WO2020245790A1

WO2020245790A1 - Anti-dmt1 or zip14 antibodies for use in the treatment of cancer

Info

Publication number: WO2020245790A1
Application number: PCT/IB2020/055317
Authority: WO
Inventors: Antonino MAIORANA; Luca RONCATI
Original assignee: Universita' Degli Studi Di Modena E Reggio Emilia
Priority date: 2019-06-07
Filing date: 2020-06-05
Publication date: 2020-12-10
Also published as: IT201900008376A1

Abstract

The present invention relates to antibodies able to bind to and inhibit the membrane proteins responsible for cellular internalisation of non-transferrin bound iron for use in the treatment of tumours.

Description

ANTI-DMT1 OR ZIP14 ANTIBODIES FOR USE IN THE TREATMENT OF CANCER

Technical field of invention

The present invention relates to antibodies with antitumoral activity that inhibit the cellular internalisation mechanism of non-transferrin bound iron.

State of the art

Iron is a basic bioelement, and the most important transition metal in all living organisms, from bacteria to human beings, due to its ability to mediate electron transfer. It is present in the haem groups of the proteins in the electron transport chain responsible for oxidative phosphorylation in the mitochondrial crests, a well-known multistage process designed to obtain energy in the form of ATP (adenosine triphosphate) through generation of a proton gradient (chemiosmosis), demand for which has considerably increased in tumour cells.

Bone marrow contains a wealth of multipotent haematopoietic stem cells which continually require iron to mature, and which resemble tumour cells in many respects, except that the latter lose their ability to differentiate. The signs and symptoms of iron- deficiency anaemia include fatigue and weakness, pale skin, pale mucosa, poor appetite, weight loss and often alopecia, all of which are also found in patients suffering from malignant tumours (with or without bleeding), and suggest its hyperconsumption in cancer patients.

Iron mainly forms compounds in the ferrous (²⁺) and ferric (³⁺) states, acting as electron donor or acceptor respectively.

After its ingestion, the gastric juices facilitate the dissociation of iron from food; haem iron directly enters the duodenal enterocytes, whereas non-haem iron is only absorbed after reduction to Fe²⁺. The brush border enzyme, duodenal cytochrome B (DcytB), therefore reduces non-haem Fe³⁺ to Fe²⁺, and this conversion is promoted by the action of ascorbic acid, which improves the bioavailability of non-haem iron. The duodenal enterocytes absorb Fe²⁺ through Divalent Metal Transporter 1 (DMT1), also known as Divalent Cation Transporter 1 (DCT1), a trans-membrane protein which is highly conserved from bacteria to human beings, and encoded by gene SLC11 A2 (Solute Carrier Family 11 member 2).

The enterocytes can release the iron absorbed into the bloodstream by means of hepcidin-sensitive ferroportin, wherein it can bind to its transport protein, transferrin, after its conversion to Fe³⁺ by ferroxidase hephaestin, giving rise to“Transferrin Bound Iron” (TBI). Each transferrin molecule can carry two Fe³⁺ ions; two particular members of the transferrin family are lactotransferrin, better known as lactoferrin, mainly present in colostrum and milk, and melanotransferrin, anchored to the membrane of melanomatous cells.

To date, however, it has been believed that the most important iron from the biological standpoint was that bound to transferrin, which is the plasma glycoprotein responsible for iron transport from the duodenum to the rest of the body. Transferrin is taken up by the cells through endocytosis mediated by the TFR1 (TransFerrin Receptor 1) or TFR2 (TransFerrin Receptor 2) receptors. TFR1 exhibits 25 times greater affinity than TFR2, and is therefore the main transferrin bond receptor.

In the past, TFR1 was used as target for the administration of numerous cytotoxic and antitumoral substances conjugated with transferrin, such as cisplatin, chlorambucil, cyclophosphamide, doxorubicin, diphtheria toxin and artemisinin. Other researchers are experimenting with new-generation antibodies against TFR1 (or TFR2) with the aim of inhibiting the endocytosis of transferrin bound iron, to obtain intracellular iron depletion.

