Anti-human B7-H6 humanized monoclonal antibody and application thereof

Abstract

The invention belongs to the technical field of biomedicine, and particularly relates to an anti-human B7-H6 humanized monoclonal antibody and application thereof. The anti-human B7-H6 humanized monoclonal antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises CDR-H1, CDR-H2 and CDR-H3, the sequence of the CDR-H1 is SEQ ID NO: 1, the sequence of the CDR-H2 is SEQ ID NO: 2, and the sequence of the CDR-H3 is SEQ ID NO: 3; the light chain variable region comprises CDR-L1, CDR-L2 and CDR-L3, the sequence of the CDR-L1 is SEQ ID NO: 4, the sequence of the CDR-L2 is SEQ ID NO: 5, and the sequence of the CDR-L3 is SEQ ID NO: 6. The anti-human B7-H6 humanized monoclonal antibody disclosed by the invention has the beneficial effects that the anti-human B7-H6 humanized monoclonal antibody disclosed by the invention can specifically recognize tumor cells, block the combination of B7-H6 and a receptor NKp30, and provide therapeutic antibody candidate molecules for immunotherapy of targeted B7-H6 molecules.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a humanized monoclonal antibody against human B7-H6 and its applications. Background Technology

[0002] B7-H6 (NCR3LG1) was first identified as a specific ligand molecule for NKp30 in the K562 cell line through functional screening in 2009, and systematic progress has been made in its gene localization, molecular characteristics, and functional studies. The B7-H6 gene is located in the 11p15.1 region of human chromosome, and its amino acid sequence is highly homologous to classic immune checkpoint molecules such as PD-L1 and B7-H3. At the protein structure level, the human B7-H6 protein is a type I transmembrane glycoprotein composed of 454 amino acids (51 kDa). It contains two extracellular domains with a total of six N-glycosylation sites: an IgSF domain (25-141, 142-245), a juxtamembrane region (246-261), a transmembrane domain (262-281), and an intracellular domain (282-454). The intracellular domain contains a Gag-MA domain (293-381) and multiple signaling motifs, such as the ITIM, SH2 binding domain, and SH3 binding motif.

[0003] B7-H6 interacts with the NK cell killing receptor NKp30 and is currently the only known membrane ligand for NKp30, possessing the function of activating the killing activity of NK cells. NKp30 is mainly expressed in peripheral blood NK cells, but studies have also indicated that CD8+ T cells, γδ T cells, ILC2 cells, and other cells with killing capabilities also express NKp30. B7-H6 is upregulated in activated monocytes, neutrophils, and stress cells, thus playing an important role in innate immunity, viral immunity, and the clearance of damaged cells. Multiple genome-wide screening studies have collectively shown that B7-H6 is a key regulatory node determining the killing efficacy of NK cells, and its deficiency can enable tumor cells to acquire significant immune evasion capabilities. Summary of the Invention

[0004] This application provides a humanized monoclonal antibody against human B7-H6 and its application. The humanized monoclonal antibody against human B7-H6 can specifically recognize human B7-H6 molecules and block the binding of B7-H6 to the receptor NKp30, thereby providing a new therapeutic antibody candidate molecule for cell immunotherapy.

[0005] The first aspect of this application provides a humanized monoclonal antibody (15G10) against human B7-H6, comprising a heavy chain variable region and a light chain variable region; The heavy chain variable region includes CDR-H1, CDR-H2 and CDR-H3, wherein the sequence of CDR-H1 is SEQ ID NO: 1, the sequence of CDR-H2 is SEQ ID NO: 2 and the sequence of CDR-H3 is SEQ ID NO: 3; The light chain variable region includes CDR-L1, CDR-L2 and CDR-L3, wherein the sequence of CDR-L1 is SEQ ID NO: 4, the sequence of CDR-L2 is SEQ ID NO: 5 and the sequence of CDR-L3 is SEQ ID NO: 6.

[0006] The sequence of CDR-H1 is SFGMH (SEQ ID NO: 1); The sequence of CDR-H2 is YISSGSSTIDYADSVKG (SEQ ID NO: 2); The sequence of CDR-H3 is GGGHWFAY (SEQ ID NO: 3); The sequence of CDR-L1 is RASDHINNWLA (SEQ ID NO: 4); The sequence of CDR-L2 is GATRLES (SEQ ID NO: 5); The sequence of CDR-L3 is QQYWSSPYT (SEQ ID NO: 6).

