WO2000064341A1

WO2000064341A1 - Angiogenesis detection

Info

Publication number: WO2000064341A1
Application number: PCT/GB2000/001565
Authority: WO
Inventors: Gino Francesco
Original assignee: Gino Francesco
Priority date: 1999-04-22
Filing date: 2000-04-20
Publication date: 2000-11-02
Also published as: AU4133500A; GB9909300D0

Abstract

The present invention relates to angiogenesis and methods and apparatus for the detection thereof. There is described, angiogenesis monitoring apparatus comprising: monitoring means adapted to receive signals output from that part of a patient's brain where activity from the neural network relating to angiogenesis occurs; analysing means for analysing the signals to determine whether the signal includes a signal indicating a neural response to cytokine stimulation. Preferably, the monitoring means receives signals of about 8Hz or more. There is also described a method of detecting angiogenesis comprising monitoring signals output from that part of a patient's brain where activity from the neural network relating to angiogenesis occurs analysing the signals to determine whether the signal includes a signal of about 8Hz or more indicating a neural response to cytokine stimulation.

Description

ANGIOGENESIS DETECTION

The present invention relates to angiogenesis and methods and apparatus for the detection thereof.

Angiogenesis is the term given to the development of new blood vessels from preexisting blood vessels in a process involving the migration and proliferation of endothelial cells from pre-existing vessels. It occurs quite normally in the embryo and wound healing and the menstrual cycle in ovaries and endrometrium, but otherwise, in adults, endothelial cells divide only rarely. However, in many diseases, including cancer, there is an increase in angiogenesis. In cancer, 98% of all soft tumours are dependant on angiogenesis for their growth and viability.

The process for a typical cell to become cancerous is as follows. Cell division occurs perhaps three billion times each day in a typical human body. With mitosis occurring at this rate, clearly a mechanism must be in place to rid the body of superfluous cells. The organism will, under normal circumstances, not allow extra cells to exist beyond its normal requirements and through a signalling method orders unwanted cells to commit suicide. This mechanism is centred around a gene identified as p53. When a cell has a propensity, for whatever reason, to become atypical, perhaps leading to a cancer, typically the p53 gene switches itself off, so that when a signal from the organism instructs the cell to die, it is ignored. The cell will thus continue to divide, with all its progeny having the same p53 gene mutation thus preventing control of these cells by the organism

At this stage, these multiplying cells are not malignant as to become so requires a blood supply to provide a food supply and to remove waste products. In the absence of a blood supply, the tumour cells simply die by necrosis. As described above, the mechanism for providing a new blood supply is angiogenesis. Whether for the benefit of the body in wound healing or to its detriment in cancer, angiogenesis uses the same vehicle to achieve the new blood supply, that of the body's neuroimmune system. When the atypical cells are ready to attempt angiogenesis, they release tumour angiogenesis factors, such as proteins called cytokines, which induce neighbouring endothelial cells to degrade their basal lamina and begin to migrate into the extra cellular matrix (ECM) towards the tumour The ECM is the barrier that the endothelial cells must overcome in order to vascularisation

Typically following 24 cell divisions, there are perhaps a million atypical cells present which would only occupy the space on top of a pin head

The released cytokines stimulate adjacent specialised cells to also release their own cytokines This in turn stimulates more cells to do likewise and so on until a significant amount of cytokine material exists These cytokines enter the bloodstream and act on the neurotransmitters which send a constant stream of messages to the brain. The brain interprets these messages and produces its own neurochemicals and signals which allows the body to produce new blood vessels at the stimulated location. The mass of atypical cells, with its own blood supply, can then develop into a viable tumour

Once a tumour mass is present, it will continually subject the host body to the process of angiogenesis to cope with its growth

The signal response of a brain's neurotransmitter to circulating cytokines may be triggered within seconds or minutes, but may be delayed for hours or even days This partly depends upon the site of neurotransmitter synthesis and/or release, the mechanism of cytokine action; whether the cytokines act directly upon the transmitter containing neuron; or whether its action is mediated via other factors or neurotransmitters. It is known that the neural response to cytokine stimulation has a unique signature and can be measured electrically at a frequency of about 8Hz or more, typically about 14Hz to about 30Hz The neural response will last at least for several days For further background information, reference is made to The Lancet 1997, 349 (suppl II) 13-15 and the Cytokines Online Pathfinder Encyclopaedia - Laboratory of Molecular Biology - Gene Centre - Ludwig-Maximilians-University of Munich http://www.copewithcytokines.de (reference 'angiogenesis')

The present invention seeks to use the presence of cytokines generated in this way and the brain neurotransmissions resulting therefrom as a means towards the early detection of pathological processes such as cancers

