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Biologically Inspired Engineering:
The Next Technology Wave
Don Ingber, MD,PhD
Founding Director, Wyss Institute
Judah Folkman Professor of Vascular Biology, Harvard Medical School & Boston Children’s Hospital
Professor of Bioengineering, Harvard School of Engineering and Applied Sciences
Hansjörg Wyss Institute for Biologically Inspired Engineering at Harvard University
Convergence of the Sciences
Life
Sciences
Engineering
Physical
Sciences
…boundaries between living and non-living systems
are beginning to break down
THE BIG CHALLENGE:
Current Drug Development Model is Broken
Computer Power Doubles Every 18 Months
Moore’s Law
Number of Medicines Invented HALVES
Every 9 Years
Eroom’s Law (Moore’s law backwards)
(Scannell et al., Nature Rev. Drug Discov. 2012; 11:191-200)
The Drug Development Model is Broken:
• Testing a single compound can cost > $2 million
• Cells cultured in dishes don’t function like in our bodies
• Animal studies take years to complete
• Innumerable animal lives are lost
• Results often don’t predict clinical responses!
• Lack of new drugs reaching patients
(Scannell et al., Nature Rev. Drug Discov. 2012)
Need better lab models that mimic
whole human organ function
Biomimetic Microsystems
• Engineer microchips containing living human cells that
reconstitute organ-level functions for drug screening,
diagnostic and therapeutic applications
• ACCELERATE drug development & REPLACE animal testing
Biomimetic
Spleen
Microchip Manufacturing offers control of
features at same size scale of living cells
Photolithographic Etching:
UV
Mask
nm to mm
Photoresist
Silicon
Cells
A Human Breathing Lung-on-a-Chip
(Dan Huh, Wyss Institute; Huh et al., Science 2010)
Alveoli (air sacs)
Measure Cell INJURY
(Reactive Oxygen Species)
SMOG Air Pollutants
Lung cells on-chip
(Nanoparticulates)
Nanoparticles
Epithelium
Endotheliu
m
Cell Injury
Inflammation
(only + Breathing Motions)
+ Breathing
Motions
WBCs
Breathing Increases Nanoparticle Absorption
Epithelium
Nanoparticles
% Absorption
Flow
Flow
Endothelium
No Stretching
Outflow
from
Lower
Channel
+ Cyclic
Stretching
0 hr
1 hr
4 hr
Prediction Tested in Whole Mouse Lung
Nanoparticles
Flow
Flow
Epithelium
Endothelium
% Absorption
Nebulizer
Ventilation
Perfusion
Prediction Confirmed in Whole Lung
Nanoparticles
Flow
Flow
Epithelium
Endothelium
% Absorption
PC
AS
AS
AS
AS
AS
PC
AS
Human Disease Model:
Pulmonary Edema (‘Fluid on the Lungs’)-on-a-Chip
Air
Membrane
IL-2
(CANCER
DRUG)
Liquid
Day 0
Air
Liquid
Liquid
Meniscus
Liquid
(Huh et al. Sci. Trans. Med., 2012)
IL-2
IL-2
Day 2
Day 3
Liquid
Day 4
Effects of IL-2 Cancer Drug on Lung Permeability
Lung
Vascular Leakage Model
Flow
Capillary
On-Chip
IL-2
FITC-inulin
Prediction
Confirmed
In Vivo
Predicting Drug Efficacy
DRUGInhibitor
A
TRPV4
Lung
DRUG
A Inhibitor
TRPV4
Flow
Capillary
IL2 + TRPV4
Inhibitor
IL-2 & Drug A
FITC-inulin
Human Disease Model On-Chip:
Pulmonary Edema (‘fluid on the lungs”)
• Human Disease Model
• Drug Toxicity Model
• Drug Efficacy Model
(Huh et al., Science 2010 & Sci. Trans. Med. 2012)
Beating Heart-on-a-chip
(Kit Parker Lab)
(Grosberg et al., Lab Chip 2011)
Structural and functional quantitation on-chip
(Wang et al., Nature Medicine 2014)
Modeling Barth Syndrome
(with human iPS cells)
Physiologial Coupling of
Heart & Lung Chips
(with Kit Parker at Wyss Institute & SEAS, Harvard)
