Modelling Pressure Induced Cell Movement and
Tumour Invasion in a Cellular Automata
Darren Hassan
This report is submitted as partial ful llment of
the requirements for the Master of Computer Science Programme of the
School of Computer Science and Software Engineering
The University of Western Australia
2010
Abstract
Inter-cellular pressure has a signi cant eect on a tumour's growth and
malignancy. When the cells in one region of a tumour proliferate more
quickly than the cells in another, due to dierences in nutrient concen-
tration, density dierences result. Density dierences lead to a build-up
of pressure that pushes cells from densely populated regions to sparsely
populated regions. This pushing eect grows the tumour outward, into
the surrounding healthy tissue. Inter-cellular pressure therefore causes
pressure-induced cell movement that promotes tumour invasion and should
thus be included in a model of tumour growth.
However, it is inherently di cult to model inter-cellular pressure in
traditional Cellular Automata (CA) tumour models. Traditional CA
tumour models are formulated on a xed, regular lattice where each
lattice site represents a cell. The restrictions imposed by the model's
lattice require a dividing cell be near an empty site in which the new cell
can be placed. It is not possible for density dierences to develop in the
bulk of the tumour, when only cells near empty sites can divide.
A CA tumour model that captures realistic inter-cellular pressure has
been developed. This was achieved by introducing local cellular density
information into the model by allowing several cells to occupy a lattice
site. Based on physical principles and porous media theory, a relation-
ship between cellular density and inter-cellular pressure is proposed that
has allowed us to model pressure-induced cell movement and the inho-
mogeneous diusion of nutrient molecules.
When applied to in vitro conditions, the model has successfully sim-
ulated tumour growth that exhibits the emergence of the typical layered
structure with expanding radius seen experimentally.
Keywords: Tumour model, mathematical model, Cellular Automata,
pressure-induced cell movement
CR classi cation: I.6.5 [Computer Methodologies]: Simulation and
Modeling - Model Development
2
Acknowledgements
I would like to acknowledge the contribution of the project's supervi-
sors, Dr Pascal Buenzli and Dr Peter Pivonka of the Engineering and
Computational Biology Group, School of Computer Science and Soft-
ware Engineering, The University of Western Australia.
3
Contents
1 Introduction 10
1.1 Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
1.2 On the Nature of Mathematical Models . . . . . . . . . . 11
1.3 Why Model? . . . . . . . . . . . . . . . . . . . . . . . . . 12
1.4 Models in Biology . . . . . . . . . . . . . . . . . . . . . . 13
1.5 Types of Mathematical Models . . . . . . . . . . . . . . 13
1.6 Thesis Motivation . . . . . . . . . . . . . . . . . . . . . . 15
2 A Biological Background to Cancer 18
2.1 Cells, Genes and DNA . . . . . . . . . . . . . . . . . . . 18
2.2 Cancer's Cause . . . . . . . . . . . . . . . . . . . . . . . 20
2.3 Malignant Tumour Development . . . . . . . . . . . . . . 21
3 Literature Review 23
3.1 Brain Tumour Growth . . . . . . . . . . . . . . . . . . . 23
3.2 Breast Tumour Growth . . . . . . . . . . . . . . . . . . . 24
3.3 Tumour Evolution . . . . . . . . . . . . . . . . . . . . . 25
3.4 Tumour Evolution and Inheritance . . . . . . . . . . . . 25
3.5 Avascular Tumour Growth . . . . . . . . . . . . . . . . . 26
3.6 Tumour Metabolism . . . . . . . . . . . . . . . . . . . . 27
4 Model Design 29
4.1 Modelling Cells . . . . . . . . . . . . . . . . . . . . . . . 30
4.1.1 A Biological Background to Cell Metabolism . . . 31
4.1.2 Modelling Cell Metabolism . . . . . . . . . . . . . 32
4.1.3 Cell Proliferation . . . . . . . . . . . . . . . . . . 33
4.2 Inter-Cellular Pressure . . . . . . . . . . . . . . . . . . . 35
4.3 Modelling Cell Migration . . . . . . . . . . . . . . . . . . 42
4.3.1 Introduction . . . . . . . . . . . . . . . . . . . . . 42
4.3.2 Modelling Pressure-Induced Cell Movement . . . 43
4.3.2.1 Displacement in 1D . . . . . . . . . . . 44
4
4.3.2.2 Displacement in 2D . . . . . . . . . . . 45
4.4 The Chemical Mass-Balance Equations . . . . . . . . . . 46
4.4.1 Glucose . . . . . . . . . . . . . . . . . . . . . . . 46
4.4.2 Oxygen . . . . . . . . . . . . . . . . . . . . . . . 48
4.5 Modelling Chemical Diusion . . . . . . . . . . . . . . . 49
4.5.1 The Adiabatic Approximation . . . . . . . . . . . 51
5 Results 52
5.1 Simulation Parameters . . . . . . . . . . . . . . . . . . . 52
5.2 Simulation Results . . . . . . . . . . . . . . . . . . . . . 53
5.3 Comparison with in vitro Experiments . . . . . . . . . . 65
6 Conclusion 67
7 Appendices 73
7.1 An Alternative Relationship Describing Cell Volume as a
Function of Pressure . . . . . . . . . . . . . . . . . . . . 73
7.2 Discretising a Continuous Chemical Concentration . . . . 79
7.3 The Adiabatic Approximation . . . . . . . . . . . . . . . 80
7.3.1 A Theoretical Analysis of the Adiabatic Approxi-
mation . . . . . . . . . . . . . . . . . . . . . . . . 80
7.3.2 A Numerical Analysis of the Adiabatic Approxi-
mation Limit . . . . . . . . . . . . . . . . . . . . 81
5
List of Tables
4.1 The metabolic threshold values. . . . . . . . . . . . . . . 33
4.2 Glucose mass-balance equation's parameters. . . . . . . . 47
4.3 Oxygen mass-balance equation's parameters. . . . . . . . 48
5.1 The simulation's parameters. . . . . . . . . . . . . . . . 52
7.1 A description of parameters describing a spherical cell's
shape as a function of pressure. . . . . . . . . . . . . . . 73
7.2 The glucose steady state time in hours. . . . . . . . . . . 82
7.3 The oxygen steady state time in hours. . . . . . . . . . . 82
6
List of Figures
2.1 The idealised avascular tumour's structure. . . . . . . . . 22
4.1 A simpli ed representation of the model, viewed from above. 29
4.2 The model's assumed lattice structure, reproduced from
[30]. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
4.3 A cell's metabolic response to chemical stimuli described
as a phase diagram. . . . . . . . . . . . . . . . . . . . . . 33
4.4 The cell volume as a function of pressure, described by
Equation (4.11). The vertical and horizontal grey lines
denote the IFP of normal tissue and intrinsic volume re-
spectively. . . . . . . . . . . . . . . . . . . . . . . . . . . 37
4.5 The regular packing of spheres in 3D. The left-hand side
describes the hexagonal close-packing formation, whereas
the right-hand side describes the face-centred cubic pack-
ing [42]. . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
4.6 The density-porosity relationship for densities greater than
the critical density. . . . . . . . . . . . . . . . . . . . . . 40
4.7 The relationship between ρ̄ and a(ρ̄). . . . . . . . . . . . 40
4.8 The average inter-cellular pressure as a function of density. 41
4.9 Migration of cells due to pressure inhomogeneities between
lattice sites. Cells in the light grey-shaded area are moved
to the dark-shaded area during the time increment 4t.
The cell counts in each lattice site are updated accordingly. 44
4.10 Migration of cells due to pressure inhomogeneities between
the sites of a 2D lattice. Cells in the light grey-shaded area
are moved to the dark grey-shaded area during the time
increment 4t. The cell count in each lattice site is updated
0
accordingly, as the −0 and
0
+0 signs suggest. . . . . . . . 45
7
5.1 The cross-sections describe the tumour's growth with the
original metabolic thresholds. Going from top to bottom,
left to right, the cross-sections describe the tumour at day
ten, twelve, fourteen, sixteen, eighteen, nineteen, twenty
and twenty-one. . . . . . . . . . . . . . . . . . . . . . . 53
5.2 The new metabolic phase diagram describing a cell's metabolic
response to chemical stimuli. . . . . . . . . . . . . . . . . 54
5.3 The tumour's composition at 10 days. . . . . . . . . . . . 55
5.4 The tumour's composition at twelve days. . . . . . . . . 55
5.5 The tumour's composition at fourteen days. . . . . . . . 56
5.6 The tumour's composition at sixteen days. . . . . . . . . 56
5.7 The tumour's composition at eighteen days. . . . . . . . 57
5.8 The tumour's composition at nineteen days. . . . . . . . 57
5.9 The tumour's composition at twenty days. . . . . . . . . 58
5.10 The tumour's composition at twenty-one days. . . . . . . 58
5.11 A density plot of the PAT layer at twenty-one days, viewed
from above; cells in the other metabolic states are not shown. 59
5.12 The change in size of the tumour's metabolic populations
with time. . . . . . . . . . . . . . . . . . . . . . . . . . . 59
5.13 The change in size of the tumour's metabolic populations
with time, plotted on a logarithmic scale. . . . . . . . . . 60
5.14 The growth in tumour radius with time. . . . . . . . . . 61
5.15 The growth in tumour radius with time, plotted on a log-
arithmic scale. . . . . . . . . . . . . . . . . . . . . . . . . 62
5.16 The change in maximum cellular density with time. . . . 62
5.17 The change in tumour radius with time, for a maximum
site density of 300 (cells)(100 µm)−3 , plotted on a loga-
rithmic scale. . . . . . . . . . . . . . . . . . . . . . . . . 64
5.18 The change in cellular population with time, for a max-
imum site density of 300 (cells)(100 µm)−3 , plotted on a
logarithmic scale. . . . . . . . . . . . . . . . . . . . . . . 64
7.1 The change in cell volume with spontaneous curvature. . 75
7.2 Cell volume as a function of pressure, according to Equa-
tion (7.8). . . . . . . . . . . . . . . . . . . . . . . . . . . 76
7.3 Cell volume as a function of pressure, according to Equa-
tion (7.8) for a large range of pressure values. . . . . . . 76
8
7.4 A comparison of the alternative approaches for describing
cell volume as a function the pressure experienced on its
membrane. The green line denotes the inverse relationship
between cell volume and pressure, described by Equation
(4.11). The blue lines denote the volume of an empty lipid
bi-layer as a function of pressure, described by Equation
(7.1). . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
7.5 The average inter-cellular pressure as a function of density,
from Equation (7.4), assuming cells deform under increas-
ing pressure. . . . . . . . . . . . . . . . . . . . . . . . . 78
7.6 A comparison of the alternative approaches for describing
average inter-cellular pressure as a function of density. The
green line denotes the pressure as a function of density for
the inverse relationship between cell volume and pressure,
described by Equation (4.11). The blue lines denote the
pressure as a function of density for an empty lipid bi-
layer, described by Equation (7.1). . . . . . . . . . . . . 78
7.7 The steady state contours for the glucose concentration eld. 83
7.8 The steady state contours for the oxygen concentration eld. 83
9
Chapter 1
Introduction
1.1 Systems
Systems are ubiquitous throughout nature, and seemingly integral to life
itself. At this moment systems are at work in your body to maintain
its homoeostatic state. Systems also result from human endeavour, some
preordained such as a mechanical clock, while others seem to emerge
independently such as the economy of a country.
A system is commonly de ned as a set of independent, interacting en-
tities that together form an integrated whole. They share several common
characteristics: a structure, inter-connectivity and behaviour. Systems
are composed of parts that are structured in an ordered way. The inter-
connectivity of the various parts of a system have a functional as well as
a structural relationship. The relationships between the individual parts
of a system result in system wide behaviour.
System complexity is the most important factor in our ability to un-
derstand and predict a system's behaviour. There is no consensus on
the de nition of a complex system. Boccara, in his 2004 book Modeling
Complex Systems [4] lists three characteristic he considers necessary for a
system to be considered complex: a system must consist of a large num-
ber of interacting agents, the system must exhibit emergent behaviour,
the emergent behaviour must not result from the existence of a central
controller. Boccara de nes emergent behaviour as the large scale eects
of locally interacting agents that are often surprising and hard to predict
even in the course of simple interactions. An example of an emergent
property would be the cooperative behaviour of ants in a colony that
results from the local interactions between its members and not from the
existence of a central controller.
10
1.2 On the Nature of Mathematical Models
Philosophically, the universe itself can be considered the most complex of
systems, where it is reasonable to suppose that everything, to a greater
or lesser degree, has a causal in uence over everything else. However,
understanding anything would be impossible from such a holistic view
point.
Wheldon, in his 1988 book Mathematical Models in Cancer Research
[41], describes the nature of a mathematical model derived from a scien-
ti c theory. He explains that the understanding of any phenomenon relies
on simpli cation, the picking out of the causal in uences essential to un-
derstanding the phenomenon. Once the dominant causal in uences have
been identi ed, they may be embodied as a set of assumptions stating
how they are believed to operate. If those assumptions form a connected
whole that deal with important questions for the subject concerned, that
collective can be described as a theory. If the theory is quantitative in na-
ture, a mathematical model can be developed describing how the causal
in uences combine to produce the phenomenon of interest.
Mathematical models of complex systems are created in a similar
way to the mathematical models of scienti c theories. In fact, the phe-
nomenon a scienti c theory seeks to explain, can be considered the be-
haviour of a complex system on some level. Understanding a complex
system relies on simplifying reality into an idealised system that can be
better understood and manipulated. An idealised system is formed by
considering only those entities that have a dominant in uence over the
system. These essential entities are usually determined through exper-
imentation. Once the essential entities have been identi ed and their
relationships understood that knowledge can be encapsulated in a set
of assumptions. If those assumptions completely describe an idealised
system and can explain the system's behaviour observed in reality, those
assumptions may be collectively known as a theory. It is possible to un-
derstand the system in a quantitative way when the theory is concerned
with measurable quantities and changes in some way, as a result of causal
in uences.
When the system is understood quantitatively, mathematical formu-
lation of the assumptions is suitable. The assumptions are said to con-
stitute a mathematical model that explains how the causal in uences
combine to produce the system's behaviour observed. A mathematical
model is therefore an exact form of a scienti c theory, with particular
11
focus on the theory's quantitative aspects. A theory in turn, is a simpli-
ed representation of a complex system that captures those entities and
their relationships essential to understanding the system's behaviour. If
the assumptions are loosely de ned, as with many complex systems, it is
possible for several models to be compatible with one scienti c theory.
