Parsortix Evaluation

 

For immediate release

22 July 2014

 

ANGLE plc

("ANGLE" or "the Company")

 

PARSORTIX EVALUATION BY THE CEP GROUP AT THE CANCER RESEARCH UK MANCHESTER INSTITUTE  

 

 

ANGLE plc (AIM: AGL), the specialist medtech company, is delighted to announce the positive evaluation of its Parsortix system by the Clinical and Experimental Pharmacology (CEP) group who are based at the Cancer Research UK Manchester Institute, part of The University of Manchester.    

 

Cancer Research UK Manchester Institute (CRUK MI), formerly known as the Paterson Institute for Cancer Research, is a leading cancer research institute spanning the whole spectrum of cancer research from investigating the molecular and cellular basis of cancer, to translational research and the development of diagnostics and therapeutics.

 

CRUK MI has approximately 300 post-doctoral scientists, clinical fellows, scientific officers, operational and technical staff, postgraduate research students and visiting fellows. It benefits from working closely with The Christie NHS Foundation Trust - the largest cancer centre in Europe, treating more than 40,000 cancer patients each year.

 

CRUK MI is one of the leading centres worldwide for research into circulating tumour cells (CTCs) and as such has a strong interest in technologies that will support CTC research and potentially lead to clinical applications for CTC analysis.

 

The CEP group at CRUK MI has now reported on its evaluation of and development work on the Parsortix CTC capture and harvest system.  CEP's initial work, as reported in July 2013, has been followed by wide ranging investigation, development and evaluation over the last year including: 

·     Blood sample pre-processing protocols

·     Removal of plasma from blood prior to Parsortix processing to assess compatibility with cell-free DNA analysis

·     Establishing the capture efficiency of Parsortix using blood samples spiked with pre-labelled cell lines

·     Assessment of the capture efficiency of Parsortix compared with an existing in market product

·     Establishing the harvest efficiency of Parsortix using blood samples spiked with pre-labelled cell lines

·     Determining the number of residual leukocytes (white blood cells) in the Parsortix harvest

·     Determining the effect of blood volume processed on the number of leukocytes in the harvest

·     Developing an optimised enrichment protocol to reduce the number of leukocytes to a minimum 

·     Determining the effect of using different cell lines on harvest efficiency

·     Fluorescent labelling of enriched samples following harvest (off-chip labelling)

·     Application of off-chip labelling protocols to clinical samples

·     Use of downstream technologies for molecular analysis at a single cell level

·     Use of the DEPArray™ on harvested cells for single cell isolation

·     Use of whole genome amplification on harvested cells in the context of clinical samples

 

The CEP group was impressed by the performance of the Parsortix system and is now planning to use the system in future clinical studies.  Key findings from their work were that:

 

1)  Standard blood preservation tubes can be used preserving the blood for up to 96 hours after blood draw before separating it with the Parsortix system

 

2)  Plasma can be removed from the blood sample without compromising the Parsortix system.  This means that the Parsortix system can be used in parallel with cell-free DNA analysis with a single patient blood sample being used for both

 

3)  Based on spiked cell experiments, the Parsortix standard separation offers "a very high level of capture" of cells (80-100%)

 

4)  Using paired spiked samples, it was shown that the capture efficiency of the Parsortix device is not significantly different from that of the existing in market product for EpCAM positive cells.  EpCAM positive cells are those which the existing in market product antibody system can capture.  In addition the Parsortix system can capture EpCAM negative cells, which cannot be captured by the existing in market product 

 

5)  Based on spiked cell experiments, the Parsortix standard separation offers "a high level of harvest" of cells (60-100%)

 

6)  A sample enriched by the Parsortix system will contain in the range of 500 - 4500 leukocytes.  The number of contaminating leukocytes appears to be independent of the volume enriched, and appears to be dependent on the donor

 

7)  The CEP group at the CRUK MI has successfully developed an optimised Parsortix separation that results in the output containing very low levels of contaminating white blood cells.  The number of leukocytes harvested following optimised protocol enrichment differs between donors and currently ranges between 69 and 178 cells in a single sample. This will allow both single CTC isolation or direct genetic analysis of the whole enriched sample representing a major step forward in CTC capture and enrichment

 

8)  The CEP group has a working protocol for cell identification staining utilising fluorochromes compatible with various downstream technologies

 

9)  Cells enriched by the Parsortix device can be subsequently subjected to detailed molecular analysis.  Cells have been successfully processed using DEPArray for single cell analysis.  Cells were then successfully subjected to whole genome amplification.