Beating Heart Chip
Breathing Lung Chip
Physiological Coupling
Between Lung & Heart Chips
Before addition
After addition of lung effluent
Doxorubicin Delivery to Lung Chip Suppresses Contractility in Linked Heart Chip
Liquid Doxurubicin
20 nM
200 nM
2000 nM
Small Airway-On-A-Chip
(unpublished work of Kambez Benam & Remi Villenave)
‘Classic’ Lung (Alveolus)-On-a-Chip
Top channel
Bottom Channel
400um
Small Airway-On-a-Chip
Small Airway:
Diameter < 2 mm
Regeneration of Airway Epithelium On-Chip
Epithelial cells
DAPI
Endothelial cells
Differentiation of Specialized Human
Bronchiolar Epithelial Cells On-Chip
Cilia (β-tubulin IV)
Mucus (Muc5AC)
Beating Cilia in the Airway-on-a-Chip
(time-lapse at 1/7th normal rate)
Top View
Visualization of Mucociliary Transport
(Real-Time Imaging)
Comparison of Airway Chip to Human Airway
‘Flu-like’ Inflammatory Response
Induced On-Chip
(Induced using Viral Mimic poly I:C)
Chemokine Production
Monocyte
Recruitment
Control
+ Poly I:C
Endothelium Influences Cytokine Response to Viral Mimic
Human COPD Patient Lung Responses
& Exacerbations are Recapitulated On-Chip
TLR4
IL8/CXCL8
Cytokine secretion
(pg/ml)
% mRNA fold change
400
*
300
200
100
0
Healthy
1.5×104
100
50
0
Healthy
COPD
Con
LPS
4×103
2×103
0
0
Healthy
Cytokine secretion
(pg/ml)
% mRNA fold change
150
M-CSF/CSF1
*
8×103
5.0×103
TLR3
*
1×104
6×103
1.0×104
COPD
200
*
2.0×104
1×104
COPD
IP10/CXCL10
**
**
1×103
Healthy
50
40
30
1×102
20
1×101
10
1×100
0
Healthy
COPD
COPD
RANTES/CCL5
*
Con
Poly I:C
**
Healthy
COPD
Peristaltic Human Gut-on-a-Chip
(Kim et al., Lab Chip 2012 & Integrative Biology 2013)
Human Intestine
Microfluidic Platform
Lumen
Intestinal Villi
Capillary
PNAS, 2007, 104:10295
Human Gut Epithelium
(Exposed to Flow + Cyclic Deformation)
Gut Chip
24 hr after seeding
+ Peristaltic-like motions
Induction of Intestinal Villi Formation
Gut Differentiation Requires Correct Mechanical Cues
Restoration of Proliferative Crypts
Barrier Function
Differentiation
Differentiation of all Intestinal Cell Lineages
Drug Metabolism
Mucus Production
Intestinal Barrier
Co-Culture with Intestinal Microbiome
Probiotic Bacteria
(Lactobacillus GG)
(Kim et al., Lab Chip 2012)
Bar, 50 μm
Gut-on-a-chip with Normal Microbiome
Top view
Side view
(8 different probiotic strains)
GFP-E. coli, F-actin, Nuclei
Mechanically Active Environment + Microbiome
Induces Small Intestine Differentiation On-Chip
Gene Expression Profile Similarities:
(Across >22,000 human genes)
Gut Chip
Gut Chip
+ VSL#3
Gut Chip
+ VSL#3
Gut Chip
Transwell
Transwell
Transwell Gut Chip Gut Chip
+VSL#3
0.97
0.98
0.99
1
Ileum Duodenum Jejunum
Organ-on-Chip Technology Pipeline
• Ongoing projects
–
–
–
–
–
–
–
–
–
–
–
–
–
–
Lung Alveolus
Lung Small Airway
Heart
Liver
Small Intestine
Large Intestine
Kidney Proximal Tubule
Kidney Glomerulus
Bone marrow
Skin
Blood-Brain Barrier
Cancer
Eye
……
Integrated Human Body-on-Chips
DARPA Multiphysiological Systems Grant
(D. Ingber, K. Parker, J. Wikswo & CFDRC)
Organ-On-Chip
INTERROGATOR
Instrument
Universal
Chip Holder
– Automated instrument
integrates multiple organs
– Designed for ease of use
“plug-and-play” approach
– Generate data to predict
human response
X 10 Organ Chips
Organ ‘Interrogator’ Prototype
Custom Enclosure
Interrogator Integrated Inside
(12 cartridge capacity)
Final Prototype
Personalized Organs on Chips
(from individuals to populations)
