Integrin alpha 10 and invasive cancer forms

Abstract

The present invention relates to a composition for the treatment of aggressive forms of cancer, including aggressive breast cancer, aggressive lung cancer, aggressive prostate cancer and aggressive pancreatic cancer as well as metastatic cancer, comprising an antibody or fragment thereof that specifically binds to integrin alpha 10 polypeptide. The present invention also relates to methods for the diagnosis and treatment of said aggressive forms of cancer. The present invention also relates to methods for predicting survival of individuals affected by said aggressive forms of cancer.

Description

Technical Field

[0001] This invention relates to compositions for treating aggressive cancers, including aggressive breast cancer, aggressive lung cancer, aggressive prostate cancer, and aggressive pancreatic cancer, as well as metastatic cancers, wherein the compositions comprise an antibody or a fragment thereof that specifically binds to an integrin α10 polypeptide. The invention also relates to methods for diagnosing and treating said aggressive cancer forms. Furthermore, the invention relates to methods for predicting the survival rate of individuals affected by said aggressive cancer forms. Background Technology

[0002] Cancer is a complex disease that progresses within a dynamically evolving extracellular matrix (ECM), which controls virtually every aspect of tumors and tumor-associated cells. The major cell adhesion receptors (integrins) of the ECM are a family of 24 transmembrane heterodimers, generated by the combination of 18 α-integrin and 8 β-integrin subunits. Different tissue types typically express a unique set of integrins on their cell surfaces. Alterations in integrin expression patterns have been linked to many types of cancer (Moschos et al., 2007). Alterations in integrin signaling involve almost every step of carcinogenesis, ranging from the shift to αβ heterodimer utilization to aberrant integrin expression, as well as constitutive activation of downstream effectors in integrin signaling and interactions with other signaling pathways.

[0003] Invasive forms of cancer are tumors that proliferate rapidly and / or quickly migrate to distant sites and other tissues; therefore, they may be invasive tumors. Furthermore, invasive forms of cancer are often associated with poorer survival prognosis.

[0004] Based on histological classification, the most common breast cancer subtype is invasive ductal carcinoma, accounting for 80% of invasive breast cancers; followed by invasive lobular carcinoma, accounting for approximately 10% (Xiaofeng et al. 2015). The expression (or lack thereof) of specific protein markers is associated with the invasiveness of breast cancer, particularly hormone receptor status [expression of estrogen receptor (ER) and progesterone receptor (PR)] and human epidermal growth factor receptor 2 (HER2) (Arpino et al. 2015; Hariri et al. 2019). Compared to other breast cancer subtypes, triple-negative breast cancer (TNBC, ER- / PR- / HER2-) is more likely to exhibit invasive behavior and is associated with poor prognosis (Rakha et al. 2007; Malorni et al. 2012). Once metastasis occurs, TNBC highly predisposes to involve key visceral organs (such as the lungs, liver, and brain), ultimately resulting in a significantly shorter median overall survival compared to other subtypes (Bianchini et al. 2016). Six subtypes of TNBC have been identified: basal-like subtypes (BL1 and BL2), immunomodulatory subtype (IM), mesenchymal subtype (M), mesenchymal stem cell-like subtype (MSL), and luminal androgen receptor subtype (LAR) (Ma et al. 2018). Therefore, developing optimal treatment strategies for early-stage TNBC is crucial to mitigating the burden of TNBC. There is overlap between histological and molecular classifications. For example, invasive ductal carcinoma can have different molecular expression profiles and thus be triple-negative, HER2-positive, luminal A, or luminal B. Triple-negative basal-like tumors are further characterized by more than 50% TP53 mutations, high proliferation due to RB1 deletion, association with BRCA-1 mutations, high aneuploidy, and unique expression of any one of cytokeratins 5, 6, or 17. Besides TNBC, other aggressive forms of breast cancer exist. Inflammatory breast cancer (IBC) is a rare and aggressive form of breast cancer. IBC tends to grow and spread rapidly, with symptoms worsening over days or even hours. Therefore, there is a need to develop novel targeted therapies for aggressive breast cancer types, and the development of such novel targeted therapies is crucial for improving related survival outcomes.

