Session 2 : Flash Talk
Activation of human gamma-delta
T cells by cancer metabolism
Benjamin Faustin,
Bordeaux University
Tumor microenvironment and immune infiltrate
IMMUNOSUPPRESSIVE
ENVIRONMENT
Day and al., Cell 2015
2
Tissue stress immunosurveillance
Lysis
Tissue γδ T lymphocytes
- epithelial surveillance
- antitumor cytotoxicity
Antitumor immune
response
- Mechanism of activation ?
3
Tissue stress immunosurveillance
SUPPRESSION
Lysis
Tissue γδ T lymphocytes
- epithelial surveillance
- antitumor cytotoxicity
Antitumor immune
response
- Mechanism of activation ?
4
Tissue stress immunosurveillance
SUPPRESSION
Tissue γδ T lymphocytes
Lysis
- epithelial surveillance
Blocking
antibodies
- antitumor cytotoxicity
Antitumor immune
response
- Mechanism of activation ?
New therapeutic limitations
- low amount of responders
- new toxicity (mostly auto-immunity)
- relapse
5
Tissue stress immunosurveillance
SUPPRESSION
Tissue γδ T lymphocytes
Lysis
- epithelial surveillance
Blocking
antibodies
- antitumor cytotoxicity
Antitumor immune
response
- Mechanism of activation ?
New therapeutic limitations
- low amount of responders
- new toxicity (mostly auto-immunity)
- relapse
6
PROJECT : boost the immunosurveillance
by soluble molecules
(colon, lung)
Cancer cell
Immune-stimulating
soluble molecules ?
stress Ags
TCR
LT γδ
1- screening of secreted molecules “Drug Discovery”
2- isolation and structural identification
3- exploration of therapeutic potential in vivo
7
PROJECT : boost the immunosurveillance
by soluble molecules
Immune-stimulating
soluble molecules ?
Metabolic stress
Inflammasomes
Aim #2:
Metabolic-dpt products
TCR
Aim #1:
Inflammasome-dpt products
LT γδ
(particularly IL-18)
8
AIM #1 : which inflammasome-dpt stimuli ?
Cancer (colon, lung)
Inflammasomes
500
IFNγ secretion
IL-18
Conditioned
supernatant
600
400
+ anti-IL18
300
200
100
0
0
IL-18R
TCR
1
10
Activation TCR
LT γδ
IFNg
cytotoxicity
Guerville et al., Oncoimmunology 2015
9
AIM #1 : which inflammasome-dpt stimuli ?
IL-18 as co-stimulator
of the immune
response in vivo ?
Cancer (colon, lung)
Inflammasomes
-Identify inflammasomes in human
cancer cell lines?
Pre-clinical model
IL-18
IL-18R
TCR
various shRNA NLRs (NLRP1, NLRP3,
NLRC4, AIM2)
-in human tumor tissue samples ?
Based on Tissue-MicroArray
130 tumors at different stages
Collab Bergonié (Bordeaux); Cordeliers (Paris)
LT γδ
xenograft model
“home-made” HT-29-Luc
IFNg
cytotoxicity
10
AIM #1 : which inflammasome-dpt stimuli ?
IL-18 as co-stimulator
of the immune
response in vivo ?
Cancer (colon, lung)
Inflammasomes
-Identify inflammasomes in human
cancer cell lines?
Pre-clinical model
IL-18
IL-18R
TCR
various shRNA NLRs (NLRP1, NLRP3,
NLRC4, AIM2)
-in human tumor tissue samples ?
Based on Tissue-MicroArray
130 tumors at different stages
Collab Bergonié (Bordeaux); Cordeliers (Paris)
LT γδ
xenograft model
In-house HT-29-Luc
INFLAMMASOME = BIOMARKER IN TRANSLATIONAL RESEARCH?
IL-18 = ADJUVANT IMMUNE RESPONSE IN VIVO ?
11
AIM #2 : overview of metabolic activities
in cancer cells
Hanahan and Weinberg Cell 2011
12
AIM #2 : overview of metabolic activities
in cancer cells
?
