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Personnel
The
Unit of Molecular and Cellular Oncology is made up from a team of
14 academic and research staff and 5 postgraduate students as follows:
Senior
Staff:
Professor Susan Watson, Professor of Pre-Clinical
Oncology, Head of Division, PRECOS Director
Dr Anna Grabowska, Lecturer, PRECOS Consultant:
Molecular biology & gene delivery
Dr. Abdolrahman Nateri, Lecturer, Cancer Genetics
Dr Rajendra Kumari, Senior Research Fellow, PRECOS
Project Manager: Cell signalling, molecular pharmacology and cancer
model development
Dr Philip Clarke, Chief Experimental Officer,
PRECOS Senior Research Officer: Immunohistology & bioimaging
Mr Andrew McKenzie, Chief Experimental Officer,
PRECOS Senior Research Officer: Pharmacology & cancer model
development
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Technical
Staff:
Amanda Tobias, Research Technician: Immunohistology and bio-imaging
Miss Jaime
Hughes, Research Technician: Molecular biology
Miss Narinder Mann, Research Technician: Tissue culture and cell
biology
PG
Students:
Eve Royal (PhD): Investigation into the role
of HIF-1alpha in the progression of gastrointestinal tumours
Yinfei Yin (PhD): Effect of gastrin on MMP/ECM interactions
in cancer invasion and metastasis
Peter Boyce (PhD): Chemically modified siRNA in the
treatment of gastrointestinal cancer
Naseem Waraich (DM): A prospective study to relate
androgen and androgen receptor expression levels to fibroblast growth
factor receptor gene and protein expression by Barrett’s Oesophagus
and malignant disorders of the oesophagus
Amanda Tobias (MPhil): Gastrin interactions which
impact upon gastric and colonic carcinogenesis
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Professor Susan Watson, Professor of Pre-Clinical Oncology, Head
of Division
Current
Research Projects
Interfering
RNA and in vivo delivery systems
Expression
of gastrin receptor isoforms
Identification
of gastrin receptor antagonists
Translational
control of gastrin and gastrin receptor expression
Role
of non-amidated gastrins in tumourigenesis
Role
of PTHrP in breast cancer
Interactions
between Barrett's Oesophagus and gastrin
Effect
of Helicobacter pylori on HB-EGF shedding
Effect
of gastrin on gastric stem cell proliferation
Role
of PTHrP and adhesion
Effect
of gastrin on pre-malignant lesions of the colon
Interests
Hormones
and cancer - gastrin
siRNA
In
vivo tumour
modelling
G-protein
coupled receptors
PTHrP
HIF-1alpha
Matrix
metalloproteinases
Chemoprophylaxis
PKB/Akt
and apoptosis
Techniques
Tissue
culture
Radioactive
labelling/RIA
Real-time
PCR
Interfering
RNA
Western
blotting
ELISA
Immunohistochemistry
Molecular
cloning
Immunofluorescence
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Dr Anna Grabowska, Lecturer
Research
Interests
Transcriptional
and translational control of expression of molecules involved in
gastrointestinal cancer progression, including investigation of
expression of the gastric hormone gastrin and its receptor in gastrointestinal
tumours. Use of interfering RNA to identify genes involved in tumorigenesis
and developing improved methods for delivery of siRNA to tumours.
Development of molecular tools to monitor signalling pathways activated
in tumours in real time.
Technical
Expertise
Molecular biology
Interfering RNA
Real time PCR
Expression cloning
Reporter systems
Immunology
Phage peptide display library technology
ELISA
Western blotting
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Dr Rajendra Kumari, Senior Research Fellow, PRECOS Project Manager
Research
Interests
Chemoprophylaxis
of GI malignancy: Chemo-prophylaxis of stomach and
bowel cancer with focus on therapeutic manipulation of gastrin and
cyclooxygenase-2 as well as screening tests based at identifying
carcinogenic strains of Helicobacter pylori funded by CORE.
Breast Cancer and PTHrP:
Breast cancer cells secrete high levels of Parathyroid hormone-related
protein (PTHrP), accounting for the high prevalence of humoral hypocalcaemia
of malignancy (HHM) in breast cancer patients. There are 3 isoforms
of PTHrP which are processed and proteolytically cleaved to yield
mature secretory peptides. These are PTH-like N-terminal, nuclear
localising mid-region peptide, osteostatin and a C-terminal peptide
of aa 141-173, which is only produced by the 173 isoform. The role
of these peptides in breast cancer growth and metastasis to bone
is being investigated.