[0005] Prostate cancer is the second most common cancer in men and the fourth most common cancer worldwide. The five-year survival rate for patients diagnosed with localized or regional cancer is close to 100%. However, the five-year survival rate for patients diagnosed with distant metastases is only 28%. Healthy prostate epithelium contains luminal epithelial cells, basal cells, and a small subset of neuroendocrine (NE) cells scattered throughout the prostate. Most prostate cancers are classified as adenocarcinomas characterized by a lack of basal cells and uncontrolled proliferation of malignant tumor cells, exhibiting features of luminal differentiation, including gland formation and expression of androgen receptor (AR) and prostate-specific antigen (PSA). Interestingly, each case of prostate adenocarcinoma also contains a small number (typically about 1%) of NE tumor cells. NE cells in adenocarcinomas share many important features with NE cells in benign prostates. For example, NE cells in both benign prostates and adenocarcinomas do not express AR and PSA compared to non-NE luminal tumor cells (Einstein et al. 2019). A minority of prostate epithelial malignancies are variant forms, including ductal adenocarcinoma, mucinous (colloid) carcinoma, signet ring cell carcinoma, and small cell (neuroendocrine) carcinoma (SCNC). Similar to NE cells in benign prostate and prostate adenocarcinoma, tumor cells in SCNC lack the expression of AR and PSA, explaining the clinical observation that, unlike adenocarcinoma, these tumors do not respond to hormone therapy that stops androgen production and inhibits AR function. Compared to most prostate cancers, SCNC is highly aggressive, often presenting as locally advanced disease or distant metastasis, and patients typically die within months of diagnosis (Saad et al. 2019).

[0006] Lung cancer is a leading cause of cancer death in both men and women worldwide (GLOBOCAN 2018). The 5-year survival rate for patients with localized lung tumors is 56%. However, the 5-year survival rate for patients diagnosed at an advanced stage and then developing distant tumors is only 5% (SEER Cancer Statistics Review, 2015). Histologically, lung cancer is classified into two main types: small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC). The majority of tumors are classified as NSCLC (approximately 85%), compared to 15% as SCLC. NSCLC comprises a heterogeneous group of tumors and is further subdivided into three main histological subtypes: adenocarcinoma, squamous cell carcinoma, and large cell carcinoma. Adenocarcinoma and squamous cell carcinoma are the predominant subtypes of NSCLC. Compared to SCLC, NSCLC is relatively insensitive to chemotherapy and radiation therapy. In recent years, more specific therapies targeting molecular subgroups have shown promising results. However, only a small percentage of patients identified by biomarkers are associated with treatment response (Bombardelli et al. 2016). Therefore, there is a need for novel biomarkers that allow for the early detection of NSCLC, SCLC, and other lung tumors, as well as novel therapeutic targets.

[0007] Pancreatic cancer can be classified into two main groups: exocrine tumors and neuroendocrine tumors. Exocrine tumors, accounting for 94% of all pancreatic cancers, originate from exocrine cells that produce digestive enzymes. This group includes ductal adenocarcinoma (the most common type of pancreatic cancer, accounting for 90% of all pancreatic cancers), acinar cell carcinoma, intraductal papillary mucinous tumors, and others. Neuroendocrine tumors (also known as islet cell tumors) account for approximately 6% of pancreatic cancers. Generally, pancreatic cancer develops in a relatively asymptomatic manner and is usually diagnosed at an advanced stage. Due to late diagnosis and poor response to chemotherapy and radiation therapy, the 5-year survival rate for pancreatic cancer patients is less than 5% (Milena et al., 2016). Therefore, there is a great need to develop new methods for the diagnosis and treatment of this aggressive disease.

[0008] Sarcomas are a broad family of cancers derived from mesenchymal cells that can differentiate in several tissue lineages: fat, muscle, fibrous, cartilage, and bone. Sarcomas account for 1% of all cancer diagnoses and cancer-related deaths. Prevalence is higher in childhood and adolescence, and in these groups, sarcomas account for 19%–21% of cancer-related deaths. This group of malignancies is particularly difficult to diagnose due to its enormous histological and molecular heterogeneity. These tumors are very rare compared to carcinomas and often grow locally with invasion of adjacent tissues. Long-term survival after surgical resection is good for localized and early-stage tumors, but there is a high risk of recurrence even after 10 to 15 years. Current treatment strategies for soft tissue sarcomas involve surgery, radiation, and chemotherapy, but these are limited by toxicity and mild reactions. The current 5-year survival rate of up to 60% reflects age, tumor type, stage, and histological grade, but in high-risk patients with metastatic tumors, survival rates drop significantly to 10%–17%. The classification of sarcoma cancer forms can be found in the WHO classification of soft tissue and bone tumors (2013).