 

In summary, work undertaken by the CEP group at CRUK MI has demonstrated a number of system benefits of the Parsortix system:

·     The system is marker independent thus does not require the use of capture antibodies to enrich CTCs. The potential advantages include ability to capture CTCs with weak cell marker expression as well as mesenchymal cells and cell clusters that may be important in dissemination and metastasis

·     The Parsortix system does not require red cell lysis, is compatible with blood preservation collection tubes, allows plasma collection from the same sample and is straight-forward to use with minimal user intervention.

 

CRUK MI intends to pursue clinical research work with the Parsortix system in colorectal cancer and pancreatic cancer.

 

Pancreatic cancer is notoriously difficult to treat and challenging to biopsy, particularly serially.  Identifying mutations in CTCs from pancreatic cancer patients to stratify treatment may have considerable utility in delivering personalised medicine.

 

The CEP group is developing methods which include using the Parsortix system aimed at genotyping CTCs from pancreatic, colorectal and lung cancer patients to facilitate an understanding of the mutations appearing in patient CTCs and how they may relate to disease progression and drug efficacy. The work is being undertaken in collaboration with consultant medical oncologists Professors Juan Valle, Mark Saunders, Fiona Blackhall (University of Manchester / The Christie NHS Foundation Trust) and Dr Claus Jorgensen who is a group leader at CRUK MI.

 

The pilot study now underway will evaluate the feasibility and potential clinical utility of routine use of the Parsortix system to provide CTC information for patients at presentation and throughout their treatment.

 

Ged Brady, Deputy and Genomics Leader within the Clinical & Experimental Pharmacology  group at Cancer Research UK Manchester Institute commented:

"The evaluation phase of our work is now successfully complete and we see great promise in the application of the Parsortix technology for harvesting CTCs for molecular analysis to enable personalised cancer care.  We are now undertaking pilot studies using the Parsortix system in both colorectal cancer and pancreatic cancer." 

 

ANGLE's Founder and Chief Executive, Andrew Newland, commented:

"It is of great importance to us that the CEP group believes in the potential of our Parsortix system to enable personalised cancer care.  Their extensive work provides us with a platform to secure clinical studies with key opinion leaders, which is the key next step towards widespread adoption of the Parsortix system in the treatment of cancer." 

 

 

 

For further information:

 

ANGLE plc	01483 685830
Andrew Newland, Chief Executive Ian Griffiths, Finance Director
Cenkos Securities Stephen Keys, Dr Christopher Golden (Nominated adviser) Andy Roberts, Christian Hobart (Sales)	020 7397 8900
Buchanan Mark Court, Fiona Henson, Sophie Cowles	020 7466 5000

 

 

 

 

Explanation of frequently used terms in connection with the Parsortix system

 

