¿Cómo puede la dieta modificar la genómica del cáncer?... · PC 1.5 up/do wn-2 +2 0 FC S cale...
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¿Cómo puede la dieta modificar la genómica del cáncer?
Gloria Pascual Angulo
Stem Cells and Cancer labInstitut de Recerca Biomèdica-Barcelona
Cancer stem cell heterogeneity within tumors
represents a therapeutic challenge
Primary TumourMetástasis
Metástasis
Metástasis
Metástasis
Molecular information about Disease Progression
Quiescent/Slow cycling population
Metastasis-Initiating CellsLong-term slow-cycling in 1ary tumor
CD36+ (fatty acid membrane transporter)
Sensitive to dietary fat levels
High Fatty Acid metabolism
Therapeutic targeting through
CD36 inhibition
Operate in many human tumors
Multi-organ metastasisPascual et al, Nature 2017
Landyani et al, Oncogene 2018
NEW QUESTIONS…
Which dietary fatty acids are pro-
metastatic?
Why are fatty acids pro-metastatic?
Is CD36+ an epigenetically-established
CD36+ cells
- Gastric cancer- Prostate cancer
- Renal cancer- Bladder cancer
- Lung SCC- Melanoma
- Breast (Luminal A/B)- Glioblastoma (intracraneal invasion)
- AML/CML chemoresistance- Melanoma therapy resistance
- Pancreatic carcinoma therapy resistance
so far…
Pascual et al., Nature 2017Benitah SA, Cell Stress (2018)
Pascual et al., DDM reviews (2018)
High fat diet boosts metastasis through CD36
PRIMARY TUMOUR
LN METASTASIS
60% Fat
CD36 RECEPTOR MUTANT CD36 RECEPTOR
The pro-metastatic effects of palmitic acid persist for a very long time
PA-treated (14 days post)
Untreated
Serial OSCC injection GFP+from primary tumours
UNTREATED PA-TREATED 14D post
Palmitic Acid pro-
metastatic memory
(epigenetic state?)
Epigenetic drivers of tumourigenesis and cancer metastasis.Seminars in Cancer Biology. 2017
Epigenetic driver of cancer metastasis?
Selection of particular epigenetic changes could provide a selective advantage for the cancer cells to successfully metastasize
Are all Fatty-Acids increasing metastatic potential of MICs?
Palmitic acid, but not linoleic or oleic, enhances the metastatic potential of MICs
Short-time effectPascual, G., Domínguez, D. et al.Manuscript under revision
CD44+
Oral squamous cell carcinoma SCC-25 pluc GFP [PLKO and shCD36]VDH15 pluc GFP [PLKO and shCD36]
Melanoma
501mel Cherry plucGFP [PLKO and shCD36]
Models
A Model to study in vivo long-term effects of diet in tumor cells and tumor progression
*
Lymph nodemetastasis
Lungmetastasis
Patient-derived xenograft
Pascual, G., Domínguez, D. et al.Manuscript under revision
Ex vivo Lung Metastasis BLI***
Primary Tumour
Lung Metastasis
Orthotopic Melanoma Model
Pascual, G., Domínguez, D. et al.Manuscript under revision
2ry Recipient- Primary Tumours
CD36+ CD36+
Oleic vs Control Palm vs Control
PC 1.5 up/down
-2
+2
0
FC Scale
CD36- CD36-
10-6
10-5
10-4
10-3
10-2
10-1
100
Response to interferon-betaRegulation of nervous system development
Regulation of apoptotic processRegulation of neuron dif erentiation
AngiogenesisNitrogen utilization
ChemotaxisEpithelium development
Regulation of transcription by RNA polymerase IIPositive regulation of RNA metabolic process
Response to lipidImmune response
VDH-15 CD36+
Biological Process Palm Diet Up
P value
VDH-15 CD36-
Biological Process Palm Diet Up
P value
10
NeurogenesisProteolysis
Neuron di erentiationPositive regulation of cellular protein metabolic process
Neuron projection morphogenesisRegulation of intracellular signal transduction
AngiogenesisResponse to lipid
