THERAPY

FIELD OF THE INVENTION

The present invention is in the field of medicine, more in particular tumor therapy and even more in particular in the monitoring of a tumor therapy.

BACKGROUND

Successful tumor therapy is dependent of the time point of the detection of the tumor and the selection of the right therapy. Identification of specific biomarkers in tissue samples of a tumor, help to determine both, the stage of malignancy and the presence of cell structures, which can serve as targets for therapy. Also, the detection of tumor-associated biomarkers in the blood is used to detect malignancies. However, expectations on biomarkers have not been met in many cases, because these are not specific enough or do not have the necessary sensitivity to be able to detect malignancies at an early stage in a reliable way. Furthermore, known biomarkers can often just be applied on specific tumor types. Therefore, biomarkers are increasingly being sought, which allow the general detection of tumors. Such a general tumor test would considerably simplify the diagnosis of a wide variety of tumors and thus significantly improve both, therapy and monitoring.

So far, small benign tumors and small malignant tumors cannot be detected with tumor markers, which are determined in blood samples. Small tumors release relatively small amounts of biomarker, which are then diluted in the blood volume, consequently no significant increase in the concentration of biomarkers in blood is measurable.

A novel method for detecting biomarkers in specific cells in the blood, which carry tumor material has solved this problem, since the tumor material in these specific cells is not diluted in the whole blood and remains in high concentration inside said cells.

This novel method is based on the principle of the immune system to detect, phagocyte and digest undesirable cell structures. Undesirable cell structures comprise intruders from outside like bacteria and parasites or even degenerated body ^' s own cells like tumor cells. During the process of eliminating undesirable cell structures, immune cells, such as monocytes, phagocyte these structures and present them. Thus, tumor cells are recognized by said immune cells, which then phagocyte and attack them. Therefore, the immune cells migrate from the blood vessels into the tumors to phagocyte tumor cells. Consequently, the immune cells enclose fragments of one or more tumor cells, which are then presented over a period of several days. The aim is to inform other immune cells what the target is. After a while, the immune cell leaves the tumor and gets back into the bloodstream containing still tumor material. A sort of immune cells, which carry tumor material in the cytoplasm are characterized by the fact that they carry the surface markers CD14 and CD16, which can be easily detected in blood samples by flow cytometry. With this method it is possible to detect these immune cells, count them and even further characterize them.

Merely recognition and counting of immune cells, such as monocytes in blood samples is not sufficient to detect tumors at an early stage, since the number of these cells is not significantly different between healthy and tumor patients. Only through the detection of tumor material in these immune cells, a population of immune cells can be identified which has a significant numerical difference between healthy and tumor patients. The detection of these immune cells is done by an elaborate method in which specific biomarkers expressed in the inside of the specific immune cells, are identified. This new technology is called "EDIM", epitope detection in monocytes. Several studies have shown that the detection of biomarkers with the EDIM technology is much more sensitive and specific than the detection of biomarkers in blood. With the EDIM technology, biomarkers can be used to detect tumors earlier and more specifically.

In search of a general biomarker for tumors, it was noticed that there is a fundamental process that is alike in all tumor cells. All forms of tumor cells have a disorder in the process of planned cell death. Only if this process is disturbed, a tumor cell can arise. Therefore, biomarkers for apoptosis disorders are of big interest. One of the known biomarkers is based on the DNaseX-enzyme, which normally performs the last step of the apoptosis and has been shown to be overexpressed in tumors. Nevertheless, the enzymatic activity of the DNase-enzyme is inhibited by the increased release of inhibitors by the tumor cells. The ApolO epitope of the DNaseX protein sequence is particularly well detectable, which therefore, can be used for diagnosis in patients with tumors.

Further studies have identified a gene which is associated with the invasive growth behavior of tumors and additionally leads to an increased cell proliferation and an inactivation of the apoptosis. Therefore, TKTL1 is a further important biomarker for tumors. The detection of biomarkers ApolO and TKTL1 in immune cells by means of blood test allows early detection of all types of carcinoma examined so far, but also of sarcomas and hematological tumors. Until now, the outcome of a tumor therapy remains unknown for some time. Therefore, there is a need in the art for a method of reliably monitoring a tumor therapy to be able to select the optimal therapeutic options for a subject with one or more tumors to an earlier time point than previously possible.

