Pa Po Va virus HPV Molecular Oncogenesis

HPV

Molecular

Oncogenesis

Pa Po Va virus

Nam Hoon Cho MD

Yonsei Univ Coll Med Dept of Pathology

Rudolf Virchow Father of Pathology

G Nikolas Pappanicaulau ndashPAP test

James Ewing ndashtumor metastasis theory

Arias-Stella- AS reaction

Azzopardi JG- One sees and recognizes only those features which have already been described

Arthur Purdy Stout- Atlas of Tumor Pathology

Stanley L Robbins- 1957 textbook 1st ed

Lesions do not arise in cadevars But the study of morphology is only one facet of pathology Pathology contributes much to clinical medicine The pathologist is interested not only in the recognition of structural alterations but also in their significance ie the effects of these changes on cellular and tissue function and ultimately the effect of these changes on the patient It is not a discipline isolated from the living patient but rather a basic approach to a better understanding of disease

and therefore a foundation of sound clinical medicine

Virchowrsquos triad

PIONEERS of PATHOLOGY

윤일선 교수 (1896-1987) 한국 최초의 (일본 연수) 병리학자1929-1945 세브란스 의전 교수-초대 주임교수

최동 교수 한국 최초의 (순수국내) 연세의대 병리학 및법의학교수- 한국 최초의 종양등록 문헌 발표

서재필 1929 병리학 전문의 취득하여 한국 최초의 병리전문의

Pioneers in Korean Pathologists

백순명 연세의대 졸업-미국 피츠버그대-유방암 조직은행-현 삼성병원 암연구소소장

Alois Alzheimer

Von Reklinghausen

인간은 생을 살려고 태어난 것이지생을 준비하려고 태어난 것은 아니다

하얀 거탑

Presentation Flow

HPV genome new highlight-

E6-E6AP-PDZ binding

E1Ê4 and E2Ê5

Overall roles of genomes

Controversial issue in cytopathology

What is phenotype with ASCUS

CIN 2 from the viewpoint of HPV

HPV testing guideline

HPV types assessed by the IARC

Monograph Working group

α β1 16 Most potent type

1 1831333539455152565859

Sufficient carcinogen

2A 68 Probably carcinogenic but limited evidence

2B 26536667707382 Possibly carcinogenic

2B 3034698597 Limited evidence

3 611

2B 58 Limited

3 Other type

Lancet Oncol 200910321-2




1 1831333539455152565859





3 611

2B 58 Limited

3 Other type


HPV 58 PPV for CIN 3+= 549

(An HJ et al Cancer 2003 971672)

HPV frequency 16185839525651

(Cho et al Am J Obstet Gynecol 2003 18856)

HPV with no CIN 372

(Hwang et al Gynecol Oncol 2003 9051)

HPV in Korean commercial sex workers47 (16 51)

(Choi et al J Med Virol 2003 71440)

HPV multiple infection in 91 and OR 318 folds for CIN 3+

(Lee et al Cancer Lett 2003198187)

Time Line of Cervical HPV

Infections And Progression

to Cervical Cancer

15 yo 30 yo 45 yo

HPV

PrecancerCancer

Age

Rate

Adapted from Schiffman amp Castle New Eng J Med 3532101-4 2005

Lifetime incidence of genital HPV infection gt80 in US

Most infections are asymptomatic and clear spontaneously

eliminating cancer risk for that infection

Persistent infection with a high-risk HPV especially HPV16

or 18 is the single most important risk factor for

progression to precancer and cancer

Life Cycle of HPV Infection

HPV Infection is usually transient8-14 months


Basement

membrane

Dermis

Stratified

squamous

epithelium Virion

Virion

Supra-

basal

cells

Basal

cellsHPV

DNA

HPV DNAreplication

AssembledVirus

HPV16 Binds

the Basement Membrane

HPV-16 Laminin-5

Mouse vaginal tract 2 hours after exposure to HPV16

(8 hours after exposure to nonoxynol-9)

Kines Thompson Lowy Schiller and Day PNAS 106 20458-63 2009

Neutralizing

L1 Antibodies

(in red)

