Virchow’s Hypothesis 1863 - Dr. Falk Pharma Cancers and Chronic Infection • EBV –nasopharyngeal cancer • H. Pylori –gastric adenocarcinoma –MALT • Clonorchis Sinesis

Virchow’s Hypothesis 1863

“lymphorecticular infiltration” of cancer reflected the origin of cancer at sites of inflammation

Barrett’s esophagus/Esophageal adenocarcinoma

Gastritis/ Gastric cancerPSC / CholangiocarcinomaViral hepatitis / Hepatocellular cancer

Colitis / Colon cancer

Gastrointestinal Cancers and Chronic Infection

• EBV–nasopharyngeal cancer

• H. Pylori–gastric adenocarcinoma–MALT

• Clonorchis Sinesis–cholangiocarcinoma

• HPV–anogenital cancer

A Generalized Scheme of Chronic InflammationA Generalized Scheme of Chronic InflammationA Generalized Scheme of Chronic Inflammation

CP1222559-2CP1222559-2

Physical injuryPhysical injuryNormal

microfloraNormal

microfloraPathogensPathogens

ToxinsToxins PAMPsPAMPs

PRRsPRRs

Host cellapoptosisHost cellapoptosis

Host cellnecrosisHost cellnecrosis Endogenous

ligandsEndogenous

ligands

ChemokinesChemokinesCytokinesCytokines

LymphocytesLymphocytes

Persistent inflammationPersistent inflammation

Anti-inflammatorycontrols


Epitheliallayer

Epitheliallayer

Geneticfactors

++ –

Macrophage

Karin, M Cell, 2006Karin, M Cell, 2006

Hallmarks of Cancer (Intrinsic Genetic Process)

Hallmarks of Cancer Hallmarks of Cancer (Intrinsic Genetic Process)(Intrinsic Genetic Process)

CP1221050-1CP1221050-1

Angiogenesis (VEGF)

Angiogenesis (VEGF)

Uncontrolled self-sufficiency in growth

(oncogenes)

Uncontrolled self-sufficiency in growth

(oncogenes)

Death signals(tumor suppressor genes p53)

Death signals(tumor suppressor genes p53)

Tissue invasionTissue invasion

Limitless replication(? stem cell)

Limitless replication(? stem cell)

Hanoham D: Cell 100:57, 2000Hanoham D: Cell 100:57, 2000

CP1221050-2CP1221050-2

Angiogenic factorsAngiogenic factorsTissue remodelingMigration, invasionTissue remodelingMigration, invasion

Growth factorsGrowth factors

Inflammatory Cells Inflammatory Cells

Genetic mutationsGenetic mutations

Survival factors

Survival factors

CytokinesCytokinesOxidative

stressOxidative

stress

Incidence of CholangiocarcinomaIncidence of CholangiocarcinomaC

umul

ativ

e in

cide

nce

ofch

olan

gioc

arci

nom

a (%

)C

umul

ativ

e in

cide

nce

ofch

olan

gioc

arci

nom

a (%

)

Years since PSC diagnosisYears since PSC diagnosisCP1042831-1

0.0

2.5

5.0

7.5

10.0

0 2 4 6

CP1221050-2CP1221050-2





Survival factors

Survival factors

CytokinesCytokinesOxidative, nitrosative

stressOxidative, nitrosative

stress

iNOS Expression is Upregulated in PSC and CCA

Normal PSC Cholangiocarcinoma

Jaiswal et al. Cancer Res, 2000

iNOS positive 0/30

iNOS positive20/20

iNOS positive 30/30

N 7

2H2N

H

O

N

N

N

H

deoxyribose

1

34

56

89

5N 7

2H2N

OH

N

N

N

deoxyribose

1

34

6

89

o

H

DeoxyguanosinedG

8-oxo-deoxyguanosine8-oxo-dG

Oxidative DNA Lesion

Oxidative Stress

ROS

8-oxodG Positive: 0/10 8-oxodG Positive: 10/10 8-oxodG Positive: 10/10

iNOS Expression in PSC and Cholangiocarcinoma is Associated with Oxidatively Damaged DNA

CholangiocarcinomaNormal PSC

Jaiswal et al, Gastroenterology,120, 190-199, 2001

p16 Point Mutations in PSC-associated Cholangiocarcinoma

Promoter/Enhancer

5’

