special approaches of tumor biology and chemotaxis orsolya láng 2014

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Special approaches of tumor biology and chemotaxis Orsolya Láng 2014.

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Page 1: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Special approaches of tumor biology and chemotaxis

Orsolya Láng

2014.

Page 2: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

TUMOR CELLS AND MIGRATION

METASTASISPRIMARY TUMOR

AngiogenesisAdhesion

Page 3: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

CELL and CELL CYCLE

Page 4: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Growth factors

Adhesion Adhesion moleculesmolecules

ChemokineChemokiness

Cell cyle Cell cyle regulatory regulatory proteinsproteins

Apoptotic Apoptotic moleculesmolecules

OncogenesOncogenes

Page 5: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

TUMOR CELL

Page 6: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

GENERAL FEATURE OF A TUMOR CELL

The hallmarks of cancer comprise six biological capabilities acquired during the multistep development of human tumors (Hanahan és Weinberg 2000)

Page 7: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

CELL KINETICS

Page 8: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Doubling time of the tumor volume (Td)

Time of the Cell cycle (Tc):Tc= Ts / Li

Ts: S phaseLi: labeling index

(proportoin of cells in S phase)

Growth fraction(GF):GF=P / (P+Q’)

P: number of the mitotic cellsQ: number of the cells in interphase

Rate of the cell loss ():= 1-Tpd / Td Tpd= *Ts/Li

Tpd: Potential tumor volume doubling timeTd: tumor volume doubling time

Lymphoma 48 h

Lung cancer 108 h Usually 15-125 h

Lymphoma 4 weeksColon adenoma 90 weeksUsually 18-200 days

Page 9: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Volume of the tumor tissue

~10 divisions =*1000 cell number increase (210 =1024)~20 divisions= 106 cells = 1 mg= 1 mm3

~30 divisions= 109 cells = 1 g= 1 cm3

~40 divisions= 1012 cells = 1 kg

1 tumor cell ~30-33,25 division=1-10 cm3

Time of the clinical symptomes / diagnosis

~27 division= 0.1cm3

Earliest time of diagnosis(Visualisation)

~40 division= 1012cellFatal

Human total cell number: 3.72 × 1013

http://www.ncbi.nlm.nih.gov/pubmed/23829164

Page 10: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

BIOLOGY OF THE TUMORPROGRESSION

Page 11: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Exogen and endogen factorsGenom instability

Activation of the oncogenesInactivation of tumorsuppressors

Local and systemic factorsinhibition acceleration

Growth rate

Ectopic survival capacity

Invasivity

De-differentation

Tumorigenesis

Epithelialcell

Hiperplasticadenoma

DisplaticCarcinoma in situ

Invasive carcinoma

Metastasis

Page 12: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Important steps of tumor progression

Transformation of the microenvironment: stromal cells,ECM components,proteolytic degradation

Induction of the angiogenesis (w/o max tumor size is 2mm)

Escaping from immune-mediated rejection Formation of metastasis

Page 13: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

MICROENVIRONMENT – STROMAL CELLS

Cell types:fibroblasts,

myofibroblasts, endothelial cells, lymphocytes, macrophagesFunction:

host defence

! MALT - B cell helps to maintain lymphomas

! Growth factors are released by the stromal cells (VEGF - angiogenesis)

http://www.nature.com/nrc/journal/v9/n4/fig_tab/nrc2618_F1.html

Page 14: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Macrophage

Cancer cell migration is controlled by paracrine loop

THE INVASIVE MICROENVIRONMENT

http://www.nature.com/nrc/journal/v9/n4/full/nrc2618.html

Macrophage

Blood vessel

Page 15: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

ANGIOGENESIS

Hypoxia formation of new vessels, proliferation of the endothelial cellsTypes: vessels

arteriovenous shunts„dead end”

/lack of smooth muscle , weak vessel wall, irregular shape(insuficient endothelial cell and basement membrane layers)/

sinuses /wall is formed by tumor cells/Venous circulation

VEGF induces angiogenesisincreases permeability

Lack of lymphatic vessels

OEDEMA, decresed blood flow

Inhibition of the VEGF pathway is a potential therapeutical tool

Page 16: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Strategies that tumors use to escape from immune-mediated rejection are:

To decrease the antigen expressionTo inhibit the immune-reactive cells:

degrade the chemoattractansdecrease their cell adhesioninhibite their phagocytotic activity

Page 17: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Angiogenesis

Local invasion

ECM

AdhesionProteolysisMigration

Intravasation

Extravasation

circulation

Metastasis

Tumor cell

Primary tumor

AdhesionProteolysisMigration

Angiogenesis

VEGFAngiogeni

nFGF

spreading

METASTATIC CASCADE

Page 18: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

INVASION

In situ carcinoma

DECREASED CELL ADHESION, INCREASED MOTILITY

ECM proteolysis

Page 19: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Angiogenesis

Local invasion

ECM

AdhesionProteolysisMigration

Intravasation

Extravasation

circulation

Metastasis

Tumor cell

Primary tumor

AdhesionProteolysisMigration

Angiogenesis

VEGFAngiogeni

nFGF

spreading

METASTATIC CASCADE

Page 20: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

CELL ADHESION

Significant change in cell-cell and cell-ECM interactionsMolecules:

selectinsintegrinsimmunoglobulin superfamily cadherins catenins

Page 21: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

SELECTINS

Cell-cel junctionsTypes:

E- endothelial cellsP- trombocytesL- leukocytes

Extracellular C-lectin domain Ca2+ dependent anchorage It binds Sialyl-Lex carbohydrates

„ROLLING”

! Tumor cells express increased amount of sialil-Lex or -Lea

Page 22: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

INTEGRINS

Transmembrane receptorsForm cell-ECM interaction8 , 14 subunites ~20 heterodimerCa2+, Mg2+ dependent anchorage„RGD” sequence is the specific substrateSignalling: outside-in – signalling

inside-out – adhesionIncreased expression of integrins promotes angiogenesis and helps to bind MMPs at the cell surfaceEXTRAVASATION, ATTACHMENT

D

G

R

Page 23: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Integrin or celladhesion regulated signalling pathways

cellproliferation

PTEN

RAC PI(3)K

CDC42

integrin

ECM

ILK

-cateninCiklin D1BAD

PKB/AKT

FAK

RASRAFMEKMAPK

GSK3

motilitygene expression

cellcycleapoptosis

SHCGRB2/SOS

Page 24: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Integrin or celladhesion regulated signalling pathways

integrin

ECM

ILK

-kateninCiklin D1BAD

PKB/AKT

FAK

cellproliferation

RASRAFMEKMAPK

GSK3

motilitygene expression

cellcycleapoptsis

SHCGRB2/SOS

PTEN

RAC PI(3)K

CDC42

Page 25: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Molecular partners of the integrins

Cytoskeletal components:actinin, talin,F- actin, filamin

Adaptors:rack 1, ICAP-1

Calcium binding proteins:CIB, calreticulin

Protein kinases:pp125FAK, p59 ILK

Membrane proteins:CD9, CD16,CD47…caveolin, urokinase-plazminogen-activator receptor

Ligands in ECM:collagen, laminin, fibronectin, fibrinogen, von

Willebrand factor, osteopontin, elastin

Page 26: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

IMMUNGLOBULIN SUPERFAMILY

has 5 Ig-like domains at the extracellular regionforms cell-cell junction interacts with integrins

VCAM - 41, PECAM - v3

takes essential part in extravasation

! ! Over expression of ICAM-1, MUC18 increased inavsion

! ! Down-regulation of VCAM-1 increased metastatic potential (faster detachment)

Page 27: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

CADHERIN

Is a transmembrane glycoproteinForms homophyl cell-cell junctionsCa2+ dependent anchorageClassical types: E- epithelial

P- placentaN- neural,

Intracellular part interacts with catenins to connect aktin filaments

! E-cadherin tumorsuppressor! Increased expression of N-cadherin invasion! N-cadherin cooperates with the FGF receptor lead to the up-

modulation of MMP-9

„Cadherin Switch”

Page 28: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

CATENINCATENIN

Is an intracellular moleculeFixes cadherins to F-actin

! -catenin binds to the APC gene product

! colon and liver cancer increased cytoplasmic and nuclear localised beta-catenin correlates with invasion and poor prognosis.