Bound to

Papillomavirus

Particle

VLP Vaccination Induces High

Titer Antibodies that Prevent

Basement Membrane Binding

Day et al Cell Host Microbe 8 260-70 2010

Basement

membrane

Dermis

Stratified

squamous

epithelium

Virion

Virion

No Infection

STOP

bull Episomal replication E1E2

bull Host integration E1E2 ORF disruption

E2-DNA dimer strong

E1-DNA hexamer weak

E2E6 real-time PCR

1episomal

gtor lt1mixed

0 integration

0 1000 2000 3000 4000 5000 6000 7000 7905 bp

E6

E1E7

E2

E4 L2E5

L1

My 0911

gp5+6+

HPV16-1

HPV16-2

HPV-induced tumorigenesis

1 integration pathway

2 episomal pathway

E1E2 promote genomic

instability through aberrant

replication of integrated

sequence

Genomic Organization of HPV

HR vs LR HPV

From Infection to Tumor InitiationIntegration 16 gt50 18 most

vs LR seldom to never

Transforming activityHR E7 immortalize at a low frequency- CDK2hg p21 p27low

vs LR E7- decreased p21 abrogation

HR E6 no transforming activity growth arrest abrogation by p53-

E6-E6AP trimeric complex vs LR E6- E6AP-p53 binding but no p53 degradation

HR E6+E7 highly efficient immortalizing proliferation and avoid

apoptosis but not tumorigenicv-rasv-fos coexpression tumorigenic

E6 telomerase ex PDZ domain [ X-ST-X-VLI]-containing protein

(p53-independent target) TNF modulation vs LR no PDZ domain

HPV E7

pRB functional inactivation

= hyperphosphorylation

=TF release

=TF activation

=G1-S restriction point entry

HPV E6

- p53 Proteolysis

- No mutation of p53

Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain

PDZ-domain protein scaffolding protein

cell polarity cell junction

E6 HRE6AP-PDZ binding induce loss of cell polarity EMT

and carcinogenesis

E2 and CK13 expression

is reverse to P16CK14ki67

Xue Y et al Cancer Res 2010705316-5325

E1Ê4 (amino-terminal E1 ORF splicing to E4 ORF)

Most divergent domain in sequence according

to HPV types

E4

L1

N terminal Binding to cytokeratin and concomitant

destabilization of cytokeratin network

Body cell cycle arrest at G2M checkpoint

L1 protein encoding

Long-acting multitasking

role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions

L1 Capsid cross-linking in oxidizing

environment of corneum

The more L1 capsid the less progressive

E5 Genomic Organization of HPV

E2

L2E56 Cys residue

bull Localized to Endoplasmic Reticulum (ER)-trafficking of cytopl memb protein

bull Weak transforming activity alone but strong in BPV E5

ndash enhancing transforming activity of E6E7 as oncogenic potential

bull EGFR (ErbB1) activity alteration through binding to vacuolar ATPase-

endosomal PH alteration- EGFR turnover alteration

bull PDGFR ligand-independent complex during tissue repair after HPV entry

bull Interfere with gap junction and alter caveolin-1

bull Anchorage-independent growth (anoikis) stimulation (HPV release)

bull Inducer of koilocytosis as cofactor with E6 in vitro (Am J Pathol2008 173682)

E2Ê5 (HPV 83 aa BPV 86 aa)

Koilocytosis by E5

Cytoplasmic vaculation-unclear reason

but contributing to fragility to make it easy

to release viral particlesAm J Pathol 2008 173682-8

E7 inhibits koilocytosis

E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-

NcCyto Cyto-ER Cyto-Nc Cyto-Nc Cyto-

Nc

Cyto-Nc

Main

action

Replication Replicat

ion

CK-disruption EGFRPDGFR

Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8

Cell cycle G2 arrest S-phase

entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal

Replication Control Most abundant

viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability

Immortalized E6E7- no tumorigenic in vitro E6E7+V-rasfos - tumorigenic

Changes in the HPV16 life cycle

during the development of cervical

cancer

CIN1 CIN2 CIN3

VS

bull CIN1 generally resemble productive lesions

bull In CIN2 and CIN3 lesions the order of life cycle events is unchanged

but the extent of E7 (dominant oncogene) expression is increased

bull Viral genome integration into cellular DNA

bull Loss of E2 leads to increased E6E7 expression

bull In cervical ca the productive stages of the virus life cycle are no longer

supported and viral episomes are usually lost

Revisited Concept

ASCUS cells caused by incomplete E4E5 activity

Abortive koilocyte

Queryocyte

Regressing koilocyte not enough cleared by immune sys

HR-LSIL

Need to monitor carefully but no overconcern

LR-HSIL

Occur rarely in single but often combined need to be further studied

CIN2

definitely heterogeneous in HPV life cycle to manifest combined

productive and proliferative phase

1

2

3

Performance characteristics

Designation ProbesprimersReaction

product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13

PCRMY0911 Dot blot Degenerate primer 450 bp 01ndash100 39

PCR PGMY0911

reverse LBA

Mixture of

consensus primers450 bp 01 27

PCR GP5+GP6+ EIA

ELISA systemConsensus primers 150 bp 05ndash10 20

PCRGP5+6+ reverse

LBAConsensus primers 150 bp 05ndash10 37

PCRSPF-PCR reverse

LiPA

Mixture of

consensus primers65 bp 01ndash10 43

Iftner T et al J Natl Cancer Inst Monogr 2003(31)80-8

The concept of HPV load

with clinical behaviorSnijders P et al J Pathol 2003 201 1ndash6

Analytical VS Clinical Sensitivity

Snijders P et al J Pathol 2003 201 1ndash6

Take Home MessagesQ1) What is the most abundant genome in LSIL

1 E6 2 E7 3 E4 4 E5

A1) 3 E4

Q2) What reacts first when infected basal cells

A2) 4 E5

Q3) What is inducing this change

A3) 4 E5

Q4) What is persistently identified in cytoplasm

A4) 3 E4

Q5) What is dominant oncogene

A5) 2 E7

Ovary Ca Represents Many Disease

bull Many ovarian ca are derived from non-ovarian tissue

bull Different ovarian histiotypes share few molecular similarities

bull Many EM hg should be reclassifed as serous ca bull Many mucinous ca should be reclassified as

