Centre for Investigative & Diagnostic Oncology

As described in Project 8, many tumours secrete a molecule which can act to stimulate growth of the tumour in a so-called autocrine loop.  One such growth factor is a genetic variant of the pregnancy hormone, human chorionic gonadotrophin (hCG), which we have recognised uses a DNA sequence not expressed by normal cells.  We will investigate the possibility that the presence of the hCG variant in the serum DNA of cancer patients and/or the abnormal protein which it encodes, can provide useful prognostic indicators and/or biomarkers to monitor therapy.

c-Met receptor is a surface marker of many tumours but is particularly related to cancer stem cells from which the tumour mass differentiates (compare Project 1).  The cells are stimulated when the c-Met binds its circulating ligand, hepatocyte growth factor (HGF).  We will examine the possibility that tumour cells (cell lines initially) express c-Met and if so, whether they secrete HGF and/or are stimulated by this growth factor (we should apply this also to the stem cell progeny produced in Project 1).

Like the hCG experiments suggested above, the presence of HGF and/or c-Met in serum should be studied along with hCG and other serum tumour biomarkers as a possible array for diagnosis and/or therapy monitoring.  Look also for possible autoantibodies and abnormal c-Met DNA.

Goals

1. To find new serum biomarkers as prognostic indicators of disease and as monitors of therapy. 
2. Look for possible new c-Met/HGF autocrine loop. 
3. Examine any surface phenotypic changes induced by autocrine stimulation.

Project team

Dr. S. Butler
Leader

Dr. S. Wen Scientific support

Dr. L. Ghali Scientific support

Beatta
(Poznan) Serum DNA

Research Assistant Cell and biomarker studies

Dr. P. Cohen (Histopath., Imperial)
Supply clinical materials

Prof. I. Cree (Warwick)
Serum samples

Look for blood biomarkers using the NALIA system (see Early lab diagnosis of cancer) to see whether they can provide early diagnosis of brain cancer, particularly glioblastoma, and also monitoring of therapy.

We will construct iron nanoparticles to target the brain tumour cells because they should permit the application of the highly economical Electrical Impedance Imaging Technology and permit thermal ablation of the tumour by application of an external energy source to targeted nanoparticles injected directly into the tumour mass.

Goals

1) Diagnostic microarray for early diagnosis and monitoring of brain tumours.
2) Economical imaging of these tumours.
3) Thermal ablation of tumours ("I'd rather have a needle than a knife").

Project team

Dr. A. Tizzard Leader

Prof. R. Bayford
Scientific support

Dr. S. Butler
Scientific support

Research Assistant
Biological testing of nanoparticles

Midatech Ltd.
Nanoparticle synthesis

Prof. A. Dalgleish, St.
Testing tumour inhibition

George's Hosp. M.Sch. Surgeon, Kings Coll. Hosp.
Provide clinical material

We have collaborated with a team at Dartmouth College in the USA who have been applying Electrical Impedance Tomography (EIT) imaging to the diagnosis of breast cancer. Our contribution has been to greatly improve image accuracy by more sophisticated mathematical analysis of the imaging data paying particular attention to the influence of breast shape. We intend to refine our algorithms even more to improve the reliability of diagnosis.

Goals

1. To help establish an accurate diagnostic system for breast cancer which is economical and does not involve the use of ionising radiation which is especially undesirable for pregnant, young and elderly women and for repeated use.

2. If skin patch delivery results are encouraging (Midatech), propose therapeutic collaboration using cytotoxic gold nanoparticles.

Project team

Dr. A. Tizzard Leader 

Research Assistant Mathematical analysis

Research Student (MSc or placement) Prototype build and testing

Background

When a normal cell divides, it produces two identical daughter cells. When a cancer stem cell divides, it produces a new cancer stem cell and a differentiated cell, which grows to form the tumour mass. The stem cells are the most important source of secondary growths and, unlike the tumour mass, are generally resistant to conventional chemotherapy (see illustration).

Early detection and eradication of the new resistant stem cells is therefore vital to increase a patient's chances of survival.

Aims

1. To develop therapies to eradicate cancer stem cells.
2. To see whether positive tests for blood stem cells indicate risk of secondary tumours (metastases) so that more aggressive treatment can be instigated.

The project

Through our research we intend to isolate and so detect any stem cells in the blood and in primary tumour tissue excised during a patient's operation. This will be done by targeting tumour cell surface markers, using a powerful antibody directed against 'c-Met' produced by the Belgian company arGEN-X. This antibody has been shown to inhibit the growth of human tumour grafts, and to kill the stem cells within them by recruiting the help of two different factors in blood, 'complement' and/or white blood cells (known as 'natural killer cells').