Exon 1 Exon 2 Exon 3

3’

8/10 3/10 None None

A

A

G

A

G

G

T

C

Patients

T

C A

G

A

G

A

G

A

G

A

G

A

GT

C

T

C

T

C A

G

A

G

A

G

A

G

CP1221050-2CP1221050-2





Survival factors

Survival factors


stressOxidative

stress

IL-6

Autocrine/paracrine pathway

Nucleus

Mcl-1

Cholangiocarcinoma cell

Kobayashi S et al, Gastroenterology 2005;128:2054-65

IL-6 Increases Expression of the Anti-Apoptotic Protein Mcl-1

NormalNormal ALCALC

PSCPSC CCACCA

70%(23/33)70%(23/33)48%(14/29)48%(14/29)

8%(2/25)8%(2/25) 20%(3/15)20%(3/15)

Mcl-1 is Overexpressed in PSC and Cholangiocarcinoma

0 2 3 (Hours)

anti-IL-6

Mcl-1

actin

CP1221050-2CP1221050-2





Survival factors

Survival factors


stressOxidative

stress

Developmental Signaling Pathways and Cancer

• Notch• Wnt• Hedgehog

CSL

Cytoplasm

Nucleus

Target Genes

Jagged

Notch

NICD

Notch Signaling Pathway

（Intracellular domain）

GolgiNotchprecursor

Sending cell

Receiving cell γ-secretase

Cholangiocyte

iNOS

Notch

Inflammation

NO

Cell proliferation AngiogenesisAnti-apoptosis

Cancer phenotype

Notch-1 Protein is Enhanced in PSC and CCA

Normal Liver PSC CCA

Normal PSC CCA(n=16) (n=12)

Sta

inin

g In

tens

ity(%

of ≥2

)

(n=15)

020

4060

80

100

Actin

250K

150K

100K

Notch-1(FL)

NICD

603B603B-iN

OS AS

*

Notch-1 Expression in Cholangiocyte is iNOS dependent

0

0.2

0.4

0.6

0.8

1

1.2

1.4

603B 603B-iNOS AS

Not

ch-1

/ 18

S ra

tio

Immunoblot analysis Real-time PCR

* Non-specific band

**** p<0.01

Cellular Mechanisms Linking Inflammation to Cholangiocarcinoma

Bile duct Cholangiocarcinoma

Inflammation

COX-2

iNOS

COX-2; Sirica AE, et al. Semin Liver Dis, 2002iNOS; Jaiswal M, et al. Gastroenterology, 2001

β-Catenin Pathway Activation in Response to PGE2ββ--Catenin Pathway Activation in Response to PGE2Catenin Pathway Activation in Response to PGE2

CP1222559-1CP1222559-1

COX-2COX-2

PGE2PGE2

EP2EP2

αsGTP

Axin

β-catenin

β-catenin

β-catenin

TcfLef

GSK-3β

AKT

γβ

PIP3PDK1

P13K

-P-P

-P-P

Geneexpressioncell growth

Geneexpressioncell growth

Science, 2005Science, 2005

CP1221050-2CP1221050-2

Angiogenic factorsAngiogenic factorsTissue remodelingand Oncogenesis

Tissue remodelingand Oncogenesis




Survival factors

Survival factors


stressOxidative

stress

CP1271074-1J Clin Inves 107(2):140, 2001

Ub-Ub-Ub-Ub-Ub-Ub

Ub-Ub-Ub-Ub-Ub-Ub

Cytoplasm

Nucleus

P P

PP

PP

PP

• Cytokines• Bacterial/viral infection• Stress

• Cytokines• Chemokines• Adhesion molecules• Receptors• Cell hB proteins, etc

• Cell proliferation• Cell growth• Cell ditherentiation

NF-κB responsive gene

P50

P50

P50

265 proteasome

IκBP65

P50

P65

P65

P65

IKK1 IKK1

IKK2IKK2

NEMO

NEMO

IκB

IKK complex

NF-κB:IκB

NF-κB

IκB

CP1220459-2CP1220459-2Karin M et al Nature Reviews in Immunology, 2005

Karin M et al Nature Reviews in Immunology, 2005

Nuclear Factor-κB has Various Tumour-PromotingFunctions Depending on Cell Type in Which it is Activated