There is consequently a strong need to identify new antitumoral agents.

Summary of the invention

The subject of the invention is antibodies able to bind to and inhibit the membrane proteins responsible for cellular internalisation of non-transferrin bound iron for use as a medicament, in particular in the treatment of tumours.

Detailed description of the invention

It has surprisingly been found that antibodies able to bind to and inhibit the membrane proteins responsible for cellular internalisation of non-transferrin bound iron (NTBI) are useful as antitumoral agents, especially in the treatment of aggressive malignant tumours.

The subject of the invention is antibodies for use as a medicament, in particular as inhibitors of the membrane proteins responsible for cellular internalisation of non-transferrin bound iron.

The invention also relates to antibodies able to bind to and inhibit the membrane proteins responsible for cellular internalisation of non-transferrin bound iron for use in the treatment of tumours.

As used according to the present invention, the term“inhibit” means the ability to reduce or block the functionality of the membrane proteins responsible for cellular internalisation of non-transferrin bound iron.

According to a preferred aspect, the membrane protein belongs to the DMT (Divalent Metal Transporter) family or the ZIP (Zinc-regulated transporter Iron-regulated transporter like protein) family and the isoforms thereof; the antibody is therefore selected from an anti-ZIP antibody, an anti-DMT antibody and combinations thereof.

According to a further preferred aspect, the membrane protein is selected from ZIP 14 (Zinc-regulated transporter Iron-regulated transporter like protein, or Human SLC39A14) and/or DMT1 (Divalent Metal Transporter); the antibody is therefore selected from an anti-ZIP 14 antibody, an anti-DMT 1 antibody and combinations thereof.

The term“antibody”, as used according to the present invention, means isolated antibodies or fragments of said antibodies, which bind specifically to the membrane proteins (transmembrane channels) responsible for cellular internalisation of non transferrin bound iron, preferably ZIP14 or DMT1.

In particular, the binding fragments can be Fab, Fab’, F(ab’)2 or Fv fragments and single-chain antibodies.

As known from the prior art, the antibodies can advantageously be, for example, polyclonal, monoclonal, chimeric and/or fully human or humanised antibodies. Anti-ZIP 14 and anti-DMTl antibodies are known in the literature and described, for example, in Liuzzi JP, Lichten LA, Rivera S, Blanchard RK, Aydemir TB, Knutson MD, Ganz T, Cousins RJ, Interleukin-6 regulates the zinc transporter Zip 14 in liver and contributes to the hypozincemia of the acute-phase response, Proc Natl Acad Sci USA, 2005 May 10;102(19):6843-8, and in Garrick MD, Dolan KG, Horbinski C, Ghio AJ,

Higgins D, Porubcin M, Moore EG, Hainsworth LN, Umbreit JN, Conrad ME, Feng L, Lis A, Roth JA, Singleton S, Garrick LM, DMT1 : a mammalian transporter for multiple metals. Biometals, 2003 Mar;16(l):41-54.

Moreover, anti-ZIP14 and anti-DMTl antibodies are available on the market, such as those manufactured by Biorbyt with the following catalogue numbers (all information is available at the corresponding links):

anti-ZIP 14: orb96723 http://www.biorbvt.com/zipl4-antibodv-7

anti-DMTl : orb 5976 http://www.biorbvt.com/dmtl-antibodv-orb5976

or by Abeam with the following catalogue numbers:

anti-ZIP14: abl06568 https://www.abcam.com/slc39al4zip-14-antibody-abl06568.html anti-ZIP14: ab 140973 https://www.abcam.com/slc39al4zip-14-antibody-abl40973.html anti-ZIP14: ab 123988 https://www.abcam.com/slc39al4zip-14-antibody-abl23988.html anti-ZIP 14: ab 133384 https://www.abcam.com/slc39al4zip-14-antibody-abl33384.html anti-ZIP14: ab219174 https://www.abcam.com/slc39al4zip-14-antibody-ab219174.html anti-ZIP14: abl91199 https://www.abcam.com/slc39al4zip-14-antibody-abl91199.html anti-DMTl : ab55735 https://www.abcam.com/dmtl-antibody-ab55735.html

anti-DMT 1 : ab 55812 https ://www. abeam . com/ dmt 1 -antibody-ab 55812.html

anti-DMTl : abl23085 https://www.abcam.com/dmtl-antibody-abl23085.html anti-DMTl : abl40977 https://www.abcam.com/dmtl-antibody-abl40977.html anti-DMTl : abl33402 https://www.abcam.com/dmtl-antibody-abl33402.html

Anti-ZIP14 and anti-DMTl antibodies are also manufactured by Abbexa, Abbiotec, Abgent, Abnova Corporation, Arigo Biolaboratories, Aviva Systems Biology Corporation, Bethyl Laboratories, Covalab, Elabscience, GeneTex, Invitrogen - Thermo Fisher Scientific, LifeSpan BioSciences, MBL International Corporation, Merck Millipore, MyBioSource, Nordic-MUbio, NovoPro, Novus Biologicals, OriGene Technologies, Osenses, PromoCell GmbH, ProSci, Proteintech, RayBiotech, Rockland Immunochemicals, Signalway Antibody and Sigma-Aldrich.

Antibodies with low immunogenicity can be generated by library-based display techniques and humanisation. Antibodies can be humanised or primatised by techniques well-known from the prior art. See, for example, Winter and Harris, Immunol. Today 14:43-46 (1993) and Wright et ah, Crit. Rev. Immunol. 12: 125-168 (1992).

The antibody can be engineered by recombinant DNA techniques to replace domains CHI, CH2 and CH3, the hinge domains and/or the framework region with the corresponding human sequence [as described, for example, in WO 92/02190 and US Patent nos. 5,530,101, 5,585,089, 5,693,761, 5,693,792, 5,714,350 and 5,777,085]

Moreover, the use of immunoglobulin (Ig) cDNA to construct chimeric Ig genes is known from the prior art [Liu et al, Proc. Natl. Acad. Sci. USA 84: 3439-43 (1987) and J. Immunol . 139: 3521-6 (1987)]. The mRNA is isolated from a hybridoma or other cell that produces the antibody and is used to produce cDNA. The desired cDNA can be amplified by polymerase chain reaction using specific primers [U.S. patent nos. 4,683,195 and 4,683,202]

Alternatively, an expression library is created and controlled to isolate the desired sequence which encodes the variable region of the antibody and consequently fuses with the sequences of the human constant region. The gene sequences of the human constant regions can be found in Rabat et al.,“Sequences of proteins of immunological interest”, N.I.H. publication no. 91-3242 (1991). The genes of the human C region are easily obtainable from known clones. The choice of isotype will be guided by the desired effector functions, such as complement fixation or activity in antibody-dependent cellular cytotoxicity. The preferred isotypes are IgGl, IgG2 and IgG4. One of the constant regions of the human light chain, kappa or lambda, can be used. The humanised chimeric antibody is then expressed by conventional methods. Expression vectors include plasmids, retroviruses, YACs, EBV-derived episomes and the like.

Antibody fragments, such as Fv, F(ab’)2 and Fab, can be prepared by cleavage of the intact protein, for example using protease or chemical cleavage. Alternatively, a truncated gene is designed. For example, a chimeric gene that encodes part of fragment F(ab’)2 can comprise DNA sequences encoding for the CHI domain and the hinge region of the H chain, followed by a translational stop codon to produce the truncated molecule.