[0007] The humanized monoclonal antibody against human B7-H6 described in this application can specifically recognize tumor cells and block the binding of B7-H6 to the receptor NKp30, providing a therapeutic antibody candidate molecule for immunotherapy targeting the B7-H6 molecule.

[0008] According to some embodiments of the humanized monoclonal antibody against human B7-H6 described in this application, the amino acid sequence of the heavy chain variable region of the humanized monoclonal antibody is as follows: EVQLVESGGGLVQPGGSLRLSCAASGFTFSSFGMHWVRQAPGKGLEWVSYISSGSSTIDYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARGGGHWFAYWGQGTTVTVSS (SEQ ID NO: 7).

[0009] According to some embodiments of the humanized monoclonal antibody against human B7-H6 described in this application, the amino acid sequence of the light chain variable region of the humanized monoclonal antibody is as follows: DIQMTQSPSTLSASSVGDRVTITCRASDHINNWLAWYQQKPGKAPKLLISGATRLESGVPSRFSGSGSGTEYTLTISSLQPDDFATYYCQQYWSSPYTFGQGTKLEIK (SEQ ID NO: 8).

[0010] The second aspect of this application provides the use of the humanized monoclonal antibody against human B7-H6 described in the first aspect of this application in the preparation of articles for detecting B7-H6 protein.

[0011] The third aspect of this application provides the use of the humanized monoclonal antibody against human B7-H6 described in the first aspect of this application in an article that blocks the B7-H6 protein.

[0012] A fourth aspect of this application provides a pharmaceutical composition comprising the humanized monoclonal antibody against human B7-H6 described in the first aspect of this application.

[0013] According to some embodiments of the pharmaceutical compositions described in this application, the pharmaceutical compositions comprise pharmaceutically acceptable carriers and / or excipients.

[0014] The fifth aspect of this application provides the use of the pharmaceutical composition described in the fourth aspect of this application in the preparation of a medicament for the prevention and / or treatment of tumors.

[0015] According to some embodiments of the application described in this application, the tumor includes one or more of the following: acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, non-Hodgkin's lymphoma, multiple myeloma, melanoma, lung cancer, colorectal cancer, kidney tumor, bladder cancer, gastrointestinal cancer, prostate cancer, liver cancer, ovarian cancer, pancreatic cancer, endometrial cancer, gastric cancer, prostate cancer, kidney cancer, cervical cancer, thyroid cancer, uterine cancer, neuroendocrine cancer, head and neck cancer, nasopharyngeal carcinoma, testicular cancer, basal cell carcinoma, squamous cell carcinoma, dermatofibrosarcoma protrusion, Merkel cell carcinoma, glioblastoma, glioma, sarcoma, mesothelioma, and myelodysplastic syndrome.

[0016] The beneficial effects of this application include: the humanized monoclonal antibody against human B7-H6 described in this application can specifically recognize tumor cells and block the binding of B7-H6 to the receptor NKp30, providing a therapeutic antibody candidate molecule for immunotherapy targeting the B7-H6 molecule. Attached Figure Description

[0017] Figure 1The humanized monoclonal antibody hu15G10 against human B7-H6 described in this application was identified by electrophoresis and Coomassie brilliant blue staining. From left to right, the structures are: Marker, recombinant antibody, chimeric antibody, and humanized antibody, where the heavy chain is H chain and the light chain is L chain.

[0018] Figure 2 This is a diagram showing the results of the humanized monoclonal antibody against human B7-H6 specifically binding to the recombinant B7-H6 protein as described in this application.

[0019] Figure 3 This image shows the results of the humanized monoclonal antibody against human B7-H6 described in this application specifically recognizing B7-H6 protein expression in cells. From left to right, the images show the flow cytometry detection of binding between hu15G10 cells and 293T-B7-H6 knockout cells, 293T-B7-H6 overexpressing cells, and K562 cells; the gray line represents the blank control group, the dashed line represents the secondary antibody control group, and the solid line represents the experimental group.