Response to oxidative stressCell chemotaxis
Immune responseCell migration
Epidermal cell di erentiationEpidermis development
0
10-5
10-1
0
10-1
5
VDH-15 CD36+Palm Diet vs Control Diet
Up-regulated Up-regulatedDown-regulatedDown-regulated
0
0 2.5 5-2.5-5
1
2
3
4
5
0 2.5 5-2.5-5
0
1
2
3
4
5
VDH-15 CD36-Palm Diet vs Control Diet
log2(Fold Change)log2(Fold Change)
-log10(P
valu
e)
-log10(P
valu
e)
PLKO-CD36+ Control DietPLKO-CD36- Control DietPLKO-CD36+ Palm DietPLKO-CD36- Palm DietshCD36-CD36- Control DietshCD36-CD36- Palm Diet
PLKO-CD36+CTD
PLKO-CD36+PALMD
PLKO-CD36-PALMD
PLKO-CD36-CTD
shCD36-CD36-PALMD
shCD36-CD36-CTD
VDH-15 Principal Component Analysis
VDH-15 Principal Component Analysis
0
1
2
3
4
-6 -3 0 3
Up-regulated Up-regulatedDown-regulatedDown-regulated
-6 -3 0 3 6
0
1
2
3
4
SCC-25 CD36+Palm Diet vs Control Diet
SCC-25 CD36-Palm Diet vs Control Diet
log2(Fold Change)
-log10(P
valu
e)
log2(Fold Change)
-log10(P
valu
e)
P value
SCC-25 CD36-
Biological Process Palm Diet Up
10-1
5
10-1
0
10-5
100
Muscle adaptationNeuron migration
Drug metabolic processNeuron di erentiation
Collagen-activated signaling pathwayNeurogenesis/Generation of neurons
Schwann cell development/di erentiation
Myelination in peripheral nervous systemPeripheral nervous system axon regeneration
Blood vessel remodelingTissue morphogenesis
Response to lipidCell migration
SCC-25 CD36+
Biological Process Palm Diet Up
10-6
10-5
10-4
10-3
10-2
10-1
100
Positive regulation of myelination
Response to xenobiotic stimulus
Synaptic signalingFibroblast growth factor production
Response to fructose
Vasculature development
Connective tissue developmentNeurogenesis
Cell migrationNeuron di erentiation
Peripheral nervous system developmentAxonogenesis
Tissue morphogenesis
p value
Palm diet also molecularly converts CD36-
into CD36+
In shCD36, CD36- cells, are not able to
respond to palm diet
Incluir Análisis más profundo del shCD36?
Incluir Análisismásprofundo de Olive Diet?
SCC-25 Principal Component Analysis
VDH-15 Principal Component Analysis
Molecular characterization of fat-exposed cancer cells- RNA Microarrays
Pascual, G., Domínguez, D. et al.Manuscript under revision
Molecular characterisation palm-exposed cancer cells
Pascual, G., Domínguez, D. et al.Manuscript under revision
Is Palmitic Acid establishing an epigenetic memory?
Fatty acid metabolism is epigenetically altered in metastatic tumors
Rinaldi et al, Cell Stem Cell (2016)Rinaldi et al, eLife (2017)
The nexus of chromatin regulation and intermediary metabolism. Nature, 2013.
Cell Line Assay Time
SCC-25(Established
OSCC)
Patient-derived OSCCs
RNA-Seq• 4 Days• 14 Days
ATAC-Seq
• 4 Days• 14 Days
h/MeDIP-Seq
ChIP-Seqs
Cell Line Histone Mark Time
SCC-25(Established OSCC)
Patient-derived OSCCs
H3K4me1
• 4 Days• 14 Days
H3K4me3
H3K27me3
H3K27ac
H3K9ac
H3K9me3
Pascual, G., Domínguez, D. et al.Manuscript under revision
H3K4me3 (i.e. promoters) shows a significant memory of palmitic acid
UNTREATED-4d
UNTREATED-14d
TREATED-4d TREATED-14d
150
729
PA-Induced Memory PCA: Condition+Factor
Principal Component #1 [94%]
Pri
ncip
al C
om
po
nen
t #2 [
2%
]
−0.6
−0.4
−0.2
0.0
0.2
−0.30 −0.25 −0.20
●●
●
●
●
●
●
●
●
●
●
●
12
3
1
2
3
1
2
3
1
2
3
Days4:UntreatedDays4:TreatedDays14:UntreatedDays14:Treated
Pascual, G., Domínguez, D. et al.Manuscript under revision
H3K4me1 (i.e. enhancers) does not retain the memory of palmitic acid
Who establishes the metastatic epigenetic memory of
dietary palmitic acid?