Hanahan and Weinberg Cell 2011
13
AIM #2 : overview of metabolic activities
in cancer cells
QUIESCENT CELLS
Glucose
Pyruvate
ADP
ATP
14
Schulze et al.; Nature review 2011
AIM #2 : overview of metabolic activities
in cancer cells
CANCER CELLS
GLYCOLYSIS
“WARBURG”
Glucose
Pyruvate
MITO CATABOLIC
ADP
FUNCTIONS
ATP
15
Schulze et al.; Nature review 2011
AIM #2 : overview of metabolic activities
in cancer cells
CANCER CELLS
GLYCOLYSIS
“WARBURG”
Glucose
Sustained survival
and proliferation
Pyruvate
MITO CATABOLIC
FUNCTIONS
16
Schulze et al.; Nature review 2011
AIM #2 : which metabolic-dpt stimuli ?
1- Glucose deprivation
REPROGRAMMING OF
CANCER METABOLISM
(switch Glucose/Galactose)
2- modulation of master reprogrammers
AICAR, Metformin (AMPK), Rapamycin (mTOR)
(reverse WARBURG)
?
TCR
LT γδ
IFNg
cytotoxicity
17
AIM #2 : which metabolic-dpt stimuli ?
1- Glucose deprivation
REPROGRAMMING OF
CANCER METABOLISM
(switch Glucose/Galactose)
2- modulation of master reprogrammers
AICAR, Metformin (AMPK)
(reverse WARBURG and AMPK activation)
TCR
IFNγ secretion (pg/ml)
7000
Activated human primary γδ T cells
(+ anti-CD3)
6000
5000
4000
3000
2000
1000
0
LT γδ
IFNg
cytotoxicity
18
AIM #2 : which metabolic-dpt stimuli ?
1- Glucose deprivation
REPROGRAMMING OF
CANCER METABOLISM
(switch Glucose/Galactose)
2- modulation of master reprogrammers
AICAR, Metformin (AMPK)
(reverse WARBURG and AMPK activation)
Ags
Co-stim
Adhesion
TCR
IFNγ secretion (pg/ml)
7000
Activated human primary γδ T cells
(+ anti-CD3)
6000
5000
4000
3000
2000
1000
0
LT γδ
IFNg
cytotoxicity
19
AIM #2 : which metabolic-dpt stimuli ?
1- Glucose deprivation
REPROGRAMMING OF
CANCER METABOLISM
(switch Glucose/Galactose)
2- modulation of master reprogrammers
AICAR, Metformin (AMPK)
(reverse WARBURG and AMPK activation)
BROAD
IMMUNE SIGNATURE
TCR
LT γδ
IFNg
cytotoxicity
20
AIM #2 : which metabolic-dpt stimuli ?
1- Identify immune1- Glucose deprivation
REPROGRAMMING OF
CANCER METABOLISM
stimulating soluble molecules
(switch Glucose/Galactose)
Fractionation of cell supernatants
2- modulation of by
master
regulators
chromatography
AICAR, Metformin (AMPK),
(mTOR)
-assay for Rapamycin
γδ T cell activation
(reverse WARBURG and AMPK activation)
(IFNγ)
-structural identification of active
molecules NMR-H1+Metabolomics
BROAD
IMMUNE SIGNATURE
2- Does promote antitumor
immune response in vivo ?
TCR
LT γδ
SOLUBLE STIMULATING MOLECULES
=
ADJUVANT OF IMMUNE RESPONSE IN VIVO ?
+ IDENTIFIED MOLECULES
xenograft model
In-house HT-29-Luc
IFNg
cytotoxicity
21
Thanks…
CNRS UMR 5164 CIRID
Bordeaux University
Julie Déchanet-Merville
Jean-François Moreau
Patrick Blanco
Myriam Capone
Florent Guerville
Charlotte Mannat
Sophie Daburon
Vincent Pitard
Bordeaux Hospital Pellegrin
-Renal Transplantation Dpt
P. Merville, L. Couzi
-Immunology and Immunogenetics
Laboratory
Rare Disease and Metabolism
Bordeaux university (MRGM, UB2)
Rodrigue Rossignol
Giovanni Benard
Emilie Obre
Flow cytometry facility
V. Pitard, S. Gonzales
INSERM VINCO
Bergonié Institute, Bordeaux
Pierre Soubeyran
Lydia Lartigue
22