secondary ca bull Serous ca hg share molecular profiles with basal-like

breast ca bull Clear cell ca share with renal CCC in sunitinib effect

bull Favoring pelvicperitoneal cancer over ovary ca

Salpingectomy best choice to prevent ovary cain case risk group

Mucinous type-exclusion Dx

Appendectomy-Best choice to manage PP

Endometriosis-High risk factor of ovary ca

Genesis and Evolution of Ovary Ca

bull Type I-low grade slow growing tumor (25) ndash Developing from borderline tumorndash Normal karyotypendash Rasmut high BRCA β-cat PTEN p53mut lack ndash PI3KCA ARID1A loss (EMCCC aw endometriosis)ndash Mucinous LG serous LG EM clear cell candash Poorly responsive to platinum-based CTx

bull Type II-rapid growing highly aggressive (75)ndash Rare Rasmut 70-80even 100 p53mut

ndash De novo Genetic instabilityndash HG serous ca HG endometrioid ca MMMT undiff sarcoma

Tumor Microenvironment-linking to Pathology-

연세의대 병리학교실

조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG

Tumor microenvironment- Plethora of interaction

Chemical factorsMechanical Factors

Cellular factors

Tumor microenvironment

Interaction Arena Terra Incognita

Extracellular matrix (ECM) Stiffness

Focal adhesion complex (FAC) Actin polymerization

Cell adhesion molecule (CAM) Integrin

Tumor resident cell Ca-associated fibroblast (CAF) Ca-associated adippocyte (CAA)

Tumor infiltrating cell Tumor-ass macropahge (TAM)

Vascular factors

Cytokine Chemokine Soluble factors Signal transduction

Interstitial fluids Endothelial cell angiogenesis

TME ldquoseed-soilrdquoampldquoPing-pongrdquo

bull Stephen Paget ldquoseed-soilrdquondash 1994 76 articles

ndash 2004 331 (3 folds1decade)

ndash 2010 4861 (15 folds half decade)

bull Post-Paget era

ndash bdquoPing-pong‟ experiments (IP Witz Cancer Res 1989)

bull BALBc 3T3 in vitroin vivo injection (ping)

bull Recultured from in vivo tumor (pong)

bull In vivo passage cells was considerably augmented as compared to that of in vitro maintained clonal ancestors

bull In vivo ldquoTME induces of pro-tumor effectrdquo

ndash Class II cancer gene (R Sager PNAS 1997)

bull Many of class II genes are regulated by TME

bull Shifting focus from DNA to RNA

Non-tumor cells in TME are different from those in normal ME

ndash Structural and functional alteration in TME

ndash Paracrine signal from tumor cell or normal cell

ndash Promalignant factors in TME

ndash Hypoxia (necrosis) in TME

ndash Low glucose concentration in TME

Question

ndashNormalization of abnormal non-tumor

constituents may reduce the malignancy

phenotype

ndashTumor reversion is indeed possible

ndashNot sufficiently cured by tumor cell

eradication alone

ndashIs there a hierarchy of interaction

Tumor microenvironment- Double-edged sword

bull Contemporary issuebull Cellular soluble factors

ndash Balance score of pro-(MCP-1 IL-6 MMP) and anti-malignant (IP-10) factors 136~ 244 (Witz IJCa 2003)

ndash No MECE

bull P-P interaction

bull Tumor heterogeneity vs signal heterogeneity

bull Cross-talk (agonistic vs antagonistic)ndash TNF family

raquo Proinflammatory cytokine as promalignant effect (since R Virchow 1863)

ndash TGF-βraquo Inhibit normal mammary epithelail cells

raquo Enhance tumor cell invasionmets

- loss of anti-proliferative role

- mutation in TGFBR

- inhibition of SMAD pathway

- suppressing anti-tumor immune response

- augmenting angiogenesis

macrophage

MCP-1TNF-α

MMP

Extracellular Matrix (ECM)

Compression buffer with structural frameCell-cell interaction regulation

Blackbox (footpath)One thing arises from all things and all things arise from

one thing (Aristotle)

Tumor migrationStiffness and destruction

No de novo synthesis

Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid

Soil researcher as in Scientific Botanist

Matrix metalloproteinase FM27 types

Integrin as heterodimer in biphasic roles

α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1

β4 - lamininβ5 β6 β8

αV

β3 - vitronectin

β2

αL

αM

αX

αD

18 α+ 8 b = 24 heterodimers

fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal

HYD-1 (RGD mimicker)decapeptide

Focal Adhesion bull Subcellular macromolecule that mediates anchorage of

ECM (lt15nm close to membrane)bull Outside-in Signal hub

Integrin-targeted therapy

TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC

Laminin-γ2 Integrin-β4

TZ IZ NZTZ IZ NZ

Laminin 5 and Igβ4 specific to IZ as invasive signature

Distinct ECM gene expression in the interface zone of invasive ductal carcinoma through