• H3K4me3 ChIP-seq - CTRL PLKO vs SETD1A KD (palmitic versus control)
Cell TypeHistone
MarkTime Point FDR FC
#Diff Peaks UP PA
#Diff Peaks DOWN PA
VDH-15 SETD1A KD H3K4me3 14 Days 0.05 1 215 116
Cell TypeHistone
MarkTime Point FDR FC
#Diff Peaks UP PA
#Diff Peaks DOWN PA
VDH-15 CTRL PLKO H3K4me3 14 Days 0.05 1 857 25
Inhibition of Setd1a prevents the H3K4me3 memory induced by dietary palmitic acid
CH
RD
L1 L
ocu
s
VDH15 CTRL UNT 14DVDH15 CTRL PA 14DSETD1A KD UNT 14DSETD1A KD PA 14D
SER
PIN
A1
Lo
cus
Inhibition of Setd1a prevents the H3K4me3 memory induced by dietary palmitic acid
VDH15 CTRL UNT 14DVDH15 CTRL PA 14DSETD1A KD UNT 14DSETD1A KD PA 14D
Inhibition of Setd1a prevents the metastatic spreading in oral cancer
Pascual, G., Domínguez, D. et al.Manuscript under revision
H3K4me3 memory induced by dietary palmitic acid
GO Biological Process
Pascual, G., Domínguez, D. et al.Manuscript under revision
NES 1.51
Nominal Pvalue 0.05NES 1.50
Nominal Pvalue 0.04
CD36+Palm vs Control Diet Enrichment
VD
H-1
5
NES 1.59
Nominal Pvalue 0.01
NES 1.57
Nominal Pvalue 0.01
NES 1.74
Nominal Pvalue 0.001
NES 3.17
Nominal Pvalue 0.0NES 1.56
Nominal Pvalue 0.0NES 1.50
CD36+Palm vs Control Diet Enrichment
CD36-Palm vs Control Diet Enrichment
NES 3.55
Nominal Pvalue 0.0NES 2.55
Nominal Pvalue 0.0
CD36-Palm vs Control Diet Enrichment
SC
C-2
5
Palm Diet-induced memory
Neural-related
Include stainings as link for the next part (trasncriptomics of neural stroma)
NES 1.51
Nominal Pvalue 0.05NES 1.50
Nominal Pvalue 0.04
CD36+Palm vs Control Diet Enrichment
VD
H-1
5
NES 1.59
Nominal Pvalue 0.01
NES 1.57
Nominal Pvalue 0.01
NES 1.74
Nominal Pvalue 0.001
NES 3.17
Nominal Pvalue 0.0NES 1.56
Nominal Pvalue 0.0NES 1.50
CD36+Palm vs Control Diet Enrichment
CD36-Palm vs Control Diet Enrichment
NES 3.55
Nominal Pvalue 0.0NES 2.55
Nominal Pvalue 0.0
CD36-Palm vs Control Diet Enrichment
SC
C-2
5
Palm Diet-induced memory
Neural-related
Include stainings as link for the next part (trasncriptomics of neural stroma)
NEURAL-RELATED SIGNATURE PALM-EXPOSED CANCER CELLS
• Transcription/Modulator Factor activity• Secreted Factors/Neurite outgrowth• Neurotransmitter release/Receptor
Activity
Is the acquisition of a neural-related signature of OSCC related with
metastasis?