Hypoxia:
The role of hypoxia in tumour cell survival has identified the transcription
factor hypoxia inducible factor-1 (HIF-1) as a key target for anti-cancer
therapeutic design. The role in tumour metastasis and mechanism
of action of inhibitors of the hypoxic pathway are being investigated.
Pre-Clinical
Tumour Modelling: Development
and validation of cancer models for pre-clinical evaluation of anti-cancer
agents.
Technical
Expertise
GPCR pharmacology (single cell, real time and plate based calcium
signalling, second messenger signalling, antagonist profiling, internalisation)
In
vitro analysis of intracellular signalling downstream of GPCRs,
RTKs and protein kinases in cancer cells (protein expression, western
blotting, fluorescent based assays, ELISAs, cell growth, apoptosis,
mechanism of action of inhibitors)
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Dr Philip Clarke, Chief Experimental Officer
Research
Interests
My primary research interests are linked to the peptide hormone
gastrin and its effects on Gastro-intestinal (GI) malignancies.
Gastrin, secreted by G cells of the antrum and duodenum under normal
physiological conditions, is involved in the endocrine control of
gastric acid secretion and mucosal growth. GI malignancies show
de novo activation of the gastrin gene resulting in production
of multiple bio-active gastrin peptides capable of exerting control
at several points in the stepwise progression to malignancy via
endocrine/paracrine/juxtacrine mechanisms. My other interests include
tumour associated angiogenesis, pre-clinical model development and
biophotonic ‘molecular’ imaging.
Technical
Expertise
Histology
– immunohistochemistry, brightfield and fluorescent microscopy,
paraffin embedded tissue and cell microarray construction
Computer aided image analysis
Biochemistry – Protein identification (electrophoresis, blotting),
purification, quantification (gel documentation)
Tissue culture.
Real time biophotonic imaging
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Mr Andrew McKenzie, Chief Experimental Officer
Research
Interests
Primary research interests primarily focusing on gastrointestinal
carcinogenesis; encompassing immunological and peptide directed
therapy, and the development of patient relevant tumour models.
Technical
expertise
Tumour
Modelling: Development of patient relevant pre-clinical models of
oncology and the metastatic cascade
Pharmacological
profiling: GPCRs and agonist-antagonist interactions, cell growth/death
and apoptosis assays
Radiopharmacological-directed
imaging and therapy
Real-time
bioluminescent imaging
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Eve Royal (PhD): Investigation into the role of HIF-1alpha in the
progression of gastrointestinal tumours
Hypoxia-inducible
factor 1 (HIF-1) is a dimeric transcription factor composed of an
inducible alpha subunit and a constitutively expressed beta subunit.
It plays an important role in allowing cellular adaptation to low
oxygen concentrations (hypoxia) and as a result the expression of
HIF-1alpha is highly regulated. HIF-1 controls the expression of
several key genes required for processes such as angiogenesis, glycolysis
and glucose transport, cell proliferation and cell motility.
HIF-1alpha is often over-expressed in cancers, due to their intratumoural
hypoxia. This leads to the initiation of tumour neovascularisation
and the encouragement of anaerobic respiration, increasing the oxygen
delivery and energy supply to the tumour and enhancing its growth
and survival.
The digestive hormone gastrin, produced in the stomach is responsible
for the stimulation of gastric acid secretion and epithelial cell
proliferation by acting at the cholecystokinin-2 (CCK2) receptor.
In colorectal cancers, gastrin has been shown to enhance the rate
of cell growth, acting in both an autocrine and endocrine manner.
It also has pro-angiogenic, pro-metastatic and anti-apoptotic roles,
similar to those produced by HIF-1 activation and therefore may
be acting, at least in part, through the induction of HIF-1alpha.
The aim of this project is to investigate the role of HIF-1alpha
in the progression of gastrointestinal tumours and the involvement
of gastrin within this process. Initial experiments investigated
the expression of HIF-1alpha protein expression within a number
of gastrointestinal carcinoma cell lines under normoxic and chemically-induced
hypoxic conditions. In general, hypoxia greatly induced the expression
of HIF-1alpha protein in all cell lines at all time points tested.
Its expression at the gene level under both normoxic and hypoxic
conditions is currently under investigation, as well as its expression
within actual tumour samples (currently being optimised in mice
subcutaneous xenografts). The next stage of the project is to examine
the effect of gastrin treatment on HIF-1 expression at both gene
and protein level, before abrogating any endogenous gastrin effect
via gastrin-specific siRNA treatment or using a CCK2-receptor antagonists/inhibitors.