Biopsy	Process by which cancer cells are removed from the tumour for molecular analysis
Capture	Process for capturing target cells from sample
Capture efficiency	Proportion of target cells captured
CD45	The CD45 antibody recognises the human CD45 antigen, also known as the leukocyte common antigen.  WBC are CD45+ whereas CTCs are CD45-.  Staining with CD45 often used as a negative confirmation that CTCs are not WBC.
Cell-free DNA	Genomic DNA found in the plasma
Cell labelling	Technique involving the staining of target cells with fluorescent and chromogenic markers for cell identification
Cell lines	Cultured cells
CE Mark	Regulatory authorisation for the sale of products for clinical use in the European Union
Circulating tumour cell	Cancer cell that is circulating in the patient blood
CTC	Circulating tumour cell
CTC labelling	CTCs are often labelled with three markers and are formally identified as CTCs if they are CK+, CD45-, DAPI+
CK	Cytokeratin
CK+	A cell positive for the presence of cytokeratin protein or mRNA with the presence of distinct cytokeratins often used to identify epithelial cells
Clinical application	Use in treating patients
Clinical samples	Patient samples usually blood
Clinical use	Use in treating patients
Cultured cells	Cultured cells grown in the laboratory from human-derived cells used for experimental work
Cytokeratin	Cytokeratins are family of intracytoplasmic cytoskeleton proteins with members  showing tissue specific expression
DAPI	A nuclear stain that is often used to identify the nucleus in a cell
DEPArray	A commercial single cell isolation system
DNA	Deoxyribonucleic acid (DNA) the molecule that encodes the genetic instructions used in the development and functioning of all known living organisms and many viruses
Downstream technologies	Technologies used to undertake molecular analysis of harvested cells after the separation has taken place
EGFR	The epidermal growth factor receptor a signalling molecule which is typically present on the cell surface and can cell activity including cell proliferation. Mutations in EGFR or deregulation have been associated with a number of cancers including ~30% of all epithelial cancers
Enrichment	Generic term for concentrating target cells or molecules in a starting heterogeneous mixture
EpCAM	The EpCAM protein is found spanning the membrane that surrounds epithelial cells, where it is involved in cell adhesion
EpCAM+ cells	Cells that express EpCAM.  CTCs can be either EpCAM+ or EpCAM-
Epithelial cells	Cells that line the surfaces and cavities of the body
Epithelial CTCs	CTCs that are epithelial often based on EpCAM+
Epithelial-mesenchymal transition	Process by which epithelial cells lose their cell polarity and cell-cell adhesion, and gain migratory and invasive properties to become mesenchymal cells. EMT is thought to occur as part of the initiation of metastasis and is often responsible for cancer progression
EMT	Epithelial-mesenchymal transition
FDA	U.S. Food and Drug Administration responsible for authorised medical products in the United States
FDA 510(k)	A 510(k) is a premarket submission made to FDA to demonstrate that the device to be marketed is at least as safe and effective, that is, substantially equivalent, to a legally marketed device that is not subject to Premarket Approval. Submitters must compare their device to one or more similar legally marketed devices and make and support their substantial equivalency claims
Genome	Genetic material of an organism. The genome includes both protein coding and non-coding sequences
Harvest	Process for recovering captured cells from the separation system to allow molecular analysis
Harvest efficiency	Proportion of target cells harvested
Harvest purity	The number of target cells (such as CTCs) in the harvest as a proportion of the WBC. The minimum purity from which downstream analysis is currently possible is 0.5%.  Analysis of one target cell therefore requires no more than 200 WBC be in the harvest
HER2	A member of the epidermal growth factor receptor (EGFR/ERBB) family. Amplification or overexpression of HER2 has been shown to play an important role in the development and progression of certain aggressive types of breast cancer. In recent years the protein has become an important biomarker and target of therapy for ~ 30% of breast cancer patients
HNV	Healthy normal volunteer
HT29	Cultured colorectal cancer cell line
In-cassette labelling or in-situ labelling	CTC labelling for cell identification undertaken inside the separation system
KRAS	A signalling molecule frequently mutated in the development of many cancers
Leukocytes	White blood cells
Liquid biopsy	Term used for the process of obtaining cancer cells (or cell-free DNA) from a blood sample.  Unlike solid biopsy, liquid biopsy is non-invasive and repeatable
Mesenchymal CTCs	CTCs generally lacking epithelial markers with mesenchymal features
Metastasis	Spread of a cancer from one site to another
Molecular analysis	Analysis of DNA, RNA and protein often used to determine the mutational status of a patient
mRNA	Messenger RNA used to direct the synthesis of proteins
Off-chip labelling	CTC labelling for cell identification of harvested cells undertaken outside the separation system
Paired samples	Two related samples often used to compare different systems
Personalised cancer care	Treating a patient individually based on their personal data often including mutational and disease status
Plasma	Pale-yellow liquid component of blood obtained following removal of cells
Pre-labelled cell lines	Cells which are labelled often with a fluorescent label to facilitate identification during analysis or enrichment
RNA	Ribonucleic acid performs multiple vital roles in the coding, decoding, regulation, and expression of genes. Together with DNA, RNA comprises the nucleic acids, which, along with proteins, constitute the three major macromolecules essential for all known forms of life
Separation	Term used for processing of a sample through the Parsortix system
Single cell analysis	Extraction of a single target cell from the harvest for analysis
Solid biopsy	Standard process for surgically excising (cutting out) cells from a solid tumour when that tumour is accessible
Spiked cell experiments	Experiments where cultured cells are added (spiked) to HNV blood to assess the capture and harvest efficiency of the system
WBC	White blood cells
WGA	Whole genome amplification
Whole genome amplification	Method for amplification of an entire genome necessary for the picogram amounts of genomic DNA present in a single cell