Epithelia Stroma
PLKO-CD36bright
PLKO-CD36bright
PLKO-CD36dim
PLKO-CD36dim
shCD36-CD36dim
shCD36-CD36dim
-6 -3 0 3 6
0
1
2
3
4
Muscle adaptation
Drug metabolic process
Collagen-activated signaling pathway
Blood vessel remodeling
SCC25 CD36bright
Palm Diet vs Control Diet
log2(Fold Change)
Up-
regulated
Down-
regulated
-log10(P
valu
e)
100
10-5
10-1
0
10-1
5
Pvalue
SCC-25 CD36bright
Palm Diet Biological Process Up
50uM
50uM
50uM
50uM
SCC-25 PLKO-Control Diet-2ry recipientNEF-LKT-14Dapi
SCC-25 shCD36-Control Diet-2ry recipientNEF-LKT-14Dapi
SCC-25 PLKO Palm Diet-2ry recipientNEF-LKT-14Dapi
SCC-25 shCD36-Palm Diet-2ry recipientNEF-LKT-14Dapi
B C
D
10-6
0
10-4
0
10-2
0
100
pvalue
Biological Process UPPPalm Diet vs Control Diet
Extracellular matrix organizationCell migration
Collagen bril organizationResponse to growth factor
Cell-matrix adhesion Supramolecular ber organization
Response to lipidNeuron di erentiation
NeurogenesisSecretion
Reg. of nervous system developmentBlood vessel remodelingSprouting angiogenesis
Actin cytoskeleton organizationResponse to steroid hormone
InnervationGliogenesis
Neuron recognitionHypothalamus cell migration
Serotonin uptake
G
6
4
2
0
-5 0
UP-RegulatedDOWN-Regulated
log2(Fold Change)
PLKO Palm vs PLKO Control
SCC-25/VDH-15
Neural-enriched stroma
5
DEGsUP/DOWN
in (F)6
4
2
0
-lo
g1
0(P
va
lue
)
shCD36 Palm vs PLKO Control
-5 0 5log2(Fold Change)
SCC-25/VDH-15
Neural-enriched stroma
-lo
g1
0(P
va
lue
)
E
F
H
Figure 3
Response to lipid
Neuron di erentiation
Neuron migration
Neurogenesis/Generation of neurons
Myelination in peripheral nervous systemPeripheral nervous system axon regeneration
Schwann cell development/di erentiation
SCC-25 Principal
Component Analysis
I
HUMAN-[CD36-]
HUMAN-[CD36+]
Mouse
Schwann celldevelopment
neuron projectionregeneration
CTD
PALMD
CTD
PALMD
PALMD
CTD
SCC-25 - H3K4me3 in vitro ChIPs GO Analysis (14D post-PA):
UP-regulated Neural-related GO terms
GO
Bio
log
ical P
ro
cess
GO
Bio
log
ical P
ro
cess
0 2 4 6 8
axonogenesisaxon guidance
cell morphogenesis involved in neuron dif erentiationglycosaminoglycan metabolic process
sprouting angiogenesisregulation of neuron projection development
chondroitin sulfate proteoglycan biosynthetic processaxon development
heart contractionspinal cord dorsal/ventral patterning
-Log (pvalue)
14D PA UP
VDH-15 - H3K4me3 in vivo ChIPs GO Analysis (2ary PTs):
UP-regulated Neural-related GO terms
0 2 4 6 8
regulation of p38MAPK cascadebranching morphogenesis of an epithelial tube
forebrain regionalizationvasculogenesis
lung developmentneuron projection morphogenesis