SUMMARY OF THE INVENTION

Based on the background above, it is the objective of the present invention to provide a method of efficiently and reliably monitoring a tumor therapy for determining the effects of a tumor therapy on a subject in need thereof, and therefore being able to select the optimal therapeutic options for said subject at an early time point (for example: already during therapy). This objective is attained by the method of monitoring a tumor therapy in a subject, the method comprising: a. detecting at least one type of monocytes in two or more samples obtained from said subject, and b. determining the level of TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof in said at least one type of monocytes.

Preferably said two or more samples are independently obtained from said subject at various time points selected from the group consisting of: a. before initiation of therapy, b. after initiation of therapy, and c. at one or more further time points during therapy.

The invention further relates to a kit for monitoring a tumor therapy, the kit comprising: a. instructions for performing the method of monitoring as disclosed herein, b. markers for monocytes comprising CD14 and CD16, c. TKTL1, ApolO, Epcam and/or other tumor specific markers, d. reaction agents selected from the group comprising reaction buffer, wash buffer, and secondary markers. e. standard data showing the correlation between the levels of TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof in said monocytes of samples taken at various time points.

Furthermore, the invention relates to a marker for TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof for use in a method for monitoring a tumor therapy.

DEFINITIONS

The term "tumor" is used in the present application and is understood to encompass malignant and non-malignant tumors, including solid tumors and hematological tumors. Solid tumors are exemplified by tumors of the breast, bladder, bone, brain, central and peripheral nervous system, colon, endocrine glands (e.g. thyroid and adrenal cortex), esophagus, endometrium, germ cells, head and neck, kidney, lover, lung, larynx and hypopharynx, mesothelioma, ovary, pancreas, rectum, renal, small intestine, soft tissue, testis, stomach, skin, ureter, vagina and vulva. In addition, it includes primary tumors in said organs and corresponding secondary tumors in distant organs (tumor metastases). Hematological tumors are exemplified by aggressive and indolent forms of leukemia and lymphoma, namely non- Hodgkins disease, chronic and acute myeloid leukemia, acute lymphoblastic leukemia, Hodgkins disease, multiple myeloma and T-cell lymphoma. Also included are myelodysplastic syndrome, plasma cell neoplasia, paraneoplastic syndromes and tumors of unknown primary site.

Herein, "tumor therapy" refers to any known therapy applied by the skilled person to combat tumors, such as immunotherapie, chemotherapy, radiation, targeted therapy and hormone therapy. Furthermore, it refers to therapies which do not affect the immune system and in which the therapy directly leads to an immunological defense reaction against the tumor. These include immunotherapies, such as, but not limited to immunotherapy with monoclonal antibodies, immunotherapy with immune checkpoint inhibitors, cancer vaccines, immunotherapy with oncoviruses, or other non-specific immunotherapies. Therapy with antibodies comprise naked monoclonal antibodies, conjugated monoclonal antibodies, or bispecific monoclonal antibodies. Naked antibodies comprise alemtuzumab, or trastuzumab; conjugated antibodies comprise radiolabeled antibodies such as Ibritumomab tiuxetan, or chemolabeled antibodies, such as Brentuximab vedotin, Ado-trastuzumab emtansine, or denileukin diftitox; bispecific monoclonal antibodies include blinatumomab. The term "tumor therapy" further relates to therapies with drugs comprising bevacizumab, cetuximab, denileukin diftitox, or further known to the person skilled in the art. Monocytes are the largest type of white blood cells. They are part of the innate immune system of vertebrates including all mammals (humans included), birds, reptiles, and fish. They are amoeboid in shape, having a granulated cytoplasm. Monocytes constitute 2% to 10% of all leukocytes in the human body. They play multiple roles in immune function.

Monocytes are produced by the bone marrow from precursors called monoblasts, which are bipotent cells that differentiated from hematopoietic stem cells. Monocytes typically circulate in the bloodstream for about one to three days and then move into tissues throughout the body. Monocytes in tissues mature to different types of macrophages depending on the anatomical location, i.e. osteoclasts, microglia cells, histiocytes, and Kupfer cells. Monocytes can move quickly to sites of infection in the tissues and divide/differentiate into macrophages and dendritic cells to elicit an immune response.

