MOLECULAR MECHANISMS INFLUENCING

THE BIOMARKER LEVELS OF ENVIRONMENTAL

GENOTOXIC EXPOSURE

MOLECULAR MECHANISMS INFLUENCING

THE BIOMARKER LEVELS OF ENVIRONMENTAL

GENOTOXIC EXPOSURE

Bernadette SchoketBernadette Schoket

Department of Molecular Environmental EpidemiologyNational Institute of Environmental Health

József Fodor National Centre for Public HealthBudapest, Hungary

Department of Molecular Environmental EpidemiologyNational Institute of Environmental Health

József Fodor National Centre for Public HealthBudapest, Hungary

IMPORTANT ISSUES IN HUMAN

BIOMONITORING

IMPORTANT ISSUES IN HUMAN

BIOMONITORING

The use of surrogate and target tissues to estimate environmental genotoxic exposure

The modulation of biomarker levels by genetic susceptibility factors

The use of surrogate and target tissues to estimate environmental genotoxic exposure

The modulation of biomarker levels by genetic susceptibility factors

AIMS OF THE STUDYAIMS OF THE STUDY

Correlation between smoking-related bulky

carcinogen-DNA adduct levels in lung tissues

and in peripheral blood lymphocytes

The influence of glutathione S-transferase

metabolic genotypes on the DNA adduct levels

induced by cigarette smoking

Correlation between smoking-related bulky

carcinogen-DNA adduct levels in lung tissues

and in peripheral blood lymphocytes

The influence of glutathione S-transferase

metabolic genotypes on the DNA adduct levels

induced by cigarette smoking

STUDY POPULATIONSTUDY POPULATION Lung cancer patients undergoing lung resection, N = 212 Lung cancer patients undergoing lung resection, N = 212

Normal bronchus

Normal peripheral lung

Peripheral blood lymphocytes

Normal bronchus

Normal peripheral lung

Peripheral blood lymphocytes

SMOKING STATUSSMOKING STATUS

Current Smokers Life-time Non-smokers

& &

Short-term Ex-smokers Long-term Ex-smokers

( 1 year of abstinence ) ( > 1 year of

abstinence )

Current Smokers Life-time Non-smokers

& &

Short-term Ex-smokers Long-term Ex-smokers

( 1 year of abstinence ) ( > 1 year of

abstinence )

‘SMOKERS’ ‘NON-SMOKERS’‘SMOKERS’ ‘NON-SMOKERS’

DETERMINATION OF BULKY CARCINOGEN-DNA ADDUCTS DETERMINATION OF BULKY CARCINOGEN-DNA ADDUCTS

32P-Postlabelling 32P-Postlabelling

METHODOLOGYMETHODOLOGY

DETERMINATION OF GENOTYPES DETERMINATION OF GENOTYPES

PCR and PCR-RFLPPCR and PCR-RFLP

GSTM1GSTT1GSTP1 Ile105Val

GSTM1GSTT1GSTP1 Ile105Val

X

Phase I Phase II

X X

OH O-conj.

DNA damage

BULKY DNA ADDUCTS IN TISSUES FROMCURRENT SMOKER LUNG CANCER PATIENTS

BULKY DNA ADDUCTS IN TISSUES FROMCURRENT SMOKER LUNG CANCER PATIENTS

Lung tumour Normal lung Normal bronchus Blood lymphocytesLung tumour Normal lung Normal bronchus Blood lymphocytes

LEVELS OF BULKY DNA ADDUCTS IN LUNG TISSUES AND PERIPHERAL BLOOD LYMPHOCYTES

LEVELS OF BULKY DNA ADDUCTS IN LUNG TISSUES AND PERIPHERAL BLOOD LYMPHOCYTES

Normal

lung(3

9)

Normal

bro

nchus(3

7)

Lymphocyt

es(25)

Normal

lung(3

2)

Normal

bro

nchus(3

2)

Lymphocyt

es(22)

0

10

20

30

Bu

lky

DN

A a

dd

uc

ts /

10

8 n

uc

leo

tid

es

Smokers Non-smokers

.

Normal

lung(3

9)

Normal

bro

nchus(3

7)

Lymphocyt

es(25)

Normal

lung(3

2)

Normal

bro

nchus(3

2)

Lymphocyt

es(22)

0

10

20

30

Bu

lky

DN

A a

dd

uc

ts /

10

8 n

uc

leo

tid

es

Smokers Non-smokers

.

CORRELATION BETWEEN BULKY DNA ADDUCT LEVELS IN NORMAL PERIPHERAL LUNG

AND BRONCHIAL TISSUES

CORRELATION BETWEEN BULKY DNA ADDUCT LEVELS IN NORMAL PERIPHERAL LUNG

AND BRONCHIAL TISSUES

• Smokers: r=0.83 P<0.0001 (N=38)

• Non-smokers: r=0.73 P<0.0001 (N=30)

• Smokers: r=0.83 P<0.0001 (N=38)

• Non-smokers: r=0.73 P<0.0001 (N=30)