The consensus sequences of the H and L J regions can be used to design oligonucleotides for use as primers to introduce useful restriction sites into the J region for subsequent linkage of V region segments to human C region segments. C region cDNA can be modified by site-directed mutagenesis to place a restriction site at the analogous position in the human sequence.

Expression vectors include plasmids, retroviruses, YACs, EBV-derived episomes and the like.

Moreover, human antibodies or antibodies of other species can be generated by techniques well-known from the prior art, such as display-based techniques including, but not limited to, phage display, retroviral display, ribosomal display and other techniques, and the resulting molecules can undergo further maturation, such as affinity maturation [Winter and Harris, Immunol. Today 14:43-46 (1993); Hanes and Pliickthun, Proc. Natl. Acad. Sci. USA 94:4937-42 (1997) (ribosomal display); Parmley and Smith, Gene 73 :305- 18 (1988) (phage display); Scott, Trends Biochem. Sci. 17:241-5 (1992); Cwirla et ak, Proc. Natl. Acad. Sci. USA 87:6378-82 (1990); Russel et ak, Nucleic Acids Res. 21 : 1081-5 (1993); Hoogenboom et ak, Immunol Rev. 130:41-68 (1992); Chiswell and McCafferty, Trends Biotechnok 10:80-4 (1992); and U.S. patent No. 5,733,743] If display techniques are used to produce non-human antibodies, said antibodies can be humanised as described above.

The antibodies according to the invention can be generated against the whole protein in one of its isoforms or against the peptide fragments thereof, for example against regions 261-291, 261-340, 1-66 of human protein DMT-1 or, for example, against region 28-58, 50-100 of human protein ZIP-14. The antibodies can be selected, by techniques known to the skilled person, on the basis of their ability to inhibit, wholly or partly, the function of the membrane proteins (transmembrane channels) responsible for cellular internalisation of non-transferrin bound iron, preferably ZIP14 and DMT1.

In oncology, histological grading, or Broder classification, measures the degree of aggression, associated with the degree of cell differentiation, of a tumour.

The most widely used tumour grading system, recognised by the World Health Organisation (WHO), comprises 4 possible grades:

^■ G1 Well differentiated (low-grade): < 25% of cells undifferentiated;

^■ G2 Moderately differentiated (intermediate-grade): < 50% of cells undifferentiated;

^■ G3 Poorly differentiated (high-grade): 50-75% of cells undifferentiated;

^■ G4 Undifferentiated (high-grade), i.e. anaplastic: > 75% of cells undifferentiated.

Grading provides an indication of the level of aggression of the tumour. Histological grading represents the degree of tumour differentiation; for example, a well-differentiated G1 tumour is generally less aggressive than a poorly-differentiated G3. The aggression progressively increases from a well-differentiated G1 form to an undifferentiated G4 form.

Malignant tumour cells can therefore have various levels of differentiation. A poorly differentiated tumour cell is defined as high-grade because it has a higher degree of non-differentiation, for example in terms of shape and size, than the tissue of origin, and therefore possesses a high degree of malignity. When a cell is undifferentiated from the tissue of origin, there is said to be anaplasia.

According to a preferred aspect of the invention, the tumour is a high-grade malignant tumour of grade G3 or G4 or combinations thereof. This type of tumour is considered aggressive.

In particular, the antibodies according to the invention can be used to treat a tumour, in particular a malignant tumour, which can be selected from tumours of the lungs, bronchi or larynx; colon/rectum; breast; pancreas; prostate; liver, gall bladder and bile ducts; ovaries; brain or other parts of the nervous system; bone marrow or blood (leukaemia); bones or joints; soft tissues (including heart and blood vessels); lymph nodes (lymphoma) and spleen; nasopharynx or oral cavity; salivary glands; pleura, pericardium and peritoneum; thyroid or thymus; oesophagus, stomach or small intestine; uterus; placenta; adrenal gland; kidneys, urinary tract or bladder; testicles; skin.