1.3 Why Model?
The main reason to create a mathematical model is to discover precisely
what phenomena ought to occur if the causal in uences taken as domi-
nant are the only ones present. If the assumptions regarding dominant
causal in uences are encapsulated in a scienti c theory, a model allows
derivation of the theory's implications. When a model captures those
aspects believed to be essential to a complex system, the model allows
discovery of the idealised system's behaviour. The implications of a sci-
enti c theory or description of an idealised system's behaviour may be
sought for many dierent reasons.
Karl Popper, in his 1963 book Conjectures and Refutations: The
Growth of Scienti c Knowledge [31], described his philosophy of science:
Critical Rationalism. Popper believed scienti c theories to be abstract
in nature, and only testable indirectly, via their implications. The truth
of a scienti c theory, he contended, could only be con rmed, but never
proved, by comparing its implications to reality. However, it is possible,
and in fact essential for a theory to be falsi able for it to be consider
scienti c. The modelling of a theory describing the workings of a complex
system is often the only way that theory's implications can be discovered.
When a theory's implications can only be discovered by a mathematical
model, the mathematical model is the only tool we have for con rming
or refuting the underlying scienti c theory.
Mathematical models are integral to the scienti c process. Science
progresses through a cyclic methodology. A theory is developed to ex-
plain a phenomenon. A mathematical model of the theory is then created.
The mathematical model allows derivation of the theory's implications,
and the experimental testing of those implications. If the implications
are found not to agree with reality, the scienti c theory is refuted in
its original form. The theory may then be abandoned, or modi ed to
improve its predictive powers before embarking on the process again.
12
1.4 Models in Biology
In biology, models can serve other roles with more medical relevance. A
medical practitioner will use models when attempting to treat a patient.
The appropriate course of action for the medical practitioner will depend
on the model used. For example, the broad theory that some diseases may
be caused by invading micro-organisms implies that attempts should be
made to kill the micro-organisms concerned. This is a recent idea, when
disease was thought to be caused by a plethora, or overabundance, in the
blood, then bloodletting would be seen as the correct course of action.
Historically, biological models have been qualitative in nature, such
as that of diseases being caused by micro-organisms. The limited use of
mathematical models in biology is in part due to the nature of biological
theories. Wheldon [41] expressed what was needed for a phenomenon to
be amenable to mathematics, when he stated 'the phenomenon of inter-
est should involve measurable quantities for which the main interest of
the subject consists in the magnitudes of the quantities or the relation-
ship between them'. Biological theories have tended to be based upon
composite quantities, such as a species, or an organism. These are enti-
ties with a myriad of dierent qualities, not amenable to a mathematical
description or manipulation.
Recently, in the eld of oncology, a biological unit fundamental to the
topic and amenable to mathematics has emerged: the cancer cell. Cancer
researchers are now asking: how do cell populations change? how does
a tumour's radius grow? Mathematics has the potential to answer these
and many other questions of this type.
1.5 Types of Mathematical Models
Mathematical models can be classi ed in many dierent ways. In the
context of this thesis, we are interested in the dichotomy between con-
tinuous and discrete mathematical models. This thesis will describe the
development of a cellular automata (CA) that models both continuous
and discrete phenomena and can therefore be considered a hybrid of the
two. An understanding of our model will be improved through an exam-
ination of models from both categories.
Continuous models deal with average quantities that have been shown
representative of phenomena by statistical and stochastic theories. Con-
tinuous models are usually formulated in terms of a system of dierential
13
equations. A dierential equation is an equation relating some function
to one or more of its derivatives of various orders. Dierential equations
occur in many areas of science and technology, whenever a determinis-
tic relation involves some continuously varying quantities (modelled by
functions) and their rates of change in space and time (expressed as
derivatives).
Dierential equations can be categorised as Ordinary dierential equa-
tions (ODEs) or Partial dierential equations (PDEs). An ODE is a rela-
tion that contains functions of only one independent variable, and one or
more of their derivatives with respect to that variable. Models consisting
of a system of ODEs, whose independent variable is time, ignore local,
spatial interactions, replacing them with uniform long-range ones. Such
models are rather a crude representation of complex systems. A PDE is
a relation that contains function of two or more independent variables,
and one or more of their derivatives with respect to those variables. Sys-
tems of PDEs can be formulated with respect to space and time, thus
enabling the modelling of local interactions over time, resulting in a more
sophisticated representation.
In contrast, discrete models deal with discrete quantities. A discrete
model of a complex system explicitly models those entities believed to
have a dominant in uence, and the causal relationships between them,
though in a simpli ed way.
CAs are a class of discrete model that were rst proposed by John
Von Neumann [27]. CAs are formulated on a spatial lattice, which can
be considered a type of graph where the lattice sites are represented by
graph nodes and the relationships between lattice sites are represented by
edges. The dynamics of the model are de ned by local rules of interaction
among neighbouring nodes, or adjacent lattice sites. The rules governing
interactions can be deterministic or stochastic. In cancer modelling, each
lattice site, of a traditional CA, represents a cell.
CAs can be used to study phenomena where agents, each following
a set of simple rules results in the sophisticated emergent behaviour of
the whole system. A CA's ability to replicate emergent behaviour makes
them suitable for modelling self organising, complex systems, such as
cancer.
One of the main advantages of an CA for modelling biological systems
is the ease with which dierent populations of cells can be modelled. By
assigning internal state variables to the sites depending on cell type a
heterogeneous population of cells can be modelled. The fact that cancer
14
arises from mutations in single cells and the ability to incorporate cell-
based data into the model makes discrete models well suited to modelling
cancer [26].
1.6 Thesis Motivation
It is our long-term aim to investigate prostatic cancer growth in bone,
by creating a CA model of the bone environment. During the last stage
of a malignant prostate tumour's development, which will be described
in more detail in Chapter 2, prostate metastases migrate to the bone via
blood vessels in the bone marrow. There are large dierences in stiness
between the mineralised bone and marrow. In-order to account for dif-
ferences in stiness we would need to model pressure. From a review of
the literature, it appears pressure is yet to be modelled by a traditional
CA, which may be due to the di culties traditional CA's have modelling
a representative cellular density. We intend to rst create a computa-
tional model of an in vitro tumour. Modelling an in vitro tumour will
allow us to ignore many of the complications involved in modelling the
bone environment, and instead focus on re ning a technique for mod-
elling pressure-induced changes in cellular density. The wealth of models
and experimental data describing in vitro tumours will provide ample
opportunity to compare and test our model's results. Opportunities to
extend the model to the bone environment will be explained to the reader
as they are encountered throughout this thesis.
A cancer cell's movement is governed by a pressure-induced force
transferred through the cells it is in contact with and chemotaxis. Chemo-
taxis is a phenomenon in which a cell directs its movement according to
chemicals within its environment. Cancer cells have been known to move
from low to high oxygen and glucose regions. However, we have decided
to ignore chemotaxis in the rst iteration of the model, and instead
focus on movement caused by pressure gradients. A pressure-induced
force, experienced by a cell on its membrane, will be related to the cellu-
lar density in its vicinity. If a cell experiences a pressure gradient across
its membrane, due to a dierence in density, the cell's movement will be
directed down that pressure gradient. Of the CA models reviewed, all of
them ignored pressure. It is inconvenient to de ne a smooth local density
in a traditional CA model as it requires averaging over a volume large
enough to comprise many cells, resulting in a loss of resolution.
Modelling cell movement due to proliferation is inherently di cult
15
in traditional CA models with a xed lattice. On a xed lattice, cell
division is taken to occur instantaneously, resulting in two cells where
there was one. One of the cells usually occupies the original site of the
dividing cell, but the other requires an extra space, and so must move
to a dierent site. Commonly, CA models, such as that by Kansal et
al. [23], place the new cell on an empty site within the dividing cell's
neighbourhood. If an empty site cannot be found the dividing cell is pre-
vented from proliferating. Rules such as this, fail to model the 'pushing'
eect caused by a buildup in cellular density. We intend to replace this
algorithmic prescription with a more realistic scheme grounded in the
physical considerations of the local pressure experienced by a cell.
We believe pressure-induced force has a dominant in uence over a
tumour's malignancy, and is therefore deserving of inclusion within our
model. I will now outline our approach, which will be described in more
detail in Chapter 4. We propose to model the tumour at the mesoscopic
scale, between that of the micro (≈
10 µm), in which individual cells are
2
modelled and that of the macro (≈ 10 mm ), in which average values for
the whole tumour are modelled. We intend to model packets of cells at
each site, where a packet is a cube of known volume and cell population.
To my knowledge, Piotrowska and Angus [30] were the rst to propose
this idea within the context of an in vitro tumour model. However, this
thesis intends to take this idea to the next level, by including the following
improvements:
(i) When modelling packets of cells, the volume over which the density
is measured becomes a model parameter. By selecting an appropriate
packet size a representative cellular density can be calculated. The in-
clusion of a representative cellular density within our model has many
bene ts. With an appropriate scheme, which will be described in Chap-
ter 4, the density of a packet of cells can be used to calculate the average
pressure experienced by the cells within the packet; information that can
then be used to model cell movement due to pressure inhomogeneities.
Modelling packets will also allow us to implicitly move cells, by subtract-
ing a cell from one packet and adding a cell to another, thus simplifying
the practicalities of cell movement.
(ii) In reality, the diusion of a substance into the tumour is aected
by the density of the tissue through which the substance is diusing.
By neglecting to model density dierences, traditional CA models have
16
assumed the homogeneous diusion of chemicals. The calculation of a
representative cellular density for a packet of cells, will enable the calcu-
lation of that packet's porosity. A packet's porosity can then be used to
modify the diusion of chemicals through that packet, with a technique
adapted from porous media transport theory. When this technique is
applied to all the packets within our model, the inhomogeneous diusion
of chemicals through the tumour can be modelled; further enhancing our
model's realism.
As with most design decisions, the modelling of packets of cells as
opposed to individual cells does have its drawbacks. The modelling of
packets of cells will result in the loss of cellular information. We will
no-longer be able to track individual cells within the model. This would
present a problem if, for instance, we were attempting to model the
tumour's sub-clonal evolution. However, at this stage, it is phenomena
at the mesoscopic scale that we are interested in, such as the tumour's
morphology, radius, and total population. These are properties that are
accessible, regardless of whether the state of every cell is known or not.
17
Chapter 2
A Biological Background to
Cancer
This chapter will discuss those aspects of cancer biology that form the
background to this thesis.
We, and all other higher species, are composed of millions of cells that
cooperate to produce a composite entity. The cells of our body dier in
their form and function depending on the role they play in maintaining a
homoeostatic state. Cooperation amongst cells is maintained by a num-
ber of checks and balances, some external to the cell, such as chemical
signals, whereas others are internal to the cell. The failure of any single
cell to cooperate does not necessarily have a systemic eect. However,
when a cell gains control over its reproductive behaviour, producing in-
de nite numbers of progeny, the survival of the organism, to which that
cell belongs, can be threatened. As will be seen, cancer is a complex dis-
ease, but at its essence it can be described as the acquisition of autonomy
by a cell, in a multi-cellular organism.
2.1 Cells, Genes and DNA
A cell can be thought of as a chemical automata, or biochemical factory,
whose behaviour is programmed by the genes it contains. The genome
refers to the entirety of a cell's genetic information. Genes provide in-
structions of the sort encountered in a computer programming language.
As with computer programs, genes may be arranged in packages to pro-
vide whole sets of complex instructions, similar to a computer program's
functions. A gene tells the cell to make a protein of a particular type, by
assembling it from its constituent parts (amino acids). The protein will
18
then participate in the biochemistry of the cell according to its chemical
nature. There are many dierent types of proteins, each with a particu-
lar function. Enzymes, for example, control the rate at which chemical
reactions occur within the cell, other proteins constitute cellular building
material, while other proteins act as signals for communication amongst
cells.
Like a computer program, genetic instructions require a physical em-
bodiment. The genetic instructions of all organisms on Earth are em-
bodied and encoded in the same way; which strongly suggests a common
ancestry for all life. The genetic instructions in all species are encoded
from small molecules (nucleotides) of four types, strung together to form
a long 'supermolecule' called deoxyribonucleic acid (DNA). It is the or-
der in which the nucleotides occur that dictates the code. Each group of
three nucleotides provides an instruction specifying a particular amino
acid. A gene is a collection of these instructions that tells the cell to put
together amino acids to form a protein.
The execution, by the cell, of a gene instruction proceeds in two
phases. In the rst phase, known as transcription, the genetic instruc-
tions are recorded in a slightly dierent chemical form that results in an-
other long molecule, called ribonucleic acid (RNA). The genetic instruc-
tions contained within DNA can be thought of as being in a high-level
programming language, while the transcription into RNA converts the
high-level language into machine code, from which the cellular machin-
ery can act more directly. In the second phase, known as translation, the
genetic instructions are executed to assemble the amino acids to form the
appropriate protein. The reading and execution of genetic instructions
is a highly complex process that has been greatly simpli ed here.
Each step of transcription and translation are prone to error, such as
when instructions are not carried out correctly. However, these errors
are insigni cant provided the genetic instructions or DNA remain intact,
the opportunity will exist to assemble the protein correctly next time the
instructions are executed. It is when the instructions have to be copied,
during cell division, that the genetic memory is at its greatest risk.
The cell cycle is a series of events that take place in a cell leading to
its division. During the rst phase of the cell cycle (interphase), the cell
enlarges by intake of the nutrients needed for cell division (mitosis) and
duplicates its DNA. During mitosis the cell splits itself into two distinct
'daughter' cells. Under ideal circumstances, the two daughter cells are
identical to each other and miniature replicas of the parent cell whose
19
ssion created them.
2.2 Cancer's Cause
Within biology, it is commonly believed that a cell's behaviour (pheno-
type) is determined by the cell's genetic instructions (genotype). Within
this context, it can be understood that corruption of a cell's DNA or geno-
type can lead to that cell having a cancerous phenotype. Speci c genes
ensure uncontrolled growth is prevented. When these genes are mutated,
cells become prone to developing a cancerous phenotype. These genes
can be broadly divided into three categories [37]: oncogenes, tumour
suppressor genes and repair genes.
A multi-cellular organism is composed of many dierent types of spe-
cialised cells that are arranged to form tissues and organs. Most tissues
and organs have a recognisable physiological function for which the cells
have adapted by evolution. Depending on their function, some of these
cells will need to be continually regenerated, such as skin (epithelial tis-
sue). Oncogenes promote the proliferation of cells in the presence of
appropriate growth signals that leads to the regulated regeneration of
such cells. When oncogenes become mutated they induce the cell to
divide continuously, irrespective of growth signals.
Tumour suppressor genes are responsible for stopping growth in nor-
mal tissue. Cell proliferation is stopped by either the repression of genes
that are essential for continuing the cell cycle (senescence), or by induc-
ing programmed cell death (apoptosis), such as when the cell's DNA has
been damaged. When tumour suppressor genes become inactivated, a
cell can proliferate uncontrollably.
