using nanostructured materials to modulate the immune system€¦ · • 1590 n/m for short versus...
TRANSCRIPT
Using Nanostructured Materials to Modulate the
Immune System
Tejal A. Desai
Professor and Chair
Dept. of Bioengineering and Therapeutic Sciences
How can material structure modulate cellular function for therapeutic purposes?
Transdermal Oral Retinal Cardiac
Can we tune material structure to modulate
fibrosis?
Fibrosis: Fibroblasts Activated by Aberrant Mechanical Tension and TGFβ
Wound Healing
Fibrosis
TGFβ
High Aspect Ratio Features Provide Anti-fibrotic Signals
Kam, et. al, Nanoletters 2013; Tissue Engineering 2014
FITC-IgG Adsorption Vinculin, F-Actin, DAPI
P(20)
3 mm
3 mm
P(1)
10 mm 10 mm
10 mm 10 mm 20 mm
20 mm
Long aspect ratio structures inhibit fibroblast activation in vitro
Allen, Ryu et al., 2016
Long Structures Decrease Fibrotic Response in vivo
Allen, J. and Ryu, J. et al, Tissue Engineering part A 2015 Submitted 8
Nanorod fabrication scheme
Zamecnik et al., ACS Nano 2017.
Scale Bar 20μm
Merge Phalloidin Nanowires DAPI
No
nan
ow
ires
Lo
w
nan
ow
ires
Hig
h
nan
ow
ires
Nanowires alter cellular morphology and actin cytoskeleton
Nanowires decrease TGFβ and collagen transcription
0
0.5
1
1.5
2
2.5
TGFbeta TBRII COL1
Fo
ld C
han
ge
RTqPCR
No nanowires + TGF
Low nanowires + TGF
High nanowires + TGF
**
**
*
*
Can we use “nanostructure” to enhance
immunotherapy?
Features vs. Challenges
Small signaling
protein
Received
interest in for
cancer &
autoimmune
disease
Short
half-life
Not bioavailable
Pleiotropic
>30% require
hospitalization
IFNγ
TNFα
IL-2
Systemic Cytokine Therapy
Endogenous cytokine capture for prolonged
& localized immune activation
Longer
half life
Applicable to
many cytokine
targets
Targeted delivery
Reduced side
effects
Tailored Immune
Response
Cytokine Capture
Strategy
Endogenous
cytokine complexes
with antibody
Localized, cell
specific
activation Cellular influx
Loose network
formation
Subcutaneous
injection
T= 0 T= minutes T= hours T= days
- Receptor A
- Receptor
B
a b c d
Nanostructures as an injectable cytokine trap
Zamecnik et al., ACS Nano 2017.
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A
b
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R
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uc
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b
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1 5 0
N W F I T C - I G G C o n j u g a t i o n
Fl
uo
re
sc
en
ce
In
te
ns
it
y * * * * p < 0 . 0 0 0 1
* * * * p < 0 . 0 0 0 1
a b
d c
Ab
Ab
Ab
e
0 . 0 0 0 . 0 5 0 . 1 0
0
1 0 0
2 0 0
3 0 0
4 0 0
5 0 0
C o n c . o f N W - A b
Co
nc
.
of
I
L-
2
Bo
un
d
to
N
W
s a t u r a t i o n o f b i n d i n g
Reduced Antibody Native Antibody
Scale Bar 20μm
Nanowires can conjugate to IgG species and sequester cytokines
Nanowires persist in vivo for >6 weeks 2 week 4 week 6 week
DAPI
F4/80
NWs
Scale bar - 100μm
Sprent et al., 2012
Can we use this strategy to activate T cells specifically and locally?