There are at least three types of monocytes in human blood:

The classical monocyte, characterized by high level expression of the CD14 cell surface receptor (CD14++ CD16- monocyte)

The non-classical monocyte shows low level expression CD14 and additional co-expression of the CD16 receptor (CD14+ CD16++ monocyte).

The intermediate monocyte with high level expression of CD14 and low level expression of CD16 (CD14++ CD16+ monocytes).

Monocyte markers comprise CD2, CDllb, CD14, CD16, CD31, CD33, CD56, CD62L, CD115, CD163, CD192, CX3CR1, CXCR3, CXC 4, CCR2, CDllb, CD16/CD32, CD31, CD43, CD44, CD45, CD62L, CD115, CX3CR1, F4/80, Grl, Ly-6C, LFA-1, VEGF, and further.

Since hardly any surface markers are known which specifically detect only monocytes, therefore the combination of surface markers is used to identify monocytes alone in the flow cytometer, for example a combination of the markers CD14 and CD16.

Macrophages engulf and digest cellular debris, foreign substances, microbes, cancer cells, and anything else that does not have types of proteins specific to healthy body cells on its surface in a process called phagocytosis. Fluman macrophages can be identified by using flow cytometry or immunohistochemical staining by their specific expression of proteins such as, but not limited to CDllb, CD14, CD16, CD40, CDllb, CD56, CD64, CD68, and CD163. Macrophages are attracted to a damaged site through chemotaxis, triggered by a range of stimuli (immune response) including pathogens and cytokines released by macrophages already at site. Macrophages phagocyting tumor cells, move away from the tumor and can be found circulating in blood. These macrophages contain debris of tumor cells.

The term "sample" as used herein refers to a sample of bodily fluid obtained for the purpose of diagnosis, prognosis, or evaluation of a subject of interest, such as a patient. Preferred test samples include blood, serum, plasma, cerebrospinal fluid, urine, saliva, sputum, and pleural effusions. In addition, one of skill in the art would realize that some test samples would be more readily analyzed following a fractionation or purification procedure, for example, separation of whole blood into serum or plasma components. The sample is obtained from the subject and subjected to the method according to the invention. In a preferred embodiment of the invention, the sample is a blood sample.

Herein the term "subject" is used to refer to an animal (e.g. a mammal, a fish, an amphibian, a reptile, a bird and an insect). In a specific embodiment, a subject is a mammal (e.g., a non-human mammal and a human). In another embodiment, a subject is a primate (e.g., a chimpanzee and a human). In another embodiment, a subject is a human. In another embodiment the subject is a human with one or more tumors.

As used herein, the term "diagnosis" refers to the identification of the disease (herein a tumor) at any stage of its development, and also includes the determination of predisposition of a subject to develop the disease. In a preferred embodiment of the invention, diagnosis of a tumor occurs prior to the manifestation of symptoms. Subjects with higher risk of developing a tumor are of particular concern. The diagnostic method of the invention also allows confirmation of the presence of a tumor in a subject suspected to have a tumor.

A "prognosis" refers to assignment of a probability that a given course or outcome will occur. This is often determined by examining one or more "prognostic indicators". These are markers, the presence or amount of which in a subject (or a sample obtained from the subject) signal a probability that a given course or outcome will occur. For example, when one or more prognostic indicators reach a sufficiently high level in samples obtained from such subjects, the level may signal that the subject is at an increased probability for eventually advancing into end-stage tumor disease.

In the context of the present invention, the term "score" refers to a value, which can be a median or mean or the 75th, 90th, 95th or 99 ^th percentile or a reference. This can be for instance also an "optimal" score. The optimal score value for a given marker is the value where the sensitivity and specificity is maximal for this marker.

The predetermined value can be established based upon comparative groups or it can be a range. The predetermined value can vary among particular reference populations selected, depending on their habits, ethnicity, genetics etc. Accordingly, the predetermined values selected may take into account the category in which an individual falls. Appropriate ranges and categories can be selected with no more than routine experimentation by the person skilled in the art.