Emulate Inc. was recently founded to commercialized the
Automated Instruments and Vascularized Microfluidic Organ Chips
that have emerged from our research effort
Disclosure Statement of Financial Interest:
I hold equity in Emulate Inc. & chair it’s Scientific Advisory Board
Programmable Nanomaterials Platform
• Create ‘smart’ medical nanotechnologies
• From Implantable Devices to INJECTABLE MEDICAL DEVICES
Bioinspired Drug Delivery for Cardiovascular Medicine
Vascular Blockage is Leading Cause of Death
Platelets ‘Activated’
by Shear Stress
•Heart Attack
•Stroke
•Pulmonary Embolism
•Atherosclerosis
•Coronary Spasm
•Peripheral Vascular Disease
•…
Shear-Targeted Drug Delivery
(Korin et al., Science 2012)
Shear-induced platelet activation
Shear
Stress
Synthetic ‘Platelet Mimetics’
Shear
Stress
Vascular Blockage-Targeted
Drug Delivery
Shear-Targeted Drug Delivery
(Korin et al., Science 2012)
Shear-induced platelet activation
Shear
Stress
Synthetic ‘Platelet Mimetics’
Shear
Stress
Vascular Blockage-Targeted
Drug Delivery
Targeting of a Clot-Busting Drug
(tPA: tissue plasminogen activator)
Removal of Artificial Clots using Shear-Targeted tPA Nanotherapeutics
0 min
1 min
60 min
In Vitro Study
Clinical Responses in an Animal Model In Vivo
(effective at 1/100th injected clinical dose of free tPA)
Targeting Pulmonary Embolism
Pulmonary
Embolism
Model
Significant Increase in Survival
SA-NT (80% Survived)
Control
(100% Dead)
Embolus
Released
Nanoparticles
Pathogen-Targeted Technology
SEPSIS:
• A major killer worldwide with few treatment options
• >30% mortality even with best antibiotic therapy and ICU care
• Bloodstream infections occur in ~10 % percent of hospital patients,
but cause of ~50% of all U.S. hospital deaths (May 18, 2014/Drugs.com)
• More than $19B spent in U.S. on treatment
Increasing Resistant Microbes
New Antibiotic Approvals
# New Antibacterial
Agents
Rapid Disease Progression
20
15
10
5
0
a
b
‘Biospleen’
Dialysis-Like Sepsis Therapeutic Device
MBL
CRD
MBL
Magnetic
nanobead
neck
Collagen
helix
IgG1
Fc
Artificial Spleen Micro-architecture
Blood
FcMBL Cleansed
(90 kDa)
Blood
MBL (650Contaminated
kDa)
Magnetic
Opsonins
arterial
Magnetic
bead
Magnetic Beads
Opsonized Pathogens
Vasculature
E.coli
sinusoid
ΔB
ΔB
Sinusoid
ΔB
External Magnets
S.aureus
c
venous
Collection Fluid
Magnetically Separated
Pathogens & Beads
Venous system
Sinusoid
slits
Lymphoid
Discard'Out'
Discard
out
Saline'In' in
Saline
Magnets
c
MAGNET'
Venous
Arterial
red-pulp
cord
Stress fiber
Waste
ΔB!
ΔB'
ΔB!
Sinusoid
Arterial
Magne+c'
Magnetic
Opsonins'
Endothelial
cells
d
Sep+c'Blood'In'
Septic
blood in
opsonin in
Cleansed'Blood'Out'
Cleansed
blood out
Peristaltic pump
Saline in
Magnetic
opsonins
Anesthesia
Saline out
for
ANALYSIS
TESTJugular
SAMPLE IN
catheters
Magnets
SAMPLE OUT
Static mixer
51
Incubation
a
b
‘Biospleen’
Dialysis-Like Sepsis Therapeutic Device
MBL
CRD
MBL
Magnetic
nanobead
neck
Collagen
helix
IgG1
Fc
Artificial Spleen Micro-architecture
Blood
FcMBL Cleansed
(90 kDa)
Blood
MBL (650Contaminated
kDa)
Magnetic
Opsonins
arterial
Magnetic
bead
Magnetic Beads
Opsonized Pathogens
Vasculature
E.coli
sinusoid
ΔB
ΔB
Sinusoid
ΔB
External Magnets
S.aureus
c
venous
Collection Fluid
Magnetically Separated
Pathogens & Beads
Venous system
Sinusoid
slits
Lymphoid
Discard'Out'
Discard
out
Saline'In' in
Saline
Magnets
c
MAGNET'
Venous
Arterial
red-pulp
cord
Stress fiber
Waste
ΔB!