cell morphogenesis involved in neuron dif erentiationregulation of angiogenesis
regulation of Wnt signaling pathwaynegative regulation of cell communication
-Log (pvalue)
UP in 2ary PALM-Diet Tumours
A
f
Cell migration
OliveControl Palm
Strategy for enrichment in mouse “tumour-neural stroma”
Neural Stem/ progenitorsInmature/Intermediate progenitorsNeuron-like cells (?)Mature neurons (X)
Glial Cellsof PNS
Glial Cellsof CNS
ModelsOral squamous cell carcinoma SCC-25 pluc GFP [PLKO and shCD36]VDH15 pluc GFP [PLKO and shCD36]
Melanoma501mel Cherry plucGFP [PLKO and shCD36]
A
Palm-
PLKO
Palm-
shCD36
Control-
shCD36
Control-
PLKO
B
PC1 (65.3%)-50 0 50 100
PALM Diet (A)
CONTROL Diet (B)
PALM Diet (B)
OLEIC Diet (B)
VDH15 Neural-enriched stroma
-20
-10
010
20
30
40
PC
2(1
1.5
%)
SCC-25 PLKO-Palm Diet
2ry Recipient/Primary tumour
GFP+
GFP-CD31-
CD31+
CD31-BV-710
GFP
Dapi-
32.7
60.0
0.46
GFP+
GFP-CD31-
CD31+
CD31-BV-710GFP
22.1
72.3
0.47
Dapi-
SCC-25 PLKO-Control Diet
CD45-PE-Cy7
Neu
ral M
ix-A
PC
APCbrightCD45low
Neural-enriched fractionGFP-CD31-
5.73
Mouse
CD45bright
CD45-PE-Cy7
Neu
ral M
ix-A
PC
Neural-enriched fractionGFP-CD31-
APCbrightCD45low
9.29
Mouse
CD45bright
MouseMouse
C
-2
0-1
01
02
00
PC1 (53.3%)
PC2 (11.6%)
PLKO- PALM
VDH- 1 5
Neural-enriched fraction
2ry recipient
E
40-20 0 20
-20
-10
010
PC1 (48.5%)
PC
2 (
13
.9%
)
PLKO- Control Diet
PLKO- Palm Diet
shCD3 6 - Control D iet
shCD3 6 - Palm Diet
PLKO- PALM
Neural-enriched stroma puri cation
Neural-enriched stroma puri cation
CD36 depletion in OSCC
SCC- 2 5
Neural-enriched fraction
2ry recipient
Supplementary Figure 10
D
F
Palm-exposed cells are able to modify the tumourneural-associated stroma in a CD36-dependent manner
Pascual, G., Domínguez, D. et al.Manuscript under revision
Palm-exposed cells are able to modify the neural stroma in a CD36-dependent manner
Pascual, G., Domínguez, D. et al.Manuscript under revision
PLKO-CD36bright
PLKO-CD36bright
PLKO-CD36dim
PLKO-CD36dim
shCD36-CD36dim
shCD36-CD36dim
-6 -3 0 3 6
0
1
2
3
4
Muscle adaptation
Drug metabolic process
Collagen-activated signaling pathway
Blood vessel remodeling
SCC25 CD36bright
Palm Diet vs Control Diet
log2(Fold Change)
Up-
regulated
Down-
regulated
-log10(P
valu
e)
100
10-5
10-1
0
10-1
5
Pvalue
SCC-25 CD36bright
Palm Diet Biological Process Up
50uM
50uM
50uM
50uM
SCC-25 PLKO-Control Diet-2ry recipientNEF-LKT-14Dapi
SCC-25 shCD36-Control Diet-2ry recipientNEF-LKT-14Dapi
SCC-25 PLKO Palm Diet-2ry recipientNEF-LKT-14Dapi
SCC-25 shCD36-Palm Diet-2ry recipientNEF-LKT-14Dapi
B C
D
10-6
0
10-4
0
10-2
0
100
pvalue
Biological Process UPPPalm Diet vs Control Diet
Extracellular matrix organizationCell migration