[0020] Figure 4 The humanized monoclonal antibody against human B7-H6 described in this application competes with the maternal mouse 15G10 monoclonal antibody for binding to the antigen.

[0021] Figure 5 This is a diagram showing the results of the humanized monoclonal antibody against human B7-H6 described in this application blocking the binding of recombinant B7H6 protein and recombinant Nkp30 protein.

[0022] Figure 6 This diagram shows the results of the humanized monoclonal antibody against human B7-H6 described in this application blocking the binding of cell surface B7H6 protein and Nkp30 recombinant protein.

[0023] Figure 7 This image shows the results of the humanized monoclonal antibody against human B7-H6 described in this application blocking the killing effect of NK-92 cells on K562 cells. Detailed Implementation

[0024] The embodiments of the present invention are described in detail below. These embodiments are exemplary and intended to explain the present invention, and should not be construed as limiting the present invention.

[0025] In this invention, the terms "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., refer to a specific feature, structure, material, or characteristic described in connection with that embodiment or example, which is included in at least one embodiment or example of the invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0026] The reagents used in the embodiments of this application, including human antibody B7-H6 (hu15G10), mouse antibody B7-H6 (15G10), PBS, 1640 culture medium, fetal bovine serum, PE-goat anti-human Fc secondary antibody, PE-mouse anti-human Fc secondary antibody, and B7-H6 antigen protein, are all commercially available products. In addition, the reagents, tools, and instruments used in the following specific embodiments, unless otherwise specifically mentioned, are all conventional reagents, tools, and instruments in the art.

[0027] The basic process of this application method is as follows: 1. Protein G affinity chromatography for separation and purification of hu15G10 protein and Lowy method for quantification. The hu15G10 monoclonal antibody was expressed in CHO eukaryotic cells, with the initial cell concentration adjusted to 2 × 10⁻⁶ cells / cells. 6 After incubating for 7 days, the expression supernatant was collected, concentrated using an ultrafiltration system, and purified using a Protein G affinity chromatography column according to the protocol provided by Phamacia. The supernatant was then quantified using the Lowy method, sterilized by antibody filtration, and aliquoted for storage at -20°C.

[0028] 2. Identification of hu15G10 heavy and light chains and preliminary determination of purity by protein electrophoresis. The purified antibody was analyzed by 12% SDS-polyacrylamide gel electrophoresis (SDS-PAGE), stained with Coomassie brilliant blue overnight, destained with destaining solution, and then imaged. A standard protein was used as a molecular weight control to compare molecular weights.

[0029] 3. ELISA identification of humanized B7-H6 antibody hu15G10 recognizing recombinant B7-H6 protein Coat ELISA plates with B7-H6 protein at a concentration of 1 μg / ml, add 100 μl to each well, and incubate overnight at 4°C. After blocking for 2 hours, the hu15G10 antibody was prepared into 15 serially diluted concentrations of 0.2ug / ml, 0.1ug / ml, 0.05ug / ml, 0.025ug / ml, 0.0125ug / ml, 0.00625ug / ml, 0.003125ug / ml, 0.0015625ug / ml, 0.00078125ug / ml, 0.00039625ug / ml, 0.0001953ug / ml, 0.00009766ug / ml, 0.00004833ug / ml, 0.00002411ug / ml, and 0.00001220ug / ml. The antibody was incubated for 1 hour, followed by incubation with secondary antibody for 1 hour. After color development with the chromogenic solution, stop solution was added, and the absorbance was measured at 450 nm using a microplate reader.

[0030] The regression analysis results showed that the EC50 of humanized 15G10 binding to B7-H6 protein was 0.008566 ug / ml.

[0031] 4. Flow cytometry identification of anti-human B7-H6 monoclonal antibodies 293T-B7-H6 knockout and overexpression cell lines and K562 cell line were selected as detection cells. Hu15G10 purified antibody was collected as primary antibody, and PE-labeled goat anti-human Fc fragment antibody was used as secondary antibody. Flow cytometry was used to detect the cells.