The benefits such an early warning may offer are obvious

If a cancer can be detected early on, the prognosis is usually better as complications which can occur with advanced cancers such as cachexia or organ failure are avoided Thirdly, even if cancer has taken hold, the present invention can be used following conventional cancer treatments to indicate the state of angiogenic activity in the body thus giving an indication as to the effectiveness of the treatment given

In its broadest sense, the present invention provides a system of monitoring angiogenesis in a patient, the system comprising an apparatus comprising monitoring means adapted to receive signals output from that part of the patient's brain where activity from the neural network relating to angiogenesis occurs and means to analyse the signals to determine whether the signal includes a signal indicating a neural response to cytokine stimulation

Typically, the monitoring means includes an electrode arrangement, the electrodes being designed to receive signals in the precise bandwidth in which the neurotransmitter transmits in response to cytokine stimulation Preferably the monitoring means receives signals of about 8Hz or more Preferably the system includes means, in response to a positive signal indicating the presence of cytokines, to advise the patient to maintain an intake of physician- prescribed angiogenic drugs

Preferably, the apparatus includes means to transmit the signal received to another location to be assessed by a physician (oncologist) and/or stored on a remote computer to maintain the medical records of the patient

Preferably, the apparatus incorporates or is adapted to be connected to a mobile (cellular or satellite) telephone or telephone connected to a land line

The apparatus may include a buffer memory to store neurotransmitter data received over a period of time prior to the data being transmitted

In one adaptation, to provide the medical practitioners with additional information, the system further includes means to monitor other physiological parameters such as blood pressure and/or provide ECG or EEG outputs, or other neurological disorders

In one embodiment, the electrode is implanted into the patient's skull and data output to a sensor by induction

In a further aspect, the present invention provides a method of detecting angiogenesis comprising monitoring signals output from that part of a patient's brain where activity from the neural network relating to angiogenesis occurs analysing the signals to determine whether the signal includes a signal of about 8Hz or more indicating a neural response to cytokine stimulation

The above and other aspects of the present invention will now be described in further detail by way of example only, with references to the accompanying figures, in which - Figure 1 illustrates the various stages of angiogenesis,

Figure 2 illustrates an International Standard 10/20 EEG System,

Figure 3 illustrates the hypothalamus area of the brain,

Figure 4 shows a typical "whole of brain" EEG, and

Figure 5 shows a typical output following conversion of EEG data

Figure 1 illustrates the various stages of angiogenesis In view (I), endothelial cells degrade basel lamina and begin to migrate, in view (n), endothelial cells form sprouts which branch and anastamose, in (in) sprouts grow towards tumour and branching increases, in (IV) sprouts finally connect with the tumour, completing angiogenesis The time taken for an angiogenesis cycle to complete is about 12 days

Tumour angiogenesis factors such as, for example, prolactin, which has been shown to have cytokine-hke activities, also act on neurotransmitters which send a constant stream of messages to the brain The signal response of a brain's neurotransmitter to circulating cytokines may be triggered within seconds, minutes, or days It is the modulatory effect of released cytokines by tumour cells on the hypothalamus where the firing rates of hypothalamic neurones have been shown to be altered, that form the basis of this invention

Following the release of angiogenic factors into the blood stream, they make use of the neuroimune network to communicate with the brain and by direct contact There is a constant neural response to angiogenic activity and this response can be measured electrically through EEG procedures as the response has its own signature Angiogenesis will occur in a pre malignant stage leading to a tumour and thereafter continually until the tumour is removed or destroyed or the host dies Angiogenic activity can be recorded using magnetic resonance or EEG

EEG is spontaneous cortical electrical activity recorded at the scalp The electrical activity of the brain behaves like any other material Changes on membrane polaπation, inhibitory and excitatory post-synaptic potential, action potentials, etc, impress voltages are conducted through the surrounding medium of CSF, meninges, skull and scalp The electrical voltages conduct up through brain tissue, enter the membranes surrounding the brain, continue on up through the skull to appear at the scalp At this point they are reduced from the millivolt range (of the membrane potential gradients and active potentials) to a few microvolts Typical values might be 20-100 microvolts for EEG with lesser values being recorded in averaged evoked potentials, perhaps 10 microvolts These potentials are easily recorded and are most often recorded from many electrodes in an arranged montage A most common standard is the International 10/20 System illustrated in Figure 2

The patient, when subjected to an EEG examination, will be relaxed and have their eyes closed The EEG equipment will be turned on and data acquisition and recording commences The area of the brain which is of interest is the hypothalamus which is shown in Figure 3 This part of the brain is the least accessible but it is this region where the hypothalamic neurones are known to be altered by the effect of released cytokines In fact there are distinctive signal changes which correspond to angiogenic activity