Nuclear FactorNuclear Factor--κκB has Various TumourB has Various Tumour--PromotingPromotingFunctions Depending on Cell Type in Which it is ActivatedFunctions Depending on Cell Type in Which it is Activated

Growth factors(such as TNF and IL-1)

Inflammatory cellInflammatory cell

Tumour cellTumour cell

p50 p65

NF-κB- binding site

Survival• BCL-XL• GADD45β• BFL1• SOD2

Classical pathway NF-κB activation

IL-1RIL-1R

IL-1IL-1TNFTNF

TNFRTNFR

Survival factors

p50 p65

NF-κB- binding site

•TNF•IL-1•IL-6•CXCL8•VEGF•CSF1

IκB

CP1271074-1J Clin Inves 107(2):140, 2001

Ub-Ub-Ub-Ub-Ub-Ub

Ub-Ub-Ub-Ub-Ub-Ub

Cytoplasm

Nucleus

P P

PP

PP

PP





P50

P50

P50

265 proteasome

κB IκBP65

P50

P65

P65

P65

IKK1 IKK1

IKK2IKK2

NEMO

NEMO

IκB

IKK complex

NF-κB:IκB

NF-κB

Colitis-Associated Cancer Is Reduced By Deleting IKKß From Intestinal Epithelial Cells

Greten et al Cell, 2004Model of AOM plus DSS tumorigenesis

GeneticmutationsGenetic

mutations

NF-κB and Tumorigenesis in ColitisNFNF--κκB and B and TumorigenesisTumorigenesis in Colitisin Colitis

CP1228720-1CP1228720-1

EnterocyteEnterocyte

Greten FR et al: Cell, 2004Greten FR et al: Cell, 2004

NF-κBintactNF-κBintact

IKKβdeficient

IKKβdeficient

CancerCancer

ApoptosisApoptosis

ANANMitosis

ABABInflammatory

cells

DEN and WT Mice DEN and Hepatocyte

IKKß Δ Mice

HCC is increased in DEN Treated Mice Deficient in Hepatocyte IKKß

Maeda et al. Cell, 2005

DEN Hepatocarcinogenesis and HepatocyeNF-κB

DEN Hepatocarcinogenesis and DEN Hepatocarcinogenesis and HepatocyeHepatocyeNFNF--κκBB

CP1228720-2CP1228720-2

HepatocyteHepatocyte

IKK-βdeficiency

IKK-βdeficiency

Growth/survivalfactors

Growth/survivalfactors

CancerCancer

Kupffercell

Kupffercell

NF-κBactivation

NF-κBactivation


mutations

cell deathcell death

DEN Hepatocarcinogenesis and Kupffer Cell NF-κB

DEN Hepatocarcinogenesis and Kupffer Cell DEN Hepatocarcinogenesis and Kupffer Cell NFNF--κκBB

HepatocyteHepatocyte

NF-κBNF-κB

CP1228720-3CP1228720-3

cell deathcell death


mutations

Kupffer cellactivationKupffer cellactivation

cancercancer

IKK-βdeficient

IKK-βdeficient

CP1271074-1J Clin Inves 107(2):140, 2001

Ub-Ub-Ub-Ub-Ub-Ub

Ub-Ub-Ub-Ub-Ub-Ub

Cytoplasm

Nucleus

P P

PP

PP

PP





P50

P50

P50

265 proteasome

IκBP65

P50

P65

P65

P65

IKK1 IKK1

IKK2IKK2

NEMO

NEMO

IκB

IKK complex

NF-κB:IκB

NF-κB

IκB

CP1266381-3CP1266381-3

Cancer Cell 11:119, 2007Cancer Cell 11:119, 2007Arrows indicate large tumorsArrows indicate large tumors