In human cells, both normal metabolic activities and environmen-
tal factors such as ultraviolet radiation can cause DNA damage. This
damage can eliminate the cell's ability to transcribe the genes that the
aected DNA encodes. Other types of damage induce potentially harmful
mutations in the cell's genome, which aect the survival of its daughter
cells. Repair genes encode for proteins that work to repair damage to the
DNA. When normal repair processes fail, and apoptosis does not occur,
irreparable DNA damage may result. A cell whose DNA is irreparably
damaged may acquire autonomy leading to unregulated cell division.
20
2.3 Malignant Tumour Development
Cancer can loosely be de ned as a condition in which inappropriate cell
proliferation leads to an excess of cells (known as a tumour), which causes
disruption of normal tissue architecture. Not all tumours are described
as cancerous. Tumours, such as a wart, that are self-limiting in growth
and whose cells stay within the tumour are said to be benign. Malignant
tumours are those that seem to have no built-in limitations to growth and
whose cells are prone to spread to other parts of the body (metastasise).
Malignant tumours are said to be cancerous.
Once a cell has gained autonomy, and begun proliferating uncontrol-
lably, a clump of cancerous cells will form over time that is initially known
as an avascular tumour. Avascular tumours are fed by the diusion of
glucose and oxygen from the surrounding vascularture. An avascular tu-
mour's size is limited by its supply of glucose and oxygen. As tumour
cells accumulate, the diusion of glucose and oxygen becomes insu cient
to sustain all the cells within the tumour mass. The tumour will then de-
velop a layered structure that depends on the distribution of glucose and
oxygen; refer to Figure 2.1 for an idealised description of an avascular tu-
mour's structure. Cells undergo un-programmed cell death (necrosis) in
the low glucose and oxygen environment at the tumour's centre. Around
the necrotic core, a layer of quiescent cells will form where there is a
su cient glucose and oxygen concentration to sustain the cells without
dividing. On the tumour's outer edge, a rim of proliferating cells will
emerge where the glucose and oxygen concentration is highest.
For an avascular tumour to progress to the next stage it must ac-
quire its own blood supply. Cells under stressful conditions release pre-
angiogenic growth factors to drive angiogenesis, the process by which
existing blood vessels are stimulated to grow from the main circulatory
system to supply tissue with blood. The accumulation of necrotic cells at
an avascular tumour's core promotes the growth of blood vessels towards
the tumour.
Once the tumour has acquired its own blood supply it is said to be in
the vascular stage of development. Once the tumour is vascularised, it
can grow larger and shed cells into blood vessels. Tumour cells can now
escape the primary tumour site via the circulatory system and set up
satellite tumours in distant parts of the body (metastases). Metastasis
is the predominant cause of mortality due to cancer.
21
The quiescent layer.
The tumour's necrotic core.
The proliferating outer rim.
Figure 2.1: The idealised avascular tumour's structure.
22
Chapter 3
Literature Review
A diverse range of CA tumour models exist. Dierences in CA mod-
els are, primarily, a result of the choices made by the modeller regarding
those aspects of tumour growth deemed essential, and therefore deserving
of inclusion within the model. Often, the modelling of one phenomenon
over another necessitates a dierent approach. A representative set of
models have been reviewed that demonstrates dierent phenomena com-
monly modelled and typical approaches taken. The reviewed models
will brie y be introduced, followed by a critical analysis of the approach
employed.
3.1 Brain Tumour Growth
Kansal et al. [23] have developed a model of brain tumour growth. Tissue
is modelled using a three-dimensional (3D) lattice. Lattice sites take
on one of four distinct states corresponding to: proliferate, quiescent,
necrotic and healthy. The tumour is initially seeded in the centre of the
lattice and then allowed to evolve in space and time.
Site transition rules are non-local. A site's state depends on its loca-
tion within the tumour, for example, sites within a given distance of the
tumour's perimeter are proliferating. The authors select input param-
eters, such as the width of the proliferating rim, that quantitatively t
experimental situations. By deciding on the tumour's structure a priori,
through their choice of input parameters, the authors are prevented from
discovering new insights into the tumour's development. CAs should be
used to model systems at the microscopic level. It can then be discovered
what aect microscopic interactions have at the macroscopic level. The
transition rules were designed to model a nutrient gradient that dimin-
ishes towards the tumour's core. However, the model does not possess
23
a nutrient parameter, the tumour's morphology could just as easily be
attributable to other factors.
It appears the authors have built a simulation to t the Gompertz
function [16], a sigmoid function in which the right-hand asymptote of
the function is approached much more gradually by the curve than the
left-hand asymptote. The modellers have forgone an opportunity to im-
prove understanding and stimulate insights into cancer's development by
deciding to build a model to t the Gompertz function. The simulation
is too speci c and simplistic to be useful beyond its original purpose.
3.2 Breast Tumour Growth
A model that adheres more closely to CA convention is that by Pi-
otrowska and Angus [30], who have developed a two-dimensional (2D)
model of an in vitro tumour. Lattice sites represent a unit cube contain-
ing a packet of homogeneous cells. Sites can exist in one of ve dierent
states: proliferate, quiescent, in either the aerobic or anaerobic metabolic
form and necrotic. A site's state depends on the local pH, glucose, oxy-
gen and metabolic waste levels. Nutrient diusion into the tumour and
waste diusion out are modelled with a numerical diusion algorithm.
The modelled tumour's composition and morphology were validated by
comparison to experimental data derived from the literature, for a range
of cells per site ratios.
The many cells per site approach can be used to scale the model to
sizes beyond what would be computationally possible otherwise. Models
employing the more typical one cell per site approach, which require
a dividing site to have a free near neighbour to place the new cell, are
restricted to modelling proliferate rims one cell thick, whereas this model
can increase the size of the proliferate rim by increasing the number of
cells per site.
The many-to-one approach comes at the expense of model resolution.
Automaton updates occur at the level of the site, rather than the cellu-
lar level, resulting in cellular information loss. The individual dierences
between cells becomes signi cant when you consider the role sub-clonal
evolution plays in tumour malignancy [28]. By neglecting to model in-
dividual cells, the authors have chosen to ignore the eects of sub-clonal
evolution.
24
3.3 Tumour Evolution
A model that does take into account the eects of sub-clonal evolution, is
that by Anderson et al. [2]. Tumour cells are modelled as discrete entities
that occupy a single lattice site on a 2D lattice. Individual tumour cells
can have dierent genotypes that results in them having dierent pheno-
types. A cell's proliferation, movement and death potential depend on
the interaction between its genotype and local environmental conditions.
The tissue environment is described with Extracellular Matrix (ECM)
concentration (host tissue), Matrix Degrading Enzyme (MDE) concen-
tration and oxygen concentration. When a cell proliferates the new cell
randomly takes on a dierent genotype, which models mutations during
cell division, thus enabling the tumour's sub-clonal evolution.
The authors were not able to quantitatively validate the model, but
were able to show that it qualitatively resembles those tumours observed
experimentally by replicating experimental situations and observing that
the simulated tumour evolved in a similar way. In a harsh environment,
characterised by either an heterogeneous ECM or low oxygen concentra-
tion, the simulated tumour develops an invasive nger-like morphology
and is composed of cells with aggressive phenotypes, characterised by
having a high proliferation potential, high oxygen consumption rate, high
MDE production rate and high movement potential. The model supports
the clonal dominance theory [24] of cancer metastasis. The clone dom-
inance theory argues that metastasis occurs when the tumour becomes
dominated by invasive (aggressive) cells under harsh environments.
The model suggests that conventional techniques for combating can-
cer, such as radiotherapy, which increase environmental hostility, are
promoting tumour evolutionary development. A better approach would
be to make the environment benign and stable so as to limit selective
pressure and retard tumour development, such is the qualitative insight
brought about by modelling.
3.4 Tumour Evolution and Inheritance
The model by Gerlee et al. [15] can be thought of as an extension of [2].
Both models investigate the eect environment has on sub-clonal evolu-
tion and tumour morphology. The most signi cant dierence between
this model and the one previously reviewed, is the modelling of a tu-
mour cell's phenotype with a feed-forward neural network. Stimuli from
25
a cell's micro-environment are processed by the neural network to calcu-
late its phenotype. When a cell proliferates there is a small probability
that its daughter cell's neural network will be slightly dierent. In the
previous model, a new cell's phenotype may bear little resemblance to
that of its parent's. The modelling of slight changes in phenotype during
proliferation, better simulates the tumour's evolution, which evolves by
minor changes from one generation to the next. Over successive genera-
tions, those phenotypes with a selective advantage grow to dominate the
tumour.
The authors had originally intended to model many dierent aspects
of tumour growth, other than those directly eecting its evolution, such
as aerobic and anaerobic metabolism. The dierent types of metabolism
are only indirectly related to the subject under investigation. The au-
thors eventually came to the conclusion that the completeness, resulting
from modelling these additional aspects was not worth the additional
complexity they incurred. The authors gathered results using a minimal
model, which only considered aerobic metabolism.
The authors can only oer supportive evidence for the model by high-
lighting experimental results that agree with its predictions, which are
similar to those of [2]: harsh environments cause the tumour to form a
ngering morphology and achieve a high level of sub-clonal diversity.
3.5 Avascular Tumour Growth
Ferreira et al. [11] have created a model of avascular tumour growth.
Nutrients diuse across a xed, regular lattice from the rst row. Lattice
sites hold one normal or necrotic cell, or many tumour cells. Nutrient con-
sumption is proportional to site population. Cells become necrotic with
a probability dependent on the nutrient per cell concentration, which
models cell death caused by nutrient starvation. Cells in the tumour's
core exhibit a higher necrotic potential than cells on the periphery.
Tumour cells divide with a probability proportional to local nutrient
per cell concentration, resulting in cells on the tumour's periphery ex-
hibiting a higher proliferation potential than cells in the core. If on the
tumour's perimeter, the new cell will randomly supplant a neighbouring
site's non-tumour cell, causing the tumour to grow, otherwise the new
cell will add to the density of the parent cell's site. Cell proliferation has
the eect of causing tumour invasion and increasing core density while a
su cient quantity of nutrients persists.
26
A tumour cell moves with a probability proportional to its local den-
sity and nutrient concentration. Cells that move, do so to a randomly
chosen neighbouring site, without regard for the density of the destina-
tion site, which could result in the unrealistic situation of a cell moving to
a more densely populated site than the site it had previously inhabited.
If the cell moves to a site containing a non-tumour cell it will supplant it,
otherwise it will exchange places with other tumour cells. The movement
rule could be improved by taking into account the density of neighbouring
sites, so that the movement potential is proportional to the density dif-
ferential between sites. As it is, cell movement has the eect of dispersing
cells from densely populated regions, albeit in an unrealistic, ine cient
manner.
The model can replicate the typical layered structure caused by nu-
trient diusion into an avascular tumour. The model is also able to
reproduce the ngering morphology described in [2, 15]. The nger-
ing morphology is caused by a phenomenon known as diusion-limited
growth. Diusion-limited growth occurs in any system in which diusion
is the primary means of transporting a substance needed for growth into
an aggregate, the tumour. Diusion-limited growth occurs because the
cells arrange themselves so as to maximise their exposure to the diusing
substance. Results were gathered from a model with no cell movement,
which leads me to believe that with cell movement the model was not
able to reproduce this phenomenon.
3.6 Tumour Metabolism
From the observations of Warburg [40, 39], it was seen that tumour cells
preferentially convert glucose to lactic acid, even under aerobic condi-
tions, which results in excess acid production; this behaviour is termed
the Warburg eect. Patel et al. [29] have developed a CA model to
study the qualitative relationship between native tissue vascular density
and avascular tumour lactic acid production and invasion.
The fabric of the model is a 2D lattice, with one cell per site. Sites
are characterised by the type of cell they contain, hydrogen ion (pH)
and glucose concentrations. Sites can contain a normal cell, tumour
cell, micro-vessel or vacant space. Micro-vessels are evenly distributed
across the lattice. Glucose enters the system by diusion across micro-
vessel walls and is consumed by normal or tumour cells. Hydrogen ions
are produced by tumour cells metabolising glucose, and are removed by
27
diusion across the micro-vessel wall.
Site pH and glucose concentration determines cell behaviour. If the
pH is below a threshold the cell dies. If the pH is above the death
threshold, but below a proliferation threshold the cell is quiescent and
if above the proliferation threshold and there exists su cient glucose
concentration, the cell will proliferate if it has a free near neighbour.
Tumour cells have a higher tolerance for pH than normal cells and are,
therefore, more likely to survive and proliferate.
The authors found that a small clone of tumour cells will develop
into a malignant tumour if the clonal phenotype alters the environment
so that it is favourable to tumour cells and hostile to normal cells. Alter-
ations in the environment are dependent on the amount of acid produced
and the density of micro-vessels. If the amount of acid produced is more
than can be removed by the micro-vessels an excess of acid will accumu-
late, creating a hostile environment for normal cells.
28
Chapter 4
Model Design
An in vitro tumour growing on an arti cial substrate will be mathemati-
cally modelled. The tumour is assumed to be growing in isolation, on an
extra-cellular matrix (ECM). The ECM is the extra-cellular part of tissue
that provides structural support to the cells. A constant concentration of
glucose and oxygen will be maintained at the ECM's perimeter. Glucose
and oxygen will diuse across the ECM and into the tumour, where it
will be consumed by the tumour's cells; as described by Figure 4.1.
The extra-cellular matrix.
The tumour.
The constant chemical concentration.
Figure 4.1: A simpli ed representation of the model, viewed from above.
During a model update, the chemical (glucose and oxygen) and cel-
lular distributions are calculated for the entire lattice from the state of
the lattice at the end of the previous update. Under an adiabatic ap-
29
proximation (see Section 4.5.1), model updates proceed by calculating
the chemical equilibrium (steady state) concentration from the bound-
ary concentration and cellular distribution. Once a lattice site's chemical
steady states is known, the metabolic state of the tumour cells at that
site are updated. The cells in a lattice site update their metabolic state
depending on the glucose and oxygen concentration found at their site.
Sites containing tumour cells in a proliferating state update their cellular
density. Eventually, cell migration will redistribute cells amongst lattice
sites so as to smooth out pressure inhomogeneities.
4.1 Modelling Cells
The model consists of a 2D, xed, regular, square lattice. Each lattice
site is assumed to be a 3D cube that may contain tumour cells. The
entire lattice represents a 3D slice through the centre of the tumour.
Figure 4.2 describes the lattice.
Figure 4.2: The model's assumed lattice structure, reproduced from [30].
Lattice site geometry will be based on the model by Piotrowska et
al. [30], i.e. having sides of length of 100 µm, resulting in a volume of
(100 µm)3 per site. Lattice sites are large enough to calculate a represen-
tative tissue density, but also small enough that interesting phenomena,
such as the tumour's structure can be observed.
