Transcript Development of in-vitro Models to Study Breast Cancer Progression

Section of Pathology and Tumour Biology
Development of laboratory models
to study Breast Cancer
Deborah Holliday
Breast Research Group
Section of Pathology & Tumour Biology
Outline
 Introduction to the cells found in breast
tissue
 Changes in breast cells during breast cancer
 Designing a model of breast cancer
 methods
 Use of the model: A tool for looking at cancer
progression
Cellular components of the normal breast
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Luminal epithelial cells:
 Milk producing cells
 Hormone responsive ER-alpha positive cells
Myoepithelial cells:
 Surround the luminal cells
 Contractile cells
Luminal and Myoepithelial cells form the glandular unit of the
breast
Fibroblasts:
 Form the structural component of the surrounding breast
tissue
 Produce proteins important for maintaining breast structure
Other cell types
 Blood vessels, fat cells, inflammatory cells
Breast Cancer Progression
Luminal cells Myoepithelial
cells
Tumour
cells
Fibroblasts
Normal breast tissue
 Cells look ordered in
appearance
Pre-invasive breast
cancer
 Tumour cells in the
centre start to grow
out of control
Invasive breast cancer
 Tumour cells escape into the
surrounding breast tissue
Ordered structure of the
tissue is lost
Pre-invasive breast cancer: Ductal carcinoma in
situ (DCIS)
 DCIS is characterised by confinement of tumour cells
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to the breast glandular unit
DCIS accounts for 40% of screen detected breast
carcinoma
25-30% of untreated DCIS will progress to invasive
carcinoma
Problems with treatment of DCIS

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Mastectomy
 Patient is cured but ? Overtreatment ?
Conservative surgery
 A proportion of tumours will recur
 Some of those will progress to invasive carcinoma
Important to define which DCIS cases
are likely to progress
A better understanding of the biology of
tumour invasion may reveal new targets
for therapies
Designing a human model of breast cancer

The model would need to include 3 major cell types involved in
breast cancer:
 Tumour cells.
 Myoepithelial cells (protector cells).
 Fibroblasts (tumour helper cells).
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Cells would need to be grown in culture conditions which resemble
those in the body:
 Able to grow in 3 dimensions rather than on a plastic Petri
dish.
 Such a model would be a valuable tool:
 To help us understand how breast cancer progresses.
 To allow us to test new drugs for therapy.
 To potentially identify new targets for future drug
development.
Methods
We isolated cells from normal breast tissue or from
breast cancer tissue and grew then in a 3 dimensional
matrix of collagen.
 By labelling our cells with different colours we were
able to identify the different cell types in our model.
 We used the model to investigate whether fibroblasts
are able to make pre-invasive lesions become invasive.
Myoepithelial
cells
Luminal cells
Fibroblasts
Collagen I
Culture media
Results
B
Day 1
Day 3
Blue: Tumour cells
Day 5
Red: Myoepithelial cells
Day 7
Green: Normal Fibroblasts
F
Green: Tumour Fibroblasts
Invasive breast
Carcinoma
Ductal carcinoma
in situ (DCIS)
Quantifying the model
Normal Fibroblasts
Normal Fibroblasts
10
Tumour Fibroblasts
Tumour Fibroblasts
number of structures per field
9
8
7
6
5
4
3
2
*
1
0
lum/myo/Nfib
mcf/myo/TAF
Summary
• We have a model which we can use to study the biology
of breast cancer
• This will help us understand how breast cancer
progresses
How we are using the model
• Different tumour cells to represent different types of breast cancer
• Different fibroblasts to understand why in some patients cancer
progresses faster than in others
• Include drugs into the model with tumour fibroblasts to see if we can
prevent ‘invasion’
– Established drugs
– New Drugs = pre-clinical drug screen
Section of Pathology and Tumour Biology
Using 3D models to study radioresistance in Breast Cancer
Laura Smith
Breast Research Group
Section of Pathology & Tumour Biology
Outline
 Radiotherapy
 Issues with radiotherapy
 What would help overcome these issues?
 The use of 3D models
Radiotherapy
 Reduces risk of the cancer coming back
 Is given to many breast cancer patients
 All patients having breast conserving surgery
 Patients having a mastectomy but at high risk
of the cancer coming back
Issues with Radiotherapy
 Unpleasant side effects
 Short term
 Long term
 Stressful regime
 Daily hospital visits 5 days/ week for 3 weeks
 Limited availability of treatment machines
 Long waiting lists in some areas
 Not all patients will benefit
 Some patients cancer will come back anyway
Overcoming these issues
 Better patient selection
 Estrogen Receptor for Tamoxifen therapy
 HER2 for Herceptin therapy
 Nothing analogous to guide radiotherapy
 Why do some cancers respond well to
radiotherapy whilst others do not?
 What factors are involved?
 Radio-sensitizing drugs?
Our Study
 It is not only cancer cells that are exposed
to radiotherapy but also the fibroblasts
 Do fibroblasts influence breast cancer cell
response to radiotherapy?
 Do fibroblasts differentially influence the
response of different breast cancer types?
Use of 3D Models
Myoepithelial
cells
Luminal cells
Untreated Fibroblasts
Collagen I
Culture media
Myoepithelial
cells
Luminal cells
Treated Fibroblasts
Collagen I
Culture media
Use of 3D Models
Myoepithelial
cells
Luminal cells Type I
Treated Fibroblasts
Collagen I
Culture media
Myoepithelial
cells
Luminal cells Type II
Treated Fibroblasts
Collagen I
Culture media
Summary
 Reduce side effects and improve quality of
life for patients that will not benefit
 Allow drs to select another type of
treatment that will work for these patients
 Reduce waiting times for those patients
that will benefit thereby increasing
survival