[0009] Therefore, there is a need for new tools that can allow for the early diagnosis of invasive tumors (such as those discussed above), as well as novel therapeutic targets and therapies that can improve their prognosis and survival. Summary of the Invention

[0010] The inventors of this invention have unexpectedly discovered that several of the most aggressive forms of cancer are characterized by high expression of integrin α10 on their cell surface. Therefore, the inventors have found that it is possible to rapidly detect the presence of an aggressive form of cancer in an individual based on the expression level of integrin α10 in cells of suspected cancerous tissue. Consequently, more specific diagnoses at earlier stages are possible compared to current diagnostic tools, leading to earlier intervention for the individuals being examined and improved prognosis.

[0011] Furthermore, the inventors of this invention have identified integrin α10 as a novel therapeutic target for both invasive and metastatic forms of cancer. In fact, they have discovered that it is possible to block the proliferation and migration of tumor cells belonging to invasive cancer forms by contacting them with an antibody or fragment thereof that specifically binds to the integrin α10 polypeptide. It is also possible to induce cell death by contacting tumor cells with a suitable antibody-drug conjugate containing an antibody or fragment thereof that specifically binds to the integrin α10 polypeptide. Moreover, the inventors have discovered that it is possible to reduce tumor growth by administering an antibody or fragment thereof that specifically binds to the integrin α10 polypeptide and thereby contacting the tumor cells with the antibody or fragment thereof.

[0012] Therefore, one aspect of this disclosure relates to an antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof specifically binds to an integrin α10 polypeptide for the treatment and / or prevention of an aggressive cancer form selected from aggressive breast cancer, aggressive lung cancer, aggressive prostate cancer, and aggressive pancreatic cancer, or metastases of any of said cancer forms.

[0013] Another aspect of this disclosure relates to an antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof specifically binds to an integrin α10 polypeptide for the diagnosis of invasive cancer, said invasive cancer being selected from invasive breast cancer, invasive lung cancer, invasive prostate cancer, and invasive pancreatic cancer, or metastatic tumors of any of said invasive cancer forms.

[0014] A further aspect of this disclosure relates to a method for treating an aggressive cancer, wherein the aggressive cancer is selected from aggressive breast cancer, aggressive lung cancer, aggressive prostate cancer, and aggressive pancreatic cancer, or wherein the aggressive cancer is a metastatic tumor, the method comprising administering to a subject in need a pharmaceutically effective amount of an antibody or an antigen-binding fragment thereof, the antibody or the antigen-binding fragment thereof specifically binding to an integrin α10 polypeptide.

[0015] One and even a further aspect of this disclosure relates to a method for detecting aggressive cancer cells in a subject, the method comprising the following steps:

[0016] a. Provide the subject with tissue suspected of containing cancer cells;

[0017] b. Analyze the presence of antigens in the tissue containing integrin α10 polypeptide or fragments thereof.

[0018] c. Determine the expression level of the integrin α10 antigen, and

[0019] d. The expression levels determined in c. will be compared with control levels.

[0020] The control level is the average expression level of the antigen observed in healthy and / or benign cells of the same tissue type as the isolated sample, wherein an expression level of the antigen higher than the control level indicates the presence of an aggressive form of cancer in the subject, and wherein the aggressive form of cancer is selected from aggressive breast cancer, aggressive lung cancer, aggressive prostate cancer, and aggressive pancreatic cancer, or metastases of any of the aggressive forms of cancer.

[0021] A further aspect of this disclosure relates to a method for detecting aggressive cancer cells in a subject, the method comprising the following steps:

[0022] a. Provide the subject with tissue suspected of containing cancer cells;

[0023] b. Optionally, analyze the presence of one or more cells with cancerous morphologies in the tissue.

[0024] c. Analyze the presence of antigens in the tissue containing integrin α10 polypeptide or fragments thereof.

[0025] d. Optionally determine the expression level of the integrin α10 antigen.

