The 26th Anti-Cancer Drug Development Forum
February 16th , 2019, Japan

Immunological resistance to Immune Checkpoint Inhibitor
Yoshihiro Ohue, MD, PhD
Laboratory HeadDivision of Cancer Immunology, National Cancer Center, Japan

1

Disclosure Information
Yoshihiro Ohue, M.D., Ph.D.
Div. of Cancer Immunology, National Cancer CenterI have no financial relationships to disclose.

Cancer Immunotherapy～the long journey to success～William Coley(1862-1936)

Paul Ehrlich(1854-1915)

2018年

3Current status of immunity checkpoint inhibitors
Activation
pathway

Inhibitory
pathway. Curr Opin Immunol 2016

Agonistic antibody

Blocking antibody

T-cell activation

No dramatic therapeutic effect except anti-PD-1 and anti-CTLA-4

48yrs, female, NSCLC (Ad) cT3N2M1b stage IVa,
EML4-ALK fusion (+)
thAt diagnosis

6 linePre anti-PD-1

Post anti-PD-1

2015.112017.22018.6

48yrs, male, NSCLC (Ad) cT1bN2M1b stage IVa,
EGFR mutaion (+)
9 th line
Pre anti-PD-1

Post anti-PD-1

Tumor microenvironment
Peripheral

Local site
Immune cell
infiltration

CD8+T cellCD4+ T cell
Regulatory T cell

MDSC/MΦ
Regulatory B cell
fibroblast
Endothelium
Lymphatic endothelium

Cause of immunological resistance to anti-tumor immunity
Mutation in IFNg signaling
Metabolite/Cytokine dysregulation
Downregulation of adhesion molecule
Oncogenic signaling
Epigenetic silencing

Lack ofinflammation

Deletion in TAP, β2M
Loss of HLA
Low TMB

Tumor intrinsic
Inadequate
pathwayantigenicity
ImmunologicalResistance
SuppressiveSuppressor
stroma fraction
immune cells
CAFs
Compensatory
Endothelium
Lymphatic endothelium
checkpoint

Treg
Breg
MDSC
TAM

PD-1, TIM-3, LAG-3, TIGIT, VISTA

Correlation of PD-L1 and Galectin-9 expression, and TIL
Analysis of each fraction
CD8P=0.01

P=0.02

P=0.02100

PD-1- TIM-3+

Apoptosis

P＝0.025040302010020

PD-1+ TIM-3+

Proliferation

P<0.01PD-1+ TIM-3-3

Differentiation

P＜0.01PD-1- TIM-3-

2025

100806040200

PD-1- TIM-3+

TIM-3

252

P＜0.01PD-1+ TIM-3+

760

TIL

P＜0.01PD-1+ TIM-3-

PD-1

22TIL

PBMC

100806040200

PD-1- TIM-3-PBMC%Annexin V %Ki-67

Gated on CD8%CD27 %frequency

Flow cytometryOhue Y, et, al. Cancer Immunol Res. 2016

T-cell apoptosis by soluble Galectin-903875150300

1113152551*60402000%Annexin V38 75 150 300
Gal-9 (pg/ml)

9Gal-9Gal-9 + Ctr-IgG

4140

Gal-9 + α-Gal-9

17Gal-9 + α-TIM-3

15Gal-9 + α-Gal-9+ α-TIM-3

17%Annexin V%Annexin VAntibody blockingNon-treat*

40%cell deathGal-9 (pg/ml)%Annexin VApoptosis by galectin-93020100038 75 150 300
Gal-9 (pg/ml)

***
***
***

50403020100Gal-9Ctr-IgG
α-Gal-9α-TIM-3－-＋－-＋＋－-＋-＋-＋-＋-＋＋ ＋Ohue Y, et, al. Cancer Immunol Res. 2016

Release of soluble Galectin-9 from tumor cells
AB

Galectin-9 expression

C****

Cell line (OU-LC-SK)

100% of Max

530102103104105010210310410510Gal-9 (pg/ml)

Intracellular

Gal-9 (pg/ml)

Surface

155Lung cancer tissue
0Afatinib-+-OU-LC-SK(EGFRwt)+PC-9(EGFRmt)

Galectin-9in clinical samples
***

2000
1800
1000
800600400
2000serumHDPt

Pleuraleffusion**** p < 0.0001

Ohue Y, et, al. Cancer Immunol Res. 2016

Involvement of immune checkpoint molecules
at the late phase of disease progression

Ohue Y, et, al. Clin Cancer Res. 201412

Tumor immune microenvironment
Antibody

CD4 T-cell
IL-4
IL-5 etc…

apoptosis

CD28

CD80/86

apoptosis

IL-2
IFN-γ etc…

TIM-3

CTLA-4B-cell

CD8 T-cell

CD8TIM-3

PD-1

Galectin-9
CD4MHC

PD-L1

MHC

APC
FcR

Tumor

Ohue Y, et, al. OncoImmunol. 2014

Comprehensive Analysis of Tumor Immunity

103

Naive Treg cells
Effector Treg cells
Non-Treg cells

0.2

0.2

IV, VIII

VICD4Treg
CD8IIIB cell0CD45RA

104

105

Fr. I:
Fr. II:
Fr. III:

61.00

FoxP3

5.5
103

33.1
104

NK cell

Myeloid cell

Acknowledgements
Dr. Hiroyoshi Nisikawa

Dr. Eiichi Nakayama Dr. Mikio Oka

Kawasaki Medical School15