Cancer cell Migration during Invasion and Metastasis

By Lokesh Patil

Overview of cancer spreading stages

Primary tumor formation Local Invasion Intravasation

Extravasation

Arrest at distant organ site

Transport and survival in through circulation

Micrometastasis formation

Metastatic colonization

Clinically detectable metastases

2

Invasion-Metastasis cascade

Tumor formation
Because of failure in DNA replication of cells causing uncontrolled division Occurs at molecular level in the cell nucleus Cellullar instability->daughter cells interacting with environment At cellular level->dynamics have longer space scale and slower time scale that molecular level. e.g, enzymatic degradation in ms whereas cell replication about a day

Fig 1 : Comparison of normal and Cancer cell division

4

Local Invasion
MMP(Matrix Metalloproteinase) driven proteolysis causes loss of BM barrier

Invasion of cancer cells into stromal compartment

Stroma becomes increasingly reactive

These Stromal cells enhance the aggressiveness of cancer cells

Fig 2: Local Invasion(marked in circle)

5

Intravasation
Entry of cancer cells into the lumina of blood vessel or lymphatic vessel Hematogenous circulation is the major mechanism for cancer cell dispersion Facilitated by molecular changes Strongly influenced by structural characteristics of tumor-associated blood vessels Weak blood vessel-endothelial cells interactions promote intravasation
Fig 3 : Tumor cell crossing the endothelial cell barrier
6

CTC(Circulating tumor cells) survival

CTCs are anchorage dependent cells CTCs(20-30um) Luminal Dia. of capillaries(~8um) Therefore CTCs spend short amount of time through circulation hence avoiding anoikis CTCs also need to avoid hemodymic shear forces and predation by immune cells. They form emboli with platelets to avoid this. Important clinical relevance
Fig 4: CTC pathway
7

CTC arrest and Extravasation

Still unknown whether specificity for sites is caused by vessel size restrictions or predetermined predilections

Fig 5: Metastasis tropism: Carcinomas originating from a particular epithelial tissue form detectable metastases in only a limited subset of theoretically possible distant organ sites. Shown here are the most common sites of metastasis for six well-studied carcinoma types. Primary tumors are depicted in red. Thickness of black lines reflects the relative frequencies with which a given primary tumor type metastasizes to the indicated distant organ site.

Tumor Metastasis: Molecular Insights and Evolving Paradigms Valastyan, Scott; Weinberg, RobertA. Cell doi:10.1016/j.cell.2011.09.024 (volume 147 issue 2 pp.275 - 292)

CTC arrest and Extravasation

Steps of Extravasation: 1) Transient Adhesion of TC(tumor cell) to EC(Endothelial cell)-involves endothelial adhesion molecules E and P selectins. This step is associated with rolling 2) This step causes even more firm adhesion 3) The TC slips through endothelial cell-cell junction

Fig 6: Simplistic mechanism of Extravasation

9

In-vitro Extravasation model- Factors affecting TC transmigration

-48 well flow migration Boyden chamber used Variables that are quantified: -PMN tethering freq is determined(which is normalized against cell flux on surface) -Total no. of tethered PMNs(polymorphonuclear neutrophils) -No. of collisions b/w TCs and tethered PMNs -Aggregation of TCs with tethered PMNs as a result of collisions -Final attachment of aggregates with EC

10

Effect of PMN, ICAM-1 and E-selectin on cell migration

Fig 7 : All cases are at flow shear stress of 0.4 dyn/cm2 C8161, WM9, WM35 are type of cancer cells from cell lines ICAM 1- Intercellular adhesion molecule (a) Effect of addition of PMN to TC cell supension (b) Effect of blocking I-CAM1 and E-selectin
Dong, C., 2011. Adhesion and Signaling of Tumor Cells to Leukocytes and Endothelium in Cancer Metastasis Studies in Mechanobiology, Tissue Engineering and Biomaterials 4: 477-521 11

Effect of fluid shear on tumor cell(TC) migration

Either shear rate() or shear stress (=) was kept constant, while other one varied by changing .

Fig 8 : (a)TC migration varies under constant shear stress but increasing shear rate [(b), (c)]migration does not change for changes in shear stress data

Conclusion:-PMN facilitated migration is affected by shear rate and not by shear stress
Dong, C., 2011. Adhesion and Signaling of Tumor Cells to Leukocytes and Endothelium in Cancer Metastasis Studies in 12 Mechanobiology, Tissue Engineering and Biomaterials 4: 477-521

Receptor-ligand mechanics
-kon governs the likelyhood of receptor to form bond with ligand on other cell

AL - available surface area for receptor on ligand bearing cell nL - no. of ligands of cell nB - no. of bonds already formed distance b/w two cells kon Association rate for receptor-ligand binding under zeroforce condition ts bond spring constant - equilibrium length
13

Mathematical model for tissue invasion

Hybrid discrete-continuum model for tissue invasion
Assumptions -Invasion triggered by peripheral cancer cell-ECM contact - Amount of molecules interacting is large enough - Cells are considered discrete particles About the model Two scale approach-Intracellular environment affects the extracellular environment Continuum part of model describes chemical-ECM interactions The discrete part models the individual cells
14

Chemical-ECM environment
Cancer cells in contact with the protein network release MMPs

ECM is modified by degradation[Fig 9 ]