[0026] The presence of one or more cells with a cancerous morphology, combined with the expression of integrin α10 antigen, indicates the presence of an invasive cancer form in the subject, wherein the invasive cancer form is selected from invasive breast cancer, invasive lung cancer, invasive prostate cancer, and invasive pancreatic cancer, or a metastatic tumor of any of the invasive cancer forms.

[0027] Another aspect of this disclosure relates to a method for diagnosing an aggressive form of cancer in a subject, the method comprising the following steps:

[0028] a. Provide the subject with tissue suspected of containing cancer cells;

[0029] b. Analyze the presence of antigens in the tissue containing integrin α10 polypeptide or fragments thereof.

[0030] c. Determine the expression level of the integrin α10 antigen, and

[0031] d. The expression level determined in c. will be compared with a control level, wherein the control level is the average expression level of the antigen; and

[0032] The expression level of the antigen being higher than the control level indicates the presence of an invasive form of cancer in the sample, wherein the invasive form of cancer is selected from invasive breast cancer, invasive lung cancer, invasive prostate cancer, and invasive pancreatic cancer, or a metastatic tumor of any of the invasive forms of cancer, thereby diagnosing the subject's invasive form of cancer.

[0033] A further aspect of this disclosure relates to a method for diagnosing an aggressive form of cancer in a subject, the method comprising the steps of:

[0034] a. Provide the subject with tissue suspected of containing cancer cells;

[0035] b. Optionally, analyze the presence of one or more cells with cancerous morphology in the tissue;

[0036] c. Analyze the presence of antigens in the sample containing integrin α10 polypeptide or fragments thereof.

[0037] d. Optionally determine the expression level of the integrin α10 antigen, and

[0038] The presence of one or more cells with cancerous morphology, combined with the expression of integrin α10 antigen, indicates the presence of an invasive cancer form in the sample, wherein the invasive cancer form is selected from invasive breast cancer, invasive lung cancer, invasive prostate cancer, and invasive pancreatic cancer, or a metastatic tumor of any of the invasive cancer forms, thereby diagnosing the subject's invasive cancer form.

[0039] A further aspect of this disclosure relates to a method for classifying triple-negative breast cancer tumor samples from a subject, the method comprising:

[0040] a. Provide the subject with breast tissue suspected of containing cancer cells;

[0041] b. Isolate breast cancer cells characterized as ER-negative, PR-negative, and HER2-negative;

[0042] c. Determine the expression level of the antigen containing integrin α10 polypeptide or fragments thereof in the isolated cells, and

[0043] d. The expression level determined in c. will be compared with a control level, wherein the control level is the average expression level of the antigen observed in healthy and / or benign breast tissue;

[0044] The expression level of the antigen in the breast cancer cells is higher than that of the control, and the expression status of ER-negative, PR-negative, and HER2-negative indicates basal-like triple-negative breast cancer or luminal triple-negative breast cancer, thereby classifying the triple-negative breast cancer tumor sample as either basal-like triple-negative breast cancer or luminal triple-negative breast cancer.

[0045] In one embodiment, the luminal triple-negative breast cancer is luminal androgen receptor triple-negative breast cancer.

[0046] A further aspect of this disclosure relates to a method for determining the prognosis of a subject's invasive cancer form, the method comprising:

[0047] a. Provide the subject with cancerous tumor tissue;

[0048] b. Analyze the presence of antigens in the sample containing integrin α10 polypeptide or fragments thereof.

[0049] c. Determine the expression level of the integrin α10 antigen.

[0050] d. The expression level determined in c. will be compared with a control level, wherein the control level is the average expression level of the antigen observed in healthy and / or benign tissues of the same tissue type as the sample;

[0051] e. When the expression level of the integrin α10 antigen is higher than the control level, a poor prognosis is determined for the aggressive cancer form.

[0052] The invasive cancer form is selected from invasive breast cancer, invasive lung cancer, invasive prostate cancer, and invasive pancreatic cancer, or a metastatic tumor of any of the invasive cancer forms.

[0053] Another aspect of this disclosure relates to a method for determining the prognosis of a subject's aggressive cancer form, the method comprising:

[0054] a. Provide the subject with cancerous tumor tissue;

[0055] b. Optionally, analyze the presence of one or more cells with cancerous morphology in the tissue;

[0056] c. Analyze the presence of antigens in the sample containing integrin α10 polypeptide or fragments thereof.