In the context of the present invention the term "score" is the ratio between whole monocytes and/or macrophages positive for CD14 and CD16, CD2, CDllb, CD14, CD16, CD16/CD32, CD31, CD33, CD40, CD43, CD44, CD45, CD56, CD62L, CD64, CD68, CD115, CD163, CD192, CX3CR1, CXCR3, CXC 4, CCR2, CDllb, CD16/CD32, CD31, CD43, CD44, CD45, CD62L, CD115, CX3CR1, F4/80, Grl, Ly-6C, LFA-1, or VEGF and said monocytes and/or macrophages which are additionally positive for TKTL1, ApolO, Epcam and/ or other tumor specific markers.

The term "level" or "expression level" in the context of the present invention relates to the level at which the biomarker is present in a sample from a subject. The expression level of a biomarker is generally measured by flow cytometry, wherein the biomarker is detected by a laser and by computational analysis a dot plot and/or a histogram can be created, wherein a score for the levels of expression can be determined.

The term "biomarker" (biological marker) and "marker" are used synonymously. The term herein relates to measurable and quantifiable biological parameters (e.g., specific enzyme concentration, specific hormone concentration, specific gene phenotype, presence of biological substances) which serve as indices for health- and physiology-related assessments, such as disease risk, disease diagnosis, etc. Furthermore, a biomarker is defined as a characteristic that is objectively measured and evaluated as an indicator of normal biological processes, pathogenic processes or responses to a therapeutic intervention. A biomarker may be measured on a sample. Biomarkers can be classified as antecedent biomarkers (identifying the risk of developing an illness), screening biomarkers (screening for disease), diagnostic biomarkers (recognizing diseases), staging biomarkers (categorizing disease severity), or prognostic biomarkers (predicting future disease course, including recurrence and response to therapy, and monitoring efficacy of therapy). A biomarker may be a protein, peptide or nucleic acid molecule.

In context of the present invention a biomarker is a biomolecule, more specifically a protein or even more specifically a protein which is expressed on the cell surface or intracellularly of immune cells. Furthermore, a biomarker is a biomolecule used to detect a further biomarker in and/or on immune cells, these comprise primary and/or secondary antibodies. The term "antibody" comprises monoclonal, oligoclonal and polyclonal antibodies and binding fragments thereof, in particular Fc-fragments as well as so called "single-chain-antibodies" (Bird R. E. et al (1988) Science 242:423-6), chimeric, humanized, in particular CDR-grafted antibodies, and dia or tetrabodies (Holliger P. et al (1993) Proc. Natl. Acad. Sci. U.S.A. 90:6444-8). The term "antibody" further comprises modified antibodies, which refers to conjugated antibodies, synthetic antibodies, bispecific antibodies, and F(ab)2-fragment antibodies, and other Fv formats. In the context of the present invention a conjugated antibody comprises conjugation with a label such as, but not limited to fluorescent dyes, a specifically introduced radioactive element, or a biotin tag. In the context of the present invention, fluorescent dyes may for example be FAM (5-or 6-carboxyfluorescein), VIC, NED, Fluorescein, Fluoresceinisothiocyanate (FITC), Phycoerythrin (PE), Peridinin-Chlorophyll-protein (PerCP), IRD-700/800, Cyanine dyes, such as CY3, CY5, CY3.5, CY5.5, Cy7, Allophycocyanin (APC), Xanthen, 6-Carboxy-2',4',7',4,7-hexachlorofluorescein (HEX), TET, 6-Carboxy-4',5'-dichloro-2',7'- dimethodyfluorescein (JOE), N,N,N',N'-Tetramethyl-6-carboxyrhodamine (TAMRA), 6-Carboxy-X- rhodamine (ROX), 5-Carboxyrhodamine-6G (R6G5), 6-carboxyrhodamine-6G (RG6), Rhodamine, Rhodamine Green, Rhodamine Red, Rhodamine 110, BODIPY dyes, such as BODIPY TMR, Oregon Green, Coumarines such as Umbelliferone, Benzimides, such as Floechst 33258; Phenanthridines, such as Texas Red, Yakima Yellow, Alexa Fluor, PET, Ethidiumbromide, Acridinium dyes, Carbazol dyes, Phenoxazine dyes, Porphyrine dyes, Polymethin dyes, and the like.