ΔB'
ΔB!
Sinusoid
Arterial
Magne+c'
Magnetic
Opsonins'
Endothelial
cells
d
Sep+c'Blood'In'
Septic
blood in
opsonin in
Cleansed'Blood'Out'
Cleansed
blood out
Peristaltic pump
Saline in
Magnetic
opsonins
Anesthesia
Saline out
for
ANALYSIS
TESTJugular
SAMPLE IN
catheters
Magnets
SAMPLE OUT
Static mixer
52
Incubation
Generic Pathogen Capture Technology
Biological Inspiration: HUMAN OPSONINS
• Natural blood proteins
• Molecular components of the Innate Immune system
• Many are LECTINS that bind to surface carbohydrates on
pathogens that are not commonly found on human cells
Engineered Human Opsonin (FcMBL)
Human Mannose Binding Lectin (MBL):
Binds diverse pathogens, targets them for
phagocytosis, stimulates immune response
MBL Carbohydrate
Recognition Domain (CRD)
neck
Engineered FcMBL:
•Binds diverse pathogens
•Lacks complement activation &
coagulation domains
• Easy purification
MBL CRD
MBL neck
Fc IgG
Complement & coagulation
activation
Collagen helices
FcMBL on
magnetic nanobead
Broad Pathogen-Binding Spectrum
S. aureus
1 mm
1 mm
(128 nm beads)
C. albicans
E. coli
1 mm
FcMBL Opsonin: Bind
> 90 Pathogens + Toxins
Fungi
Gram negative
Gram positive
Viruses
Parasites
Aspergillus spp
Acinetobacter baumanii*
Burkholderia cepacia,
Bacterioides fragilis
Bacillus subtilis
“Dengue”
Cryptosporidium
Blastomyces
Chlamydia trachomatis,
Clostridium neoformans, Clostridium
difficile, perfringens
“Ebola”
Leishmania
Candida, albicans,
glabrata, guilliermondii,
krusei, parapsilosis,
tropicalis
Escherichia coli*
Enterobacter aerogenes*
Enterobacter cloacae (abx)*
EBV
Malaria
Cryptococcus
Haemophilus inf b
Listeria monocytogenes*
Hep B, C
Schistosoma
Fusarium spp.
Helicobacter pylori
Mycobactrium avium
HIV
Trypanosoma
Mucor spp.
Klebsiella oxytoca,
K. pneumonia (abx)*
Mycobacterium tuberculosis, leprae
HSV 1, 2, CMV
Saccharomyces
Legionella pneumophila
Nocardia farcinica, P. acnes
“ Influenza A”
Pneumocystis jirovecii
(carinii)
Neisseria meningitidis B,C,
Neisseria gonorrhoeae
Staphylococcus
aureus MSSA*, MRSA*
S. epidermidis*,
Marburg
LPS
Pseudomonas aeruginosa*,
S. marcescens (abx)*
S. maltophilia*
Streptococcus pyogenes
Group A,
RSV
LTA
SARS-CoV
Ricin
Salmonella typhi, paratyphi,
typhimurium (abx)*
Shigella flexneri
Mycoplasma
Yersinia pseudotuberculosis
M. pneumoniae, M. hominis, M. orale
West Nile
Toxins
Rat Sepsis Model
Peristaltic
pump
Catheterized
Rat
Microfluidic
Separator
Syringe pump
injecting MBL1
beads & Heparin
Static
mixer
Spleen-on-a-Chip
Cleansing of Blood Pathogens in Rats
Spleen chip
+ no beads
Control
Spleen Chip
• Rats treated with mf-DLT device had consistently lower levels of
S. aureus pathogens at all time points (n=5) (p<0.001)
• 90% of pathogens removed from human whole blood within 1 hr (n=4)
Cytokine Levels (pg/mL)
Reduction of Cytokine Levels In Vivo
120
SAIP No treatment
100
80
SAIP Biospleen
60
40
20
0
GM-CSF IFN-G
IL-1A
IL-4
IL-6
Blood Cleansing Increases Survival
in Rats with Endotoxemia
LPS intensity (a.u.)