Screening by ECM cDNA array and validated by real-time RT-PCR and WBKim amp Cho Am J Pathol 2011

Integrin β4Laminin-γ2

Rela

tive inte

nsity

Expression of integrin β4 in fibroblasts only when cocultured directly with invasive cancer in vitro

ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture

Igβ4 switching in invasion

Integrin β4Integrin α6

In Situ Proximity Ligation Assay (PLA)Red dots denote regions of signal amplification

consistent with a6β4 integrin interactions

MDA-MB-231

MDA-MB-231 Integrin a6β4

Integrin a6β4

Secondary antibodies (PLA probe MINUS and PLA probe PLUS)

Ligase is added and theoligonucleotides will hybridize to the two PLA probes and join to a closed circle if they are in close proximity

Amplification solution and fluorescently labeled oligontis added together with polymeraseAs a primer for a rolling-circle amplification (RCA) reaction

Ig a6 (rabbit)Ig b4 (rat or mouse)

Red dots amplification-a6β4 integrin interactions

In Situ Proximity Ligation Assay (PLA)- Ig a6β4

On Submission Park and Cho 2012

Polarized morphology in Cell Migration

bullIntegrin-cytoskeleton linkage 4-fold difference between the front and rear Thatrsquos Rapid and intermittent (Schmidt 1993 JBC)

bull Migration rate depends on1 stimulus gradient2 locomotion speed

(integrin-cytoskeleton)

3 directional persistence time (inverse relation to speed)

Desmoplasia

CAFaSMA

Correlation of Cell Migration amp Tissue Invasion

Cell biology Asymmetric polarized morphology Kinetic fluctuation in receptor-ligand binding Concentration gradient- independent

Cell migration Cell 1996 84359-69 (Bible for migration)

Tissue biology Early onset of microinvasion step-wise model Diagnostic useful universal kit BM disruption or formation with

different composition

Clinical biology TNM stage reassessment Aggressive non-invasive cancer Biomarker to predict invasion or metastasis

Plasticity of tumor invasion-more than cell level

Tumor-cell invasion and migration Friedl amp Wolf Nat Rev Cancer

2003 3362-74

Integrin recruitement

Heterogeneous TMEHomogeneous TME

The stressful ME of intraductal niche promote genetic instability

5-25 layers thick around comedo necrosis

A simplified overview of signalling networks regulating epithelialndashmesenchymal transitions (EMT)

EMT InducerLoss of cell junction

ECM Remodelling

Polyak amp Weinberg Nat Rev Cancer 2009 9265-73

TF

Cell migration in Pathology조직에 항상 적용되지 않는다

bull Early transient phenomenon

ndash Actin polymerization SMA+ more eosinophilic

ndash Collective invasion에서도 asymmetric polarity Be careful with TMAbull 대부분 EMT process는 아님

bull EMT를 조직에서 확인하기는 어려움

ndash Keep aware of aggressive non-invasive ca

ndash What is EMT in Pathologybull TsF more sensitive

bull cell junction molecule more specific

G-protein (GTPase)

bull Outside-in signaling

bull G-protein coupled receptor (GPCR)-950개

ndash Molecular switch (GTP-on GDP-off)

ndash Heterotrimeric (large) G-protein αβγ ndash GPCR-

dependent

bull Gα cAMP (1994 Alfred Gilman amp M Rodbell Nobel pr)

ndash Heterodimeric homologous Ras G-protein (small) α( Gαs Gαio

Gαq11 Gα1213 ) 10 types ( Ras Rho Rab Rap Arf Ran Rheb Rad Rit Miro)

Motility factors triad in cellular level

Cdc42 controls the cell polarity and the

formation of filopodia and nascent focal

complexes (shown as yellow dots)

Rho influences cell adhesion

assembly and maturation in

addition to controlling stress fiber

formation and contractile activity

Rac1 primarily controls actin

assembly and adhesion in the

lamellipodium

Chemokinebull 48 types (8~10 kD)

bull Four cystein residue forming 3-D shape

bull G-protein receptor

(γ-2) (β-28) (α-17)

(δ-1)

CXCR-1 CXCL-6 CXCL-8 (IL-8) - neutrophilCXCR-2 CXCL-1~ 7 - neutrophilCXCR-3 CXCL-9 ~11 - T-cellCXCR-4 CXCL-12 (SDF-1) - hematopoietic cells

endotheial cells neuronCXCR-5 CXCL-13 -B-cellCXCR-6 CXCL-16CXCR-7 CXCL-12 (SDF-1) -memory B-cell

CCR-1~11 CCR-11 (scavenger receptor)-no signal

XCR-1 XCL-1~2CX3CR1 CX3CL-1

TAM

AMD3100

Pro-inflammatory Pro-angiogenic

Newly Identified zone-Interface Zone resorted from Normal Zone

Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1

Matrisomics using ECM array in Tumor and Normal ME

Matrix stiffness inducing Igβ4

Kim and Cho BCR 2012

IZ as EMT playground

IZ as migratory zone

Scratching assay ORIS invasion assayChoi and Cho BBRC 2012Gao and Cho Oncogene 2011