Also comprised are immunoglobulins like proteins that are selected through techniques including, for example, phage display to specifically bind to the molecule of interest contained in a sample. In this context the term "specific binding" refers to antibodies raised against the molecule of interest or a fragment thereof. An antibody is considered to be specific, if its affinity towards the molecule of interest or the aforementioned fragment thereof is at least 50-fold higher, preferably 100-fold higher, more preferably at least 1000-fold higher than towards other molecules comprised in a sample containing the molecule of interest. It is well known in the art how to make antibodies and to select antibodies with a given specificity.

The term "correlation", as used herein in reference to the use of specific markers, refers to comparing the presence or amount of the biomarker(s) in a subject to a specific time point, to its presence or amount to another specific time point. The subject is one known to suffer from, or known to be at risk of, a given condition; or one to be free of a given condition. The level of the biomarker in the sample is said to have been correlated with a diagnosis; i.e. the level of the biomarkers can be used by the person skilled in the art to determine whether the subject suffers from a specific type diagnosis and respond accordingly. Alternatively, the level of the biomarker can be compared to a biomarker level known to be associated with a good outcome (e.g., the absence of disease, etc.). In preferred embodiments, a profile of the level of the biomarker is correlated to a particular outcome of a tumor therapy.

The parameters monitored by the method as disclosed herein may include effectiveness of the therapy and impact of the therapy on the subject.

In the present invention, the term "monitoring" denotes the observation of the state or progression of a subject's medical condition by measuring the level of a certain diagnostic marker or markers for said medical condition at various points of time.

Herein, the term "beneficial outcome" refers to the outcome a subject may experience after tumor therapy, which leads to an improvement in at least one sign of a tumor, which comprise tumor shrinkage, decrease in growth rate or suppression of tumor growth. Further, a reduction of the number of tumor cells, reduction of tumor size, inhibition of tumor cell infiltration into peripheral organs, retarded, slowed or stopped cell infiltration, inhibition of tumor metastasis or slowed tumor metastasis, prevention or delayed recurrence of tumor. Also, it may mean that the disease is no longer progressive. Further, it refers to the relieve of one, more, or even all symptoms associated with tumors.

As used herein, a "kit" is a packaged combination optionally including instructions for use of the combination and/or other reactions and components for such use.

All other terms are, unless explicitly defined otherwise, used as understood by those skilled in the art.

DETAILED DESCRIPTION

The present invention relates to a method for monitoring a tumor therapy in a subject, the method comprising: a. detecting at least one type of monocytes in two or more samples obtained from said subject, and

b. determining the level of TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof in said at least one type of monocytes.

In a preferred embodiment the said two or more samples are independently obtained from said subject at various time points selected from the group consisting of: a. before initiation of therapy, b. after initiation of therapy, and c. at one or more further time points during therapy.

The invention further relates to a method for monitoring a tumor therapy, wherein said monocytes are selected from the group comprising monocytes positive for CD14 and CD16, CD2, CDllb, CD14, CD16, CD16/CD32, CD31, CD33, CD40, CD43, CD44, CD45, CD56, CD62L, CD64, CD68, CD115, CD163, CD192, CX3CR1, CXCR3, CXC 4, CCR2, CDllb, CD16/CD32, CD31, CD43, CD44, CD45, CD62L, CD115, CX3CR1, F4/80, Grl, Ly-6C, LFA-1, or VEGF. In a preferred embodiment the monocytes are CD14 and CD16 positive.

In one embodiment the method of monitoring a tumor therapy, wherein the two or more samples comprise a first and a second sample, wherein the first sample is obtained before initiation of the therapy and the second sample is obtained 3 to 10 days after initiation of therapy. Preferably, the second sample is obtained 4 to 8 days after initiation of therapy, more preferably 5 to 7 days after initiation of therapy, and ideally 6 days after initiation of therapy. Preferably, the first sample is obtained 1 to 14 days before initiation of therapy, more preferably 1 to 7 days, even more preferably 1 to 3 days. Ideally the first sample is obtained directly before initiation of therapy.