30
A lattice site may contain tumour cells of several type. Tumour cell
types are based on the cell's phenotype (behaviour) and metabolic state.
Cells can be proliferating, quiescent or necrotic. Cells are further cate-
gorised by their metabolic state, which can be either aerobic or anaerobic.
When the phenotype and metabolic categories are taken together, there
are ve dierent tumour cell types: proliferating aerobic (PAT), prolifer-
ating anaerobic (PANT), quiescent aerobic (QAT), quiescent anaerobic
(QANT) and necrotic.
An introduction to cell metabolism that forms the background to
modelling cells is presented below.
4.1.1 A Biological Background to Cell Metabolism
Within a human cell, energy is stored and released in the form of adeno-
sine triphosphate (ATP), which is converted from glucose, fats and amino
acids. ATP is the body's fundamental unit of energy storage. ATP's
breakdown powers muscle contraction, the generation of electrical signals,
the pumping of ions and other molecules against concentration gradients,
and much more.
The vast majority of energy used in the human body is derived from
glucose. Energy from glucose is released during glucose oxidation [9]. Ox-
idation is chemically de ned as the removal of electrons from a molecule.
In oxygen rich conditions, aerobic glucose oxidation progresses through
three stages. In oxygen poor environments, anaerobic glucose oxidation
is halted after the rst stage.
Almost all cancer cells preferentially consume glucose and produce
lactic acid under aerobic conditions, a behaviour termed the Warburg
eect [40, 39]. The Warburg eect is believed to aid the rapid prolifera-
tion of cancer cells, by increasing a cell's uptake of the nutrients needed
for proliferation [18, 7] and by altering the cell's local environment, by
making it hostile to native tissue [14]. It is common for CA tumour
models to dierentiate between aerobic and anaerobic metabolism. The
main physiological dierence between these two forms of metabolism is
the number of ATP molecules produced per glucose molecule. Anaerobic
metabolism produces only two ATP molecules per molecule of glucose,
whereas aerobic metabolism produces more than thirty ATP molecules
per molecule of glucose.
Stage one, glycolysis, occurs within the cytoplasm of cells. The ly-
sis, or degradation, of a glucose molecule into two C3 fragments yields
31
pyruvate, accompanied by a reduction of N AD+ to NADH. N AD+ , or
nicotinamide adenine dinucleotide, acts catalytically by being continually
re-oxidised to N AD+ and reduced to NADH, and in doing so transfers
electrons to the mitochondria.
Anaerobic glycolysis occurs in the absence of oxygen. For glycolysis
to proceed, NADH must be recycled back to N AD+ . When a cell's
oxygen supply is insu cient, the capacity of its mitochondria to recycle
NADH back to N AD+ is inadequate. NADH can then be re-oxidised by
reducing pyruvate to produce lactic acid. Anaerobic glycolysis produces
two ATP molecules per glucose molecule [9].
When the oxygen supply is plentiful, aerobic glycolysis occurs and the
cell's metabolism enters the second stage of glucose oxidation. Stage two,
the citric acid cycle or Krebs cycle (named after its discoverer), occurs
within the cell's mitochondria. Aerobic glycolysis produces pyruvate and
NADH in the cytoplasm. To be further oxidised, the pyruvate must
enter the mitochondria. A molecule of pyruvate from the cytoplasm is
converted in the mitochondria by pyruvate dehydrogenase and the citric
acid cycle to three molecules of CO2 , during this process three molecules
of N AD+ and one molecule of FAD are reduced. At this point, two
molecules of ATP have been created in glycolysis and two in the citric
acid cycle per molecule of glucose, it is in the nal stage that the majority
of ATP is created.
The nal stage of glucose oxidation, the electron transport system,
occurs in the inner mitochondrial membrane. The electron transport
system is a hierarchy of electron carriers of dierent energy potentials.
Starting from NADH and FADH, electrons move from one carrier to
the other down the energy gradient until they are eventually passed to
oxygen, resulting in the formation of water. The energy released during
electron transport is ultimately trapped as ATP. The glucose oxidation
process can result in the synthesis of more than thirty molecules of ATP
from one molecule of glucose [9].
4.1.2 Modelling Cell Metabolism
A modelled cell responds to chemical stimuli by changing its metabolic
state, in a way similar to that implemented by Piotrowska et al. [30]. A
tumour cell's metabolic phase space is depicted in Figure 4.3. Metabolic
states are delineated by the metabolic thresholds described in Table 4.1.
32
Glucose concentration @molH100 ΜmL3 D
7. ´ 10-16
PANT
6. ´ 10-16
5. ´ 10-16
QANT
4. ´ 10-16 PAT
3. ´ 10-16 Βpant
Γqant
2. ´ 10-16 Necrotic
Γpant
1. ´ 10-16 QAT
Γpat
0
4. ´ 10-17 5. ´ 10-17 6. ´ 10-17 7. ´ 10-17 8. ´ 10-17 9. ´ 10-17 1. ´ 10-16
Oxygen concentration @molH100 ΜmL3 D
Figure 4.3: A cell's metabolic response to chemical stimuli described as
a phase diagram.
Parameter Value Units Ref.
βpat 8.3 × 10−17 (mol)(cell)−1 (s)−1 [13]
γpat 18 × 10−17 (mol)(cell)−1 (s)−1 [13]
γpant 52 × 10−17 (mol)(cell)−1 (s)−1 [13]
γqant 43 × 10−17 (mol)(cell)−1 (s)−1 [13]
Table 4.1: The metabolic threshold values.
A cell's metabolic state depends on where its lattice site's chemical
environment falls in Figure 4.3. The most important distinction to draw,
is between those cells that are alive and those that are necrotic. Referring
to Figure 4.3, a cell becomes necrotic when its local glucose and oxygen
concentration is below the proliferate aerobic oxygen (βpat ) and quies-
cent anaerobic glucose (γqant ) thresholds. Once necrotic, tumour cells
cease to consume chemicals and proliferate, however, their mass still ex-
ists within the model; and plays an important role in creating pressure
inhomogeneities.
4.1.3 Cell Proliferation
Cells in a proliferating state add to the cellular density of their lattice
site at each model update. The model's proliferation scheme is based on
the mass-balance equation [10, 3]. Within our model, the cellular density
of a lattice site is de ned as:
33
Ncells
ρ= (4.1)
Vsite
Where:
• ρ is the cellular density of a lattice site, with units of (cells)(100 µm)−3 .
• Ncells is the number of cells at the lattice site, with units of (cells).
• Vsite is the volume of the lattice site, with units of (100 µm)3 .
The change in cellular density of a lattice site can be described by a PDE
as:
∂ρ
=Pρ (4.2)
∂t
Where P is the proliferation rate. Additional terms, such as those de-
scribing the migration of cells into and out of a lattice site, can be added
to Equation (4.2) without loss of generality. Equation (4.2) has a con-
tinuous analytical solution of:
ρ(t) = ρ(0) e(P t) (4.3)
A cell's proliferation rate is commonly characterised by the time T needed
for a population to double, known as the doubling time. In the absence
of additional terms in Equation (4.2), the proliferation rate P can be
related to T by:
Ln(2)
P (T ) = (4.4)
T
A lattice site's cellular density will be updated at discrete time intervals
while the site's porosity is greater than zero. The porosity of tissue
is de ned as the fractional volume of tissue not occupied by cellular
material:
Vcells
0≤φ=1− ≤1 (4.5)
Vsite
Where:
• φ is the tissue porosity, which is dimensionless.
• Vcells is the total tumour cell volume at a lattice site, with units of
(100 µm)3 .
A discrete approximation of Equation (4.2) can be found with an explicit
scheme as:
34
ρ(t) + 4t P ρ(t) , φ > 0
ρ(t + 4t) = (4.6)
ρ(t) , φ=0
During the time (cellular update) interval 4t, an additional 4t P ρ(t)
cells are added to the site. When the porosity of a lattice site equals
zero, cellular material is completely occupying the site's volume, leaving
no space for additional cells.
In discretising Equation (4.2), we have assumed the cellular update
interval 4t, is small in comparison to the typical cellular time scale of
the model, the doubling time T. Numerical experiments were conducted
to ascertain a cellular update interval that balances stability with e -
ciency. The cellular update interval has to be short enough to calculate
a stable cellular density, but long enough to simulate tumour growth in
a reasonable amount of time. As 4t → 0, the stability of our numer-
ical scheme improves and the discrete approximation converges to the
continuous solution.
The proliferation scheme can be extended to model sub-populations of
cells with dierent proliferation rates. The modelling of sub-populations
of cells can be achieved with a separate discrete approximation for each
sub-population. Consider sub-population ρ1 and ρ2 , their progression
could be modelled as:
ρ1 (t + 4t) = ρ1 (t) + 4t P1 ρ1 (t) (4.7)
and
ρ2 (t + 4t) = ρ2 (t) + 4t P2 ρ2 (t) (4.8)
For now, we have assumed all proliferating cells have the same doubling
time. There are, however, grounds for modelling a cell's doubling time
dependent on its metabolic state, due to the diering amount of ATP
produced in the anaerobic and aerobic states, an extension that will be
considered for the planned in vivo model.
4.2 Inter-Cellular Pressure
We want to nd a phenomenological relationship between a lattice site's
cellular density and the average pressure experienced by the site's cells
on their membrane, a quantity we will term the average inter-cellular
35
pressure. This aim forms the core of this thesis, as it is the average
inter-cellular pressure that will drive cell movement away from densely
populated regions into sparsely populated regions, thus promoting the
tumour's invasion of healthy tissue. It is possible to relate Equation
(4.1) and (4.5) as:
Ncells Vcells 1−φ
ρ= = = (4.9)
Vsite Vcell Vsite Vcell
Where Vcells is the total volume of cellular material at the lattice site,
given by:
N
Xcells
Vcells = Vcelli = Ncells Vcell (4.10)
i=1
Where Vcell is the average volume of a single cell at the lattice site. Some
preliminary observations can now be made by rearranging Equation (4.9).
Assuming that a cell's shape deforms under the pressure experienced
on its membrane, it can be observed that as the ratio Vcells /Vsite → 1,
Vcell → 1/ρ. When the cells within a lattice site are at full contact, so that
the site's volume is completely occupied by cellular material, geometric
considerations tell us that the average volume of a cell is equal to the
inverse of the cellular density or the site volume divided by the number
of cells. Also, as the ratio Vcells /Vsite → 0, Vcell → 0, the average volume
of an individual cell goes to zero.
It is now possible to relate a site's cellular density to its average
inter-cellular pressure. The average volume of a site's cells is inversely
proportional to the average inter-cellular pressure. We considered two
alternatives for describing a cell's volume as a function of the pressure
experienced on its membrane. The simpler relationship, which was used
in the experiments described in Chapter 5, will be described here, refer to
the appendices of a description of the alternative relationship. A simple
inverse relationship between average cell volume and pressure:
α
Vcell (p) = (4.11)
p
Where:
• Vcell is the average volume of a site's cells, with units of (100 µm)3 .
• p is the average inter-cellular pressure, with units of (kg)(100 µm)−1 (s)−2 .
• α is a constant, with units of (kg)(100 µm)2 (s)−2 .
36
Average inter-cellular pressure can be retrieved from the inverse of Equa-
tion (4.11).
In sparsely populated lattice sites, the probability of a cell coming
into contact with other cells is low, thus the majority of the pressure
experienced on a cell's membrane will be caused by the interstitial uid
pressure (IFP) present in the ECM. Within our model the IFP of nor-
mal tissue (p0 ) is assumed constant and equal to 7 mmHg (millimetres
−1 −2
of mercury) or 0.09333 (kg)(100 µm) (s) , from Jain's review of the
transport of molecules in the tumour interstitium [21]. Cells are assumed
to form the intrinsic shape of a sphere under the IFP of normal tissue, as
this is the lowest pressure the cell will experience. The intrinsic shape of
a cell is assumed to have a radius of 10 µm [1], from which the intrinsic
volume (Vcell0 ) can be calculated as 0.00419 (100 µm)3 . The constant α,
in Equation (4.11) can now be found algebraically from our understand-
ing of a cell's volume under IFP, as:
α = Vcell0 p0 = 0.00039 (kg)(100µm)2 (s)−2 (4.12)
It is now possible to solve Equation (4.11) and described cell volume as
a function of pressure, as in Figure 4.4.
0.12
0.10
Cell volume @H100 ΜmL3 D
0.08
0.06
0.04
0.02
0.00
0.00 0.02 0.04 0.06 0.08 0.10
Pressure @kg H100 ΜmLHsL2 D
Figure 4.4: The cell volume as a function of pressure, described by Equa-
tion (4.11). The vertical and horizontal grey lines denote the IFP of
normal tissue and intrinsic volume respectively.
In sparsely populated lattice sites, where the pressure is assumed
constant and equal to the IFP (p = p0 ), relation (4.9) speci es how the
37
porosity depends on the density as:
φ(ρ) = 1 − Vcell (p0 ) ρ (4.13)
As the pressure is constant, the cell volume is also constant and equal to
the intrinsic cell volume.
Vcell0 = Vcell (p0 ) (4.14)
However, in densely populated lattice sites, a cell has a high proba-
bility of being in contact with other cells, thus the majority of the force
experienced on its membrane will be due to the cells it is in contact with.
Within densely populated regions, pressure is expected to increase with
density as the probability of a cell coming into contact with other cells
increases.
In-order to describe the average inter-cellular pressure of densely pop-
ulated sites we will start by considering an idealised tumour, composed of
impermeable cells, which are perfectly spherical and uniform in size. The
cells are imagined to be growing within an ECM. Our idealised tumour is
analogous to spheres packed in 3D. As the cells within a site proliferate,
the cellular density increases until a maximum is reached. Figure 4.5
describes the two regular packing arrangements that achieve a minimum
porosity of ≈ 0.26, according to the Hales proof of the Kepler conjecture
[17].
Figure 4.5: The regular packing of spheres in 3D. The left-hand side
describes the hexagonal close-packing formation, whereas the right-hand
side describes the face-centred cubic packing [42].
38
By assuming cells are spheres of uniform size, we can determine the
range of densities for which the average pressure experienced by the cells
in a site is equal to the IFP, and for which Equation (4.13) provides a
valid description. It is assumed that the cells within a site will form an
irregular arrangement. It is known that the irregular packing of spheres
can achieve a minimum porosity of 0.36 [33], which we will term the
critical porosity (φc ). The close packing of cells will therefore occur when
Vcells /Vsite ≈ 0.64. The density at which all the cells within a site are
in contact with their neighbours, which we will term the critical density
(ρc ), can now be found as approximately 153 (cells)(100 µm)−3 .