Tumor senescenceHypoxic signature EMT

CSC-nicheInvasion signature

Why Interface zone

Quiscent zone

EMTInvasion signatureAnoikis-resistance

Quiscent Normal

PAST

PRESENT

NEAR FUTURERECUR ZONE

FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4

Focal Adhesion Tumor-dependent

microenvironment

IZ is permissive for tumor invasion

IF zone implication in practice

bull Potential candidate for molecular margin

ndash How far in tumor extent is safebull Near Normal (overlap with TZ) vs Far normal

bull Normal is no more

bull Molecular margin within 5 mm from the tumor margin even in case of no tumor should be removed for Recurrence-zero

bull Interaction mechanismndash Lymphangiogenesis

bull Drug-delivery huddle ndash ECM target moelcule to lessen stiffness

IZ-signature using MALDI-Imaing MSMS

IgHA2

Kang amp Cho JPR 2012

Sampling in TME studyNormal is no more Tumor is too more

bull Tumor periphery vs Far normal to tumor

bull Highlight the Interface zone between TM amp NL ndash Cell invasionmigration zone

ndash Cancer stem cellEMT enriched zone

bull Molecular marginndash How far in tumor extent is safe

bull Near Normal vs Far normal

bull Interface zone (IZ) within 5mm circumferential belt

bull IZ ME is much more similar to TZ ME than far normal IZ in ECM array

bull Molecular margin within 5 mm from the tumor margin even in case of no tumor should be removed (Am J Pathol 2011 178373)

ndash for Recurrence-zero

Take Home Message

Do not always force to match data between in vitroin vivohuman samples-blood tissue

Human samples are too heterogeneous to simplify and apply in vitro data or hypothesis

Rudolf Virchow Father of Pathology

G Nikolas Pappanicaulau ndashPAP test

James Ewing ndashtumor metastasis theory

Arias-Stella- AS reaction

Azzopardi JG- One sees and recognizes only those features which have already been described

Arthur Purdy Stout- Atlas of Tumor Pathology

Stanley L Robbins- 1957 textbook 1st ed

Lesions do not arise in cadevars But the study of morphology is only one facet of pathology Pathology contributes much to clinical medicine The pathologist is interested not only in the recognition of structural alterations but also in their significance ie the effects of these changes on cellular and tissue function and ultimately the effect of these changes on the patient It is not a discipline isolated from the living patient but rather a basic approach to a better understanding of disease

and therefore a foundation of sound clinical medicine

Virchowrsquos triad

PIONEERS of PATHOLOGY






Alois Alzheimer

Von Reklinghausen


하얀 거탑

Presentation Flow


E6-E6AP-PDZ binding

E1Ê4 and E2Ê5









1 1831333539455152565859





3 611

2B 58 Limited

3 Other type





1 1831333539455152565859





3 611

2B 58 Limited

3 Other type






HPV with no CIN 372








to Cervical Cancer

15 yo 30 yo 45 yo

HPV

PrecancerCancer

Age

Rate











Basement

membrane

Dermis

Stratified

squamous

epithelium Virion

Virion

Supra-

basal

cells

Basal

cellsHPV

DNA

HPV DNAreplication

AssembledVirus

HPV16 Binds


HPV-16 Laminin-5




Neutralizing

L1 Antibodies

(in red)

Bound to

Papillomavirus

Particle





Basement

membrane

Dermis

Stratified

squamous

epithelium

Virion

Virion

No Infection

STOP



E2-DNA dimer strong

E1-DNA hexamer weak

E2E6 real-time PCR

1episomal

gtor lt1mixed

0 integration

0 1000 2000 3000 4000 5000 6000 7000 7905 bp

E6

E1E7

E2

E4 L2E5

L1

My 0911

gp5+6+

HPV16-1

HPV16-2



2 episomal pathway




sequence


HR vs LR HPV











HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message








Alois Alzheimer

Von Reklinghausen


하얀 거탑

Presentation Flow


E6-E6AP-PDZ binding

E1Ê4 and E2Ê5









1 1831333539455152565859





3 611

2B 58 Limited

3 Other type





1 1831333539455152565859





3 611

2B 58 Limited

3 Other type






HPV with no CIN 372








to Cervical Cancer

15 yo 30 yo 45 yo

HPV

PrecancerCancer

Age

Rate











Basement

membrane

Dermis

Stratified

squamous

epithelium Virion

Virion

Supra-

basal

cells

Basal

cellsHPV

DNA

HPV DNAreplication

AssembledVirus

HPV16 Binds


HPV-16 Laminin-5




Neutralizing

L1 Antibodies

(in red)

