developments in liquid biopsy in oncology · center of experimental medicine, department of tumor...

Simon Joosse ([email protected])

University Medical Center Hamburg-Eppendorf

Center of Experimental Medicine, Department of Tumor Biology

Developments in liquid biopsy in

oncology

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Joosse et al. (2014) EMBO Mol Med

Metastatic cascade – Tumor cell dissemination

CTCs = Circulating Tumor Cells

DTCs = Disseminated Tumor Cells

EMT = epithelial-mesenchymal transition

MET = mesenchymal-epithelial transition

DTC

Extravasation

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Circulating tumor markers as liquid biopsy

Elazezy & Joosse (2018) Comput Struct Biotechnol J

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Alix-Panabières & Pantel (2016) Cancer Discovery

Aims of liquid biopsy

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Circulating tumor cell (CTC) enrichment


Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


CellSave Preservative Tube Enrichment of EpCAM positive cells and staining for Keratin and CD45

Cytoplasm Nucleus Cell membrane Composite

CD45+ membrane

Keratin-PEpositive

DAPI

positive

CD45-APC

negative Tumor cell

Leukocyte Tumor cells

CTC enrichment - CellSearch System (FDA-cleared)

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


CTC enrichment - in vivo capture

Saucedo-Zeni et al. (2012) Int J Oncol

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Gorges et al. (2016) Clin Cancer Res

in vivo capture of CTCs in lung cancer patients

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO




corr=0.36, p=0.003

p<0.0001

p=0.0013 p=0.127

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO




- Improved capture of CTCs in lung cancer

- Molecular characterization

- Predictive value

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO



Screening and early

detection of cancer

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


ctDNA for early detection


Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Fernandez-Cuesta et al. (2016) EBioMedicine

Krimmel et al. (2016) PNAS

Challenges of early detection of cancer

TP53 mutation detection in cfDNA of ovarian

cancer patients and control subjects

TP53 mutation detection in cfDNA of

lung cancer patients and control subjects

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Challenges of early detection of cancer

Detection of leukemia mutations in non-cancer subjects

Genovese et al. (2014) NEJM

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO



Risk for metastatic

relapse (prognosis)

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Bone-marrow aspirates

from upper iliac crests

Breast cancer (M0): 199/552 (36%)Braun et al. NEJM 2000

Bone-marrow

Ficoll gradient

Mononuclear cells

Keratin staining Tumor cell screening

adapted from: Pantel & Brakenhoff (2004) Nat Rev Cancer

Prostate cancer (pM0N0): 86/193 (44.6%)Köllermann et al. JCO 2008

Nonmalignant

disease: 2/191 (1%)

Prognostic value of Disseminated Tumor Cells (DTCs)

DTCs correlate with relapse

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Prognosis of liquid biopsy in advanced stage cancer - CTCs

De Bono et al. (2008) Clin Cancer Res

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Prognosis of liquid biopsy in early stage cancer - CTCs

Breast cancer

Bladder cancer

Head & neck cancer

Testicular germ cell cancer

Colorectal cancer

EpCAM+/K+ CTCs are associated

with unfavorable prognosis in

early stage cancer

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Prognosis of liquid biopsy in early stage breast cancer - ctDNA

Neoadjuvant

chemotherapy SurgeryPost-

surgeryStandard follow-up

Baseline

Post-surgery

Mutation tracking

Garcia-Murillas et al. (2015) Sci Transl Med

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO



Therapeutic targets

and resistance

mechanismsDo not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Identification of therapeutic targets and resistance mechanisms- ER in breast cancer

Johnston Dowsett (2003) Nat Rev Cancer

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Babayan et al. (2013) PLoS One

ER K DAPI CD45 Merge

Ove

rall

su

rviv

al

p = 0.033

HR: 7.38

CTC+

CTC-

Therapy relevant markers

expressed in CTCs

Identification of therapeutic targets and resistance mechanisms- ER in breast cancer

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Identification of therapeutic targets and resistance mechanisms- PD1-PDL1

Pardoll et al. (2012) Nat Rev Cancer

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Identification of therapeutic targets and resistance mechanisms- PD1-PDL1

Mazel et al. (2015) Mol Onc

PD-L1 expression found on CTCs

in 11/16 patients

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


CTC without HER2 gene amplification

CTC with HER2 gene amplification

SK-BR-3

Red: CK

ICCGreen: HER2

FISH

Red: Cen17

FISH



SK-BR-3



SK-BR-3

Red: CK

ICCGreen: HER2

FISH

Red: Cen17

FISH

Riethdorf et al. (2010) Clin Cancer Res

DETECT-III study:

Anti-HER2 therapy in metastatic

breast cancer patients with

HER2-neg primary tumors but

HER2-pos CTC

Identification of therapeutic targets and resistance mechanisms- HER2 in breast cancer

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Cayrefourcq et al. (2015) Cancer Res

AACR Award for most cited article

in Cancer Research 2015

Characterization through cell line

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


• Different substrates can be used as liquid biopsy

(e.g., CTCs, cfDNA, exosomes)

• Liquid biopsy can be used

• To stratify cancer patients

• To monitor therapy response

• For early detection

• For characterization of cancer heterogeneity

• Challenging in symptom-free subjects

• Liquid biopsy can be used for personalized treatment

Conclusion / Take home message (1)

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Understanding the biology of

metastasis

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Cancer heterogeneity and liquid biopsy

Alix-Panabières & Pantel (2010) Clin Cancer Res



Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Circulating tumor markers as liquid biopsy

Joosse & Pantel (2015) Cancer Cell

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Tumor DNA

Reference DNA

Chromosome

Cop

y n

um

be

r va

lue

Joosse et al. (2007) BMC Cancer

Array CGH Next Generation Sequencing

DNA fragments

Molecular characterization of single tumor cells

Babayan et al. (2016) Oncotarget

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


NG

S

Single SKBR3 cell #1

Chromosome 8 Chromosome 17

aC

GH

NG

S

SKBR3 pool (reference)

NG

S

Chromosome 8 Chromosome 17

Single SKBR3 cell #2

Single cell vs genomic

r2 = 0.91 Babayan et al. (2016) Oncotarget

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO



Metastatic cascade – Tumor cell dissemination




MET = mesenchymal-epithelial transition

DTC

Extravasation

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Breast Cancer

Prostate cancer

Lung cancer (NSCLC)

Gastric cancer

Esophageal cancer

Colorectal cancer

Pancreatic cancer

Head and neck cancer

20-40

20-30

40-60

35-60

30-40

20-30

10-20

20-30

Tumor type Detection rate (%)

Pantel & Brakenhof (2004) Nat Cancer Rev

Bone marrow is a reservoir for DTCs

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Croucher et al. (2016) Nat Rev Cancer

Bone metastasis

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Mohme et al. (2017) Sci Rep

CTC release through cement augmentation of vertebral metastases

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Joosse and Pantel (2013) Cancer Research

What drives tumor dissemination?


Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


vs.

Classification of early stage breast tumors

according to DTC status

Expression profiling on micro-dissected

tumor tissue

Bioinformatic analysis

and validation

of identified genes

DTC- DTC+

RAI2→

ER+ breast tumors

Drivers of tumor cell dissemination

Werner et al. (2015) Cancer Discovery

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Epithelial-mesenchymal transition: tumor cell dissemination

Esposito and Kang (2015) Cancer Discovery

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO



What enables cell survival?


Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Giuliano et al. (2018) Cancer Res

Intravasation of tumor cells

Single cell dissemination Cell cluster dissemination

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Aceto et al. (2014) Cell

CTC Clusters have higher metastatic potential compared to single CTCs

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Aceto et al. (2014) Cell

CTC Clusters have higher metastatic potential compared to single CTCs

Prostate cancer patientsBreast cancer patients

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO



What plays a role in extravasation of CTCs?


Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Extravasation of tumor cells

Follain et al. (2018) Dev Cell

Caudal Plexus

Head Intravascular injection

of tumor cells (2.103)

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Follain et al. (2018) Dev Cell

Extravasation of tumor cells

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO



What determines the metastatic location?


Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Extracellular vesicles (EVs) carry a tumor’s instructions


Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Exosomes determine organotropic metastasis

Hoshino et al. (2015) Nature

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


Genomic progression in colorectal cancer metastasis

Joosse et al. (2018) Clin Chem

Chromosomal

aberration

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO


• Liquid biopsy may be used to study cancer biology:

• Intravasation

• CTC survival

• Extravasation

• Outgrowth of metastasis

• Tumor heterogeneity

Conclusion / Take home message (2)

Do not d

uplicate

or d

istrib

ute w

ithout

permiss

ion from

auth

or and E

SO

developments in liquid biopsy in oncology · center of experimental medicine, department of tumor...

Documents