Founded in July 2008, Metavectum is located as an independent company on the premises of the DESY (Deutsches Elektronen-Synchrotron) a major research facility of the Federal Republic of Germany. Metavectum’s main businesses are human medical molecular analysis and the synthesis of ultra-pure substances for use in complementary medicine.
In the team of Metavectum molecular biologists and biochemist collaborate with laboratory physicians.
Analysis and validation of assays and reagents are in line with the guidelines of the federal medical association (Bundesärtzekammer, Rili-BÄK) and the international MIQE standards. Equipment used in the biomolecular analysis is CE certified.
Metavectum is part of a network of oncologists, surgeons, radiologists, pathologists, specialized physicians and specialist clinics, to ensure the direct implementation of the results is possible.
The business model covers the following areas:
- Metavectum Prevention Test
- Metavectum Tumor Test
Metavectum Prevention Test
With the Metavectum Prevention Test, a weakening, malfunction or disease is diagnosed at an early stage by means of a novel array analysis of the overall metabolism. A therapy suggestion is derived from the results and submitted to the physician. Deviations in metabolism that could lead to disease are detected at a very early stage so that appropriate measures can be taken.
The test is also used during and after tumor therapy to obtain information about the onset of metabolic changes that indicate a recurrence or an early indication of a metabolic slide into cachexia, for example.
For analytical purposes, equipment is available that is normally only found in larger pharmaceutical companies (600 MHz NMR coupled with mass spectrometer and HPLC). These data at the low molecular level (metabolome) are coupled with data from the levels of gene expressions (transcriptome) and proteins (proteome) to give a detailed picture of the overall metabolism.
Metavectum Tumor Test
Tumors and metastases develop a very individual profile, which varies from person to person. Consequently, the treatment of the tumor must also be individualized and the variety of therapeutic methods (radiotherapy, surgery, chemotherapy, complementary therapies, immunotherapy, etc.) should be used in concerted interaction.
The Metavectum Tumor Test is used to determine the individual metabolic activity of tumor-specific receptors and proteins (“Achilles heel” of the tumor) at the level of gene expressions in the transcriptome and gene mutations in the genome. The analyses are performed at the written request of the treating physician, and a therapy proposal is derived from the results and submitted to the physician. The test system determines the optimal drugs for each patient from the total number of approved tumor drugs.
Both gene expression and gene mutation data are correlated in the test. The reason for this is that currently, depending on the tumor, only about 2-35% of all tumors reveal clinically relevant mutations that allow drug inhibition of corresponding receptors.
It must be pointed out that cultivation of tumor cells and incubation of the cells with various drugs as a proof of effect is not performed, the procedure does not provide reproducible results. The method used here is based solely on the evaluation of relevant gene expression on an mRNA basis and the determination of clinically relevant mutations. DNA or mRNA is extracted from the isolated tumor cells.
On the one hand, resected tissue is used as test material, which is sent to Metavectum on dry ice (-80 ºC) or in paraffin by the surgeon/oncologist after the patient has given his written consent. Since tumor biopsy contain on average only 10-20% pure tumor material in addition to connective tissue and precancerous tissue, the samples are first sent to the pathologist (in this case Pathologie Grandweg, Hamburg). The pathologist describes the type and status of the tumor and indicates its exact location in the tissue. On the basis of the pathological report, the tumor cells are isolated from the biopsy at Metavectum and examined.
In parallel or as a mono analysis, circulating tumor cells (CTC) are isolated from the patient’s whole blood (“liquid biopsy”). CTCs are the link between the primary tumor and metastases and provide information on which therapeutic agents can be used to treat the metastases.
The analysis of at least 10 tumor markers ensures that the isolated cells are indeed circulating tumor cells and not endothelial or epithelial cells that are continuously released into the bloodstream by organs or the vascular system and feign large numbers of circulating tumor cells as false positives. Normally, between 5 and 120 CTC are found in one milliliter of blood.
DNA and mRNA are now extracted from the tumor biopsy and the CTC. Using real-time PCR (qPCR), the expressions of tumor-relevant genes in the transcriptome are quantified in the tumor RNA (biopsy) and compared with the corresponding expressions of the circulating tumor cells (liquid-biopsy) and with reference material. Therapy-relevant mutations (BRAF, EGFR, MET, KRAS, NRAS, PTEN, MEK, etc.) are determined in the DNA of the tumor tissue and, for comparison, in the free circulating DNA (cfDNA, cell free DNA) and correlated with the results of the gene expression analysis.
In the final metavectum-report a comprehensive picture of the individual tumor metabolism is obtained and thus indications of the drug targets for optimal therapy. The expression levels of the CTC also allow the aggressiveness of the tumor to be estimated. CTC mutation analysis also reveals whether many different clones have formed, complicating therapy.
In gene expression analysis, the expression or formation of certain receptors that the tumor uses for its purposes is mapped to the corresponding approved drugs. For example, if topoisomerase II (TOPOIIA1) is highly expressed relative to non-tumor tissue, it concludes that use of the appropriate inhibitor (> anthracyclines) is advisable. If the tumor instead uses topoisomerase I (TOPOI) for its growth, topotecan must now be used as the corresponding TOPOI inhibitor. In this case the use of anthracyclines would be ineffective, associated with severe side effects and the development of resistance. This can then only be combated at very high cost. Resistance and certain transport pathways that the tumor can use to eject drugs from the cell play an important role.