Collagen bril organizationResponse to growth factor
Cell-matrix adhesion Supramolecular ber organization
Response to lipidNeuron di erentiation
NeurogenesisSecretion
Reg. of nervous system developmentBlood vessel remodelingSprouting angiogenesis
Actin cytoskeleton organizationResponse to steroid hormone
InnervationGliogenesis
Neuron recognitionHypothalamus cell migration
Serotonin uptake
G
6
4
2
0
-5 0
UP-RegulatedDOWN-Regulated
log2(Fold Change)
PLKO Palm vs PLKO Control
SCC-25/VDH-15
Neural-enriched stroma
5
DEGsUP/DOWN
in (F)6
4
2
0
-lo
g1
0(P
va
lue
)
shCD36 Palm vs PLKO Control
-5 0 5log2(Fold Change)
SCC-25/VDH-15
Neural-enriched stroma
-lo
g1
0(P
va
lue
)
E
F
H
Figure 3
Response to lipid
Neuron di erentiation
Neuron migration
Neurogenesis/Generation of neurons
Myelination in peripheral nervous systemPeripheral nervous system axon regeneration
Schwann cell development/di erentiation
SCC-25 Principal
Component Analysis
I
HUMAN-[CD36-]
HUMAN-[CD36+]
Mouse
Schwann celldevelopment
neuron projectionregeneration
CTD
PALMD
CTD
PALMD
PALMD
CTD
SCC-25 - H3K4me3 in vitro ChIPs GO Analysis (14D post-PA):
UP-regulated Neural-related GO terms
GO
Bio
log
ical P
ro
cess
GO
Bio
log
ical P
ro
cess
0 2 4 6 8
axonogenesisaxon guidance
cell morphogenesis involved in neuron dif erentiationglycosaminoglycan metabolic process
sprouting angiogenesisregulation of neuron projection development
chondroitin sulfate proteoglycan biosynthetic processaxon development
heart contractionspinal cord dorsal/ventral patterning
-Log (pvalue)
14D PA UP
VDH-15 - H3K4me3 in vivo ChIPs GO Analysis (2ary PTs):
UP-regulated Neural-related GO terms
0 2 4 6 8
regulation of p38MAPK cascadebranching morphogenesis of an epithelial tube
forebrain regionalizationvasculogenesis
lung developmentneuron projection morphogenesis
cell morphogenesis involved in neuron dif erentiationregulation of angiogenesis
regulation of Wnt signaling pathwaynegative regulation of cell communication
-Log (pvalue)
UP in 2ary PALM-Diet Tumours
A
f
Cell migration
Functional RelationHuman Palm/OSCC-Neural-enriched Stroma
Pascual, G., Domínguez, D. et al.Manuscript under revision
0.1
0.3
0.2
0.4
1.0
No
rma
lise
d P
ho
ton
u
x 1
0E
6
PLKO
CTD
shEGR2
PALM CTD PALM CTD PALM
#38_9 #40_9
CTD PALM
shGAL
#73_4
n.s.
*
n.s.n.s.
PLKO
CTD
shEGR2
PALM CTD PALM CTD PALM
#38_9 #40_9
CTD PALM
shGAL
#73_4
No
rma
lise
d P
ho
ton
ux 1
0E
5
0.5
1.5
1
2
2.5 Lung MetBLI/Tumour size
LN MetBLI/Tumour size
*
n.s.
n.s.
**
0
25
50
75
100
PLKO
CTD
shEGR2
PALM CTD PALM CTD PALM
#38_9 #40_9
CTD PALM
shGAL
#73_4
fre
qu
en
cy o
f m
eta
sta
ses (
%)
MetMet-free
Met Frequency
*
n.s.
n.s.n.s.