ANANMitosis

ABABInflammatory

cells

TNF-R1N

C

Fas TRAIL-R1/2N

C

N

C

Extracellular Cysteine-rich Domain

Transmembrane Domain

Cytoplasmic Death Domain

The Fas DISCThe Fas DISC

FADDFADD

ActiveActiveCaspase 10Caspase 10

FasL/CD95LFasL/CD95L

ProPro--Caspase 10Caspase 10

FADDFADD

ActiveActiveCaspase 8Caspase 8

ProPro--Caspase 8Caspase 8

Fas/CD95Fas/CD95

DE

DD

ED

DD

DD

DE

DD

DDD

DE

DD

D�

�

DE

DD

ED

DDDD

DEDDED

Death DomainDeath Domain

Death Death EffectorEffector DomainDomain

CP1266381-1CP1266381-1

Cancer Cell 11:119, 2007Cancer Cell 11:119, 2007

Tune

l(LP

S 10

hr)

Tune

l(LP

S 10

hr)

NEMOLPC-KONEMOLPC-KO NEMO/FADDLPC-KONEMO/FADDLPC-KO

0

200

400

600

800

1,000

CP1266381-2CP1266381-2

Cancer Cell 11:119, 2007Cancer Cell 11:119, 2007

NEMOLPC-KONEMOLPC-KO NEMO/FADDLPC-KO

NEMO/FADDLPC-KO

Serum ALTSerum ALTSerum ALT

P<0.05P<0.05

NF-κB in Different Cancer ModelsNFNF--κκB in Different Cancer ModelsB in Different Cancer Models

CP1262985-1CP1262985-1

MM SS

NFκBhi


CP1262985-2CP1262985-2

MM SS

NFκBhi

Tonic signalsTonic signals

Inflammatorysignals

Inflammatorysignals

IKKNFκB

Kupffer cells


CP1262985-3CP1262985-3

IKKNFκB

Kupffer cells

CancerCancer

TNFIL-6TNFIL-6

MM SS

JNKhi

IKKNFκB

JNKhi

ROShi

A Generalized Scheme of Chronic InflammationA Generalized Scheme of Chronic InflammationA Generalized Scheme of Chronic Inflammation

CP1222559-2CP1222559-2

Physical injuryPhysical injuryNormal

microfloraNormal

microfloraPathogensPathogens

ToxinsToxins PAMPsPAMPs

PRRsPRRs

Host cellapoptosisHost cellapoptosis

Host cellnecrosisHost cellnecrosis Endogenous

ligandsEndogenous

ligands

ChemokinesChemokinesCytokinesCytokines

LymphocytesLymphocytes

Persistent inflammationPersistent inflammation



Epitheliallayer

Epitheliallayer

Geneticfactors

++ –

Macrophage

Karin, M Cell, 2006Karin, M Cell, 2006

CP1221050-2CP1221050-2





Survival factors

Survival factors


stressOxidative

stress

Tumor Associated Macrophages Tumor Associated Macrophages (TAM)(TAM)

MacrophageMacrophage

Common functionsCommon functionsAngiogenic factors

Tissue remodeling and repairGrowth and migration factors

Angiogenic factorsTissue remodeling and repairGrowth and migration factors

MacrophogenesisMacrophogenesis MetastasisMetastasis

Blood vesselBlood vessel

Tumor cells

Tumor cellsNormal

epithelialcells

Normalepithelial

cells

Ductal invasionDuctal invasion Tumor invasionTumor invasion

CP1221406-1CP1221406-1

Condeelis, J et al Cell, 2006

Contribution of Macrophages to MetastasisContribution of Macrophages to MetastasisContribution of Macrophages to Metastasis

CP1225113-1CP1225113-1Wang et al: Trends in cell biology 15:141, 2005Wang et al: Trends in cell biology 15:141, 2005

BloodvesselBloodvessel

ParacrinechemotaxisParacrine

chemotaxis

EGFEGF

CSF-1CSF-1CSF-1CSF-1

EGFEGF

CCCC

The Cancer EcosystemThe Cancer EcosystemThe Cancer Ecosystem

CP1224433-1CP1224433-1

NeoangiogenesisNeoangiogenesis

TILTIL

TAMTAM

APCAPC

Connective tissue

Connective tissue

FibroblastsFibroblasts

CP1224433-2CP1224433-2

CP1224433-3CP1224433-3

Dead snakeDead snake

Documents

Virchow’s Hypothesis 1863 - Dr. Falk Pharma Cancers and Chronic Infection • EBV –nasopharyngeal cancer • H. Pylori –gastric adenocarcinoma –MALT • Clonorchis Sinesis