According to a further preferred aspect of the invention, the tumour is selected from multiple myeloma and plasma cell leukaemia, high-grade surface osteosarcoma, Merkel cell carcinoma, choriocarcinoma, anaplastic thyroid carcinoma, medullary carcinoma, high-grade pleomorphic sarcoma, undifferentiated sarcoma, anaplastic large-cell lymphoma, diffuse large-cell lymphoma, acute myeloid leukaemia, acute lymphoblastic leukaemia, dedifferentiated salivary duct carcinoma, poorly differentiated invasive ductal carcinoma, invasive lobular carcinoma, neuroendocrine carcinoma (lung), poorly differentiated squamous cell carcinoma (uterus, bronchi, oral cavity, thymus, oesophagus), high-grade stromal sarcoma, high-grade endometrial carcinoma, poorly cohesive carcinoma (signet-ring cell type), poorly differentiated adenocarcinoma (pancreas, lung, colon/rectum, oesophagus, small intestine), sarcomatoid carcinoma, oncocytic carcinoma, malignant mesothelioma, nodular melanoma, glioblastoma, anaplastic seminoma, neuroblastoma, undifferentiated nasopharyngeal carcinoma, high-grade gastrointestinal stromal tumour, poorly differentiated cholangiocarcinoma, high-grade serous or mucinous cystadenocarcinoma, and poorly differentiated acinar adenocarcinoma.

According to a further preferred aspect of the invention, the high-grade malignant tumour of grade G3 or G4 or combinations thereof is a solid tumour or a haematological neoplasm.

The antibodies according to the invention have proved effective in vitro in the treatment of high-grade malignant tumours of grade G3 or G4 or combinations thereof which, in particular, express, and preferably overexpress, the ZIP membrane protein in any isoform, in particular ZIP 14, even in over 50% of the neoplastic cellularity, and/or the DMT membrane protein in any isoform, in particular DMT1, even in over 50% of the neoplastic cellularity.

Said antibodies bind to the membrane proteins responsible for cellular internalisation of non-transferrin bound iron, in particular DMT1 and/or ZIP14, which has surprisingly been found to be used by high-grade malignant tumour cells of grade G3 or G4 or combinations thereof, to support their accelerated metabolism. In this way, an intracytoplasmic iron deficiency is created in the tumour cells, leading to a metabolic blockage and inhibition of the proliferation of the tumour cells.

Expression and overexpression of ZIP membrane proteins in any isoform, in particular ZIP 14, and DMT in any isoform, in particular DMT1, can be determined by immunohistochemical techniques known from the prior art, in tumours wherein the ZIP membrane protein in any isoform, in particular ZIP14, and/or the DMT membrane protein in any isoform, in particular DMT1, is expressed, and preferably overexpressed, in over 50% of the tumour cells. Treatment with the antibodies according to the present invention induces inhibition of tumour cell proliferation, thus effectively treating the tumour (Roncati L., Diagnostic, Prognostic and Predictive Immunohistochemistry in Malignant Melanoma of the Skin, Klin Onkol 2018; 31(2): 152-155).

The antibodies according to the present invention can also be used as antitumoral agents in combination with agents able to bind to and inhibit/block the transferrin receptor responsible for internalisation of transferrin bound iron, anti-TFRl and/or anti-TRF2, or with other chemotherapy and immunotherapy agents or in combination with radiotherapy.

According to a preferred aspect, the antibodies according to the present invention, namely anti-ZIP, preferably anti-ZIP 14, or anti-DMT, preferably anti-DMTl, or combinations thereof, can be used in combination with agents able to inhibit/block the receptor responsible for internalisation of transferrin bound iron, namely anti-TFRl, in high-grade malignant tumours of grade G3 or G4 or combinations thereof, which overexpress membrane protein TFR1 in over 50% of the neoplastic cellularity.

The invention also relates to a method of treating a tumour in a patient which comprises administration to said patient of an effective dose of an antibody as defined above.