For densities above the critical density, we expect there to be an
interplay between a cell changing its volume, but maintaining its shape
and a cell changing its shape with a constant volume, under increasing
pressure on its membrane. We expect that a cell's shape would change
rst at the critical density, due to the uneven pressure distribution across
its membrane. When spherical cells rst come into contact it will be at
a small point across their surfaces. As the contact area enlarges, due
to deformation of the cell's shape, the pressure distribution will even
out, which will cause the cell's volume to shrink. Then as the cells
shrink the contact point will become small again, thus increasing the
pressure. The relationship between cell volume, shape and pressure could
be investigated by constructing a cell structural model that included such
phenomena as cell membrane and cytoskeleton, however such a model
would be beyond the scope of this project. At the site level, we would
expect the porosity to converge to zero at a slower rate for densities
greater than the critical density, as described in Figure 4.6.
The change in porosity for densities greater than the critical density
can be described by a relationship of the form:
φρ̄ (ρ) = a(ρ̄)(1−Vcell0 ρ) − a(ρ̄)(1−Vcell0 ρc ) − φc
(4.15)
Where:
• ρ̄ is the site density at which the porosity equals zero, with units
of (cells)(100 µm)−3 .
The term a(ρ̄) ≥ 1 is chosen such that φρ̄ (ρ̄) = 0. The change in a(ρ̄)
with ρ̄ is described in Figure 4.7.
39
1.0
0.8
HΡc , Φc L
0.6
Porosity Φ
0.4
0.2 HΡ, 0L
0.0
0 100 200 300 400 500 600 700
Density Ρ @cells H100 ΜmL3 D
Figure 4.6: The density-porosity relationship for densities greater than
the critical density.
3
aHΡL
0
0 200 400 600 800 1000 1200 1400
Figure 4.7: The relationship between ρ̄ and a(ρ̄).
Referring to Figure 4.7, as the value of ρ̄ decreases, the value of the
term a(ρ̄) increases to in nity at the critical density. For the experiments
described in Chapter 5, ρ̄ was chosen to be near the upper bound of the
−3
densities recorded experimentally by [13], ρ̄ = 691 (cells)(100 µm) .
Cells will eventually die due to the pressure experienced on their mem-
40
brane and be converted into necrotic material, which is uid in nature.
Under pressure, the necrotic material would be forced out of high density
regions if it has channels through which it can drain. The necrotic ma-
terial would then be replaced by surrounding cells. When the pressure
reaches the range of pressures for which cells become necrotic, we would
expect to see a further slowing in the decline of porosity and perhaps
even an increase in porosity with density, this is a modi cation that will
be considered for the planned in vivo model.
Cell volume as a function of density can be found from Equation (4.9)
as:
1 − φ(ρ)
Vcell (ρ) = (4.16)
ρ
The average inter-cellular pressure as a function of cellular density can
be found by substituting Equation (4.16) into the inverse of Equation
(4.11), which yields:
α αρ
p(ρ) = = (4.17)
( 1−φ(ρ)
ρ
) 1 − φ(ρ)
The density-pressure relationship is described in Figure (4.8), according
to the density-porosity relationship described by Equation (4.15).
0.30
0.25
Pressure @kg H100 ΜmLHsL2 D
0.20
0.15
0.10
0.05
0.00
0 100 200 300 400 500 600 700
Density Ρ @cells H100 ΜmL3 D
Figure 4.8: The average inter-cellular pressure as a function of density.
Referring to Figure 4.8, the average inter-cellular pressure in a site
is constant and equal to the IFP for densities less than the critical den-
41
sity. Once the density equals the critical density, Average inter-cellular
pressure increases in a curve until the density equals ρ̄, at which point
the porosity equals zero. Average inter-cellular pressure then increases
linearly for densities greater than ρ̄.
Having described the average inter-cellular pressure as a function of
density, it is now possible to model pressure-induced cell movement.
4.3 Modelling Cell Migration
4.3.1 Introduction
Within our model, a cell's movement is driven by a pressure force trans-
ferred through the cells it is in contact with. Cell movement due to
pressure is assumed independent of cell's metabolic state. A pressure
force will act on cells in a region where the cellular density is above the
critical density. A cell's velocity due to pressure will be assumed propor-
tional to the pressure gradient across its membrane (see Equation (4.21)
to (4.25)), expressed as:
vp = −ωp ∇p (4.18)
Where:
• vp is a cell's velocity induced by a pressure gradient, with units of
(100 µm)(s)−1 .
• p is the average inter-cellular pressure, with units of (kg)(100 µm)−1 (s)−2 .
• ∇p is the pressure gradient along which the cell is moving.
• ωp is a proportionality factor with units of (100 µm)2 (s)(kg)−1 .
For now we have chosen to ignore cell movement due to a chemotatic
attraction to higher concentrations of glucose and oxygen. However, we
anticipate modelling chemotaxis induced movement in the planned in
vivo model. Unlike pressure induced cell movement, chemotatic move-
ment would be dependent on a cell's metabolic state. A cell would need
to be alive for it to move. Generally, a cell's velocity due to chemotaxis
would be proportional to the chemical concentration gradient across its
membrane. Glucose-induced chemotaxis can be expressed as:
vg = ωg ∇g (4.19)
42
Where:
• vg is a cell's velocity induced by a glucose concentration gradient,
with units of (100 µm)(s)−1 .
• g is the glucose concentration within a site, with units of (mol)(100 µm)−3 .
• ∇g is the glucose concentration gradient along which the cell is
moving.
• ωg is a proportionality factor with unit of (100 µm)4 (mol)(s)−1 .
Chemotaxis due to oxygen would be similar to Equation (4.19). We
then anticipate combining the separate equations describing the dierent
velocity components, to form a single equation describing changes in cell
density due to movement, as:
∂ρ
= ∇ · (ρ v) (4.20)
∂t
Where:
• ∇ is the gradient of the velocity components, with units of (100 µm)−1 .
• v = v p + vg + vo is the sum pressure, glucose and oxygen velocity
components respectively, with units of (100 µm)(s)−1 .
4.3.2 Modelling Pressure-Induced Cell Movement
Below, the details of the cell migration algorithm are described. Cell
migration is the result of an inhomogeneous pressure eld that results
from dierences in cellular density. The inhomogeneous pressure eld is
assumed to generate a driving force that gives rise to a cellular velocity.
An object of volume V, surface S, embedded in a pressure eld p(r)
experiences a total force:
Z
F =− dσ n(r) p(r) (4.21)
Where n(r) is an outward-pointing unit vector normal to the surface and
dσ a line integral. If the pressure eld can be extended to the region
occupied by the object, the divergence theorem can be used to express
the force as a volume integral:
Z
F =− d3 r ∇p(r) (4.22)
43
For small objects, Equation (4.22) is approximated by:
F ≈ −V ∇p(r) (4.23)
For an object experiencing a force F in a viscous uid, the equation of
motion reads:
m v̇ = F − λ v (4.24)
If friction is strong or the object's mass small, inertial eects are negligible
(terminal velocity is reached very quickly) resulting in:
1
v= F (4.25)
λ
Where v is the cell's velocity, λ is viscous drag coe cient and F the
pressure force experienced by a cell. A cell's velocity is, therefore, pro-
portional to the pressure gradient experienced across its membrane. Ad-
ditional terms, such as those describing velocity driven by chemotactic
attraction can be added to Equation (4.25) without loss of generality.
4.3.2.1 Displacement in 1D
Δx Δx
pi pi+1 pi + pi+1
i i+1 i i+1
Figure 4.9: Migration of cells due to pressure inhomogeneities between
lattice sites. Cells in the light grey-shaded area are moved to the dark-
shaded area during the time increment 4t. The cell counts in each lattice
site are updated accordingly.
Within our model, pressure is an average quantity over lattice sites. Pres-
sure inhomogeneities, therefore, only occur between neighbouring sites.
Consider a patch of cells that occupies an area overlapping two neigh-
bouring lattice sites, site i and site i+1 (Figure 4.9). During the small
time increment 4t, these cells are displaced by a distance 4x ≈ v 4t.
Equation (4.23) and (4.25) can be combined to describe the displacement
of our patch of cells between neighbouring sites:
44
h2
4x ≈ − (pi+1 − pi ) 4t (4.26)
λ
Where h is the length of a lattice site, and pi+1 and pi are the average
pressure experienced by cells in site i+1 and i respectively. Knowing the
displacement of our patch of cells, enables us to calculate the change in
cellular density of the two sites straddled by our patch of cells. In 1D,
this method can be extended to a strip of lattice sites.
4.3.2.2 Displacement in 2D
pi,j+1 pi+1,j+1 pi,j+1 pi+1,j+1
+ + +
j+1 j+1
+
Δr
j j
pi,j pi+1,j pi,j pi+1,j
i i+1 i i+1
Figure 4.10: Migration of cells due to pressure inhomogeneities between
the sites of a 2D lattice. Cells in the light grey-shaded area are moved
to the dark grey-shaded area during the time increment 4t. The cell
0 0
− and 0 +0 signs
count in each lattice site is updated accordingly, as the
suggest.
Now consider a patch of cells occupying an area overlapping four neigh-
bouring lattice sites, sites (i, j), (i+1, j), (i, j +1) and (i+1, j +1) (Figure
4.10). From Equation 4.21, we can calculate the pressure-induced force
exerted along the i and j axis as:
h2
Fi = −e1 [(pi+1,j − pi,j ) + (pi+1,j+1 − pi,j+1 )] (4.27)
2
and
h2
Fj = −e2 [(pi,j+1 − pi,j ) + (pi+1,j+1 − pi+1,j )] (4.28)
2
Where e1 and e2 are unit vectors along the i and j axes respectively.
Again, the pressure force induces a velocity component along the i and
j axis as:
45
1
vi = Fi (4.29)
λ
and
1
vj = Fj (4.30)
λ
During the time increment 4t, the displacement of our patch of cells in
the x and y directions can be calculated as:
4x = vi 4t (4.31)
and
4y = vj 4t (4.32)
which can be combined to form the vector:
! !
4x vi
4r = = 4x e1 + 4y e2 ≈ 4t (4.33)
4y vj
Knowing the displacement of our patch of cells in 2D allows us to calcu-
late the change in cellular density of the lattice sites it straddles. This
method can be applied to the lattice to model cellular migration for the
entire tumour.
4.4 The Chemical Mass-Balance Equations
The glucose and oxygen concentration of a lattice site dictates the metabolic
state of the site's cells. A site's glucose and oxygen concentrations are
described by the following mass-balance equations.
4.4.1 Glucose
The glucose mass-balance equation describing the rate of change of glu-
cose entering and leaving a lattice site:
∂g X
= ∇(Dg ∇ g) − γσ ρσ g + γv ρv (4.34)
∂t
σ∈{pat,pant,qat,qant,dt}
Refer to Table 4.2 for a description of the equation's parameters. The
rst term on the right-hand side of the equation describes the change in
46
Description Symbol Value Units
Glucose diusion parameters
Concentration gradient ∇ (100 µm)−1
Apparent diusion coe cient Dg (100 µm)2 (s)−1
Concentration g (mol)(100 µm)−3
Scaled tumour cell glucose consumption/production rates
Proliferate aerobic γpat
¯ 1.9 × 10−4 (100 µm)3 (cell)−1 (s)−1
Proliferate anaerobic γpant
¯ 1.9 × 10−4 (100 µm)3 (cell)−1 (s)−1
Quiescent aerobic γqat
¯ 1.9 × 10−4 (100 µm)3 (cell)−1 (s)−1
Quiescent anaerobic γqant
¯ 1.9 × 10−4 (100 µm)3 (cell)−1 (s)−1
Dead γ¯dt 0 (100 µm)3 (cell)−1 (s)−1
Blood vessel γv (mol)(cell)−1 (s)−1
Cell densities
Proliferate aerobic ρpat (cell)(100 µm)−3
Proliferate anaerobic ρpant (cell)(100 µm)−3
Quiescent aerobic ρqat (cell)(100 µm)−3
Quiescent anaerobic ρqant (cell)(100 µm)−3
Dead ρdt (cell)(100 µm)−3
Blood vessel ρv (cell)(100 µm)−3
Table 4.2: Glucose mass-balance equation's parameters.
glucose concentration due to the diusion of glucose into and out of a
site, this term will be discussed in more detail in Section 4.5.
The second term on the right-hand side describes the cellular con-
sumption of glucose. Note, the chemical consumption term takes on a
dierent form from that common to most CA tumour models, such as
[30, 15, 29], which consist of a consumption rate multiplied by a cellular
density. We reason that for a cell to consume glucose, glucose must be
present in the cell's local environment, which justi es inclusion of the
local glucose concentration as a factor in the consumption term.
The form of our consumption term necessitates we adapt the reported
consumption rates (in the literature), for our consumption de nition.
Conventional models vary the consumption rate depending on the cell's
metabolic state, to account for their lack of local glucose concentration in
the consumption term, so as to simulate a decrease in consumption going
from the edge of the tumour to its core. Due to the form of our consump-
tion term, the glucose consumption rate is proportional to the glucose
concentration, which more closely resembles reality, where tumour cells
will attempt to consume glucose at the highest possible rate, regardless
of metabolic type. The form of our consumption term is more akin to a
chemical reaction than that of [30, 15, 29], which are more algorithmic
in nature.
47
The third term on the right-hand side of Equation (4.34) describes the
production of glucose from a blood vessel within the model. The glucose
production term has more relevance to the anticipated in vivo model,
and has been included here for the completeness of our description. The
production term was set to zero in the experiments conducted for Chapter
5.
4.4.2 Oxygen
The oxygen mass-balance equation describing the rate of change of oxy-
gen entering and leaving a lattice site:
∂o X
= ∇(Do ∇ o) − βσ ρ o + βv ρv (4.35)
∂t
σ∈{pat,pant,qat,qant,dt}
Description Symbol Value Units
Oxygen diusion parameters
Concentration gradient ∇ (100 µm)−1
apparent diusion coe cient Do (100 µm)2 (s)−1
Concentration o (mol)(100 µm)−3
Scaled tumour cell oxygen consumption/production rates
Proliferate aerobic β¯pat 2.4 × 10−3 (100 µm)3 (cell)−1 (s)−1
Proliferate anaerobic ¯
βpant 2.4 × 10−3 (100 µm)3 (cell)−1 (s)−1
Quiescent aerobic β¯qat 2.4 × 10−3 (100 µm)3 (cell)−1 (s)−1
Quiescent anaerobic ¯
βqant 2.4 × 10−3 (100 µm)3 (cell)−1 (s)−1
Dead β¯dt 0 (100 µm)3 (cell)−1 (s)−1
Blood vessel βv (mol)(cell)−1 (s)−1
Table 4.3: Oxygen mass-balance equation's parameters.
Refer to Table 4.3 for a description of the equation's parameters.