To determine whether HIF-1 plays any role in the cellular effects
mediated by gastrin, HIF-1 activity will also be blocked via signal
pathway inhibitors or HIF-1alpha siRNA.
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Yinfei Yin (PhD): Effect of gastrin on MMP/ECM interactions in cancer
invasion and metastasis
Invasion
and metastasis are important stages in cancer development and associated
with a poor prognosis for patients. This project aims to explore
the role of the growth hormone gastrin in this process through the
investigation of downstream pathways triggered by over-expression
of gastrin or its receptor in gastrointestinal (GI) cancer. Pilot
genomics data obtained within the Division of Pre-Clinical Oncology
has highlighted changes in expression of a set of genes (including
specific matrix metalloproteinases and their inhibitors, integrins
and matrix components) in gastrin-expressing GI cancer cells. The
changes observed potentially increase the invasive and metastatic
potential of such cells. Initial studies will focus on extending
these findings at the gene and protein level by examination of a
range of gastrointestinal cell-lines. The project will go on to
investigate the biological roles of these genes through the use
of molecular cloning, gene knock-down, and cellular attachment and
invasion assays.
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Peter Boyce (PhD): Chemically modified siRNA in the treatment of
gastrointestinal cancer
Small
interfering RNA (siRNA) are a class of 19-25 nucleotide long dsRNA
molecules which are involved in the RNA interference pathway. Initially
discovered as part of post-transcriptional gene silencing in plants
(Hamilton and Baulcombe, 1999), these molecules are now of great
interest due to the gene knockdown effect in mammalian cells. This
project aims to investigate the effect of chemical modifications
of siRNA molecules on their efficacy in vitro and in
vivo, and whether chemical modifications can increase the viability
of siRNA as a therapeutic treatment. Another aspect of this project
involves investigating the Sonic Hedgehog (Shh) pathway in gastrointestinal
cancer and the downstream cellular effects of siRNA mediated knockdown
of components of this pathway.
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Naseem Waraich (DM): A prospective study to relate androgen and
androgen receptor expression levels to fibroblast growth factor
receptors gene and protein expression by Barrett’s Oesophagus
and malignant disorders of the oesophagus
There
are approximately 3000 new cases of oesophageal adenocarcinoma (OAC)
per year in the UK and the incidence rate has increased by around
50% over the last 10 years. Men are more frequently affected than
women with a ratio of 7:1. This project investigates the possibility
of a role for androgens in the development of OAC. We have recently
shown androgen receptor expression by immunohistochemistry in the
stromal component of human OAC sections and that testosterone treatment
increases Fibroblast growth factor-8 expression by AR-expressing
fibroblasts. Production of such growth factors may promote the growth
of oesophageal cancer cells.
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Amanda Tobias (MPhil): Gastrin interactions which impact upon gastric
and colonic carcinogenesis
Gastrin,
a polypeptide hormone, secreted by G-cells in the stomach, is a
promoter of cancer cell growth both in vitro and in
vivo. Gastrin acts through cholecystokinin type two receptors
(CCK-2R) and the expression of both the gastrin gene and the CCK-2R
are closely linked to the development, progression and invasion
of cancer cells. Anti-gastrin immunogen (G17DT) consists of a synthetic
peptide from the N-terminal of human G17, attached to a carrier
protein, Dipheria Toxoid (DT). G17DT induces an antibody response
which neutralises the target hormone.
Studies have included investigating the effect of G17DT on the development
of pre-cancerous changes in the stomachs of transgenic InsGas mice,
which over express the amidated form of gastrin (G17) under control
of a rat insulin I promoter, and go on to develop glandular atrophy
and invasive gastric cancer. The effect of G17DT has also been studied
in the APC[Min] mouse model, in this model the loss of
the normal APC allele in the intestinal epithelium precedes adenoma
formation and the mice go on to develop numerous polyps in their
small intestine.
Future work will look at the effect of infecting the above models
with Helicobacter pylori, a carcinogenic bacterium linked
to gastric cancer, and then treatment with G17DT.
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For further
information, please do not hesitate to contact us: |
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UMCO
Tel: +44 (0)115 8231135
Fax: +44 (0)115 8231137
Email: jane.mcclelland@nottingham.ac.uk
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