[0057] d. Optionally determine the expression level of the integrin α10 antigen and compare the determined expression level with a control level, wherein the control level is the average expression level of the antigen observed in healthy and / or benign tissues of the same tissue type as the sample;

[0058] e. When a combination of one or more cells with a cancerous morphology and the expression of the integrin α10 antigen is present in the tissue, and / or

[0059] When the expression level of the integrin α10 antigen is higher than the control level, a poor prognosis is determined for the aggressive cancer form.

[0060] The invasive cancer form is selected from invasive breast cancer, invasive lung cancer, invasive prostate cancer, and invasive pancreatic cancer, or a metastatic tumor of any of the invasive cancer forms.

[0061] A further aspect of this disclosure relates to a method for preventing the metastasis of a primary invasive cancer selected from invasive breast cancer, invasive lung cancer, invasive prostate cancer, and invasive pancreatic cancer, the method comprising administering to a patient in need a therapeutically effective amount of an antibody or an antigen-binding fragment thereof, wherein the antibody is specific for integrin α10 polypeptide.

[0062] A further aspect of this disclosure relates to a method for inhibiting integrin α10-mediated signaling in at least one cancer cell, the method comprising contacting the at least one cancer cell with an effective amount of an antibody or antigen-binding fragment thereof that is specific to an integrin α10 polypeptide, wherein the at least one cancer cell is selected from invasive breast cancer cells, invasive lung cancer cells, invasive prostate cancer cells, invasive pancreatic cancer cells, and metastatic tumor cells.

[0063] One or even further aspect of this disclosure relates to a method for inhibiting the cellular function of at least one cancer cell, the method comprising contacting the at least one cancer cell with an effective amount of an antibody or antigen-binding fragment specifically targeting the integrin α10 polypeptide, wherein the at least one cancer cell is selected from invasive breast cancer cells, invasive lung cancer cells, invasive prostate cancer cells, invasive pancreatic cancer cells, and metastatic tumor cells.

[0064] One or even further aspect of this disclosure relates to the use of an antibody or an antigen-binding fragment thereof in the manufacture of an agent for the treatment and / or prevention of invasive cancers selected from invasive breast cancer, invasive lung cancer, invasive prostate cancer, and invasive pancreatic cancer, or metastatic tumors of any of these cancer forms, wherein the antibody is specific for integrin α10 polypeptide. Attached Figure Description

[0065] Figure 1 Integrin α10 is expressed by invasive ductal carcinoma (IDC) cells in breast cancer tissue. Immunohistochemical analysis using an antibody against integrin α10 shows that in breast cancer tissue, integrin α10 is specifically and strongly expressed on IDC cells (arrows) and a few cells in the stroma (asterisks) (A). Integrin α10 expression is weaker in morphologically unaffected benign breast tissue (B).

[0066] Figure 2 Integrin α10 is expressed in triple-negative breast cancer tumor tissue. Immunohistochemical analysis using an antibody against integrin α10 shows that integrin α10 is specifically and strongly expressed on triple-negative breast cancer cells (arrows).

[0067] Figure 3 Integrin α10 is expressed in cells of undifferentiated pleomorphic sarcoma tissue. Immunohistochemical analysis using an antibody against integrin α10 shows that integrin α10 is specifically and strongly expressed on undifferentiated pleomorphic sarcoma cells (arrows) in undifferentiated pleomorphic sarcoma tissue.

[0068] Figure 4Integrin α10 is expressed by aggressive cancer cell lines. Immunofluorescence staining of integrin α10 in T47D and BT549 cells (invasive breast cancer, monolayer culture) and in PC-3 cells (invasive prostate cancer, spheroid culture). Confocal images of T47D cells (A), BT549 cells (B), and PC-3 cells (C) show staining of integrin α10 on the cell membrane.