In addition to researching detection, we will grow the stem cells and test the survival of the progeny cultured again with the c-Met antibody. As a back-up, we will also test with tiny gold nanoparticles developed by Midatech Ltd – coated with epithelial growth factor or folic acid to get the particles into the cells – alongside small drug inhibitors of c-Met or conventional chemotherapy drugs. We will compare the effects on newly-generated stem cells and any differentiated tumour cells.

We will also compare the stem cells with their parent tumour cells for their ability to penetrate a human skin model, as an indicator of metastatic potential.

Dr. T. Lund, Research Associate – Isolation and killing of stem cells

Dr. Song Wen, Research Associate – Growing and identifying cancer stem cells

Clinician, Imperial NHS Trust & St. George's Hospital – Provision of clinical material

Polyoma virus infection can transform normal into cancer cells and this has been an important model system in deciphering the basic mechanisms underlying all human cancers.  The polyoma viral oncogene, MT, acts as an analogue of an activated growth factor receptor. Mutated receptors play a part in generating most cancers, so understanding MT's function will help decipher how these receptors work during cancer formation and hopefully provide clues to reversing this action. We have recently identified a nanocluster containing MT on the surface of polyoma virus-transformed cells that is essential for its cancer forming action.  We plan to purify these MT containing complexes and identify the constituents using our mass spectrometry facilities.  This is an essential prerequisite to searching for novel therapeutic inhibitors.

Goals

    1) Identify the components of the virally-induced cancer-forming complex

    2) Use the information to devise new therapeutic agents.

Project team

Prof. S. Dilworth
Leader

Research Assistant Analysis of complex

Background

Colorectal cancer is one of the most common forms of cancer. While the primary tumour can be surgically removed or ablated by radiotherapy, neither procedure is able to eliminate secondary metastatic growths at distant sites. Therapy with cytotoxic drugs (chemotherapy) can hunt down these metastases but there are most unpleasant side-effects due to the drugs attacking non-tumour sites.

The project

We aim to deliver cytotoxic drugs to the tumours selectively by attaching the drugs to minute gold particles, or 'nanoparticles' (5 million = 1 cm). The nanoparticles will targeted specifically at the surface of the cancer cell and the drug released intracellularly, exactly in the location for it to be effective (see cancer targeting illustration above).

This will have the double advantage of being highly successful at killing the cancer cells while causing minimal side-effects, as the non-tumour sites are left alone.

Project team

Professor I M Roitt – Lead

Dr T Lund – Cellular targeting of gold nanoparticles

Midatech Ltd  – Tuning gold nanoparticles

Connective tissue factors in breast cancer

Despite improvements in the screening and treatment, for 30% of women diagnosed with invasive breast cancer these tumours will spread to other tissues. Tumour growth and spreading is caused by a series of growth factors. The activity of these factors is brought about by interactions with a series of heparin-like molecules termed glycosaminoglycans (GAGs), found in connective tissue. Some of these heparin-like molecules encourage growth factors, whereas others appear to block growth factor effects. Dr Hills and his team have identified the presence of several of these GAGs in breast cancer cells along with the expression of the growth factors to which they bind. In addition, they have identified mechanisms by which GAGs may control key features of tumour development. They are currently investigating how subtle changes in the structure of these molecules alter their effect on tumour progression.

Goals

    1) See whether detection of large quantities of certain GAGs detect early stage breast cancer.

    2) Identify structural differences to determine whether a heparin-like molecule promotes or inhibits  tumour growth.

Project team

Dr Frank Hills
Project leader 

Research Associate In vitro lab studies

Research Associate Molecular characterisation and nanoparticle conjugation

Histopathologist
Cancer tissues

Project

Professor Richard Bayford and colleagues developed and patented a straightforward method for immunological detection of biomarkers, in particular the tumour growth factor chorionic gonadotrophin (see Autocrine tumour growth factors).  The gold electrode is coated with an agent which can specifically bind to a selected cancer biomarker in patient's blood so when the serum is added, the layer coating the electrode is increased in thickness and there is a change in electrical resistance which can be readily measured. The device can incorporate several electrodes, each of which can detect a different cancer biomarker.  The simplicity of the device means that it could readily be used in a doctor's surgery or a public pharmacy, or even in a room adjoining a clinic to give both doctor and patient a rapid diagnosis enabling earlier instigation of therapy and relieving the patient of an agonising wait for the results.

Goals

1)  Development of a compact, economical device for early detection and/or monitoring therapy of cancer suitable for Point of Care use.

2)  Commercialisation of the device for use in cancer, allergy and even heart disease.

Project team

Prof. R. Bayford Team Leader

Dr. A Tizzard Technical support

Research Associate Biophysics development

U.C.L. Engineer Hardware development

Nottingham University Provision of sera

Background

Tumour cells release unique breakdown components into the blood; known as biological markers (biomarkers), these can be used as to diagnose cancer. The immune system, confronted with these unusual biomarkers, often reacts to make antibodies (autoantibodies) against them. Professor Robertson's group at Nottingham University have shown that the presence of these autoantibodies can signal the early onset of cancer. A company based on their findings measures several of these antibodies in the serum of patients as a diagnostic service, each measurement being a separate test.