Medical-molecular biological laboratory testing by means of qPCR and mutation analysis for the optimization of a therapy decision are in principle routinely practiced both in Germany and worldwide.
In tumor therapy, the four best-known biomarkers ERBB2/Her2neu (e.g. drug Herzeptin: yes or no), PGR, ER1 and KI67 are routinely analyzed for gene expression by immunohistochemistry in almost every oncology practice and clinic. Determination of EGFR and KRAS mutations etc. are mandatory for the use of certain drugs.
Metavectum has expanded the panel of biomarkers or the determination of gene expressions to more than 90 expressions. This enables Metavectum to identify drug targets for all currently approved drugs for the therapy of solid tumors and to make individual therapy suggestions based on this information. This allows the physician to target the therapy which also saves costs. The procedure and the selection of gene expressions are based on clinical studies, which will be gladly shared upon request.
Apart from the four markers mentioned above, the vast majority of tumor drugs are currently used without appropriate testing, which explains the relatively low cure rate and the increased incidence of metastases and resistant cells. The physician is forced to try out several therapy regimens on a trial and error basis. This is associated with considerable side effects (toxicity) for the patients and with considerable additional costs, which could be avoided.
The Metavectum analytical system can then be used to monitor the success of the therapy.
The data also provide important information for long-term prognosis with regard to the occurrence of recurrences and metastases. If the molecular biological analyses after surgical or radiotherapeutic removal of the primary tumor show that the aggressiveness of the tumor and the probability of recurrence are low, low-dose therapy can be initiated or, in individual cases, medication can be dispensed with altogether. However, this necessitates biannual monitoring.
The method is also suitable for intermittent monitoring and therapy adjustment, e.g. if tumors or metastases have been brought into an operable form by individual chemotherapy, but residues are likely on the basis of molecular biological data.
The cost of the Metavectum Tumor Test is usually covered by private health insurance if a request is made by the oncologist.
Circulating tumor cells (CTC)
There are a number of companies worldwide that focus on circulating tumor cells and attempt to obtain medically usable results from their analysis. The majority of these companies simpley quantify CTCs before and after chemotherapy, from the point of view that a reduction should be detectable if the therapy is successful.
In recent studies, however, it has been shown that the number of CTCs is fundamentally subject to temporal fluctuations: CTC are not equally distributed over all blood vessels of the body, CTC are replenished from epithelial tissue and bone marrow, CTC disintegrate due to intravasation and extravasation, EMT transitions alter the CTC barcode used for identification, the immune system destroys CTC in the bloodstream according to activity, CTC join together to form clusters.
This makes clear that dealing with circulating tumor cells is a complex field and requires an extensive periphery of equipment and methods to arrive at a medically usable finding for the therapist.
The difficulties in dealing with CTC can be well illustrated by a graph taken from a review published in 2013 [Lit: Bin Hong et al.; Theranostics 2013, Vol. 3, p 377-394]. The picture shows the occurrence and development of CTC in the blood vessels and in the body up to metastases.
The primary tumor releases single cells and clusters that enter the bloodstream under EMT (epithelial–mesenchymal transition) junctions. There, the bulk of CTC is driven into apoptosis and necrosis, either by pre-damage to the cell structure during detachment from the primary tumor or during invasion into the vasculature or by the immune system. During apoptosis, the cell contents are released (CTMat = circulating tumor material, CTDNA = circulating tumor DNA), which include low molecular weight substances, proteins and DNA.
Surviving CTC can now leave the bloodstream again and establish metastases at a suitable site or they associate in the bloodstream to form clusters (CTM). These clusters act as conglomerates integrating other blood components such as erythrocytes and leukocytes and also contain the above-mentioned CTDNA and CTMat. These cell clusters (CTMs) are transported further by the bloodstream, settle in capillaries due to their size, and form micrometastases and later metastases in this location.
This demonstrates that certain techniques and methods are essential for investigations of CTC and their derivatives in order to arrive at a medically relevant conclusion. These include:
- Mutation analysis in the genome
- Gene expression analysis in the transcriptome
- Protein qualification and quantification in the proteome
- Analysis of low-molecular substances (amino acids, etc.) in the metabolome
- Immunohistochemistry on individual CTC
In order to analyze the chain of primary tumor, CTC, secondary products and metastases from the point of view of a therapeutically relevant statement, Metavectum links information from the genome, the transcriptome, the proteome and the metabolome, since these four levels have been shown to influence each other mutually.
This also means that the individual levels (genome, proteome, metabolome) are able to ignore, block or even reverse work instructions or information from another level. For example, the work instruction arriving in the transcriptome to induce apoptosis of a cell is sometimes reversed to its opposite at the proteome level.
For these reasons, it is necessary to look at all levels if valid conclusions are to be drawn.
For the analyses, Metavectum uses the following methods and equipment, among others:
- Hybridization techniques
- Next Generation Sequencing
- PCR and qPCR
- Mutation analysis: Pyrosequencing
Proteome and metabolome analysis
- 2D-gel, FPLC, HPLC, GC
- NMR spektroskopy (600 MHz NMR)
- Mass spectrometry
- 2D and 3D cell culture
- Immunofluorescence microscopy
- Single cell analysis
All analyses are performed according to the international MIQE Guidelines. All instruments are CE-certified.