[0032] 5. Identification of the hu15G10 antigen-binding epitope Coat ELISA plates with B7-H6 protein at a concentration of 1 μg / ml, add 100 μl to each well, and incubate overnight at 4°C. After blocking for 2 hours, the biotinylated mouse antibody concentration was diluted to 2 μg / ml. The hu15G10 antibody was prepared into 15 gradient dilutions: 20 μg / ml, 10 μg / ml, 5 μg / ml, 2.5 μg / ml, 1.25 μg / ml, 0.625 μg / ml, 0.3125 μg / ml, 0.1563 μg / ml, 0.07813 μg / ml, 0.03906 μg / ml, 0.01953 μg / ml, 0.009766 μg / ml, 0.004883 μg / ml, 0.002411 μg / ml, and 0.001221 μg / ml. The two were mixed in a 1:1 ratio and added to the wells. After incubation for 1 hour, the mixture was incubated with streptavidin-HRP for 1 hour. After color development, stop solution was added, and absorbance was measured at 450 nm using a microplate reader.

[0033] The results showed that the IC50 was 0.5627 ug / ml.

[0034] 6. ELISA detection of hu15G10 blocking the binding of recombinant B7H6 and Nkp30 proteins. Coat ELISA plates with B7-H6 protein at a concentration of 1 μg / ml, add 100 μl to each well, and incubate overnight at 4°C. After blocking for 2 hours, biotinylated Nkp30-hFc protein was prepared to a concentration of 20 ug / ml, and hu15G10 antibody was prepared to a gradient of 14 concentrations: 20 ug / ml, 10 ug / ml, 5 ug / ml, 2.5 ug / ml, 1.25 ug / ml, 0.625 ug / ml, 0.3125 ug / ml, 0.1563 ug / ml, 0.07813 ug / ml, 0.03906 ug / ml, 0.01953 ug / ml, 0.009766 ug / ml, 0.004883 ug / ml, and 0.002411 ug / ml. The two were mixed at a 1:1 ratio and added to the wells. The mixture was incubated overnight at 4°C, followed by incubation with streptavidin-HRP for 1 hour. After color development, stop solution was added, and absorbance was measured at 450 nm using a microplate reader.

[0035] The results showed that the IC50 was 0.05104 ug / ml.

[0036] 7. Flow cytometry analysis showed that hu15G10 blocked the binding of cell surface B7H6 protein and Nkp30 recombinant protein.

[0037] 293T-B7-H6 overexpressing cells were added to flow cytometry tubes, 10W per tube, along with 20ug / ml of biotinylated Nkp30-hFc protein and different concentrations (20ug / ml, 6.8ug / ml, 2ug / ml, 0.68ug / ml, 0.2ug / ml) of hu15G10. The cells were incubated for 2 h, stained with streptavidin-PE, and analyzed by flow cytometry.

[0038] 8. Hu15G10 can block the killing effect of NK-92 cells on K562 cells.

[0039] After staining target cells K562 cells with calcein, 1W of cells were seeded into each well of a 96-well plate. Effector cells NK-92 were added at an effector-to-target ratio of 10:1. Different concentrations of hu15G10, m15G10, IgG, or PBS were added, and the cells were co-cultured for 7 hours. Fluorescence values ​​were detected, and the killing effect was calculated.

[0040] The specific embodiments of the present invention will be described in further detail below with reference to the accompanying drawings and examples.

[0041] Example 1 The anti-B7-H6 monoclonal antibody hu15G10 recognizes the B7-H6 molecule. The purified hu15G10 SDS-PAGE electrophoresis showed the position and size of the heavy and light chains of the human B7-H6 antibody as follows: Figure 1 As shown.

[0042] Sequencing confirmed that the CDR region of the human B7-H6 antibody heavy chain contains CDR-H1, CDR-H2, and CDR-H3, and their sequence information is as follows: CDR-H1: SFGMH (SEQ ID NO: 1); CDR-H2: YISSGSSTIDYADSVKG (SEQ ID NO: 2); CDR-H3: GGGHWFAY (SEQ ID NO: 3); The amino acid sequence of the heavy chain variable region (VH) of the humanized monoclonal antibody hu15G10 is as follows: EVQLVESGGGLVQPGGSLRLSCAASGFTFSSFGMHWVRQAPGKGLEWVSYISSGSSTIDYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARGGGHWFAYWGQGTTVTVSS (SEQ ID NO: 7).