The inventors surprisingly found, that if the level of TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof in said monocytes is increased after initiation of therapy, a beneficial outcome is associated to said therapy.

In one embodiment, a beneficial outcome can be associated to the therapy, if the level of TKTL1, ApolO, Epcam and/or other tumor specific markers in said monocytes after initiation of the therapy is at least 1.1 times, preferably 1.2 times, more preferably 1.3 times, even more preferably 1.4 times higher than the level of TKTL1, ApolO, Epcam and/or other tumor specific markers in said monocytes before initiation of the therapy.

The inventors further found, that typically the level of TKTL1, ApolO, Epcam and/ or other tumor specific markers or fragments thereof in said monocytes having a peak five to seven days after initiation of therapy and then continuously decreasing until day 9-11, where the expression levels of TKTL1, ApolO, Epcam and/or other tumor specific markers are similar to their expression levels before initiation of therapy.

In a preferred embodiment of the invention the level of TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof is determined in one or more samples obtained at various timepoints after initiation of therapy and are compared to the level of TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof in one or more samples obtained before initiation of the therapy. If the level of TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof in samples obtained after initiation of therapy are significantly higher than in samples obtained before initiation of therapy, a beneficial outcome is associated to said therapy.

In an alternative embodiment of the invention the level of TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof in a first sample obtained before initiation of therapy is compared to the level of TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof in a second sample obtained after initiation of therapy. If the level of TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof in the second sample is significantly higher than the level of TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof in the first sample, a beneficial outcome is associated to said therapy. The method according to the invention involves comparing the level of said markers in a subject, which has a tumor or is suspected to have a tumor and will initiate tumor therapy. The predetermined score can take a variety of forms. It can be a single score: This can be for instance a median or mean or the 75th, 90th, 95th or 99 ^th percentile or a reference. This can be for instance also an "optimal" score. The optimal score value for a given marker is the value where the sensitivity and specificity is maximal for this marker.

The predetermined value can be established based upon comparative groups or it can be a range. The predetermined value can vary among particular reference populations selected, depending on their habits, ethnicity, genetics etc. Accordingly, the predetermined values selected may take into account the category in which an individual falls. Appropriate ranges and categories can be selected with no more than routine experimentation by the person skilled in the art.

In one alternative embodiment the invention relates to the method of monitoring a tumor therapy, wherein if the levels of TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof in said samples are above a determined score, a beneficial outcome is associated to said therapy.

In one embodiment of the invention the relative amount of positive monocytes and/or macrophages is multiplied by factor 10 resulting in a "sore" - for a better handling and representation of the data.

In an alternative embodiment the determined relative amount of positive monocytes and/or macrophages score is multiplied by factor 10 resulting in a score of TKTL1 is between 1 and 1000, preferably between 50 and 500, more preferably between 90 and 150 and ideally between 110 and 126, and/or the score of ApolO is between 1 and 1000, preferably between 50 and 500, more preferably between 100 and 160 and ideally between 120 and 140 In a further embodiment of the invention, the determined score of TKTL1 and ApolO is added and is between 2 and 2000, preferably between 100 and 600, more preferably between 200 and 300 and ideally between 230 and 260. Especially preferred is a determined score of TKTL1 of 118, a determined score of ApolO of 130 and/or a determined score of TKTL and ApolO added of 245.

The method according to the invention involves tumor therapies selected from the group consisting of: a. immunotherapy,

b. chemotherapy,

c. radiation,

d. targeted therapy, and

e. hormone therapy.

In one embodiment of the invention the tumor therapy is a therapy which does not affect the immune system and in which the therapy directly leads to an immunological defense reaction against the tumor. These include immunotherapies, comprising immunotherapy with monoclonal antibodies, immunotherapy with immune checkpoint inhibitors, cancer vaccines, immunotherapy with oncoviruses, or other non-specific immunotherapies. Therapy with antibodies comprise naked monoclonal antibodies, conjugated monoclonal antibodies, or bispecific monoclonal antibodies. Naked antibodies comprise alemtuzumab, or trastuzumab; conjugated antibodies comprise radiolabeled antibodies such as Ibritumomab tiuxetan, or chemolabeled antibodies, such as Brentuximab vedotin, Ado-trastuzumab emtansine, or denileukin diftitox; bispecific monoclonal antibodies include blinatumomab. The term "tumor therapy" further relates to therapies with drugs comprising bevacizumab, cetuximab, denileukin diftitox, or further known to the person skilled in the art.