Professor Roitt and his team have developed a generic system in which each serum undergoes multiple tests (multiplexing) in a single well of a 96-well plate containing an array of spots, each of which functions as an individual test (see illustration). The system was patented and published in Clinical Chemistry (2008), and a company, NALIA Systems Ltd, formed. 

Aims

1. To show that the NALIA array provides a very economical alternative to the current Nottingham system for multiple autoantibody tests diagnosing early cancer.

2. To improve the sensitivity and specificity of tests for early cancer diagnosis by recognition of novel biomarker patterns. This will enable earlier therapy, improving the outcome for patients.

3. To seek commercial exploitation of the findings to support further research.

The project

The Nottingham group are keen to collaborate with our Middlesex team who, when funding is available, will create arrays using the NALIA system to combine the various autoantibody tests into a single multiplexed operation.

Subsequently, we will try to identify new autoantibodies. The search for tumour-related small metabolites on biomarkers in serum and urine by mass spectroscopy (metabolomics) is a relatively unexplored area; Dr Shah is highly experienced in the use of this technology and has the sophisticated equipment needed here in the University.

By adding results from arrays for the original tumour components with those from autoantibody arrays and metabolomics, plus analysis of the raised DNA in patients' blood (with a close colleague, Professor I. Cree, Head of Pathology at Warwick University Hospital), we would accumulate a very large number of different results for each patient. This would allow identification of novel patterns by a process called 'neural network analysis'; these should provide new, highly sensitive and accurate diagnosis of many different types of cancer at an early stage. Starting therapy then will greatly improve prospects for patients.

Project team

Dr S. Butler – Project leader

Dr A. Shah – Metabolomics (Mass Spec)

Professor I.M. Roitt – Scientific support

Dr Y. Yatsenko – Postgraduate Research Assistant

Tumours of the skin are particularly accessible to imaging. We will initiate these studies together with a Consultant Dermatologist who in the normal course of investigation, would biopsy suspected growths.

Targeting tumour cells with drug-coated gold nanoparticles (see Colorectal cancer therapy) would seem to be an attractive proposition, given their accessibility to delivery through skin patches, a technology which Midatech has developed. This would obviate the need for systemic infusion. We will construct nanoparticles coated with a tumour-seeking ligand and a chemotherapeutic drug linked in such a way as to be released within the intracellular environment. These nanoparticles with different ligands would be tested on cell lines, cells derived from operation specimens, and in 3-dimensional skin models in vitro using a technique employed by Dr. Lucy Ghali. We will also investigate application of a recent publication showing that gold nanoparticles themselves would penetrate skin when applied topically and allow simultaneous entry of a co-administered protein. Clearly, we should look at the possibility of delivery of a co-administered independent cytotoxic drug.

Goals

1)  Verifying the applicability of a probe for skin cancer imaging.
2)  Delivery of topical therapeutic gold nanoparticles to skin tumours.
3)  Ultimately, extend the unconventional delivery of gold nanoparticles to breast cancer, and  inhalation for lung cancer.

Project team

Dr. L. Ghali
Leader

Dr. Dong Li Histological assessment

Ph.D. student Producing and testing gnp on cells and reconstructed skin in vitro

Dermatology MD student Testing probe

Dr. A. Agaiby Skin imaging and provision of operation sample

Dermatology: Consultant Northwick Pk.

Dr. Stephen Butler, together with Professor Ray Iles, made the fascinating discovery that many cancer cells secreted a variant of the human pregnancy hormone, chorionic gonadotrophin (hCG), which bizarrely acted as a growth promoter for the very cells that produced it – we call it an autocrine growth promoter. As might be expected, antibody raised by vaccination with hCG neutralised the growth-promoting properties of this factor.  Professor Roitt & Dr. Lund and their colleagues, while at U.C.L., constructed a mutant of hCG which not only produced more effected antibodies than the native protein, but also essentially eliminated one of the undesirable side-effects which was to cross-react with another important hormone, luteinizing hormone.

We now wish to develop an exciting, new, potentially very high effective vaccine platform to improve the ability of the hCG mutant to generate an excellent antibody response.  Together with Midatech Ltd., a company with expertise in the production of minute gold nanoparticles (see also Colorectal cancer therapy) we intend to attach these particles coated with agents essential for boosting the antibody response to the hCG.

Goals

1.  To produce a novel highly effective vaccine platform to generate cancer growth inhibiting antibodies.
2. To arrange Clinical Trials of the vaccine in patients with bladder cancer.
3. To show that the platform may produce effective vaccines for another hormone, gonadotrophin- releasing hormone (GnRH), important for controlling prostate cancer and may provide a more general vaccine formulation.

