Reinvigorating Antitumor Immunity in Renal Cell Carcinoma with
Nanoparticulate STING Agonists
Assistant Professor
Department of Chemical & Biomolecular Engineering
Department of Biomedical Engineering
Vanderbilt-Ingram Cancer Center
Vanderbilt University
Kidney Cancer Research Summit 2019
John T. Wilson, Ph.D.
Overcoming the Nonimmunogenic TME: A Grand Challenge in Immuno-Oncology
complete response non-immunogenic “cold” TME
CD8+ T cells PD-L1 IFN-α/β
T cell chemokines MHC-I
TAMs/MDSCs M2,Th2 cytokines
TAM /
M2-MΦ
tumor
iDCMDSC
CD86low
MHC-Ilow
IFN-α/βlow
?
complete responseimmunogenic “hot” TME
CD8+ T cells PD-L1 IFN-α/β
T cell chemokines MHC-I
TAMs/MDSCs M1,Th1 cytokines
M1-MΦ
tumor
mDCMDSC
CD86hi
MHC-Ihi
IFN-α/βhi
T cell
CD8+
IFN-γ
GzmB
engineered materialsmonths
ove
rall
surv
ival
engineering
immunology medicine
immune prodrugs
cancer vaccinesinnate immunity
drug combos
Shae+, Nature Nano. (2019)
Jacobson+, Biomat Sci (2018)
Garland+, CMBE (2019)
Qui+, Biomaterials (2018)
Sevimli+, ACS Biomat (2017)
Wilson+, AAPS J (2015)
Palmer+, Bioconj Chem (2018) Elion+, Cancer Res. (2018)
Motzer et al.; Lancet Oncol; 2019 (https://doi.org/10.1016/S1470-2045(19)30413-9)
Endogenous DNA Sensing via the cGAS-STING Pathway Stimulates Anti-Tumor Immunity
N
N
NH2
N
NO
OH O
P-O
O
O
O
OH O
P O-
O
N
N
N
NH
O
NH2
2’
3’
G
cGAMP
Chen e
t al, N
at. Im
munolo
gy,
2016
Key Drug Delivery Barriers
• Not “drug-like” – inefficient cytosolic delivery
• Rapidly cleared (t1/2~1-2 min)
• Minimal delivery to tumors / lymphoid organs
• Current clinical trials (Phase I/II)
• Intratumoral administration
• Many patients are ineligible
“hot” tumor
Delivering a “Smart” STING: pH-Responsive Polymersomes for cGAMP Delivery
pH sensormembrane-
interactive
alkyl chain
PEG corona
[mPEG]2kD-b-[DEAEMA0.55-c-BMA0.35-c-PDSMA0.1]5kD
bilayer
crosslinking
Shae D, […], Wilson JT; Nature Nanotechnology, 14(3):269-278 (2019)
0.1 1 10 100 10000
10
20
30
40
50
Diameter (nm)
Fre
qu
en
cy (
%) pH 7.4
pH 6.6
pH 5.8
100 nm 7.4 7.0 6.6 6.2 5.8
0
20
40
60
80
pH
% h
em
oly
sis
Delivering a “Smart” STING: pH-Responsive Polymersomes for cGAMP Delivery
encapsulation of cGAMP in “smart” nanoparticles enhances activity ~250-600 fold
100 101 102 1030.0
0.2
0.4
0.6
cGAMP (nM)
IFN
-I R
esp
on
se (
AU
)
100 101 102 1030.0
0.1
0.2
0.3
Dose (nM)IF
N-I
Resp
on
se (
AU
)
100 101 102 1030.0
0.1
0.2
cGAMP (nM)
IFN
-I R
esp
on
se (
AU
)
EC50:
cGAMP: 31,000 nMSTING-NP: 67 nM
EC50:
cGAMP: 21,000 nMSTING-NP: 36 nM
EC50:
cGAMP: 55,000 nMSTING-NP: 224 nM
THP-1 monocyte RAW 264.7 mΦ B16 melanoma
Free cGAMP STING-NP
(crosslinked)
STING-NP
(non-crosslinked)
PEG-DB Mix
(cGAMP + NP)
Shae D, […], Wilson JT; Nature Nanotechnology, 14(3):269-278 (2019)
STING-NPs Improve the Pharmacokinetic Properties of Systemically Administered cGAMP
0 1 2 3 40
1000
2000
3000
Time (hr)
cG
AM
P n
g/m
L
STING-NPcGAMP
t1/2 ≅ 0.03 h
t1/2 ≅ 1.3 h
AUC ≅ 850 ng-h/mL
AUC ≅ 5500 ng-h/mL
0.25 1 4 80
2
4
6
8
time (hr)
% ID
per
gra
m t
um
or
liver
spleen
kidney
lung
0.25 1 4 80.0
0.5
1.0
1.5
2.0
Time (hr)
% ID
per
gra
m t
issu
e
0.25 1 4 80
5
10
15
20
Time (hr)
% ID
per
gra
m t
issu
e
0.25 1 4 80
5
10
15
20
Time (hr)
% ID
per
gra
m t
issu
e
0.25 1 4 80
2
4
6
8
Time (hr)
% ID
per
gra
m t
issu
e
STING-NPcGAMP
STING-NPs Reprogram the Tumor Immune Microenvironment in Renca Model of RCC
PBS
STIN
G-N
P
0.0
0.1
0.2
0.3
0.4
Ifnb1/Hmbs
**
PBS
STIN
G-N
P
0
2
4
6
8
Cxcl10/Hmbs *
PBS STING-NP0
200
400
600
CD
8+ T
cells / m
g t
um
or
PBS STING-NP
****
PBS STING-NPs0
500
1000
1500
CD
4+ T
cells m
g-1
PBS
STING-NPs
***
PBS STING-NP0
200
400
600
CD
8+ T
cells / m
g t
um
or
PBS STING-NP
****PBS STING-NP
STING-NPs Open a Therapeutic Window for Systemic Administration of cGAMP
d0 d7 d10 d3
10 μg cGAMP
(intravenous)
100 μg αPD-L1
(intraperitoneal)
10 20 30 40 500
25
50
75
100
Day
Perc
en
t su
rviv
al (%
)
PBS aPD-L1 STING-NP STING-NP+aPD-L1cGAMP
****
***
10 15 200
300
600
900
1200
1500
1800
Days Post-Tumor Inoculation
Tu
mo
r V
olu
me (
mm
3)
******
free cGAMP
PBS
Legend
Legend
Legend
10 20 30 40 500
25
50
75
100
Day
Perc
en
t su
rviv
al (%
)
PBS aPD-L1 STING-NP STING-NP+aPD-L1cGAMP
****
***
Renca (SubQ)
(~70 mm3)
Acknowledgements
www.wilsonlabvanderbilt.com
current: Kyle Becker, Max Jacobson, Frances Knight,
Carcia Carson, Jessalyn Baljon, Lihong Bishop,
Moe Wehbe, Kyle Garland, Christian Palmer, Lucinda
Pastora, Taylor Sheehy
former: Dan Shae, Sema Sevimli, Feng Qiu Discovery Grant Program
@WilsonLabVU
collaborators
Jeff Rathmell, PhD
Kathryn Beckermann, MD, PhD
Manuel Ascano, PhD; Justin Balko, PhD; Rebecca
Cook, PhD; Craig Duvall, PhD; Doug Johnson, MD;
Sebastian Joyce, PhD; Young Kim, MD, PhD; Ann
Richmond, PhD; Julie Rhoades, PhD; Scott Guelcher,
PhD; Gary Sulikowski, PhD; Plamen Christov, PhD;
Anna M. Pyle, PhD (Yale); Venkata Krishnamurthy,
PhD (AstraZeneca)