Bound to

Papillomavirus

Particle





Basement

membrane

Dermis

Stratified

squamous

epithelium

Virion

Virion

No Infection

STOP



E2-DNA dimer strong

E1-DNA hexamer weak

E2E6 real-time PCR

1episomal

gtor lt1mixed

0 integration

0 1000 2000 3000 4000 5000 6000 7000 7905 bp

E6

E1E7

E2

E4 L2E5

L1

My 0911

gp5+6+

HPV16-1

HPV16-2



2 episomal pathway




sequence


HR vs LR HPV











HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



Alois Alzheimer

Von Reklinghausen


하얀 거탑

Presentation Flow


E6-E6AP-PDZ binding

E1Ê4 and E2Ê5









1 1831333539455152565859





3 611

2B 58 Limited

3 Other type





1 1831333539455152565859





3 611

2B 58 Limited

3 Other type






HPV with no CIN 372








to Cervical Cancer

15 yo 30 yo 45 yo

HPV

PrecancerCancer

Age

Rate











Basement

membrane

Dermis

Stratified

squamous

epithelium Virion

Virion

Supra-

basal

cells

Basal

cellsHPV

DNA

HPV DNAreplication

AssembledVirus

HPV16 Binds


HPV-16 Laminin-5




Neutralizing

L1 Antibodies

(in red)

Bound to

Papillomavirus

Particle





Basement

membrane

Dermis

Stratified

squamous

epithelium

Virion

Virion

No Infection

STOP



E2-DNA dimer strong

E1-DNA hexamer weak

E2E6 real-time PCR

1episomal

gtor lt1mixed

0 integration

0 1000 2000 3000 4000 5000 6000 7000 7905 bp

E6

E1E7

E2

E4 L2E5

L1

My 0911

gp5+6+

HPV16-1

HPV16-2



2 episomal pathway




sequence


HR vs LR HPV











HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



Presentation Flow


E6-E6AP-PDZ binding

E1Ê4 and E2Ê5









1 1831333539455152565859





3 611

2B 58 Limited

3 Other type





1 1831333539455152565859





3 611

2B 58 Limited

3 Other type






HPV with no CIN 372








to Cervical Cancer

15 yo 30 yo 45 yo

HPV

PrecancerCancer

Age

Rate











Basement

membrane

Dermis

Stratified

squamous

epithelium Virion

Virion

Supra-

basal

cells

Basal

cellsHPV

DNA

HPV DNAreplication

AssembledVirus

HPV16 Binds


HPV-16 Laminin-5




Neutralizing

L1 Antibodies

(in red)

Bound to

Papillomavirus

Particle





Basement

membrane

Dermis

Stratified

squamous

epithelium

Virion

Virion

No Infection

STOP



E2-DNA dimer strong

E1-DNA hexamer weak

E2E6 real-time PCR

1episomal

gtor lt1mixed

0 integration

0 1000 2000 3000 4000 5000 6000 7000 7905 bp

E6

E1E7

E2

E4 L2E5

L1

My 0911

gp5+6+

HPV16-1

HPV16-2



2 episomal pathway




sequence


HR vs LR HPV











HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message






1 1831333539455152565859





3 611

2B 58 Limited

3 Other type





1 1831333539455152565859





3 611

2B 58 Limited

3 Other type






HPV with no CIN 372








to Cervical Cancer

15 yo 30 yo 45 yo

HPV

PrecancerCancer

Age

Rate











Basement

membrane

Dermis

Stratified

squamous

epithelium Virion

Virion

Supra-

basal

cells

Basal

cellsHPV

DNA

HPV DNAreplication

AssembledVirus

HPV16 Binds


HPV-16 Laminin-5




Neutralizing

L1 Antibodies

(in red)

Bound to

Papillomavirus

Particle





Basement

membrane

Dermis

Stratified

squamous

epithelium

Virion

Virion

No Infection

STOP



E2-DNA dimer strong

E1-DNA hexamer weak

E2E6 real-time PCR

1episomal

gtor lt1mixed

0 integration

0 1000 2000 3000 4000 5000 6000 7000 7905 bp

E6

E1E7

E2

E4 L2E5

L1

My 0911

gp5+6+

HPV16-1

HPV16-2



2 episomal pathway




sequence


HR vs LR HPV











HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message






1 1831333539455152565859





3 611

2B 58 Limited

3 Other type






HPV with no CIN 372








to Cervical Cancer

15 yo 30 yo 45 yo

HPV

PrecancerCancer

Age

Rate











Basement

membrane

Dermis

Stratified

squamous

epithelium Virion

Virion

Supra-

basal

cells

Basal

cellsHPV

DNA

HPV DNAreplication

AssembledVirus

HPV16 Binds


HPV-16 Laminin-5




Neutralizing

L1 Antibodies

(in red)

Bound to

Papillomavirus

Particle





Basement

membrane

Dermis

Stratified

squamous

epithelium

Virion

Virion

No Infection

STOP



E2-DNA dimer strong

E1-DNA hexamer weak

E2E6 real-time PCR

1episomal

gtor lt1mixed

0 integration

0 1000 2000 3000 4000 5000 6000 7000 7905 bp

E6

E1E7

E2

E4 L2E5

L1

My 0911

gp5+6+

HPV16-1

HPV16-2



2 episomal pathway




sequence


HR vs LR HPV











HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message





to Cervical Cancer

15 yo 30 yo 45 yo

HPV

PrecancerCancer

Age

Rate











Basement

membrane

Dermis

Stratified

squamous

epithelium Virion

Virion

Supra-

basal

cells

Basal

cellsHPV

DNA

HPV DNAreplication

AssembledVirus

HPV16 Binds


HPV-16 Laminin-5




Neutralizing

L1 Antibodies

(in red)