0 10 20 300

10

20

30 n=19r=0.25P=0.30

Smokers

Bulky adducts/108 nucleotides

Normal lung

Blo

od

lym

ph

oc

yte

sB

ulk

y a

dd

ucts

/10

8 n

ucle

oti

des

0 10 20 300

10

20

30 n=17r=0.55P=0.023

Non-smokers

Bulky adducts/108 nucleotides

Normal lung

Blo

od

lym

ph

oc

yte

sB

ulk

y a

dd

ucts

/10

8 nu

cle

oti

des

0 10 20 300

10

20

30 n=19r=0.25P=0.30

Smokers

Bulky adducts/108 nucleotides

Normal lung

Blo

od

lym

ph

oc

yte

sB

ulk

y a

dd

ucts

/10

8 n

ucle

oti

des

0 10 20 300

10

20

30 n=17r=0.55P=0.023

Non-smokers

Bulky adducts/108 nucleotides

Normal lung

Blo

od

lym

ph

oc

yte

sB

ulk

y a

dd

ucts

/10

8 nu

cle

oti

des

0 10 20 300

10

20

30 n=19r=0.31P=0.20

Smokers

Bulky adducts/108 nucleotides

Normal bronchus

Blo

od

lym

ph

oc

yte

s

Bu

lky a

dd

ucts

/10

8 n

ucle

oti

des

0 10 20 300

10

20

30 n=16r=0.55P=0.026

Non-smokers

Bulky adducts/108 nucleotides

Normal bronchus

Blo

od

lym

ph

oc

yte

s

Bu

lky a

dd

ucts

/10

8 n

ucle

oti

des

0 10 20 300

10

20

30 n=19r=0.31P=0.20

Smokers

Bulky adducts/108 nucleotides

Normal bronchus

Blo

od

lym

ph

oc

yte

s

Bu

lky a

dd

ucts

/10

8 n

ucle

oti

des

0 10 20 300

10

20

30 n=16r=0.55P=0.026

Non-smokers

Bulky adducts/108 nucleotides

Normal bronchus

Blo

od

lym

ph

oc

yte

s

Bu

lky a

dd

ucts

/10

8 n

ucle

oti

des

CORRELATION BETWEEN DNA ADDUCT LEVELSIN LUNG TISSUES AND LYMPHOCYTES

CORRELATION BETWEEN DNA ADDUCT LEVELSIN LUNG TISSUES AND LYMPHOCYTES

FOR SMOKERS AND NON-SMOKERSFOR SMOKERS AND NON-SMOKERS

Positiv

e (7

4)

Null (6

6)

Positiv

e (3

1)

Null (4

1)0

10

20

30

DN

A a

dd

uct

s /

108

nu

cleo

tid

es

Positiv

e (1

01)

Null (4

0)

Positiv

e (4

5)

Null (2

6)0

10

20

30

DN

A a

dd

uct

s /

108 n

ucl

eoti

des

Pos&Pos

(46)

Null&Null

(13)

Pos&Pos

(16)

Null&Null

(11)

0

10

20

30P=0.058

DN

A a

dd

uct

s /

108 n

ucl

eoti

des

Positiv

e (7

4)

Null (6

6)

Positiv

e (3

1)

Null (4

1)0

10

20

30

DN

A a

dd

uct

s /

108

nu

cleo

tid

es

Positiv

e (1

01)

Null (4

0)

Positiv

e (4

5)

Null (2

6)0

10

20

30

DN

A a

dd

uct

s /

108 n

ucl

eoti

des

Pos&Pos

(46)

Null&Null

(13)

Pos&Pos

(16)

Null&Null

(11)

0

10

20

30P=0.058

DN

A a

dd

uct

s /

108 n

ucl

eoti

des

GSTM1 AND GSTT1 GENOTYPES AND BULKY DNA ADDUCT LEVELS IN BRONCHUS

GSTM1 AND GSTT1 GENOTYPES AND BULKY DNA ADDUCT LEVELS IN BRONCHUS

GSTM1GSTM1 GSTM1&GSTT1GSTM1&GSTT1GSTT1GSTT1SmokersSmokers Non-smokersNon-smokersSmokersSmokersNon-smokersNon-smokersSmokersSmokersNon-smokersNon-smokers

THE EFFECT OF GSTP1 Ile105Val GENOTYPEON BULKY DNA ADDUCT LEVELS IN BRONCHUS

THE EFFECT OF GSTP1 Ile105Val GENOTYPEON BULKY DNA ADDUCT LEVELS IN BRONCHUS

0

10

20

30

.

DN

A a

dd

uct

s /

108

nu

cleo

tid

es

0

10

20

30

.

DN

A a

dd

uct

s /

108

nu

cleo

tid

es

*

*

P=0.012

SmokersSmokers Non-smokersNon-smokers

MAIN CONCLUSIONSMAIN CONCLUSIONS The dose of exposure and the metabolic capacity of

the tissues influence the correlation between target

and surrogate tissue

Genetic polymorphisms of xenobiotic-metabolising

enzymes may affect the biologically effective dose of genotoxic exposure

Implications for human biomonitoring

low dose - high dose exposure surrogate - target tissue metabolic genotypes - biomarker levels

The dose of exposure and the metabolic capacity of

the tissues influence the correlation between target

and surrogate tissue

Genetic polymorphisms of xenobiotic-metabolising

enzymes may affect the biologically effective dose of genotoxic exposure

Implications for human biomonitoring

low dose - high dose exposure surrogate - target tissue metabolic genotypes - biomarker levels

PARTICIPANTS OF THE STUDYPARTICIPANTS OF THE STUDY

National Institute of Environmental Health, Budapest, HungaryBernadette Schoket

Erika Győrffy

Lívia Anna

National Center for Epidemiology, Budapest, HungaryJudit Segesdi

János Minárovits

Zoltán Győri

National Institute of Pulmonology, Budapest, HungaryIbolya Soltész

Szilárd Kostic

Attila Csekeő

National Institute of Environmental Health, Budapest, HungaryBernadette Schoket

Erika Győrffy

Lívia Anna

National Center for Epidemiology, Budapest, HungaryJudit Segesdi

János Minárovits

Zoltán Győri

National Institute of Pulmonology, Budapest, HungaryIbolya Soltész

Szilárd Kostic

Attila Csekeő