[0043] The light chain CDR region of the human B7-H6 antibody contains CDR-L1, CDR-L2, and CDR-L3, and their sequence information is as follows: CDR-L1: RASDHINNWLA (SEQ ID NO: 4); CDR-L2: GATRLES (SEQ ID NO: 5); CDR-L3: QQYWSSPYT (SEQ ID NO: 6); The humanized monoclonal antibody hu15G10 has the following amino acid sequence in its light chain variable region (VL): DIQMTQSPSTLSASSVGDRVTITCRASDHINNWLAWYQQKPGKAPKLLISGATRLESGVPSRFSGSGSGTEYTLTISSLQPDDFATYYCQQYWSSPYTFGQGTKLEIK (SEQ ID NO: 8).

[0044] B7-H6 protein was coated onto ELISA plates at a concentration of 1 μg / ml. Hu15G10 antibody at concentrations of 0.2, 0.1, 0.05, 0.025, 0.0125, 0.00625, 0.003125, 0.0015625, 0.00078125, 0.00039625, 0.0001953, 0.00009766, 0.00004833, 0.00002411, and 0.00001220 μg / ml was added and reacted for 1 h. Secondary antibody was incubated for 1 h. After color development, stop solution was added, and absorbance was measured at 450 nm using a microplate reader. The results are as follows: Figure 2 As shown.

[0045] from Figure 2 As can be seen from the regression fitting analysis, the results show that humanized 15G10 can bind to B7-H6 protein, and the EC50 of binding is 0.008566ug / ml.

[0046] 293T-B7-H6 knockout and overexpressing cells and K562 cells in logarithmic growth phase were incubated with hu15G10 and corresponding isotype control IgG at 4℃ for 40 min. After washing, the corresponding secondary antibodies were added and incubated at 4℃ for 20 min. After washing twice with PBS buffer, the cells were resuspended and the expression of B7-H6 on the cell surface was detected by flow cytometry. The results are as follows: Figure 3 As shown.

[0047] from Figure 3 As can be seen from this study, hu15G10 can recognize and bind to B7-H6 molecules on the cell surface.

[0048] Example 2 Humanized B7-H6 monoclonal antibody competes with maternal mouse antibody for recognition of antigenic epitopes. To identify the differences in antigen recognition sites between human B7-H6 monoclonal antibody hu15G10 and maternal mouse B7-H6 monoclonal antibody m15G10, B7-H6 protein was coated onto ELISA plates at a concentration of 1 μg / ml. Hu15G10 antibody at concentrations of 20, 10, 5, 2.5, 1.25, 0.625, 0.3125, 0.1563, 0.07813, 0.03906, 0.01953, 0.009766, 0.004883, 0.002411, and 0.001221 μg / ml was added and incubated for 1 h. Biotin-labeled mouse 15G10 was then added, and the reaction proceeded for another 1 h. Secondary antibody was then added, and the absorbance was measured at 450 nm using a microplate reader. The results are as follows: Figure 4 As shown.

[0049] from Figure 4As can be seen, human hu15G10 can block the binding of the B7-H6 monoclonal antibody m15G10 to B7-H6, indicating that human hu15G10 and the mouse parent antibody recognize the same site of the B7-H6 molecule, and the IC50 is 0.5627 ug / ml.

[0050] Example 3 Humanized B7-H6 monoclonal antibody blocks the binding of B7-H6 protein to its receptor molecule NKP30. B7-H6 protein was coated onto an ELISA plate at a concentration of 1 μg / ml. Hu15G10 antibody was added at concentrations of 20, 10, 5, 2.5, 1.25, 0.625, 0.3125, 0.1563, 0.07813, 0.03906, 0.01953, 0.009766, 0.004883, and 0.002411 μg / ml, respectively. After reaction, biotinylated Nkp30-hFc protein was added, and the plate was incubated with streptavidin-HRP for 1 h. The absorbance was measured at 450 nm using a microplate reader. The results are as follows: Figure 5 As shown.