Cont
rol D
iet v
s Pa
lm D
iet
pv<0.05
Palm-induced mouse
neural signature
FC (f
old
chan
ge)
NES -1.57
Nominal pvalue 0.03
NES -1.50
Nominal pvalue 0.03PLKO-Palm
Human_Neural-related UPshGAL-Palm
Up-regulatedFC 1.5 pvalue<0.05
11 49655
FC 1.5
PLKO-Palm
Human_Neural-related UP
shEGR2-Palm
Up-regulated
FC 1.5 pvalue<0.05
5 52461
FC 1.5
11 58755
NES -1.49Nominal pvalue 0.008
NES -1.76Nominal pvalue 0.003
shGAL-Palm Diet vs PLKO-Control Diet
shEGR2-Palm Diet vs PLKO-Control Diet
Gene
Exp
ress
ion
Figure 4
A VDH-15 - H3K4me3 Set1 KD ChIPs GO Analysis (14D post-PA):
0 1 2 3 4 5
regulation of actin cytoskeleton organizationpeptidyl-tyrosine modif cation
modulation of excitatory postsynaptic potentialregulation of phospholipase C acitvity
positive regulation of excitatory postsynaptic potentialpositive regulation of synaptic transmissionpositive regulation of neuron di erentiation
glycosaminoglycan biosynthetic processcell di erentiation in hindbrain
regulation of dendritic spine development
-Log (pvalue)B VDH-15 - 1ary Diets Tumour BLI
(CTRL pLKO.1 vs :)DK 1teS
Ph
oto
n f
lux
BLI LNmet - Week 3 post-IT
CTRLPLKO.1 CTRL Diet CTRLPLKO.1 PALM Diet 10e5
10e8
10e5
10e7
LNmet Visualization:
02 1064 1066 1068 1061 107
5.0 107
1.0 108
1.5 108
2.0 108n.s.
*
CTRL PLKO.1 CTRL DIET
CTRL PLKO.1 PALM DIET
Set1 KD CTRL DIET
Set1 KD PALM DIETn.s.
*
DOWN in 14D post-PA Set1A KDvs14D UNTR Set1A KD
#38_9
#40_9
Ne
uro
n M
igra
tio
nO
lig
od
en
dro
cy
te
De
ve
lop
me
en
t
5
4
3
2
1
0
-5
-4
-3
-2
-1
PL
KO
sh
CD
36
#98_
99
sh
EG
R2
#4
0_
9
sh
EG
R2
#3
8_
9
sh
GA
L#7
3_
4
shGAL-Palm Diet vs PLKO-Control Diet
shEGR2-Palm Diet vs PLKO-Control Diet
Sy
mp
ath
eti
c N
erv
ou
sS
ys
tem
De
ve
lop
me
nt
Gli
al
Ce
ll P
roli
fera
tio
n
C D E
F
G
H
I
J K
L
A
A
A
A
CTRL Diet PALM DietSet1A KDSet1A KD
CHRDL1
Set1A-regulated Neural genes
H3K4me3 Peak Visualization:
CTRL pLKO.1 UNTREATED post 14 Days
CTRL pLKO.1 PA-TREATED post 14 Days
Set1A UNTREATED post 14 Days
Set1A PA-TREATED post 14 Days
GRIP2
Functional Study
0.1
0.3
0.2
0.4
1.0
Norm
alis
ed P
ho
ton
ux 1
0E
6PLKO
CTD
shEGR2
PALM CTD PALM CTD PALM
#38_9 #40_9
CTD PALM
shGAL
#73_4
n.s.
*
n.s.n.s.
PLKO
CTD
shEGR2
PALM CTD PALM CTD PALM
#38_9 #40_9
CTD PALM
shGAL
#73_4
Norm
alis
ed
Ph
oto
n
ux 1
0E
5
0.5
1.5
1
2
2.5 Lung MetBLI/Tumour size
LN MetBLI/Tumour size
*
n.s.
n.s.
**
0
25
50
75
100
PLKO
CTD
shEGR2
PALM CTD PALM CTD PALM
#38_9 #40_9
CTD PALM
shGAL
#73_4
fre
qu
en
cy o
f m
eta
sta
se
s (
%)
MetMet-free
Met Frequency
*
n.s.
n.s.n.s.