Equation (4.35) is similar in form to Equation (4.34). The rst term
on the right-hand side describes the diusion of oxygen into and out
of a lattice site, which will be discussed in Section 4.5. The second
term describes the cellular consumption of oxygen, while the third term
accounts for the production of oxygen from a blood vessel located within
the model and was again set to zero for the experiments performed in
Chapter 5.
As before, the oxygen consumption rate is taken to be constant ir-
respective of a cell's metabolic state. Indeed, oxygen diuses through a
cell's membrane without the need for active uptake on the cell's behalf.
48
Cells consume oxygen at a rate proportional to their local concentration.
This behaviour is re ected in the oxygen consumption term (the second
term on the right-hand side) of Equation (4.35). Again, this is in contrast
to conventional CA tumour models, such as those by [30, 15, 29], which
vary a cell's oxygen consumption rate depending on its metabolic state.
4.5 Modelling Chemical Diusion
The diusion of a chemical species is described by the rst term on the
right-hand side of Equation (4.34) and Equation (4.35), for an apparent
diusion coe cient. The apparent diusion coe cient (D ), can be de-
ned phenomenologically as the proportionality factor between the ux
(J ) and the gradient of the density (∇n) of a diusing species:
J = −D ∇n (4.36)
In a medium with several phases, we can either consider each phase
separately, or speak of averaged quantities over a representative volume
element large enough to comprise these phases, yet small enough to pre-
serve media/tissue heterogeneity. Within our model, a chemical species
can either diuse through the ECM or tumour phase. We will consider
the average quantities of a chemical diusing through a lattice site, which
can contain either ECM or tumour cells.
The porosity of tissue, is de ned as the fractional volume of the tis-
sue in which the diusing substance can move. Molecular transportation,
under diusion, through tissue, depends on the porosity of the tissue. If
porosity is high, a molecule can diuse further, within a certain time,
than when porosity is low. We imagine an idealised tumour, made of
impermeable cancer cells living in an ECM. Chemical molecules are as-
sumed to diuse through the ECM, but not through the cells. Molecules
can be captured by a cell and metabolised.
Tortuosity (τ ) is the property of a curve describing its twists. Within
our model, we are concerned with the tortuosity of a chemical molecule's
path between cells, which is related to the tissue's porosity. Tortuosity af-
fects the distance a molecule can diuse within a certain time. The more
tortuous (twisted) the path a molecule takes, the shorter the straight line
distance it will travel under diusion, within a given time and the lower
the eective diusion of the species to which it belongs.
In 2D, the tortuosity of a curve can be de ned as the ratio of the
49
distance (d) between the end points of a curve, and (l ) the length of a
curve [8], more formally:
d
τ= ≤1 (4.37)
l
In 3D, for spherical cells, the tortuosity can be related to the porosity by
a technique borrowed from porous media transport theory [8], as:
p
τ= φ (4.38)
The tortuosity is a proportionality factor relating the apparent diusion
coe cient to the diusion coe cient of the uid phase (D0 ), which cor-
responds to diusion in the absence of cells. Within our model, the ECM
is assumed to be free of cells and therefore has the diusion coe cient
of the uid phase. The diusion coe cient of the uid phase is related
to the apparent diusion coe cient by:
D = τ D0 (4.39)
It is now possible to calculate a lattice site's diusion coe cients as a
function of its density. A lattice site's porosity can be calculated as a
function of its density from Equation (4.13) for sparsely populated lattice
sites and Equation (4.15) for densely populated lattice sites. A lattice
site's tortuosity can then be found from Equation (4.38). A lattice site's
diusion coe cients can be related to the diusion coe cients in the
uid phase and tortuosity by Equation (4.39), thus allow us to calculate
a chemical specie's ux through the lattice site as:
J = −τ D0 ∇n (4.40)
When applied to the entire lattice, this technique allows us to model the
inhomogeneous diusion of a chemical species. The continuous equations
describing the inhomogeneous diusion of glucose and oxygen were dis-
cretised on to the lattice using the Forward Euler method in time [5],
refer to the Appendices for a description of their discretisation. Once dis-
cretised, the diusion equation were solved by the method of Successive
Overrelaxation (SOR) [32].
Note that if the cells are assumed not to deform under pressure, but
remain spherical, the porosity will not reach zero with increasing density
in either 2D or 3D. However, in 2D the tortuosity will become zero at the
maximum packing density when a path no-longer exists between the cells
50
because all the cells are in contact, thus causing the ux to equal zero. In
3D the tortuosity will not reach zero as a path always exists around the
cells even when all the cells are in contact, which can be seen in Figure
4.5. In 3D, a ux will exist allowing the transportation of molecules
between the cells, albeit at a restricted rate. Although our model is
composed on a 2D lattice, lattice sites represent a 3D slice through the
tumour. A ux will, therefore, exist even when all the cells in a lattice
site are in contact.
4.5.1 The Adiabatic Approximation
Chemical elds are, in principle, time dependent and therefore described
by a PDE, which determines their spatio-temporal evolution. However,
as chemical molecules are negligible in mass compared to the mass of a
typical cell, they reach their equilibrium state on a much shorter time
scale than a cellular distribution. Cell doubling time is of the order of 22
hours [13, 22, 12], whereas the inter-vessel diusion time for lactic acid
and glucose is only of the order of 1-10 seconds [29] (with a mean inter-
vessel distance 10−2 cm and lactic acid diusion coe cient of 1.08 × 10−5
(cm)2 (s)−1 ).
The dierence in representative time scales allows us to assume that a
chemical concentration eld will have reached equilibrium (steady state)
between cellular updates. As it is the cells and their evolution that we
are interested in, this observation allows us to ignore the chemical eld's
evolution and directly calculate its steady state, upon which the next
cellular update can be based.
This assumption has also been made by Patel et al. [29] to greatly im-
prove their model's e ciency, they termed it the adiabatic approximation.
The adiabatic approximation, however, has certain limits beyond which
it cannot be used, refer to the Appendices for a study of the adiabatic
approximation.
51
Chapter 5
Results
5.1 Simulation Parameters
Tumour growth was simulated from a single cell at the centre of the
lattice. The lattice consisted of 101 × 101 sites, corresponding to a total
area of ≈ 1 cm2 . The tumour is assumed to grow within an extra-cellular
matrix (ECM), a circular region with a radius of 49.5 (100 µm), see Figure
4.1. The simulation terminated when the tumour grew to the edge of the
ECM. Constant concentrations of glucose and oxygen were maintained
outside of the ECM. The simulation's parameter can be found in Table
5.1.
Parameter name Symbol Value Units Ref.
Chemical parameters
−12
Glucose boundary concentration 2.8 × 10 (mol)(100 µm)−3 [13]
Oxygen boundary concentration 3.5 × 10−14 (mol)(100 µm)−3 [13]
Scaled glucose consumption rate γ̄ 1.9 × 10−4 (100 µm)3 (cell)−1 (s)−1
Scaled oxygen consumption rate β̄ 2.4 × 10−3 (100 µm)3 (cell)−1 (s)−1
ECM parameters
IFP p0 0.9333 (kg)(100 µm)−1 (s)−2 [21]
Glucose diusion coe cient Dg 0.91 (100 µm)2 (s)−1 [6]
Oxygen diusion coe cient Do 0.182 (100 µm)2 (s)−1 [36]
Tumour parameters
Mitotic cycle time 22 hours [13]
Cellular update interval 11.25 minutes
Maximum site density ρ̄ 691 (cells)(100 µm)3 [13]
Cell migration parameters
Viscous drag coe cient λ 10 (kg)(s)−1
Table 5.1: The simulation's parameters.
52
5.2 Simulation Results
Initially, the simulation was conducted with the metabolic threshold val-
ues described in Table 4.1. We found that is was not possible to maintain
a stable quiescent layer with the original metabolic thresholds, which can
be seen in Figure 5.1.
Cellular density [(cells)/(100 micrometer)3]
Cellular density [(cells)/(100 micrometer) ]
160
3
PAT 200
PANT
140
PAT
120 150
100
80 100
60
40 50
20
0
10 20 30 40 50 60 70 80 90 100 10 20 30 40 50 60 70 80 90 100
Lattice site [100 micrometer] Lattice site [100 micrometer]
Cellular density [(cells)/(100 micrometer)3]
Cellular density [(cells)/(100 micrometer) ]
3
250 350
PANT PANT
PAT 300 PAT
200
250
150 200
100 150
100
50
50
10 20 30 40 50 60 70 80 90 100 10 20 30 40 50 60 70 80 90 100
Lattice site [100 micrometer] Lattice site [100 micrometer]
Cellular density [(cells)/(100 micrometer)3]
Cellular density [(cells)/(100 micrometer) ]
3
600 600
Dead Dead
PANT PANT
500 PAT 500 PAT
QANT QANT
400 400
300 300
200 200
100 100
10 20 30 40 50 60 70 80 90 100 10 20 30 40 50 60 70 80 90 100
Lattice site [100 micrometer] Lattice site [100 micrometer]
Cellular density [(cells)/(100 micrometer)3]
Cellular density [(cells)/(100 micrometer) ]
3
700 700
Dead Dead
600 PANT 600 PANT
PAT PAT
500 QANT 500 QANT
400 400
300 300
200 200
100 100
10 20 30 40 50 60 70 80 90 100 10 20 30 40 50 60 70 80 90 100
Lattice site [100 micrometer] Lattice site [100 micrometer]
Figure 5.1: The cross-sections describe the tumour's growth with the
original metabolic thresholds. Going from top to bottom, left to right, the
cross-sections describe the tumour at day ten, twelve, fourteen, sixteen,
eighteen, nineteen, twenty and twenty-one.
Referring to Figure 5.1, the quiescent layer (yellow bar) is seen to form
on day eighteen, then vanish on day nineteen, only to re-emerge again
on day twenty. It was felt that the oscillating behaviour of the quiescent
layer was due to the narrow range of glucose concentrations for which cells
are quiescent (see Figure 4.3). To improve the quiescent layer's stability,
the γqant threshold was lowered to be less than the γpant threshold by
53
an order of magnitude, so that γqant = 5.2 × 10−17 (mol)(cell)−1 (s)−1 ;
Figure 5.2 describes the new metabolic phase diagram.
Glucose concentration @molH100 ΜmL3 D
7. ´ 10-16
6. ´ 10-16 PANT
5. ´ 10-16
PAT
4. ´ 10-16
Βpant
3. ´ 10-16 Γqant
QANT
-16 Γpant
2. ´ 10
Γpat
1. ´ 10-16 QAT
Necrotic
0
4. ´ 10-17 5. ´ 10-17 6. ´ 10-17 7. ´ 10-17 8. ´ 10-17 9. ´ 10-17 1. ´ 10-16
Oxygen concentration @molH100 ΜmL3 D
Figure 5.2: The new metabolic phase diagram describing a cell's
metabolic response to chemical stimuli.
With the new metabolic thresholds, the tumour reached the edge
of the ECM, i.e. a radius of ≈ 5 mm, after 21.28125 days of growth,
which were simulated in approximately 129 minutes. The model was
implemented in the C++ programming language. The simulations were
conducted on an Intel Core 2 Duo CPU (2.40 GHz, with 4 GB RAM)
running a Ubuntu 9.04 virtual machine.
The tumour achieved a maximum site density of 626 (cells)(100 µm)3 ,
which, according to Equation (4.15), equates to a minimum porosity
of 0.03613. The corresponding apparent glucose and oxygen diusion
coe cient can be calculated from Equation (4.38) and Equation (4.39)
as 0.17297 (100 µm)2 (s)−1 and 0.03459 (100 µm)2 (s)−1 respectively. The
ratio of the diusion coe cient in the uid phase and tumour, for both
glucose and oxygen is 5.26, which, according to Figure 7.7 and 7.8, is
within the limits of the adiabatic approximation.
We will begin by analysing the tumour's growth at a microscopic level,
which will be followed by a macroscopic interpretation. The following
gures describe the composition of a strip of lattice sites, running through
the centre of the lattice, for the period between the tenth and twenty-
rst day of growth. The bars in the plots correspond to the cellular
density and composition of a lattice site. The bar's magnitude describes
54
the lattice site's cellular density and its colour the metabolic state of the
cells located at that lattice site.
Cellular density [cells/(100 micrometer)3]
160
PAT
140
120
100
80
60
40
20
10 20 30 40 50 60 70 80 90 100
Lattice site [100 micrometer]
Figure 5.3: The tumour's composition at 10 days.
During the rst ten days of growth, the relatively small tumour has
remained in a proliferate aerobic (PAT) state. After ten days, the tumour
has a radius of 2.5 (100 µm).
Cellular density [cells/(100 micrometer)3]
180
PANT
160 PAT
140
120
100
80
60
40
20
10 20 30 40 50 60 70 80 90 100
Lattice site [100 micrometer]
Figure 5.4: The tumour's composition at twelve days.
After twelve days, the tumour has developed a proliferate anaerobic
(PANT) core surrounded by a rim of PAT cells. Due to a density and
pressure build-up and pressure-induced cell movement, the tumour has
become large enough to cause the oxygen concentration in the centre of
the tumour to drop below the βpat threshold (see Fig. 5.2), thus causing
the cells at the tumour's core to transition to a PANT metabolic state.
55
Cellular density [cells/(100 micrometer)3]
250
PANT
PAT
200
150
100
50
10 20 30 40 50 60 70 80 90 100
Lattice site [100 micrometer]
Figure 5.5: The tumour's composition at fourteen days.
After fourteen days, the tumour has grown to form a dome shaped
density, centred on the origin of the tumour, with a radius of approxi-
mately 8 (100 µm).
Cellular density [cells/(100 micrometer)3]
350
PANT
300 PAT
250
200
150
100
50
10 20 30 40 50 60 70 80 90 100
Lattice site [100 micrometer]
Figure 5.6: The tumour's composition at sixteen days.
Referring to Figure 5.6, after sixteen days, the tumour's dome shaped
density is more exaggerated and its radius has grown to approximately
14 (100 µm). Unlike conventional CA tumour models, cells in the bulk
of the tumour are still proliferating: the height of the 'dome' is seen
to increase from fourteen to sixteen days. This increase 'pushes' cells
outwards, causing the tumour to grow in radius. The tumour's radius
grows, primarily, because of the internal proliferation of its bulk.
56
Cellular density [cell/(100 micrometer)3]
600
Dead
PANT
500 PAT
QANT
400
300
200
100
10 20 30 40 50 60 70 80 90 100
Lattice site [100 micrometer]
Figure 5.7: The tumour's composition at eighteen days.
After eighteen days of growth, a dead (necrotic) core has emerged
at the tumour's centre, surrounded by a layer of quiescent anaerobic
(QANT) cells. The density distribution curve has developed a depression
at the tumour's core, due to the inability of the necrotic and QANT cells
to divide, while cells at the rim of the necrotic core are still dividing.