[0051] from Figure 5 It can be seen that the humanized B7-H6 monoclonal antibody blocks the binding of B7-H6 protein to its receptor molecule NKP30 protein, and the IC50 value is 0.05104ug / ml.

[0052] In addition, 293T-B7-H6 overexpressing cells were added to flow cytometry tubes, 10 μg / ml per tube, along with 20 μg / ml biotinylated Nkp30-hFc protein and different concentrations (20 μg / ml, 6.8 μg / ml, 2 μg / ml, 0.68 μg / ml, 0.2 μg / ml) of hu15G10. The cells were incubated for 2 h, stained with streptavidin-PE, and analyzed using flow cytometry. The results are shown below. Figure 6 As shown.

[0053] from Figure 6 It can be seen that the humanized B7-H6 monoclonal antibody can block the binding of NKP30 protein to B7-H6 on the cell membrane surface.

[0054] Example 4 Hu15G10 can block the killing effect of NK-92 cells on K562 cells. K562 target cells were stained with calcein and seeded at 1000g per well in 96-well plates. NK-92 effector cells were added at a target-effect ratio of 10:1. Different concentrations of hu15G10, m15G10, IgG, or PBS were added, and the plates were co-cultured for 7 hours. Fluorescence values ​​were detected, and the killing effect was calculated. Results are shown below. Figure 7 As shown from Figure 7It can be seen that both the Hu15G10 antibody and its murine maternal antibody can significantly inhibit the killing effect of NK cells on tumor cells.

[0055] Although the above embodiments have been shown and described, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Any changes, modifications, substitutions and variations made to the above embodiments by those skilled in the art are within the protection scope of the present invention.

Claims

1. An anti-human B7-H6 humanized monoclonal antibody, characterized in that, comprises a heavy chain variable region and a light chain variable region; the heavy chain variable region comprises CDR-H1, CDR-H2 and CDR-H3, the sequence of CDR-H1 is SEQ ID NO: 1, the sequence of CDR-H2 is SEQ ID NO: 2, and the sequence of CDR-H3 is SEQ ID NO: 3; the light chain variable region comprises CDR-L1, CDR-L2 and CDR-L3, the sequence of CDR-L1 is SEQ ID NO: 4, the sequence of CDR-L2 is SEQ ID NO: 5, and the sequence of CDR-L3 is SEQ ID NO:

6.

2. The humanized monoclonal antibody against human B7-H6 according to claim 1, characterized in that, The amino acid sequence of the heavy chain variable region of the humanized monoclonal antibody is SEQ ID NO:

7.

3. The humanized monoclonal antibody against human B7-H6 according to claim 1, characterized in that, The amino acid sequence of the light chain variable region of the humanized monoclonal antibody is SEQ ID NO:

8.

4. Use of the humanized monoclonal antibody against human B7-H6 according to any one of claims 1-3 in the preparation of a preparation for detecting B7-H6 protein.

5. Use of the humanized monoclonal antibody against human B7-H6 according to any one of claims 1-3 in the preparation of a preparation for blocking B7-H6 protein.

6. A pharmaceutical composition, characterized by, The pharmaceutical composition comprises the humanized monoclonal antibody against human B7-H6 according to any one of claims 1-3.

7. The pharmaceutical composition of claim 6, wherein, The pharmaceutical composition comprises a pharmaceutically acceptable carrier and / or excipient.

8. Use of the pharmaceutical composition according to claim 6 or 7 in the preparation of a drug for preventing and / or treating tumors.

9. Use according to claim 8, characterized in that, The tumors include one or more of acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, non-Hodgkin's lymphoma, multiple myeloma, melanoma, lung cancer, colorectal cancer, kidney tumor, bladder cancer, gastrointestinal cancer, prostate cancer, liver cancer, ovarian cancer, pancreatic cancer, endometrial cancer, stomach cancer, prostate cancer, kidney cancer, cervical cancer, thyroid cancer, uterine cancer, neuroendocrine cancer, head and neck cancer, nasopharyngeal cancer, testicular cancer, basal cell skin cancer, squamous cell skin cancer, dermatofibrosarcoma protuberans, Merkel cell carcinoma, glioblastoma, glioma, sarcoma, mesothelioma, and myelodysplastic syndrome.

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