Cont
rol D
iet v
s Pa
lm D
iet
pv<0.05
Palm-induced mouse
neural signature
FC (f
old
chan
ge)
NES -1.57
Nominal pvalue 0.03
NES -1.50
Nominal pvalue 0.03PLKO-Palm
Human_Neural-related UPshGAL-Palm
Up-regulatedFC 1.5 pvalue<0.05
11 49655
FC 1.5
PLKO-Palm
Human_Neural-related UP
shEGR2-Palm
Up-regulated
FC 1.5 pvalue<0.05
5 52461
FC 1.5
11 58755
NES -1.49Nominal pvalue 0.008
NES -1.76Nominal pvalue 0.003
shGAL-Palm Diet vs PLKO-Control Diet
shEGR2-Palm Diet vs PLKO-Control Diet
Gene
Exp
ress
ion
Figure 4
A VDH-15 - H3K4me3 Set1 KD ChIPs GO Analysis (14D post-PA):
0 1 2 3 4 5
regulation of actin cytoskeleton organizationpeptidyl-tyrosine modif cation
modulation of excitatory postsynaptic potentialregulation of phospholipase C acitvity
positive regulation of excitatory postsynaptic potentialpositive regulation of synaptic transmission
positive regulation of neuron di erentiationglycosaminoglycan biosynthetic process
cell di erentiation in hindbrainregulation of dendritic spine development
-Log (pvalue)B VDH-15 - 1ary Diets Tumour BLI
(CTRL pLKO.1 vs :)DK 1teS
Ph
oto
n f
lux
BLI LNmet - Week 3 post-IT
CTRLPLKO.1 CTRL Diet CTRLPLKO.1 PALM Diet 10e5
10e8
10e5
10e7
LNmet Visualization:
02 1064 1066 1068 1061 107
5.0 107
1.0 108
1.5 108
2.0 108n.s.
*
CTRL PLKO.1 CTRL DIET
CTRL PLKO.1 PALM DIET
Set1 KD CTRL DIET
Set1 KD PALM DIETn.s.
*
DOWN in 14D post-PA Set1A KDvs14D UNTR Set1A KD
#38_9
#40_9
Ne
uro
n M
igra
tio
nO
lig
od
en
dro
cy
te
De
ve
lop
me
en
t
5
4
3
2
1
0
-5
-4
-3
-2
-1
PL
KO
sh
CD
36
#9
8_
99
sh
EG
R2
#4
0_
9
sh
EG
R2
#3
8_
9
sh
GA
L#
73
_4
shGAL-Palm Diet vs PLKO-Control Diet
shEGR2-Palm Diet vs PLKO-Control Diet
Sy
mp
ath
eti
c N
erv
ou
sS
ys
tem
De
ve
lop
me
nt
Gli
al
Ce
ll P
roli
fera
tio
n
C D E
F
G
H
I
J K
L
A
A
A
A
CTRL Diet PALM DietSet1A KDSet1A KD
CHRDL1
Set1A-regulated Neural genes
H3K4me3 Peak Visualization:
CTRL pLKO.1 UNTREATED post 14 Days
CTRL pLKO.1 PA-TREATED post 14 Days
Set1A UNTREATED post 14 Days
Set1A PA-TREATED post 14 Days
GRIP2
Neural-Enriched Stroma
Pascual, G., Domínguez, D. et al.Manuscript under revision
Overall survival
Overall survival
• Metastatic stem cells depend on CD36 to metastasize
• Palmitic Acid promotes the function of MICs through CD36
• Palmitic acid elicits a metastatic epigenetic memory in MICs: Setd1a and H3K4me3
• CD36 and palmitic acid induce innervation of tumors
• Metastatic cells directly reshape tumour innervation
• Epigenetic PA-induced tumor innervation facilitates multi-organ metastasis
CONCLUSIONS
The Stem Cells and Cancer Lab
Diana Dominguez (PhD)Uxue Urricelqui (PhD)
Paloma Solá (PhD)Valentina Zinna (PhD)
Guiomar Solanas (Research Associate)Gloria Pascual (Research Associate)
Alexandra Avgustinova (Postdoc)Magdolna Djurec (Postdoc)
Carmelo Laudana (Bioinformatician)Andrés Castellanos (Research Assistant)
Thanks: IRB Administration + Scientific Services
Collaborators for this projectLuciano di Croce (CRG, Barcelona)
Holger Heyn (CNAG, Barcelona)Ali Shilatifard (Feinberg School of Medicine, USA)
Ramin Shiekhatar (University of Miami)
Salvador Aznar-Benitah
¿Cómo puede la dieta modificar la genómica del cáncer?
Gloria Pascua Angulo
Stem Cells and Cancer labInstitut de Recerca Biomèdica-Barcelona