The mass of PAT and PANT cells has become large enough to cause
the glucose concentration in the centre of the tumour to drop below the
γqant threshold, thus causing the cells at the tumour's core to become
necrotic. As we move from the centre of the tumour to its edge, the
glucose concentration increases. The QANT layer has formed where the
glucose concentration is between the γqant and γpant thresholds.
Cellular density [cells/(100 micrometer)3]
600
Dead
PANT
500 PAT
QANT
400
300
200
100
10 20 30 40 50 60 70 80 90 100
Lattice site [100 micrometer]
Figure 5.8: The tumour's composition at nineteen days.
Referring to Figure 5.8, unlike the previous simulation, with the origi-
nal metabolic thresholds, the quiescent layer has maintained a consistent
57
width of ve lattice sites between day eighteen and nineteen.
Necrotic and QANT cells are seen to occupy a lattice site on the edge
of the necrotic core. QANT cells have been pushed into the necrotic
1
site .
Cellular density [cells/(100 micrometer)3]
600
Dead
PANT
500 PAT
QANT
400
300
200
100
10 20 30 40 50 60 70 80 90 100
Lattice site [100 micrometer]
Figure 5.9: The tumour's composition at twenty days.
The nal composition of the tumour can be seen in Figure 5.10.
Cellular density [cells/(100 micrometer)3]
700
Dead
600 PANT
PAT
500 QANT
400
300
200
100
10 20 30 40 50 60 70 80 90 100
Lattice site [100 micrometer]
Figure 5.10: The tumour's composition at twenty-one days.
After the emergence of the necrotic core, the PAT and PANT layers
grow out across the lattice maintaining a relatively consistent width,
approximately 2 and 15 (100 µm) respectively. While the cellular density
of the PANT layer steadily increases, the PAT rim maintains a relatively
consistent cellular density around the critical density.
1 Smaller bars have been superimposed over larger bars, thus obscuring some of
the larger bar.
58
100
200
80
150
60
40 100
20
50
20 40 60 80 100
Figure 5.11: A density plot of the PAT layer at twenty-one days, viewed
from above; cells in the other metabolic states are not shown.
Figure 5.11 describes the extreme of the tumour's spatial expansion
across the lattice. It can be seen that the tumour has a circular shape,
due to the constant concentration of glucose and oxygen on the lattice
perimeter in this simulation; as expected the tumour's growth is not
biased in any one direction.
We will now analyse the tumour morphology at a macroscopic level,
to ascertain what eect the tumour's microscopic evolution had on the
overall system.
3.5 ´ 106
QANT
6
3.0 ´ 10 Dead
PANT
2.5 ´ 106
Population @cellsD
PAT
6
2.0 ´ 10 Total
1.5 ´ 106
1.0 ´ 106
500 000
0
0 5 10 15 20
Time @daysD
Figure 5.12: The change in size of the tumour's metabolic populations
with time.
The QANT and PAT populations grow relatively slowly in comparison
59
to the PANT and dead populations that exhibit rapid growth. At the
end of the simulation, the dead population has grown to be greater than
the PANT population.
106 QANT
Dead
PANT
PAT
Population @cellsD
4
10 Total
100
1
0 5 10 15 20
Time @daysD
Figure 5.13: The change in size of the tumour's metabolic populations
with time, plotted on a logarithmic scale.
Referring to Figure 5.13, the PAT population size increases exponen-
tially until, approximately, day twelve (obscured by the blue line). The
end of the PAT population's exponential growth corresponds with the
creation of cells in a PANT state. The PANT population then grows
exponentially, from (approximately) day thirteen to eighteen. The end
of the PANT population's exponential growth corresponds with the cre-
ation of cells in a QANT and dead state.
The tumour exhibits an exponential growth rate in its cellular popu-
lation. The growth rate slows at approximately eighteen days, with the
development of a necrotic core. The tumour achieves a maximum cellular
population of approximately 4.8 × 106 , after twenty-one days of growth.
60
50
40
Tumour radius @H100 ΜmLD
30
20
10
0
0 5 10 15 20
Time @daysD
Figure 5.14: The growth in tumour radius with time.
Referring to Figure 5.14, at the start of the simulation all the cells
are located in the central lattice site that was seeded with the original
cell. While the central site's density is less than the critical density, the
pressure is not su cient to push cells into neighbouring sites, thus all
the cells remain in the central site; the tumour has a radius less than our
lattice resolution of 100 µm. When the density of the central site reaches
the critical density the tumour begins to grow out across the ECM.
61
50
Tumour radius @H100 ΜmLD
20
10
1
10 15 20
Time @daysD
Figure 5.15: The growth in tumour radius with time, plotted on a loga-
rithmic scale.
Referring to Figure 5.15, the tumour's radius is seen to exhibit ex-
ponential growth for the rst seventeen days, before slowing to linear
growth.
600
Density @HcellsLH100 ΜmL3 D
500
400
300
200
100
0
0 5 10 15 20
Time @daysD
Figure 5.16: The change in maximum cellular density with time.
Referring to Figure 5.16, initially, the maximum cellular density de-
scribes the density of the central site. The maximum cellular density
grows exponentially for the rst seven days, which has been checked by
62
plotting the change in maximum cellular density with time on a logarith-
mic axis ( gure not included). The point at which the maximum cellular
density equals the critical density, the seventh day, corresponds with the
point at which the tumour radius begins to grow beyond the central site,
see Figure 5.14.
After exceeding the critical density, the maximum cellular density
grows slowly. The large dierence in pressure between the central site
and its neighbours causes most new cells, created in the central site, to
be pushed into its neighbouring sites. Cell movement acts to homogenise
the pressure. As pressure dierences equalise, less cells are pushed out of
the central site and the maximum cellular density begins to grow more
rapidly.
Growth in the maximum cellular density then slows at, approxi-
mately, eighteen days corresponding to the onset of cells in a quiescent
and necrotic state. The tumour eventually achieves a maximum cellu-
lar density of 626 (cells)(100 µm)−3 , which is below the maximum site
density (ρ̄) of 691 (cells)(100 µm)−3 . An additional experiment was per-
formed, to observe the simulated tumour's behaviour on reaching the
maximum site density.
The new maximum site density (ρ̄) was set to equal 300 (cells)(100 µm)−3 ,
all other variables were the same as in the previous experiment. A density
of 300 (cells)(100 µm)−3 is on the lower bound of the range of tumour
densities recorded experimentally by [13].
63
50
Tumour radius @H100 ΜmLD
20
10
1
10 15 20 25
Time @daysD
Figure 5.17: The change in tumour radius with time, for a maximum site
−3
density of 300 (cells)(100 µm) , plotted on a logarithmic scale.
106
105
Population @cellsD
104
1000
100
10
1
0 5 10 15 20 25
Time @daysD
Figure 5.18: The change in cellular population with time, for a maximum
−3
site density of 300 (cells)(100 µm) , plotted on a logarithmic scale.
Some sites achieve the maximum site density at approximately four-
teen days, which has been seen by plotting the maximum site density
with time ( gure not included). On reaching the maximum site den-
sity, the growth rate of both the tumour's radius, Figure 5.17, and the
tumour's cellular density, Figure 5.18, transitions from exponential to
64
linear growth.
5.3 Comparison with in vitro Experiments
In the later stages of tumour growth, once a nutrient concentration gra-
dient has formed, the simulated tumour's morphology resembles those
seen in experimental histologies [38], and described by Figure 2.1. The
tumour is composed of a necrotic core, surrounded by cells in a quiescent
state, with proliferating cells located on its rim.
The tumour's morphology was also compared on quantitative aspects,
with data found in the literature. Freyer and Sutherland [13], report on
assays of multicellular spheroid of the EMT6/Ro mouse mammary tu-
mour cell line. Many of the model parameters' values were referenced
from Freyer and Sutherland, such as maximum site density, medium nu-
trient concentrations and the mitotic cycle time. It is therefore appropri-
ate to compare the simulated tumour's morphology with those reported
in [13].
Freyer and Sutherland report that after ve days of growth, the av-
erage diameter of their tumour spheroid grew linearly, at a rate of 75
µm per day [13, Page 519]. In comparison, our tumour's diameter grew
linearly after approximate thirteen days (Fig. 5.15), at a rate of approxi-
mately 400 µm per day. The larger growth rate of our tumour has meant
we are not able to quantitatively replicated Freyer and Sutherland's nd-
ing, but by demonstrating a linear growth rate in diameter, our model
has been able to qualitatively replicate the behaviour of those tumours
observed experimentally.
The average number of cells per spheroid, as a function of time, were
also described in [13]. The number of cell per spheroid as a function of
time, plotted on a logarithmic scale, indicates that the growth rate of
the cellular population goes from exponential growth to linear growth
at, approximately, twelve days [13, Fig. 1B]. Again, our tumour's larger
growth rate has caused its cellular population to exceed those of Freyer
and Sutherland. Our model was, however, able to replicate the transition
from an exponential growth rate in cellular population to a linear growth
rate, which is con rmed by Figure 5.18.
Freyer and Sutherland also report data regarding the emergence of
a necrotic core. They report that necrosis rst developed in tumour
spheroids with a diameter of 400 µm, but it was not until the tumour
had achieved a diameter of 1000 µm that the necrotic fraction constituted
65
a signi cant portion. Further growth of their tumour spheroids resulted
in a large increase in the relative amount of necrotic material [13, Page
521]. In contrast, our tumour's diameter was approximately 5000 µm
when a signi cant necrotic core rst developed (see Figure 5.7). Like
those tumours studied by Freyer and Sutherland, the relative amount
of necrotic material then steadily increased until it eventually came to
dominate the tumour, see Figure 5.12.
Our model has been able to qualitatively replicate many of the phe-
nomena observed experimentally, such as a linear growth rate in diame-
ter, a transition from exponential to linear growth in cellular population
and the growth of a necrotic core that eventually dominates the tumour.
66
Chapter 6
Conclusion
The aim of this project was to develop an approach to modelling pressure-
induced cell movement and tumour invasion. When the cells in one region
of a tumour proliferate more quickly than the cells in another, density
dierences result. Density dierences lead to a build-up of pressure that
pushes cells from densely populated regions to sparsely populated regions.
This pushing eect grows the tumour outwards into the surrounding
healthy tissue, thus promoting tumour invasion.
It is inherently di cult to model pressure-induced cell movement and
proliferation in a traditional CA tumour model. Traditional CA tumour
models are formulated on a xed, regular lattice, where each lattice site
represents a cell. The restrictions imposed by the model's structure re-
quire a dividing cell be near an empty site in which the new cell can be
placed. It is not possible for density dierence to develop, except at the
tumour's boundary, when only cells near empty sites can divide.
In this project, a CA tumour model that captures realistic pressure-
induced cell movement and tumour growth has been developed. By allow-
ing several cells to occupy one lattice site, we have been able to introduce
local cell density information into a CA model. Based on physical prin-
ciples and porous media theory, a relationship between pressure and cell
density has been proposed that has allowed us to model pressure-induced
cell movement and the inhomogeneous diusion of nutrient molecules.
Applying this model to in vitro conditions, we have simulated tumour
growth that qualitatively replicates experimentally observed phenomena,
such as the emergence of the typical layered structure with expanding
radius. However, quantitative aspects of simulated tumour growth were
found not to re ect reality with the current model parameter estimates.
The simulated tumour grew much faster than those observed by Freyer
and Sutherland [13], having a larger growth rate in diameter and cellular
67
population. Performing a parametric study to ascertain what eect vary-
ing the model's parameters has on the simulated tumour's growth would
provide a more quantitative understanding of the mechanisms underlying
the model, and enable us to identify a set of parameters that will cause
the simulated tumour's growth to better re ect reality.
Designing the model, we neglecting to model chemotaxis induced cell
movement and the production of hydrogen ions, as a by-product of cell
metabolism. Chemotaxis directly eects a cell's movement and hydrogen
ion concentration (pH) eects a cell's viability. We intend to incorporate
these factors into our model, as they both in uence tumour growth.
This model constitutes the rst stage of a larger goal to create a CA
model of solid tumour growth in the bone environment, which can be
used to simulate prostatic tumour growth in bone. The present model
will need to be adapted to investigate prostatic tumour growth in bone,
by modelling pressure pro les in patterns found in the bone environment.
It would then be possible to seed the model with prostate metastases and
simulate tumour growth under varying conditions.
It is expected that hard tissue, such as bone, will constrain a tu-
mour's growth, causing a buildup of inter-cellular pressure within the
tumour. Under such circumstances, inter-cellular pressure may have a
dominant in uence over the tumour's growth, and therefore needs to be
taken into account. It is hoped that this CA tumour model will become
an invaluable tool for studying solid tumour growth.
68
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72
Chapter 7
Appendices
7.1 An Alternative Relationship Describing
Cell Volume as a Function of Pressure
Cell volume, as a function of the pressure experienced on its membrane,
has been investigated theoretically on robust grounds. The surface of
a cell is formed by lipids that constitute the membrane bi-layer, the
shape of a cell is therefore determined by its membrane's shape [43]. The
common understanding of the energetics of membrane shaping are largely
based on the seminal Helfrich theory of membrane bending elasticity
[19], from which Helfrich was able to describe a spherical cell's shape
as a function of pressure, refer to Table 7.1 for a descriptions of the
parameters.
k c c0 2 2λ
p(R) = ( − c0 − ) (7.1)
R R k c c0
Description Symbol Value Units Ref.
−1 −2
Osmotic pressure p (kg)(100 µm) (s) [35]
Cell radius R (100 µm) [35]
Bending moduli kc 20 kB T J [34]
Spontaneous curvature c0 1/R0 (100 µm)−1 [34]
Surface tension λ 0 (J)(100 µm)−1 [20]
Scaled Boltzmann constant kB 1.38 × 10−15 (kg)(100 µm)2 (s)−2 (K)−1 [35]
Room temperature T 298 K [35]
Spontaneous radius R0 (100 µm)
Table 7.1: A description of parameters describing a spherical cell's shape
as a function of pressure.
The membrane composition controls the non-deformed unstressed
73
shape of the membrane, which is referred to as the 'spontaneous shape',
which is characterised by the spontaneous curvature. The spontaneous
curvature of the whole membrane depends on the spontaneous curvature
of the inner and outer membrane monolayers [43].
Note, Equation 7.1 describes the osmotic pressure across a cell's mem-
brane, which is the dierence between the external and internal cell pres-
sure. By using this equation to describe only the external pressure, we
are making the implicit assumption that the internal pressure is zero,
which is a valid assumption if the internal cell pressure is very low.
