IMPLICACIÓN BIOLÓGICA DE LA

CTCS Y LOS ÁCIDOS

NUCLEICOS CIRCULANTES

Paloma Bragado Domingo, PhD

SEAP-IAP-Reunión De Primavera

“Biopsia Líquida: Un Nuevo Reto Para

El Patólogo Y El Oncólogo

CANCER IS A SYSTEMIC AND HETEROGENEOUS

DISEASE

DTCs at secondary organs:Metastasis precursor cells

Adapted from Alberter, B et al., (2016) Expert review of Molecular Diagnostics, 16:1,25-38.

Tum

or m

ass

Primary tumor Minimal residual disease

Disease free period/resistance to therapyLate DTCs Disseminated tumor cell dormancy

metastasis

CANCER AS AN EVOLUTIONARY DISEASET

umor

mas

s

Surgery and therapy

Timeyears Months years decadesEarly

DTCs

Late DTCs Disseminated tumor cell dormancy

Bragado P, Nat Rev Cancer. 2014

ZZ

Z

ZZ

Z

ZZ

Z

G0-G1 arrest

Survival

CLINICAL EVIDENCES FOR TUMOR DORMANCY

Klein, C. A.. Curr. Opin. Genet. Dev. 21, 42–49 (2010).

Aguirre-Ghiso, J. A. Nature Rev. Cancer 7, 834–846 (2007)

Wolfgang Janni et al. Clin Cancer Res 2011;17:2967-2976

WHAT ARE THE MOLECULAR

MECHANISMS THAT REGULATE DTCS

DORMANCY?DORMANCY?

CAN MICROENVIRONMENTAL SIGNALS

REGULATE DTCS DORMANCY?

HEp3-GFP

HEp3-GFP Primary tumor + surgery

~2weeks

~4weeks

HEP3 MINIMAL RESIDUAL DISEASE MODEL

73

81

24

5,65,6

14 Lungs

Lymph Nodes

bone Marrow

Liver

Spleen

Others

DTCs incidence

DTCs?

Lungs

Bone Marrow

29.15 ± 9.1

(55.8± 17.3)*104 VimentinP-H3DAPI

p27CKDAPI

GFP/DAPI

DTCs

P-H3CK

DAPI

Bragado P, et al., (2013) Nat Cell Biol.;15(11):1351-61.

BM-D

QUIESCENCE OF BM DTCS DERIVED CELLS

DEPENDS ON P38 ACTIVATION AND DEC2 INDUCTION

BM-HEp3

In vitro expansion

Lu

ng

sB

on

e M

arro

w

DTCs

20 Generations

20 Generations

15% Efficiency In vivo

α

DEC2

p53

Low expression

High Expression

Fold change: -5.6 +2.6

PT

-3

Lu

-7

BM

-3

Lu

-6

PT

-4

BM

-D

4T1

4T07

F3I

I

Lu-HEp3Lung DTCs 100% tumorigenic in vivo

BM DTCs 73% dormant in vivo

Lu

ng

s

100% Efficiency

Bragado P, et al., (2013) Nat Cell Biol.;15(11):1351-61.

% o

f p

27+

cells 100

8060

*

TGFβ2

:mAbPI-IgG α-TGFβ2

BM CM

Bone Marrow

P38 ,TGF 2 AND TGF R3 REGULATE DTCS

DORMANCY

α β β

Control

SB203580

% o

f p

27+

6040200

Si scr Si p38α

*

Lungs

Bragado P, et al., (2013) Nat Cell Biol.;15(11):1351-61.

Dormancy inducing factors: P-p38, TGFβ2, TGFβR3, DEC2, p27, NR2F1, SOX9

TGF 2 / TGF 1 RATIO DICTATES DISSEMINATED

TUMOUR CELL FATE IN TARGET ORGANS THROUGH TGF R3 AND P38 / SIGNALLING

α ββ

ββ

Can we use the expression of this dormancy inducing factors to predict relapse and stratify

patients?

DORMANCY SIGNATURES PREDICT METASTASIS IN

OESTROGEN RECEPTOR POSITIVE BREAST CANCER

Kim RS, et al., PLoS One. 2012;7(4):e35569

DTCS FROM PATIENTS WITH NO EVIDENCE OF DISEASE

WERE SIGNIFICANTLY ENRICHED FOR DORMANCY-RELATED GENES

Chéry L et al. Oncotarget. 2014 Oct 30;5(20):9939-51.

Sosa MS et al., Nat Commun. 2015 Jan 30;6:6170.

BONE MARROW AS A RESERVOIR FOR DTCS: A SPECIAL

SOURCE FOR LIQUID BIOPSY DTC MONITORING TO TREAT

DORMANT RESIDUAL DISEASE

Adapted from Pantel K, Alix-Panabières C. Bonekey Rep. 2014 Nov 19;3:584.

Aguirre-Ghiso, J et al., 2013 Mar;19(3):276-7.

DTCS VS CTCS

� Do CTCs express dormancy markers?DTCs Limitations:• Repeated sampling of BM or

LN is difficult• DTCs reflect only the

situation in the BM• However the reflect the

”history” of tumour cell dissemination

� Do dormant DTCs disseminate and become CTCs?

CTCs advantages:• Easy access• Minimal invasiveness• Minimal Risk• Provide information on

metastatic cells derived from various distant sites

• However, they represent only a snapshot of tumour cell dissemination

MICRO-RNAS AND DORMANCY

� Do Patient DTCs also overexpress miR 190, miR 580 and miR 588?

� Can we detect those miR in blood?

� Would their expression correlate with the dormancy phase?dormancy phase?

Laboratory of signaling and metastasisJulio Aguirre Ghiso•Yeriel Estrada •Alvaro Avivar Valderas •Soledad Sosa•Alfred Adomako•Kathryn Harper Molecular and Translational •Kathryn Harper•Falguni Parikh•Nina Linden•Veronica Calvo•Denis Schewe•Aparna Ranganathan•Wen Huei-Chi•David Cannan

Molecular and Translational oncology Pere Gascon•Raul Alonso•Gemma Fuster•Mario Mancino •Patricia Fernandez •Arantzazu Zubeldia•Estel Enreig •Anna Lopez•Elisabet Ametller