in lean mass are seen in all or subset of
muscles
(*), refers to significance relative to its respective
control at each time point, All p values < 0.05.
Effects of myostatin inhibition on skeletal
muscle mass can occur under conditions of
microgravity
*
*
*
*
60. Next Steps
• Continue analysis of samples and data
• Use data to further understand effects of microgravity on skeletal muscle
• Use data to assist drug discovery efforts to find drugs to combat muscle-
wasting diseases
63. 3D Printing and Biology Makes Headlines
Modern medicine: Lab-grown genitals,
spray-on skin ... - NBC News
3-D printed livers offer glimpse into the
future ... - NBC News
Father and Son Produce Leather via
Bioprinting - VegNews Magazine
Bioprinting cartilage into people
is doctor's goal
New bioprinting technique creates
thicker, healthier tissue
New Bioprinting Method Borrows Ancient
Technique
3D bioprinting could spark ethical debate
3D Printing Reshapes Healthcare
“ team of cardiovascular scientists has
announced it will be able to 3D print a
whole heart from the recipients' own cells
within a decade."
64. Reality of Organ Shortage*
• 120,206 People on transplant list in the USA
• 16,445 organs transplanted in first 6 months of 2016
• 7,764 organ donors in the first 6 months of 2016
• For 2016, 1 person is added to list every 10 minutes
• For 2016, 22 people die waiting each day
All Data current as of 2:30 pm EDT 07/12/2016 as reported by the
United Network of Organ Sharing (UNOS)
65. What if One Day?
• Personalized Medicine was more than a better prescription?
• Stem Cell Therapy moved from the Wall Street Journal and
USA Today to Main Street even if it had to go to space first?
• A child born with a congenital heart defect had only one
surgery then grew up to be the best third baseman on his/her
little league team instead on just watching sports?
• Third Party Payers realized it is cheaper to fix a problem right
the first time that prolong the wrong treatment?
66. What is Bioprinting?
Cells
Polymers
Image File
Image Processing
Additives
Organs
Tissues
3D
Constructs
Image Rendering
BioInk
Dispensing Pen
Transfer Language Bioprinter
Pre-Fabricated
Systems
Structural
Components
67. Development Team
•Project management (prime contractor), spaceflight hardware
development, integration and operation. Space medical solutions
provider. Tissue bioprinter production management/supervision.
•Pioneer in the Direct-Write 3D tissue printing field. Has
expertise in Bioink and currently has a patent protected delivery
system with the highest resolution and broadest dynamic range
in terms of bioink capability. Supplier of delivery systems and
control software integration partner.
72. Technology Milestones*
• Month 8/15 – Parabolic/suborbital flights to
test enabling technologies in microgravity.
Key parameters include viscosity, cavitation
potential / pressure gradient, cell viability,
density gradient and thermal gradients.
• Month 28 - ISS Technology Demonstration.
Print a microvascularized tissue patch and
return it for terrestrial evaluation.
• Month 36 - ISS Experiment. Print a beating
heart patch within bioreactor to mature the
tissue. Tissue to contain arteriole, capillary
bed and venule structures. Tissue will be
perfused to demonstrate the ability for use
as a transplant.
*no NASA or CASIS funding or commitments have been implied or received in these forward looking statements