Lymph
Node
N
an
ow
ir
es
A
b N
an
ow
ir
es
A
b o
nly
0
1 0
2 0
3 0
4 0
%K
i6
7+
N a t u r a l K i l l e r
* * * p < 0 . 0 0 0 1
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ow
ir
es
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b N
an
ow
ir
es
A
b o
nly
0
2 0
4 0
6 0
8 0
%C
D1
22
+
C D 8
* * p < 0 . 0 0 5 * p < 0 . 0 5
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an
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es
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b N
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ow
ir
es
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b o
nly
4 0
5 0
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%C
D2
5+
T r e g
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b N
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ow
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es
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b o
nly
0
1 0
2 0
3 0
4 0
5 0
%K
i6
7+
N a t u r a l K i l l e r
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ow
ir
es
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b N
an
ow
ir
es
A
b o
nly
0
2
4
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8
%C
D1
22
+
C D 8
* p < 0 . 0 0 3
* p < 0 . 0 5
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an
ow
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es
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b N
an
ow
ir
es
A
b o
nly
4 0
4 5
5 0
5 5
6 0
6 5
%
CD
25
+
T r e g
Skin IL-2 S4B6
Strong signal Weak signal
CD122
+ CD25
CD8 Memory
T cells, NK
Cells
Regulatory T
Cells
CD122
S4B6 antibody-conjugated wires locally activate NK andd CD8 Cells in vivo
IL-2 JES6-1
Strong signal Weak signal
CD122
only
Tregs NK and
effector T
cells
CD25
JES6-1-NWs locally activate Tregs and inhibit Teffs in the skin
N
an
ow
ir
es
A
b N
an
ow
ir
es
A
b o
nly
4 0
5 0
6 0
7 0
8 0
E f f e c t o r T C e l l s
%
Pa
re
nt
p < 0 . 0 5
N
an
ow
ir
es
A
b N
an
ow
ir
es
A
b o
nly
0
2 0
4 0
6 0
R e g u l a t o r y T C e l l s
%
Pa
re
nt
p < 0 . 0 5
N
an
ow
ir
es
A
b N
an
ow
ir
es
A
b o
nly
0
1 0
2 0
3 0
C D 8 T C e l l s
%
Pa
re
nt
N
an
ow
ir
es
A
b N
an
ow
ir
es
A
b o
nly
5 0
6 0
7 0
8 0
9 0
C D 4 T C e l l s
%
Pa
re
nt
IL-2 JES6-1
Strong signal Weak signal
CD122
only
Tregs NK and
effector T
cells
CD25
JES6-1 NWs have little effect in the draining lymph nodes
N
an
ow
ir
es
A
b N
an
ow
ir
es
A
b o
nly
8 0
8 5
9 0
9 5
1 0 0
E f f e c t o r T C e l l s
%
Pa
re
nt
N
an
ow
ir
es
A
b N
an
ow
ir
es
A
b o
nly
6
8
1 0
1 2
1 4
R e g u l a t o r y T C e l l s
%
Pa
re
nt
N
an
ow
ir
es
A
b N
an
ow
ir
es
A
b o
nly
0
1 0
2 0
3 0
4 0
C D 8 T C e l l s
%
Pa
re
nt
N
an
ow
ir
es
A
b N
an
ow
ir
es
A
b o
nly
6 0
6 5
7 0
7 5
8 0
8 5
9 0
C D 4 T C e l l s
%
Pa
re
nt
Disease Model –K5-TGO-DO11 Autoimmune Skin Disease
• K5-TGO-DO11 transgenic mouse that exhibits antigen specific immune response to OVA
• OVA under control of tetracycline promotor in keratinocytes,
• Leads to acute dermatitis and influx of CD4’s into the skin
Hypothesis – local augmentation of Treg activation before antigen is turned on will ameliorate disease phenotype
Rosenblum et al, Nature, 2011
B
la
nk
, -
D
is
ea
se
A
b-
N
W
, -
D
is
ea
se
B
la
nk
, +
D
is
ea
se
A
b-
N
W
, +
D
is
ea
se
0
2 0
4 0
6 0
8 0
T e f f ( D O 1 1 S u b s e t ) % C T L A - 4 +
%
Pa
re
nt
B
la
nk
, -
D
is
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se
A
b-
N
W
, -
D
is
ea
se
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nk
, +
D
is
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se
A
b-
N
W
, +
D
is
ea
se
0
2 0
4 0
6 0
8 0
T e f f ( D O 1 1 S u b s e t ) % K i 6 7 +
%
Pa
re
nt
B
la
nk
, -
D
is
ea
se
A
b-
N
W
, -
D
is
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se
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la
nk
, +
D
is
ea
se
A
b-
N
W
, +
D
is
ea
se
7 0
8 0
9 0
1 0 0
T r e g ( D O 1 1 S u b s e t ) % C T L A - 4 +
%
Pa
re
nt
* * *
B
la
nk
, -
D
is
ea
se
A
b-
N
W
, -
D
is
ea
se
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la
nk
, +
D
is
ea
se
A
b-
N
W
, +
D
is
ea
se
0
2 0
4 0
6 0
8 0
1 0 0
T r e g ( D O 1 1 S u b s e t ) % K i 6 7 +
%
Pa
re
nt
* * *
Ab-NWs selectively activate antigen specific Tregs - but not effector cells - in the skin
Decreased epithelial hyperplasia and myeloid infiltrate observed in vivo
No Treatment (Blank wires) Treated (AB-NWs)
“Nanostructured” implants for improved wound healing: Stents and Vascular Grafts
Lee et al, Nanoletters 2014; ACS Biomaterial Science, 2016
Injected Microstructures preserve and improve cardiac output after MI
Le LV et al., Biomaterials 2018
Harnessing micro- and nano-topographical cues for therapy
Smooth
Muscle
Cells
Endothelial
Cells
Epithelial
Cells
Fibroblasts
Acknowledgements
• NIH • NSF • JDRF • Zambon Ltd • Al Mann Institute • CIRM • Eli Lily • Abbvie • Santen • Gates Foundation
The Therapeutic Micro and
Nanotechnology Laboratory
at UCSF
Characterizing mechanics of fibers
Nanoindentation:
• 1590 N/m for short versus 750 N/m for long microfibers (** p < 0.01, n ≥ 12)
• constant prescribed displacement rate of 10 nm/s
with Julia Greer at Caltech