Department of Immunology- Metformin Club -Conducted by Heiichiro UDONO MD, Ph.D., Professor

Okayama University Graduate School of Medicine,
Dentistry and Pharmaceutical Sciences

To eat Glucose or Fatty Acids,that is a problem for Antitumor
Immunity in our body!
Tumor infiltrating CD8+T lymphocytes(CD8TILs) areboosted by eating glucose, while exhausted by eatingfatty acids.
•As tumor cells eat up glucose due to Warburg Effectin tumor microenvironment, CD8TILs has nothing buteating fatty acids. This is a reason why immune system
in cancer patients is weak, which allows rapid tumor
growth.
•We need a Game Changer that can change metabolic
imbalance between CD8TILs and tumor cells.
•Metformin, an old but widely prescribed antidiabetes drug, is a candidate for this, because it
reprograms metabolism in immune cells.
•

We dissect the underlying mechanism behind
Metformin-induced antitumor Immunity.
•

Underlying mechanism of metformin-dependent anti-tumor immunity

OXPHOS
Glucose

MDSCs

Warburg effectMetformin +(anti-PD-1Ab)

Tumor growth
M2CD8 TexhaustTreg

Metabolicreprogrammingscramble for glucose

CD8 T
rebootGlycolysis

M1Tumor regression

Metformin-induced metabolic change reactivates tumor immunity

Figure 2

Metformin

A ( 10 )BCMethA
MethA for Immune cells & Tumor cells
Flow
Cytometry
analysis
5MethA

Glycolysis

1Single cell
Uehara et al. Int Immunol. 2019preparation

Mean tumor volume (mm3)miceCollect
Tumor tissue

Survival rate

02040

60

80 100

2.52F

100

1.51

Iso
αPD-1

No ne
αPD-1
Glc
αPD-1+Glc80

None
αPD-1+Glc

MethA
p-S6

Glut-1

Ki6760RNAseq analysis for tu40200.5

p=0.03p=0.010001 5 9 13 17 21 25 30 (days)G

p=0.0920(%)

No ne
αPD-1
Glc
αPD-1+Glc

Ki67

p=0.002

EImmunohistochemistry
MethA
MethA
3p-S6

Glut-140

5 9 14 18 22 26 30 (days)

***
***

Tumor
inoculation

Anti-tumor effect

None
Met+Glc20( 103)

020

4060

80 1002群間の発現変動（Volcanoplot）
Cell Sorting&値はCountRNAseq&cpm）
singlecell RNAseq
CD45+のData（
per million：
Hcont vs PD-1: Nrf2

WT : Nrf2 f/+ - Gzmb cre
KO : Nrf2 f/f - Gzmb cre( 103)

WT× 1.3
× 1/1.3

KO : Nrf2( 103)

MO52

No ne (WT)
Met+Glc (WT)No ne (KO)
Met+Glc (KO)

1.5

Contで上昇0.5・ 抗⽣ 物質に対する 反応

f/f

MO51

4H2で上昇・ 免疫反応・ 接着

0.5

Gap junction

1.5

No ne (WT)DNA replication
Ribosome
αPD-1+Glc (WT)No ne (KO) Autoimmune thyroid disease
Graft-versus-host disease
αPD-1+Glc (KO)1

Allograft rejectioncont vs P/E

VVP>0.293 min=2（ ： cpmが2以下の遺伝⼦ が4サン プ ル以上ある 遺伝⼦ を 除く ）9 13・12072
17genes
21⇒10913
25 (days)
01 5 9 13 17 21 25(days)
genes
Highly Confidential
5/1
厳秘開 ⽰ 範 囲 ∶ 社内
管 理 部 署 ∶ オンコロジー研究センター
作 成 ⽇ ∶ 2020年11⽉00⽇

J
WT : Nrf2 f/+ - Gzmb creIso
None
Met + αPD-1 + Glc

Glut-1

・ 翻訳伸⻑
厳秘Highly Confidential

KMO5p-S6

Ki67

Leone et al., Science 366, 10

Viral myocarditis

Phagosome

0.5

・ RNA合成経路の抑制

・ Clamp up 0.2（ 0.2以下の値は0.1と し て代⼊ ） で今後解析を 実施。

501

5None Met aPD-1 Met +aPD-1

・ ⽩ ⾎ 球移⾏・ 免疫反応913 17 21 25 (days)

KO : Nrf2 f/f - Gzmb creIsoNone
Met + αPD-1 + Glc

MO5Glut-1p-S6・ 免疫反応・ 細胞活性化開 ⽰ 範 囲 ∶ 社内
管 理 部 署 ∶ オンコロジー研究センター
作 成 ⽇ ∶ 2020年11⽉00⽇

KAT2A
SIR1MAXMYC
SIN3A
SIN3B
KAT8
KDM5A
KAT5
NFYA
ETS2
MYCN
E2F4
STAT2
STAT1
IRF1
IRF8
NFKB1
RELB
IRF2

Glutamine blockade induces divergent
programs to overcome tumor immune

MetabolicProfilingby Seahorse 00001

GAGE-KEGG

WT : Nrf2 f/+ - Gzmb cre: Nrf2 f/f - Gzmb creLongevity regulating pathway
I M M U NKOOME TA B OLI S M
Estrogen signaling pathway( 103)
MO5

No ne (WT)
Met+αPD-1+Glc (WT)No ne (KO)Met+αPD-1+Glc (KO)

1.5Cont vs EP4

I- Gzmb cre- Gzmb cre

***
***

P<0.22f/+

***
***

1

***
***

Mean tumor volume (mm3)Anti-tumor effect

60001

D80

Mean tumor volume (mm3 )TAMs(M1)

controlNoneMet
Glc
Met+Glc

ECAR (mpH/min)

OxPhosOxPhosSurvival rate

Glycolysis

2** **MDSCs3

Mean tumor volume (mm3)Treg

4IsoMet100***

CD8TEM(%)

controlMet
GlcMet+Glc*TAMs(M2)

Mean tumor volume (mm3 )3Metabolic reprogramming

/1

Ki67

NoneReferences
Metformin induces CD11b+-cell-mediated growth inhibition of an osteosarcoma: implications formetabolic reprogramming of myeloid cells and antitumor effects.
International Immunology, Volume 31, Issue 4, April 2019, Pages 187–198,
https://doi.org/10.1093/intimm/dxy079
Metformin prevents Peritoneal dissemination via Immune-suppressive cells in the tumor microenvironment.
Anticancer Research, September 2019, vol. 39 no. 9 4699-4709.
doi: 10.21873/anticanres.13652Metformin mediates protection against Legionella Pneumonia through activation of AMPK andMitochondrial reactive oxygen species.
J Immunol, January 15, 2018, 200 (2) 623-631;
DOI: https://doi.org/10.4049/jimmunol.1700474
Attenuation of CD4+CD25+ regulatory T cells in the tumor microenvironment by metformin, a type 2 diabetes drug.
EBioMedicine, volume 25, November 2017, Pages154-164
https://doi.org/10.1016/j.ebiom.2017.10.009
Immune-mediated antitumor effect by type 2 diabetes drug, metformin.
PNAS, February 10, 2015 112 (6) 1809-1814; first published January 26, 2015;
https://doi.org/10.1073/pnas.1417636112

Professor: Heiichiro Udono (udono@ cc.okayama-u.ac.jp)
Department of Immunology
Okayama University Graduate School of M edicine, Dentistry and Pharmaceutical Science
2-5-1 Shikata-cho, kita-ku, Okayama 700-8558, JapanI prefer Glucose (Sugar) !

O92

Biomedical Engineering Student

19/09/2019
1