Targeting Prostate Cancer Metabolism Robert Gillies,

et al. H. Lee Moffitt Cancer Center and Research Institute; Tampa FL USA;

Robert.Gillies@Moffitt.org

Frontiers in Urologic Oncology!

Disclosures

•  Nothing to disclose relevant to this presentation.!

Objec.ves•  High Grade Prostate Cancers exhibit a Warburg

Effect (aerobic glycolysis); possibly related to PTEN loss!

•  Aerobic glycolysis leads to acidification of the tumor microenvironment, which can be neutralized with oral buffers!

•  Oral buffers neutralize acidity and reduce spontaneous and experimental metastases; and can retard emergence of cancer in TRAMP models.!

•  This is related to buffers reducing the evolutionary fitness of aggressive cells in favor of more benign cell types within the tumor.!

Pasteur Effect is acute; Warburg Effect is hardwired

pHe=6.74 pHi=7.28

β− NTP

α− NTP γ− NTP

3-APP GPC

PME

P i

Chemical Shift, d (ppm) -30 -20 -10 0 10 20 30

Elevated glycolysis leads to tumor acidosis!

H2N-CH2-CH2-CH2-P=O

O

OH

pKa = 6.91

H2N-CH2-CH2-CH2-P=O

O

OH

pKa = 6.91

Resolution = 8 x 8 x 8 mm3 Gillies, et al. NMRiB, 1994

In vivo 31P spectrum of MCF-7/s tumor in SCID mouse!

pHex=6.84!

pHin=7.32!

pHe mapping with MRSI of IEPA

DCE of MDA-435 pHe map of MDA-435

6.8

6.4

Extra

cellu

lar p

H

Van Sluis et al., MRM, 2001

FITC-pHlipimagingofperi-tumoralacidity(MDA-mb-231inDWC)

5X, 45 min

(Tumor)!

(Stroma)!

Estrellaetal“AcidityGeneratedbytheTumorMicroenvironmentDrivesLocalInvasion”CancerResearch,2013

pHlip = pH low insertion peptide, which undergoes an acid catalyzed transition causing membrane insertion

Day 7 Day 14

A

6.6

pH

6.6

7.4

270º

180º

90º

0º

C

B

6.836.83

7.15

7.15

7.26

7.26

7.27

7.267.26 7.15

7.04

7.02

6.73

6.65

6.78

7.15

0º

270º

180º

90º

B

6.836.83

7.15

7.15

7.26

7.26

7.27

7.267.26 7.15

7.04

7.02

6.73

6.65

6.78

7.15

0º

270º

180º

90º

B

D

Estrella et al Canc Res (2013)

Both growth and acidity were

inhibited with 200 mM bicarbonate Tx!

Local Invasion is associated with an acidic pH!

It’sallaboutthatbase…NaHCO3raisesTumorpH

Chemical Shift (ppm)-30-20-100102030

β-NTP

α-NTPγ-NTP

3-APP

GPC

PME

Piδ (ppm)

182022242628

B.

A.

+ bicarbcontrol

Bicarb Control p!pHe 7.4 ± 0.06 7.0 ± 0.11 <0.005!

pHi 7.0 ± 0.06 7.1 ± 0.09 n.s.!

MDA-mb-231 br. ca.

Differen.alSensi.vity(PC3mareverysensi.ve,B16reresistant)

Robey et al., Canc. Res., 2009

BufferTherapyinhibitsinvasionandmetastasis(PC3mandTRAMP)

Arig Ibrahim-Hashim

BicarbraisespHandprevents1otumorsifgivenearly

Ibrahim-Hashim, Urology, 2012!

BicarbTreatTRAMPmice:Lowdoseearlyorhighlateà

nomets

Ibrahim-Hashim et al., Cancer Research 2017

C2/C3cellphenotypesinTRAMPtumors

C2:!-glycolytic!-invasive!

-high CA-IX!-high GLUT1!

!C3:!

-oxidative!-not invasive!

!Ibrahim-Hashim et al., Cancer Research 2017

BufferTxselectsforlessaggressiveC3phenotype

Ibrahim-Hashim et al., Cancer Research 2017

Glycoly.cPhenotypeassociatedwithPTENloss

P T E N -P 8 P T E N -C a P 80

5

1 0

1 5

G ly c o ly s is

PP

R(p

Mo

les

/min

/ug

pro

tein

)

****

P T E N -P 8 P T E N -C a P 80

1

2

3

G ly c o ly t ic R e s e rv e

PP

R(p

Mo

les

/min

/ug

pro

tein

) ****

P T E N -P 8 P T E N -C a P 80

2

4

6

8

A T P L in k e d O C R

OC

R (

pM

ole

s/m

in/u

g p

rote

in) ****

P T E N -P 8 P T E N -C a P 80

5

1 0

1 5

2 0

M ito c h o n d r ia l R e s e rv e C a p a c ity

OC

R (

pM

ole

s/m

in/u

g p

rote

in)

****

Model

Eco-Evolu.onarySolidTumorModel

  Hybrid cellular automata: Discrete cellular populations, continuous chemical fields.

  Focused on tumour-mE interactions   Evolving tumour phenotypes

Mark Robertson-Tessi

normal.ssuecellsbloodvessels

nocells(ECM)

Tumorcells

Robertson-Tessi, M. et al. Cancer Research. 2015 Apr 15;75(8):1567-79.

Eco-Evolu.onarySolidTumorModel

Robertson-Tessi, M. et al. Cancer Research. 2015 Apr 15;75(8):1567-79.

SteeringProstateCancerEvolu.on

5! 1!

3!4!

5

4

1

3

Untreated Low dose bicarbonate given early

Low dose bicarbonate given late High dose bicarbonate given late

CAmodelpredictsC2àC3withbicarbtreatment

5 No Treatment

Treated Early 200 1

Treated Late 400 4

3 Treated Late 200

Summary

•  Tumorsarecomprisedofcellspecieswithdifferentphenotypes.–  Glycoly.càmoreaggressive(pioneer)–  Oxida.veàmorebenign(engineer)

•  FitnessofpioneercanbereducedbyneutralizingtumorpH.

•  NeutralizingtumorpHreducesmetastases(insomesystems).

”Oneshouldmakethingsassimpleaspossible…butnotsimpler.“

AlbertEinstein

Acknowledgements•  Moffi`

– BobGatenby– ArigIbrahim-Hashim– DomAbrahams– VeronicaEstrella– PedroEnriquez-Navas– MehdiDamaghi–  ShonaghRussell– MarkRobertson-Tessi(IMO)–  SandyAnderson(IMO)

!•  NIH

•  U54CA143970;R01CA077575;U01CA143062;R01CA189295;R01CA190105;R01CA187532;P30CA076292-13