Project team

Dr. S. Butler
Lead role

Prof. I.M. Roitt Scientific support

Ph.D. student Antibody analysis

Prof. N. Porakishvili Mutant production (Westminster Uni.)
Vaccine testing (Georgia, Tbilisi)

Midatech Ltd. Nanoparticle production

New papilloma virus-dependent tumours

It is well established that infection with certain strains of the papilloma virus is responsible for the majority of cases of cervical cancer. Drs. S. Wen & L. Ghali have demonstrated that they can identify an involvement of the virus in a given tumour by immunohistochemical staining (revealing a molecule in a tissue section through binding of an antibody linked to a colour-producing moiety) of paraffin embedded tumour tissue and by DNA analysis. The team has preliminary evidence that the virus can be detected in a minority of breast cancer patients from Austria and Nigeria and they wish to follow this up with a larger well-defined population, looking also to probe the possibility of there being a genetic predisposition to this subsection of breast and other cancers (e.g. major histocompatibility complex genes).

Goal

Identify tumour types not previously linked to papilloma virus.

Project team

Dr. S. Wen
Leader 

Dr. L. Ghali Scientific support

Dr. Dong Li Immunohistochemistry

Dr. P. Cohen (Histopath., Imperial)
Supply clinical materials

Project B: Prostate cancer

Infections with high-risk human papilloma viruses (HPVs), causatively linked to cervical cancer, might also play a role in the development of prostate cancer due to the proximity in the type of the cervical transitional zone epithelium and prostate epithelium per se in terms of the histological characteristics and the embryologic origin.

Infection with a number of human polyomaviruses  has been described (BKV, JCV, KIPyV, MUPyV & MCPyV) and some have been associated with tumour development, for example, it has been suggested that BK is associated with brain tumours, adenocarcinomas, and prostate carcinomas. As is the case for all polyomaviruses, BKV encodes two proteins that deregulate the ability of the cell to control its growth, namely 'large' and 'small' T antigens. BKV, like JCV and the similar simian virus, SV40, is recognised as a tumour virus because of its ability to cause changes in cell growth and tumour formation in cultured cells and animal models, respectively. BKV and JCV oncogenically transform non-permissive rodent cells in culture and immortalize permissive human cells alone or in the presence of other oncogenes. 

We will examine prostate cancer operation specimens for evidence of viral infection and relate this to the clinical status of the patient. 

Goals

1) To establish the prevalence of infection with human papillomaviruses and polyomaviruses in prostate cancer.

2) To find out if there is any correlation between viral load or genome expression and the clinical grading on the hyperplasia or tumour.

Project team and indicative budget

Dr. Christopher Ring
Leader

Dr. Mahmoud Naase
Scientific support

Ph.D. student
Analyse clinical samples for viral infection

Dr. R. Persad (Bristol)
Provide clinical samples

Background and aim

Conventional imaging to screen for cancer typically uses MRI – a vastly expensive procedure not necessarily available outside main centres, particularly in second and third world countries. We aim to develop a low cost device especially for screening for rectal and prostate cancers, and for monitoring treatment.

The project

Led by expert in the field Professor Bayford, we are developing a new probe using Electrical Impedance Tomography (EIT) – a very low cost imaging system that can be used to detect abnormal tissue such as a tumour.

With EIT, a series of electrodes are placed over the area to be imaged, inducing minute electric currents. The pattern of the currents is then mapped by sophisticated mathematical analysis to reveal any abnormal tissue, such as a tumour. 

A portable probe based on this principle has been developed for detection of cervical cancer by Zilico Ltd., a small firm in Sheffield. We are adapting this idea to create a small circular inflatable device consisting of a ring of electrodes with an inbuilt camera; this will fit into the anus to detect any overt or underlying rectal cancer growths.

During development the accuracy of the system will be checked against conventional imaging with MRI. Once developed, the device will be adapted for the detection of prostate cancer.

Project team

Professor R. Bayford – Project leader

Dr A. Tizzard – Technical support

Research Associate – Testing device in lab and clinic

Professor A. Demosthenes – Design of electrodes (UCL)

A.N.Other – Design of hardware (UCL)

Mr G Buchanan – Provide patients (Imperial)

Radiologist – Parallel imaging MRI (Imperial normal NHS assessment)

Zilico Ltd. – Technical support

Project

The strategy for targeting colorectal cancers described in Colorectal cancer therapy will be applied to targeting prostate cancers using gold nanoparticles and liposomes.

Goal

Efficient targeting of drugs to cancer cells without the unpleasant systemic effects of the free drug. 

Project team

Prof. Ivan Roitt Project Leader

Dr. Torben Lund  Research Fellow

Dr. Scarlet Wang Research Associate