Change in adjacent stroma configuration

Stimulates cells to migrate via chemotaxis and haptotaxis

Protein network-cell interaction causing mitosis via GF absorbtion in degraded ECM

15

Chemical-ECM environment
Equations governing enzymes interactions with adjacent stroma
E(x,t) Conc. Of MMPs/uPAs(Urokinase plasminogen actovator) 2 M(x,t)- Density of ECM Where N(x, t) is the no. of cells at time t in surrounding neighbourhood such that

A(x,t) Density of degraded ECM, in which cells can absorb GFs B(x) - ball of radius , centered at xi 1 - Instantaneous local enzyme production
2- Diffusion of enzymes in surrounding environment
16

Chemical-ECM environment

Fig 9 : Change in ECM density as it is degraded by a single cell placed on a petri dish
17

Chemical-ECM environment

Fig 10 : Plot of concentration profile overtime of chemoattractants and GFs as they are released from the ECM
18

Cell modelling
Cells are assumed as free interacting particles in 2d space Two cells interact with each other according to a potential energy function(represents cell-cell adhesion) Potential energy of cell-cell bond at time t is given as h->capacity to bond

Ignacio Ramis-Conde, Mark A.J. Chaplain, Alexander R.A. Anderson, Mathematical modelling of cancer cell invasion of tissue, Mathematical and Computer Modelling, Volume 47, Issues 5-6, March 2008, Pages 533-545, ISSN 0895-7177, 19 10.1016/j.mcm.2007.02.034.

Intercellular Adhesion as a Potential function

Fig 11 : (Left)Interaction energy between two cells separated by a distance x(scaled relative to radius of avg cancer cell. (Right) Interaction energy between two cells in a 2D domain
Ignacio Ramis-Conde, Mark A.J. Chaplain, Alexander R.A. Anderson, Mathematical modelling of cancer cell invasion of tissue, Mathematical and Computer Modelling, Volume 47, Issues 5-6, March 2008, Pages 533-545, ISSN 0895-7177, 20 10.1016/j.mcm.2007.02.034.

Modelling cell movement

-Cells movement is governed by potential function
-In absence of any interaction with ECM motion is decided solely by

- Cells are assumed to move at constant speed - In case of cell-ECM interactions, cell movement is also affected by chemoattractant gradients

21

Cell mitosis
Two causes-Autocrine stimulus and cell-GF interaction

R(xi , t) Probability function for mitosis rate P0 Mitosis rate caused solely by autocrine stimulus
22

Tumor growth

Fig 12:Evolution of cancer cells as they invade the ECM

23

References
[1] Tumor Metastasis: Molecular Insights and Evolving Paradigms Valastyan, Scott; Weinberg, RobertA. Cell doi:10.1016/j.cell.2011.09.024 [2] http://www.oncoursesystems.com/school/webpage.aspx?id=10845583&xpage=789375 [3] http://theoncologist.alphamedpress.org/content/14/11/1070/F1.expansion [4] http://www.springerimages.com/Images/Biomedicine/1-10.1007_s12307-008-0007-2-4 [5] Dong, C., 2011. Adhesion and Signaling of Tumor Cells to Leukocytes and Endothelium in Cancer Metastasis Studies in Mechanobiology, Tissue Engineering and Biomaterials 4: 477-521 [6] Ignacio Ramis-Conde, Mark A.J. Chaplain, Alexander R.A. Anderson, Mathematical modelling of cancer cell invasion of tissue, Mathematical and Computer Modelling, Volume 47, Issues 5-6, March 2008, Pages 533-545, ISSN 0895-7177, 10.1016/j.mcm.2007.02.034. [7] Subra Suresh, Biomechanics and biophysics of cancer cells, Acta Biomaterialia, Volume 3, Issue 4, July 2007, Pages 413-438, ISSN 1742-7061, 10.1016/j.actbio.2007.04.002. [8] Yangjin Kim, Magdalena A. Stolarska, Hans G. Othmer, The role of the microenvironment in tumor growth and invasion, Progress in Biophysics and Molecular Biology, Volume 106, Issue 2, August 2011, Pages 353-379, ISSN 0079-6107, 10.1016/j.pbiomolbio.2011.06.006. [9] Simone Spaderna, Otto Schmalhofer, Falk Hlubek, Geert Berx, Andreas Eger, Susanne Merkel, Andreas Jung, Thomas Kirchner, Thomas Brabletz, A Transient, EMT-Linked Loss of Basement Membranes Indicates Metastasis and Poor Survival in Colorectal Cancer, Gastroenterology, Volume 131, Issue 3, September 2006, Pages 830-840, ISSN 0016-5085, 10.1053/j.gastro.2006.06.016. [10] Brian Elenbaas, Robert A. Weinberg, Heterotypic Signaling between Epithelial Tumor Cells and Fibroblasts in Carcinoma Formation, Experimental Cell Research, Volume 264, Issue 1, 10 March 2001, Pages 169-184, ISSN 0014-4827, 10.1006/excr.2000.5133. [11] N. BELLOMO. ON THE FOUNDATIONS OF CANCER MODELLING: SELECTED TOPICS, SPECULATIONS, AND PERSPECTIVES. Mathematical Models and Methods in Applied Sciences Vol. 18, No. 4 (2008) 593646c World Scientific Publishing Company [12] http://topnews.us/content/221000-pancreatic-cancer-and-circulating-tumor-cells-linked-each-other

24

25