The following examples further illustrate the invention.

EXAMPLES

Example 1 - Evaluation of expression of receptors ZIP14 and DMT1 in tumour tissue sections

Biopsy or surgical samples were selected from 46 patients of both sexes and all ages who died of histologically aggressive tumours within 5 years of diagnosis, despite chemo- or radiotherapy, as summarised in Table 1 below:

Table 1

The samples, after being fixed in formaldehyde and embedded in paraffin wax, were subjected to haematoxylin/eosin (HE) staining, histochemical staining for Fe²⁺ (Turnbull staining) and Fe³⁺ (Peris staining), and immunohistochemical characterisation. After deparaffinisation, hydration, endogenous peroxidase blocking and antigen unmasking, the tissue sections were incubated for 30 minutes at room temperature with anti-TFRl (clone DF1513, Santa Cruz Biotechnology Inc., Dallas, TX, USA), anti-TFR2 (clone 9F8 1C11, Santa Cruz), anti-DMTl (polyclonal, Biorbyt LLC, San Francisco, CA, USA) and anti- ZIP14 (polyclonal, Biorbyt). A secondary biotinylated antibody was applied, and the product of staining was detected with avidin-biotin complex, counterstained with haematoxylin. The reaction was detected with a complex polymer suitable for Roche- Ventana automated Stainers, with DAB (3,3’-diaminobenzidine tetrahydrochloride) as chromogen. The immunohistochemical results for each neoplastic sample were compared with the adjacent normal tissue.

The same validation criteria as already employed to evaluate Pd-Ll (Programmed death-Ligand 1) expression in cancer by means of immunohistochemistry [Garon EB, Rizvi NA, Hui R, et al. Pembrolizumab for the treatment of non-small-cell lung cancer. N Engl J Med. 2015;372:2018-28] were also used, as follows:

• none (-): no staining, or membrane staining of any extent, was observed in a number of tumour cells <1%, namely a Tumour Proportion Score (TPS) <1%; • low (+): membrane staining of any extent in a number of tumour cells ranging from > 1% to <49%, or a TPS ranging from > 1% to <49%;

• high (++): membrane staining of any extent in a number of tumour cells > 50%, or a TPS > 50%.

The results are summarised in Table 2 below:

Table 2

In almost all cases, therefore (45 cases out of 46: 97.8%), increased expression of protein ZIP14, responsible for internalisation of non-transferrin bound iron, was observed in over 50% of the neoplastic cellularity, and non-overexpression thereof was found in no cases (0%).

DMT1 was overexpressed in 43.5% of cases, the percentage of cellularity involved varying case by case. In 24.0% of cases there was increased expression of protein DMT1 in over 50% of the neoplastic cellularity.

In 43.5% of cases there was also increased expression, in over 50% of the neoplastic cellularity, of membrane protein TFR1, which can therefore be considered as a valid molecular target, simultaneously with blockage of the pathway of non-transferrin bound iron.

Example 2 - Test of efficacy of tumour-cell proliferation inhibition

The behaviour of the anti-ZIP 14 and anti-DMTl antibodies was then evaluated on a neoplastic human cell line, in particular on a multiple myeloma human cell line (i.e. a high-grade haematological neoplasm), wherein very intense overexpression of ZIP 14 was found in over 50% of the neoplastic cellularity (increased expression of DMT1, TFR1 and TFR2 below 50% of the neoplastic cellularity).

In detail, 1000 pL of multiple myeloma human cell line RPMI 8226 (Sigma-Aldrich), consisting of about 600,000 cells maintained at 37°C and at a 5% CO2 concentration, in suspension with RPMI 1640, glutamine (2 mM) and foetal bovine serum (10%), was placed in contact for 12 consecutive hours with 20 pL of anti-ZIP 14 antibody (Biorbyt - orb96723), obtaining a reduction of proliferating cells in the proportion of 17: 1 compared with the untreated cell line, detected by EdU red fluorescence (5-ethynyl-2'- deoxyuridine).