Bound to

Papillomavirus

Particle





Basement

membrane

Dermis

Stratified

squamous

epithelium

Virion

Virion

No Infection

STOP



E2-DNA dimer strong

E1-DNA hexamer weak

E2E6 real-time PCR

1episomal

gtor lt1mixed

0 integration

0 1000 2000 3000 4000 5000 6000 7000 7905 bp

E6

E1E7

E2

E4 L2E5

L1

My 0911

gp5+6+

HPV16-1

HPV16-2



2 episomal pathway




sequence


HR vs LR HPV











HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message






Basement

membrane

Dermis

Stratified

squamous

epithelium Virion

Virion

Supra-

basal

cells

Basal

cellsHPV

DNA

HPV DNAreplication

AssembledVirus

HPV16 Binds


HPV-16 Laminin-5




Neutralizing

L1 Antibodies

(in red)

Bound to

Papillomavirus

Particle





Basement

membrane

Dermis

Stratified

squamous

epithelium

Virion

Virion

No Infection

STOP



E2-DNA dimer strong

E1-DNA hexamer weak

E2E6 real-time PCR

1episomal

gtor lt1mixed

0 integration

0 1000 2000 3000 4000 5000 6000 7000 7905 bp

E6

E1E7

E2

E4 L2E5

L1

My 0911

gp5+6+

HPV16-1

HPV16-2



2 episomal pathway




sequence


HR vs LR HPV











HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message




Basement

membrane

Dermis

Stratified

squamous

epithelium Virion

Virion

Supra-

basal

cells

Basal

cellsHPV

DNA

HPV DNAreplication

AssembledVirus

HPV16 Binds


HPV-16 Laminin-5




Neutralizing

L1 Antibodies

(in red)

Bound to

Papillomavirus

Particle





Basement

membrane

Dermis

Stratified

squamous

epithelium

Virion

Virion

No Infection

STOP



E2-DNA dimer strong

E1-DNA hexamer weak

E2E6 real-time PCR

1episomal

gtor lt1mixed

0 integration

0 1000 2000 3000 4000 5000 6000 7000 7905 bp

E6

E1E7

E2

E4 L2E5

L1

My 0911

gp5+6+

HPV16-1

HPV16-2



2 episomal pathway




sequence


HR vs LR HPV











HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



HPV16 Binds


HPV-16 Laminin-5




Neutralizing

L1 Antibodies

(in red)

Bound to

Papillomavirus

Particle





Basement

membrane

Dermis

Stratified

squamous

epithelium

Virion

Virion

No Infection

STOP



E2-DNA dimer strong

E1-DNA hexamer weak

E2E6 real-time PCR

1episomal

gtor lt1mixed

0 integration

0 1000 2000 3000 4000 5000 6000 7000 7905 bp

E6

E1E7

E2

E4 L2E5

L1

My 0911

gp5+6+

HPV16-1

HPV16-2



2 episomal pathway




sequence


HR vs LR HPV











HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



Neutralizing

L1 Antibodies

(in red)

Bound to

Papillomavirus

Particle





Basement

membrane

Dermis

Stratified

squamous

epithelium

Virion

Virion

No Infection

STOP



E2-DNA dimer strong

E1-DNA hexamer weak

E2E6 real-time PCR

1episomal

gtor lt1mixed

0 integration

0 1000 2000 3000 4000 5000 6000 7000 7905 bp

E6

E1E7

E2

E4 L2E5

L1

My 0911

gp5+6+

HPV16-1

HPV16-2



2 episomal pathway




sequence


HR vs LR HPV











HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message







Basement

membrane

Dermis

Stratified

squamous

epithelium

Virion

Virion

No Infection

STOP



E2-DNA dimer strong

E1-DNA hexamer weak

E2E6 real-time PCR

1episomal

gtor lt1mixed

0 integration

0 1000 2000 3000 4000 5000 6000 7000 7905 bp

E6

E1E7

E2

E4 L2E5

L1

My 0911

gp5+6+

HPV16-1

HPV16-2



2 episomal pathway




sequence


HR vs LR HPV











HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message





E2-DNA dimer strong

E1-DNA hexamer weak

E2E6 real-time PCR

1episomal

gtor lt1mixed

0 integration

0 1000 2000 3000 4000 5000 6000 7000 7905 bp

E6

E1E7

E2

E4 L2E5

L1

My 0911

gp5+6+

HPV16-1

HPV16-2



2 episomal pathway




sequence


HR vs LR HPV











HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



HR vs LR HPV











HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



HPV E7



=TF release

=TF activation


HPV E6

- p53 Proteolysis


Both TSG are

impaired by

HPV E6E7

E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



E6 and PDZ domain




and carcinogenesis






to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message








to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message





to HPV types

E4

L1




L1 protein encoding


role of E4 genome

HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



HPV E4

functions

cytokeratin

disruption

Virions





E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message




E2

L2E56 Cys residue











Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



Koilocytosis by E5





E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message




E5 and E6

induce

koilocyte

E7 inhibits

koilocyte

Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



Sequence of events

of HPV genome

E1 E2 E4 E5 E6 E7 L1 L2

Location Cyto-Nc

shuttling

Cyto-


Nc

Cyto-Nc

Main

action


ion


Anoikis-

resistance

P53

disruption

pRb mut

P16 ovex

Ki67 ovex

Major

capsid

Minor

capsid

Sequence

of events

5 6 () 2

(92a-a)