in melanoma increased cytoplasmic and nuclear beta-catenin is currently emerging as a marker for good prognosis

Page 29: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Angiogenesis

Local invasion

ECM

AdhesionProteolysisMigration

Intravasation

Extravasation

Circulation

Metastasis

Tumor cell

Primary tumor

AdhesionProteolysisMigration

Angiogenesis

VEGFAngiogeni

nFGF

spreading

METASTATIC CASCADE

IntegrinscadherinsSelectins

CAM

Page 30: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

PROTEOLYSISComponents of the basement membrane(BM) and ECM: IV collagen, laminin, proteoglycanesTumorcells (stromal cells) secrete proteasesCathepsinMatrix metalloproteinase (MMP)Plazmin, tPA ,Urokinase (plasminogen activator inhibitor 1&2)Tissue inhibitor of metalloproteinases

INVASION

Page 31: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

MATRIX METTALLOPROTEINASESMATRIX METTALLOPROTEINASES (MMP)(MMP)

Page 32: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

MOLECULAR STRUCTURE OF THE MATRIX MOLECULAR STRUCTURE OF THE MATRIX METTALLOPROTEINASESMETTALLOPROTEINASES

SUBSTRATE OF TIMP

Nature Reviews Cancer 2, 161-174 (March 2002)http://www.nature.com/nrc/journal/v2/n3/full/nrc745.html

Page 33: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

MMP/TIMP EXPRESSION IN BREAST MMP/TIMP EXPRESSION IN BREAST CANCERCANCER

Nature Reviews Cancer 2, 161-174 (March 2002)

Page 34: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

MMP – TUMORPROGRESSION?!?

Nature Reviews Cancer 2, 161-174 (March 2002)

Page 35: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Angiogenesis

Local invasion

ECM

AdhesionProteolysisMigration

Intravasation

Extravasation

Circulation

Metastasis

Tumor cell

Primary tumor

AdhesionProteolysisMigration

Angiogenesis

VEGFAngiogeni

nFGF

spreading

METASTATIC CASCADE

IntegrinscadherinsSelectins

CAM

MMP/TIMPCathepsin

Plasminogen

Page 36: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

MIGRATORY MECHANISMS IN TUMOR

Small-cell lung cancer

Page 37: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

FORMS OF MIGRATORY ADAPTATION

http://www.nature.com/nrc/journal/v3/n5/full/nrc1075.html

Page 38: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

2D –3D MIGRATIONS

http://www.nature.com/nrc/journal/v3/n5/full/nrc1075.html

Page 39: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

STEPS OF 3D MIGRATION

1. Pseudopod protrusion2. Formation of focal contact3. Focal ECM proteolysis4. Actomyosin contraction5. Detachment

Page 40: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Cell-cell interactions visualized in tumorigenesis

Page 41: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Angiogenesis

Local invasion

ECM

AdhesionProteolysisMigration

Intravasation

Extravasation

Circulation

Metastasis

Tumor cell

Primary tumor

AdhesionProteolysisMigration

Angiogenesis

VEGFAngiogeni

nFGF

spreading

METASTATIC CASCADE

IntegrinscadherinsSelectins

CAM

MMP/TIMPCathepsin

Plasminogen

AMF/gp78AutotaxinHGF/c-MET

Page 42: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

!! Tumor markers e.g. cytokeratin, mucin

HEMATOGENIC DISSEMINATION

http://www.nature.com/nrc/journal/v4/n6/fig_tab/nrc1370_F3.html

Page 43: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

Blood flow patterns can predict the specific regions of metastases in approximately two-thirds of cancers

TYPICAL SITE OF METASTASIS

Page 44: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

LOCALISATION OF THE METASTASIS

Page 45: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

CHEMOKINES – TISSUE SPECIFIC LOCALISATION

http://www.readcube.com/articles/10.1038/nrc865

Page 46: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014
Page 47: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

adhesion

moti

lity

?

Page 48: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

EXTRAVASATION

?

Attachment

Migration

Page 49: Special approaches of tumor biology and chemotaxis Orsolya Láng 2014

http://www.nature.com/nrc/journal/v12/n1/fig_tab/nrc3180_F2.html#figure-title