Reinvigorating Antitumor Immunity in Renal Cell Carcinoma ... · 09/12/2019  · Reinvigorating Antitumor Immunity in Renal Cell Carcinoma with Nanoparticulate STING Agonists Assistant

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)