1

(16kDa83a-a)

4

(18kDa

150a-a)

3

(13 kDa

98 a-a)

7 8


entry

Oncogeni

c

suggested Key role

(HR vs LR)

Key role

(HR vs LR)

Prime

goal


viral product

Lateral spread

Infected basal

cells

proliferation in

viral entry

Koilocytosis

Immortalize

but not

alone

Permissive

milieu

Immortalize

Dominant

oncogene

Cross-

linking

VLP-

vaccine

Shape

and

stability




cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message





cancer

CIN1 CIN2 CIN3

VS








Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



Revisited Concept


Abortive koilocyte

Queryocyte


HR-LSIL


LR-HSIL


CIN2



1

2

3



product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message





product

Analytical

sensitivity fg

detectable

types

Hybridi

zation

HC2 HPV DNA

assay

Mixture of

RNA probes

DNARNA

hybrids25ndash75 13


PCR PGMY0911

reverse LBA

Mixture of


PCR GP5+GP6+ EIA


PCRGP5+6+ reverse


PCRSPF-PCR reverse

LiPA

Mixture of








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message








1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message






1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message




1 E6 2 E7 3 E4 4 E5

A1) 3 E4


A2) 4 E5


A3) 4 E5


A4) 3 E4


A5) 2 E7


















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message




















조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message













조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message









조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message





조 남훈

Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



Cell cycle

Hallmarks of Cancer

apoptosis

TME TME

TSG



Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message





Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message




Cellular factors








Vascular factors







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message







bull Post-Paget era















Question



phenotype



eradication alone





ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message






ndash No MECE








- mutation in TGFBR




macrophage

MCP-1TNF-α

MMP







Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message









Proteoglycan

Heparan sulfate

Chondroitin

sulfate

Keratan sulfate

Non-proteoglycan

Fiber

Fibronectin

Collagen-14 types

Elastin

Laminin

Hyaluronic acid




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message





α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message




α1

α2

α3

α4

α5

α6

α7

α8

α9

α10

α11

β7

αE

β1


αV

β3 - vitronectin

β2

αL

αM

αX

αD


fibronectincollagen

Outs

ide-

in s

ignal

Insid

e-out

sig

nal





TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message






TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message




TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



TZ

1 2 3 4 5

NZIZ

DCIS

Laminin-γ2

GAPDH

Integrin β4

TZ

1 2 3 4 5

NZIZ

IDCA

Tum

or

Interface

5 mm

Normal 1 2 3 4 5

Rela

tive inte

nsity

Laminin-γ2

1 2

TZ IZ NZTZ IZ NZ

Laminin-α3

4 5

TZ IZ NZTZ IZ NZ

Integrin β4

1

TZ IZ NZ

3

TZ IZ NZ

2

TZ IZ NZ

3

TZ IZ NZ

DCIS IDC

Integrin α6

GAPDH

C

B

DCIS IDC


TZ IZ NZTZ IZ NZ





Rela

tive inte

nsity


ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message




ACAF

Integrin

β1

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

BCAF

Integrin

β4

Events

InF NBF

SF

MM

CF

7

CM

MD

A-

MB

-231

CM

CCAF InF NBF

Fibroblast(FITC+)

Inte

grin

β1

MC

F

-7

MD

A-M

B-

231

DCAF InF NBF

Fibroblast(FITC+)

Positiv

e

(MD

A0M

B-2

31

)

Negative

(Fib

robla

st)

MC

F-7

MD

A-M

B-

231

Inte

grin

β4

Control

Integrin β1

Integrin β4

GAPDH

EMCF7 MDA-MB-231

Co

culture





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message






MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message





MDA-MB-231


Integrin a6β4













Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message











Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message








Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



Desmoplasia

CAFaSMA









2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message











2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message





2003 3362-74







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message







ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message





ECM Remodelling


TF









G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message











G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



G-protein (GTPase)





dependent














lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message













lamellipodium




(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message






(γ-2) (β-28) (α-17)

(δ-1)





TAM

AMD3100



Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message




Matrix stiffness

Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message



Min

Avg

NZ-2

TZ-3

NZ-1

NZ-3NZ-4

IZ-2IZ-3

TZ-1

IZ-1









Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message








Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message







Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message





Why Interface zone

Quiscent zone


Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message




Quiscent Normal

PAST

PRESENT


FUTURE

Rho-GTP

ROCK

WNT

ECM stiffness

Ig β4


microenvironment










IgHA2












Take Home Message











IgHA2












Take Home Message




IgHA2












Take Home Message













Take Home Message



Take Home Message



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