Ordinarily an EEG is performed over a period of time which would not normally exceed 25 minutes Clean signals are dependant on low scalp/electrode impedance Differential amplifiers and filters, which also utilise a "common" electrode, are used to remove noise

Figures 4 shows a paper print snapshot of a typical EEG graph which covers a "whole of brain" area There are typically 22 channels although only two channels may be required to record activity from the hypothalamus region

In order to interpret this EEG data, software tools are available to allow for data reduction and frequency spectrum analysis This can be used on Fourier transform and wavelet transform Figure 5 shows a typical output of an EEG data conversion, using this method Another approach involves the setting up of a neural-network based on algorithms to recognise angiogenic activity Compared with spectrum analysis, a neural network, when properly trained by the input of much data, can give an immediate answer that angiogenesis is occurring, or not as the case may be, even though the underlying relationship of converted EEG data may not be known One may not known how the results are reached as there are not clues about the underlying physical model, but similar results can be obtained.

Whilst a conventional EEG apparatus will generally be used, a miniaturised purpose designed device has also been proposed. The device includes electrodes, is battery powered, and includes an integrated communication device which records the signal for digital conversion and downloaded to a host computer which analyses the data to decide whether angiogenesis is occurring or not

The benefits to this type of technology include

Everyone in the world may be subjected to a tumour growth sometime in their life Whilst there are great strides in diagnosis media, they all cost money to perform and there would be a reluctance for individuals to continually subject themselves to a test The present invention is very cheap, in the context of using existing EEG apparatus, almost to the point of nearly being free

If the user underwent a 5 minutes examination every 10 days using this method, any angiogenic activity may be identified. This is not a procedure to determine the start or development of a tumour, but is a test that alerts a person through a managed computer that angiogenesis is taking place and may require further investigation by his physician

All cancer tests must operate within a proof positive/negative regime which must, for obvious reasons, be exacting. This is not a test for cancer therefore this criteria does not strictly apply, but clearly, if there is a cancer present and it is developing, then there will be a continual angiogenic process occurring which allows the user multiple chances of detecting a tumour. This is not the case for any other type of cancer test.

[Footnote to Figure 2]

The 10-20 system of electrode placement is based on the relationship between the location of an electrode and the underlying area of cerebral cortex. Each point on this figure to the left indicates a possible electrode position. Each site has a letter (to identify the lobe) and a number or another letter to identify the hemisphere location. The letters F, T, C, P and O stand for Frontal, Temporal, Central, Parietal and Occipital. (Note that there is no "central lobe", but this is just used for identification purposes.) Even numbers (2,4,6,8) refer to the right hemisphere and odd numbers (1,3,5,7) refer to the left hemisphere. The z refers to an electrode placed on the midline. Also note that the smaller the number, the closer the position is to the midline.

The "10" and "20" refer to the 10% or 20% interelectrode system.

Nasion - point between the forehead and nose. Inion - Bump at back of skull.

Claims

1. Angiogenesis monitoring apparatus comprising: monitoring means adapted to receive signals output from that part of a patient's brain where activity from the neural network relating to angiogenesis occurs; analysing means for analysing the signals to determine whether the signal includes a signal indicating a neural response to cytokine stimulation.

2. Monitoring apparatus as claimed in claim 1 wherein the monitoring means includes an electrode, the electrode being designed to receive signals in the precise bandwidth in which the neurotransmitter transmits in response to cytokine stimulation.

3. Monitoring apparatus as claimed in claim 2 wherein the monitoring means receives signals of about 8Hz or more.

4. Monitoring apparatus as claimed in any one of claims 1 to 3 wherein the monitoring means includes an electroencephalograph.

5. Monitoring apparatus as claimed in any one of claims 1 to 3 further including means, in response to a positive signal indicating the presence of cytokines, to advise the patient to maintain an intake of physician-prescribed angiogenic drugs.

6. Monitoring apparatus as claimed in any preceding claim further including transmitter means to transmit the signal received to another location to be assessed by a physician (oncologist) and/or stored on a remote computer to maintain the medical records of the patient.

7. Monitoring apparatus as claimed in claim 6 wherein the transmitter further including means enables connection to a mobile (cellular or satellite) telephone or telephone connected to a land line.

8. Monitoring apparatus as claimed in any preceding claim further comprising buffer memory to store neurotransmitter data received over a period of time prior to the data being transmitted.

9. Monitoring apparatus as claimed in any preceding claim further including means to monitor other physiological parameters such as blood pressure and/or provide ECG or EEG outputs.

10. A method of detecting angiogenesis comprising monitoring signals output from that part of a patient's brain where activity from the neural network relating to angiogenesis occurs analysing the signals to determine whether the signal includes a signal of about 8Hz or more indicating a neural response to cytokine stimulation.