Another 1000 pL of multiple myeloma human cell line RPMI 8226, also consisting of about 600,000 cells maintained at 37°C and at a 5% CO2 concentration, in standard suspension with RPMI 1640, glutamine (2 mM) and foetal bovine serum (10%), was placed in contact for 12 consecutive hours with 25 pL of anti-DMTl antibody (Biorbyt - orb5976), obtaining a reduction of proliferating cells in the proportion of 2: 1 compared with the untreated cell line, again detected by EdU red fluorescence.

These results are therefore attributable to metabolic, and consequently proliferative blockage of the neoplastic cells due to lack of uptake of the non-transferrin bound iron present in the serum of the culture medium.

Claims

1. Antibody inhibitor of ZIP membrane proteins in any isoform or DMT membrane proteins in any isoform, responsible for cellular internalisation of non-transferrin bound iron, for use as a medicament.

2. Antibody inhibitor of ZIP membrane proteins in any isoform or DMT membrane proteins in any isoform, responsible for cellular internalisation of non-transferrin bound iron, for use in the treatment of high-grade malignant tumours.

3. Antibody for use according to claim 2, wherein the ZIP and/or DMT membrane protein is expressed in over 50% of the neoplastic cellularity.

4. Antibody for use according to claim 3, wherein the ZIP and/or DMT membrane protein is overexpressed in over 50% of the neoplastic cellularity.

5. Antibody for use according to claims 1-4, wherein the membrane protein is ZIP14.

6. Antibody for use according to claims 1-4, wherein the membrane protein is DMT1.

7. Antibody for use according to claims 1-6, selected from an anti-ZIP14 antibody or an anti-DMTl antibody or combinations thereof.

8. Antibody for use according to claims 2-7, wherein the tumour is grade G3 or G4 or combinations thereof.

9. Antibody for use according to claims 2-8, wherein the tumour is selected from a tumour of the lungs, bronchi or larynx; colon-rectum; breast; pancreas; prostate; liver, gallbladder and biliary tract; ovaries; brain or other districts of the nervous system; bone marrow or blood (leukaemia); bones or joints; soft tissues (including heart and blood vessels); lymph nodes (lymphoma) and spleen; nasopharynx or oral cavity; salivary glands; pleura, pericardium and peritoneum; thyroid or thymus; oesophagus, stomach or small intestine; uterus; placenta; adrenal gland; kidney, urinary tract or bladder; testicles; skin.

10. Antibody for use according to claims 2-9, wherein the tumour is selected from multiple myeloma and plasma cell leukaemia, high-grade surface osteosarcoma, Merkel cell carcinoma, choriocarcinoma, anaplastic thyroid carcinoma, medullary carcinoma, high-grade pleomorphic sarcoma, undifferentiated sarcoma, anaplastic large cell lymphoma, diffuse large cell lymphoma, acute myeloid leukaemia, acute lymphoblastic leukaemia, dedifferentiated salivary duct carcinoma, poorly differentiated invasive ductal carcinoma, invasive lobular carcinoma, neuroendocrine carcinoma (lung), poorly differentiated squamous cell carcinoma (uterus, bronchi, oral cavity, thymus, oesophagus), high-grade stromal sarcoma, high-grade endometrial carcinoma, poorly cohesive carcinoma (signet-ring cell type), poorly differentiated adenocarcinoma (pancreas, lung, colon/rectum, oesophagus, small intestine), sarcomatoid carcinoma, oncocytic carcinoma, malignant mesothelioma, nodular melanoma, glioblastoma, anaplastic seminoma, neuroblastoma, undifferentiated nasopharyngeal carcinoma, high-grade gastrointestinal stromal tumour, poorly differentiated cholangiocarcinoma, high-grade serous or mucinous cystadenocarcinoma, and poorly differentiated acinar adenocarcinoma.