(Kang et al., Nature Medicine 2014)
Survival (%)
100
50
0
0
1
2
3
4
5
0.8
0.6
0.4
0.2
0.0
Lung
Time (h)
•
•
LPS (1.4 x107 EU/mL) injected IV into rats (n=6)
100% of control animals die by 4.5 hrs vs. 85% survival
LPSDLT Device
with
therapy
−
+
+
−
+
Biospleen
+
Inspiration from the Non-Medical World
Non-Stick
Slippery Liquid-Infused Porous Surfaces (SLIPS)
(Joanna Aizenberg Lab)
Winner of 2012 R&D Technology Award
Medical SLIPS
Tethered Liquid Perfluorocarbon (TLP) Fig. 1
A
Liquid
Perfluorocarbon
(LP)
Blood
Blood
Substrate
Substrate
q
Sliding Angle
Tethered
Perfluorocarbon
(TP)
t=5 s
are silanized with Tethered Perfluorocarbon (TP), then
B FDA approved indwelling devices
0 sec
2 sec
5 sec
1.
sterilized and stored > 1 year
1.
Prior to use, an FDA-approved Liquid Perfluorocarbon (e.g. Perfluorodecalin; PFD) is added
1. Control
The TP retains the LP and repels blood
TLP Coating for Medical Devices
(Leslie et al., Nature Biotechnology 2014)
• TLP works on SMOOTH FDA-approved Medical Materials
• TLP-treated arteriovenous (AV) shunt functioned for 8 hrs in a pig without heparin
Arterial
PU
Venous
PC
PVC
It’s Just the Tip of the Wave…
Injectable Engineered Heart Valve
Kit Parker (Wyss Institute Harvard) and Simon Hoerstrup (U. Zurich/Wyss Translation Center)
-
290,000 heart valve replacements annually worldwide
Bioprostheses
Mechanical Valves
( 3x by 2050; Yacoub et al., 2005)
-
Current replacements materials are suboptimal
 lack of growth / regeneration
 surface thrombogenicity / Immunogenicity
Engineer scalable nanofiber based, seamless
semilunar cardiac valves using automated RJS
 Fast, inexpensive and regenerative
 Easily manufacturable / off-the-shelf
 Biomimetic mechanics in vitro / in vivo
 in situ remodeling & regeneration
Rotary Jet Spinning
(RJS)
Robbins et al., 2009
Robbins et al., 2009
Biomimetic Mechanics In Vitro
Injectable Engineered Heart Valve (JetValve)
Kit Parker (Wyss Institute Harvard) and Simon Hoerstrup (U. Zurich/Wyss Translation Center)
After implantation
Before implantation
Follow up 1 MONTH in vivo:
aSMA
JetValve
3
2
2
2
1
1
MG
Von Kossa
3
H&E
 The thin and fully loaded heart leaflet shows cellular infiltration, remodeling, and endothelialization
after 1 month in vivo implantation at the pulmonary valve position
Bioinspired Robotics
• Develop biologically inspired robots that move and adapt
like living organisms
• Create AUTONOMOUS SELF-ASSEMBLING MACHINES
Normal Embryogenesis
(Karlstrom & Kane;
http://zfin.org)
Bioinspired Programming Algorithms
Kasper Stoy (U. Southern Denmark &
Wyss Visiting Scholar) &
Radhika Nagpal (Wyss Institute)
Bioinspired Robotics
Rob Wood
Radhika Nagpal
L. Mahadevan
Kit Parker
George Whitesides
Conor Walsh
Autonomous Construction
Autonomous Control
Automated “POP-UP” Manufacturing
Wyss Core Faculty
Donald Ingber
Kevin Parker
Joanna Aizenberg
George Church
2008
L. Mahadevan
Pamela Silver
James Collins
Peng Yin
Radhika Nagpal
David Mooney
David Edwards
William Shih
Conor Walsh
Ary Goldberger
Robert Wood
George Whitesides
Jennifer Lewis
Neel Joshi
2009
2012
2013
ATT Staff Add Broad Industrial Experience:
CELL BIOLOGY
ENGINEERING
PHARMACOLOGY
VIROLOGY
MODELING
MEDICINE
PHYSIOLOGY
IMMUNOLOGY
MATERIALS
SOFTWARE
wyss.harvard.edu