Assuming that cells are spherical vesicles that conserve their spheri-
cal shape, but are allowed to modify their radius and therefore volume,
according to the pressure experienced on their membrane allows us to
determine a cell's volume as a function of pressure. The volume of a
sphere, as a function of its radius:
4 π R3
Vsphere (R) = (7.2)
3
Resolving Equation (7.1) for R and combining with Equation (7.2), yields
a relationship for cell volume as a function of pressure:
1
Vcell (p) = p (7.3)
(β1 + β12 + β2 p)3
Where:
β4
• β1 = √
3
β3
, with units of (100 µm)−1 .
• β2 = √1
( 3 β3 )2
, with units of (J)(100 µm)−1 .
q √
• β3 = 3 4π
3
( 2 kc c0 )3 , with units of (J)3/2 (100 µm)−3/2 .
√ c0
2 k c c0 + k λc
• β4 = 2
2 c 0
, with units of (J)1/2 (100 µm)−3/2 .
The pressure as a function of cell volume can be found from the inverse
of Equation (7.3) as:
−1 −1
V 3 (V 3 − 2 β1 )
p(Vcell ) = cell cell (7.4)
β2
In-order to solve Equation (7.3) it is necessary to calculate the sponta-
neous curvature (c0 ). As only the spontaneous curvature is unknown,
it is, in principle, possible nd the spontaneous curvature algebraically.
However, we found that no real solutions for c0 exist for the intrinsic
volume.
74
1.4 ´ 10-10
Cell volume @H100 ΜmL3 D 1.2 ´ 10-10
1. ´ 10-10
8. ´ 10-11
6. ´ 10-11
4. ´ 10-11
2. ´ 10-11
0
0 2000 4000 6000 8000 10 000
c0 @H100 ΜmL-1 D
Figure 7.1: The change in cell volume with spontaneous curvature.
Indeed, referring to Figure 7.1, it can be seen that the curve describing
cell volume as a function of c0 never reaches the intrinsic cell volume
(Vcell0 ) of 0.00419 (100 µm)3 , no real solutions of c0 exist. It appears
that Equation (7.1) describes the shape of a empty lipid bi-layer, not one
containing cellular material, as the intrinsic shape is much smaller than
that of a cell.
Alternatively the shape of a cell can be described phenomenologically.
Equation (7.3) describes a cell's volume as a function of β1 , β2 and p. We
wish to nd values for β1 and β2 , so that Vcell (p0 ) = Vcell0 , the cell volume
equals the intrinsic cell volume for the IFP of normal tissue. This can
be achieved by solving Equation (7.3) for β2 , and then substituting the
intrinsic cell volume (Vcell0 ) for Vcell and IFP of normal tissue (p0 ) for p:
−2 −1
(V 3 − 2 β1 Vcell30 )
β2 (β1 ) = cell0 (7.5)
p0
Since β2 ≥ 0 (see Eq. (7.3)), the range of acceptable values for β1 can be
found from Equation (7.5) as:
−2 −1
Vcell30 − 2 β1 Vcell30 > 0 (7.6)
which can be solved as:
−1
V 3
β1 < cell0 = 3.10175 (100 µm)−1 (7.7)
2
The change in cell volume with β1 can be found by substituting Equation
75
(7.5) into Equation (7.3):
1
Vcell (p) = p
2
(7.8)
(β1 + β1 + β2 (β1 ) p)3
Figure (7.2) and (7.3) describe how cell volume changes with pressure
when β1 is in the range 1 − 3, the most dramatic change in cell volume
occurs when β1 = 1.
0.0050
Β1 = 1
0.0048
Cell volume @H100 ΜmL3 D
Β1 = 2
0.0046
0.0044
Β1 = 3
0.0042
0.0040
0.080 0.085 0.090 0.095 0.100
Pressure @kg H100 ΜmLHsL2 D
Figure 7.2: Cell volume as a function of pressure, according to Equation
(7.8).
0.12
0.10
Cell volume @H100 ΜmL3 D
0.08
Β1 = 1
0.06
0.04
0.02 Β1 = 2
Β1 = 3
0.00
0.00 0.02 0.04 0.06 0.08 0.10
Pressure @kg H100 ΜmLHsL2 D
Figure 7.3: Cell volume as a function of pressure, according to Equation
(7.8) for a large range of pressure values.
76
0.12
Cell volume @H100 ΜmL3 D 0.10
0.08
0.06
0.04
0.02
0.00
0.00 0.02 0.04 0.06 0.08 0.10
Pressure @kg H100 ΜmLHsL2 D
Figure 7.4: A comparison of the alternative approaches for describing
cell volume as a function the pressure experienced on its membrane.
The green line denotes the inverse relationship between cell volume and
pressure, described by Equation (4.11). The blue lines denote the volume
of an empty lipid bi-layer as a function of pressure, described by Equation
(7.1).
Referring to Figure 7.4, it can be seen that the two equations de-
scribing a cell's volume as a function of pressure, Equation (4.11) and
Equation (7.8), are similar when β1 = 1 for Equation (7.8).
The average inter-cellular pressure as a function of cellular density
can be found by substituting Equation (4.16) into Equation (7.4), which
yields:
2 1
( 1−φ
ρ
)− 3 − 2 β1 ( 1−φ
ρ
)− 3
p(ρ) = (7.9)
β2
Figure 7.5 describes how average inter-cellular pressure changes with
pressure when β1 is in the range 1 − 3, the most dramatic change in
average inter-cellular pressure occurs when β1 = 3.
77
7
Β1 = 3
Pressure @kg H100 ΜmLHsL2 D 6
2
Β1 = 2
1
Β1 = 1
0
0 200 400 600 800 1000 1200 1400
Density @cells H100 ΜmL3 D
Figure 7.5: The average inter-cellular pressure as a function of density,
from Equation (7.4), assuming cells deform under increasing pressure.
2.0
Pressure @kg H100 ΜmLHsL2 D
1.5
1.0
0.5
0.0
0 200 400 600 800 1000 1200 1400
Density @cells H100 ΜmL3 D
Figure 7.6: A comparison of the alternative approaches for describing
average inter-cellular pressure as a function of density. The green line
denotes the pressure as a function of density for the inverse relationship
between cell volume and pressure, described by Equation (4.11). The
blue lines denote the pressure as a function of density for an empty lipid
bi-layer, described by Equation (7.1).
78
7.2 Discretising a Continuous Chemical Con-
centration
Below, our method of discretising a continuous chemical concentration
on to the model lattice will be discussed. The method will be described
for the glucose concentration, which is the same for that of the oxygen
concentration. When the apparent glucose diusion coe cient (Dg ) is
invariant with position, such as for an inhomogeneous cellular density,
the continuous glucose concentration is expressed as:
∂g
= ∇ · (Dg ∇ g) (7.10)
∂t
or
∂g(x, y, t) ∂ ∂g(x, y, t) ∂ ∂g(x, y, t)
= Dg (x, y) + Dg (x, y) (7.11)
∂t ∂x ∂x ∂y ∂y
Using the Forward Euler method in time [5], the derivative on the
left-hand side can be approximated by:
∂g(x, y, t) g(x, y, t + 4t) − g(x, y, t)
≈ (7.12)
∂t 4t
Using the central dierence approximation for the second derivatives,
taken from [25, Eq. 4.23], the rst term on the right-hand side of Equa-
tion (7.11), can be expressed as:
h
∂
∂x
Dg (x, y) ∂g(x,y,t)
∂x
≈ i
1 Dg (xi+1 ,y)+Dg (x,y) g(xi+1 ,y,t)−g(x,y,t) Dg (x,y)+Dg (xi−1 ,y) g(x,y,t)−g(xi−1 ,y,t)
4x
(( 2
)( 4x
)) − (( 2
)( 4x
))
Simplifying:
∂
∂x
Dg (x, y) ∂g(x,y,t)
∂x
≈ (0.5 (Dg (xi+1 ,y)+Dg (x,y))
(4x)2
(g(xi+1 ,y,t)−g(x,y,t))
−
(0.5 (Dg (x,y)+Dg (xi−1 ,y)) (g(x,y,t)−g(xi−1 ,y,t))
(4x)2
Equation (7.11) can be discretised as:
g(x, y, t + 4t) =
4t
g(x, y, t)+ (4x)2 [((0.5 (Dg (xi+1 , y) + Dg (x, y)) (g(xi+1 , y, t) − g(x, y, t)))−
((0.5 (Dg (x, y) + Dg (xi−1 , y)) (g(x, y, t) − g(xi−1 , y, t)))] +
4t
(4y)2
[((0.5 (Dg (x, yj+1 ) + Dg (x, y)) (g(x, yj+1 , t) − g(x, y, t))) −
((0.5 (Dg (x, y) + Dg (x, yj−1 )) (g(x, y, t) − g(x, yj−1 , t)))]
79
Press et al. [32] describe a heuristic stability criterion of:
(4x)2
4t ≤ mini (7.13)
2 (0.5 (Dg (xi+1 , y) + Dg (x, y))
or
(4y)2
4t ≤ minj (7.14)
2 (0.5 (Dg (x, yj+1 ) + Dg (x, y))
7.3 The Adiabatic Approximation
7.3.1 A Theoretical Analysis of the Adiabatic Ap-
proximation
Below, the theoretical basis of the adiabatic approximation is explored.
Glucose is chosen as our representative chemical concentration for this
discussion, equally oxygen or any other chemical consumed or produced
by tumour cells could have been used. The spatio-temporal evolution of
glucose is described by Equation (4.34), where the consumption rate γσ
is the same for all cell types. Dividing Equation (4.34) by γσ yields:
1 ∂g Dg γv
=∇ ∇ g − ρσ g + ρv (7.15)
γσ ∂t γσ γσ
Assuming γσ and γv are of the same order of magnitude, the terms on
the right-hand side will be of order 1 whereas the term on the left-hand
side will be of order 1/γσ . If is it assumed that γσ 1, then the change
in glucose concentration with respect to time will be negligible, so that:
Dg γv
0=∇ ∇ g − ρσ g + ρv (7.16)
γσ γσ
Thus, the glucose concentration eld will instantaneously reach a steady
state in our model when the tumour cell consumption rate is high.
Let us now assume that the glucose diusion coe cient is low, such
that Dg 1, which will occur when the cellular density is high. Now,
only the glucose production and consumption terms will be of order 1.
The glucose concentration and diusion terms will be of order 1/γσ , if
we again assume γσ 1:
γσ ρ g = γv ρv (7.17)
When the cellular density is high, the diusion into and out of a
lattice site with respect to time will be negligible, as will be the change
80
in glucose concentration. Glucose consumption will be equal to glucose
production, if glucose is not produced within a lattice site, cells within
that lattice site will not consume any glucose and will eventually become
necrotic. This nding supports the hypothesis that an avascular tumour's
necrotic core is caused by its increased cellular density.
Even with a small diusion coe cient, diusion into a lattice will
occur given a long enough time interval. However, when the time interval
is increased so that it approximates that of the cellular time scale, the
adiabatic approximation will be invalidated, because we can no-longer
assume the chemical concentration has reached equilibrium.
7.3.2 A Numerical Analysis of the Adiabatic Ap-
proximation Limit
The theoretical analysis of the adiabatic approximation does not apply
to a necrotic tumour. Necrotic tumours do not consume nutrients and
therefore have a consumption rate of zero. A numerical analysis enabled
us to quantitatively assess what eect a necrotic tumour's cellular density
and radius has on the time taken for a glucose and oxygen concentration
eld to reach equilibrium.
The numerical analysis was conducted on a square lattice with con-
stant chemical concentration at the perimeter and zero concentration
within the tumour growth region. A tumour composed of necrotic tissue
was modelled at the lattice centre. Over a series of tests, the tumour's
cellular density was exponentially increased while the tumour's radius
was incrementally increased. The increase of a lattice site's cellular den-
sity was realised through a decrease in its diusion coe cients. The
time take for the chemical concentration elds to reach equilibrium were
recorded for each test.
As a necrotic tumour does not consume glucose or oxygen, given a
long enough time, a homogeneous concentration of glucose and oxygen
would form across the entire lattice. To reduce simulation time, a chemi-
cal steady state is de ned as a state where the chemical concentration of
any lattice site does not change by more than 10−6 % from one iteration
to the next.
Table 7.2 and 7.3 describes the experiment's results for the glucose
and oxygen concentration elds respectively.
81
Tumour radius [100 um]
Dg0 /Dg 5 10 15 20 25 30
1 1.46 1.46 1.46 1.46 1.46 1.46
2 1.52 1.67 1.88 2.11 2.32 2.5
4 1.64 2.14 2.78 3.42 4 4.47
8 1.93 3.14 4.56 5.93 7.14 8.13
16 2.58 5.12 7.92 10.6 12.96 14.88
32 3.96 8.88 14.17 19.19 23.6 27.2
64 6.65 15.85 25.6 34.81 42.91 49.5
Table 7.2: The glucose steady state time in hours.
Tumour radius [100 µm]
Do0 /Do 5 10 15 20 25 30
1 6.26 6.26 6.26 6.26 6.26 6.26
2 6.51 7.16 8.05 9.01 9.89 10.61
4 7.04 9.12 11.79 14.45 16.81 18.73
8 8.26 13.28 19.09 24.68 29.6 33.61
16 10.98 21.4 32.72 43.43 52.83 60.48
32 16.66 36.55 57.5 77.17 94.37 108.38
64 37.59 64.04 101.77 136.91 167.52 192.46
Table 7.3: The oxygen steady state time in hours.
Where Dg0 /Dg and Do0 /Do are the ratio of the glucose and oxygen
diusion coe cients in the uid phase (in the absence of cells) to the
diusion coe cients within the tumour. Figure 7.7 and 7.8 graphically
describe how the time taken for the glucose and oxygen concentration
elds to reach equilibrium changes with the tumour's density and radius.
82
The Glucose Steady State Time for a Necrotic Tumour
30 45
40
Tumour radius [100 micrometers] 25
35
20 30
25
15 20
15
10
10
Steady state time [h]
5 5
1 2 4 8 16 32 64
D0/D1
Figure 7.7: The steady state contours for the glucose concentration eld.
The Oxygen Steady State Time for a Necrotic Tumour
30 180
160
Tumour radius [100 micrometer]
25
140
20 120
100
15 80
60
10
40
Steady state time [h]
5 20
1 2 4 8 16 32 64
D0/D1
Figure 7.8: The steady state contours for the oxygen concentration eld.
Figure 7.7 and 7.8 describes an upper-bound on the adiabatic approx-
imation assumption that underlies the modelling of chemical diusion.
Provided the time for a chemical eld to reach equilibrium does not ex-
ceed the model's representative time, the mitotic cycle time (cell doubling
time), the adiabatic approximation will hold and enable us to ignore a
chemical concentration eld's evolutions and directly calculate its steady
state.
83
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