In a preferred embodiment of the invention the tumor therapy is an immune therapy, and ideally an immunotherapy with oncoviruses.

In another preferred embodiment of the invention the tumor therapy is a targeted therapy. All targeted tumor therapies are comprised. Especially preferred is a targeted therapy using HER2, PSMA, Beta-HCG, CEA, PSA, CA 125, CA 15-3, CA 19-9, CA 72-4, Calcitonin, CgA, CYFRA 21-1, NSE, Protein S100, PD-L1 and/or other checkpoint inhibitors as the target.

The method according to the invention is also for use in medical decision making for an individual subject tumor therapy. In one embodiment of the invention, the medical decision is made after evaluating if the subject who underwent a tumor therapy according to the invention shows a beneficial outcome or not.

The monocytes of interest according to the present invention are detected by means of flow cytometry. Further the level of TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof in said monocytes is determined by using a method selected from the group comprising flow cytometry, and ELISA.

In certain embodiments, the markers for TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof exhibit at least about 70% sensitivity, more preferably at least about 80% sensitivity, even more preferably at least about 85% specificity, still more preferably at least about 90% specificity, and most preferably at least about 95% specificity. In particularly preferred embodiments, both the sensitivity and specificity are at least about 75% more preferably at least about 80%, even more preferably at least about 85%, still more preferably at least about 90%, and most preferably at least about 95%. The term "about" in this context refers to +/- 5% of a given measurement.

The invention further relates to a kit for monitoring a tumor therapy, the kit comprising: a. instructions for performing the method of monitoring as disclosed herein, b. markers for monocytes comprising CD14 and CD16,

c. marker for TKTL1, ApolO, Epcam and/or other tumor specific markers, d. reaction agents selected from the group comprising reaction buffer, wash buffer, and secondary markers.

e. standard data showing the correlation between the levels of TKTL, ApolO, Epcam and/or other tumor specific markers or fragments thereof in said monocytes of samples taken at various time points.

Furthermore, the invention relates to a marker for TKTL1, ApolO, Epcam and/or other tumor specific markers or fragments thereof for use in a method for monitoring a tumor therapy.

The invention further relates to a method of treating a tumor patient in need thereof, wherein the tumor therapy is initiated or altered depending on the outcome of the method of monitoring according to the present invention. FIGURE LEGENDS

Figure 1 shows the ApolO score in a treatment cycle of a patient. Figure 2 shows the TKTL1 score in a treatment cycle of a patient.

Figure 3 shows the cumulated score of ApolO and TKTL1 in a treatment cycle of different patients.

The results of some working examples of the invention are shown in figures 1-3. The level of TKTL1 and ApolO in CD14 and CD16 positive monocytes were measured by flow cytometry. The score is achieved by calculating the ratio of the level of the CD14 and CD16 positive monocytes to the level of said monocytes which are additionally TKTL1 and/ or ApolO positive. This ratio is multiplied by factor 10. Figure 1 shows the ApolO scores in a treatment cycle of a patient, who is suffering from cancer. The scores are obtained from samples of a patient, which were taken at days 1, 2, 3, 4, 5 and 6 of the therapy.

All measurements resulting in scores above the red border line shows positive ApolO levels.

Figure 2 shows the TKTL1 scores in a treatment cycle of a patient, who is suffering from cancer. The scores are obtained from samples of a patient, which were taken at days 1, 2, 3, 4, 5 and 6 of the therapy.

All measurements resulting in scores above the red border line shows positive TKTL1 levels. Figure 3 shows the cumulated scores of ApolO and TKTL 1 in a treatment cycle of different patients, who is suffering from cancer. The scores are obtained from samples of seven different patients, which were taken at days 1, 2, 3, 4, 5 and 6 of the therapy.

Figure 3 also shows, that in some patients, the score starts decreasing at day 6 of the therapy.