[0069] Figure 5Integrin α10 is expressed in invasive cancer cell lines in monolayers, particularly in spheroid cultures. AD: Analysis of integrin α10 expression in breast cancer cells in monolayers (A and B) and spheroid structures (mammary globules) (C and D). (A) As determined by flow cytometry scatter plots, the number of integrin α10-positive cells in the invasive breast cancer cell lines BT549 and T47D was higher than that in the less invasive cancer cell lines 184A1, HCC1428, and MDA-MB-231, which showed a low number of integrin α10-positive cells. (B) Summary of the percentage of integrin α10-positive cells from each cell line in n = 5 independent experiments. (C) Flow cytometry scatter plots showed a high increase in integrin α10 expression in the triple-negative cell lines MDA-MB-231 and BT549 grown in mammary globules compared to monolayer cultures, while 184A1, HCC1428, and T47D showed only a slight increase or no increase in integrin α10 expression compared to monolayer culture conditions. (D) Summary of the percentage of integrin α10-positive cells from each cell line in n = 5 independent experiments. EH: Analysis of integrin α10 expression in prostate cancer cells in monolayers (E and F) and globular structures (prostate globules) (G and H). The most aggressive prostate cancer cell line PC-3 showed the highest integrin α10 expression. Globular culture conditions significantly increased integrin α10 expression in PC-3 cells, but not in the less aggressive cell lines 22Rv1 or DU145. Representative flow cytometry images illustrate the proportion of integrin α10-positive cells in each cell line cultured in monolayers (E) and spheroids (G). A summary of the mean proportion of integrin α10-positive cells from different experiments cultured in monolayers (F) and spheroids (H). Data represent mean ± SD for n ≥ 3. IJ: Analysis of integrin α10 expression in pancreatic cells in monolayers (I) and spheroid structures (J). Spheroid culture conditions significantly increased integrin α10 protein expression levels in the most invasive cell line MiaPaCa-2 (Grade III), but not in the less invasive cell lines BxPC-3 and AsPC-1 (Grade II). KL: Analysis of integrin α10 expression in lung cancer cells in monolayers (K) and spheroid structures (L). Integrin α10 protein expression was highly increased in invasive lung cancer cell lines grown in spheroids (K) compared to monolayer culture conditions (L). M: Analysis of integrin α10 expression in sarcoma cells in monolayers and globular structures (M). Globular culture conditions significantly increased integrin α10 expression in undifferentiated pleomorphic sarcoma cancer cells.

[0070] Figure 6Integrin α10 (ITGA10) is expressed in aggressive cancer cell lines in monolayers, particularly three-dimensional (3D) cultures (spheres) (mRNA analysis). AB: Integrin α10 expression is increased in breast cancer cells cultured in breast spheres. ITGA10 expression is highest in the most aggressive cancer cell lines, such as triple-negative breast cancer: BT549, MDA-MB-231, and lowest in the least aggressive cell line T47D, especially when cancer cells are cultured in spheres. CD: Integrin α10 expression is increased in prostate cancer cells cultured in prostate spheres. ITGA10 expression is highest in the most aggressive cell line PC-3 and lowest in the least aggressive cell line 22RV1, especially when cancer cells are cultured in spheres. EF: Integrin α10 expression is increased in the invasive pancreatic cell line grade III. Spherical culture conditions significantly increased integrin α10 mRNA levels in the most invasive cell lines MiaPaCa-2 and PANC-1 (Grade III), but not in the less invasive cell lines BxPC-3 and AsPC-1 (Grade II). Data represent mean ± SD for n ≥ 3.

[0071] Figure 7 High ITGA10 expression was associated with poor overall survival in patients with different aggressive cancer indications. Overall survival curves for ITGA10 gene expression in patients with different aggressive cancers are shown. Patients were divided into high (solid line) and low (dashed line) ITGA10 expression groups based on the median cutoff value of survival analysis and log-rank test. The p-value represents the log-rank test for the difference in overall survival. For all indicated cancer forms, the high ITGA10 expression group had lower survival compared to the low ITGA10 expression group.

[0072] Figure 8Monoclonal integrin α10 antibody blocks adhesion of invasive cancer cells to collagen. AD: Triple-negative breast cancer cells. BT549 cells were cultured in monolayers (A and B) and mammary globules (C and D). Cells were pre-incubated for 30 min with monoclonal anti-integrin α10 blocking antibody mAbα10 (5 μg / ml) or IgG2a isotype control antibody (5 μg / ml). Cells were then allowed to adhere to culture dishes coated with type I collagen (A and C), type IV collagen (B and D), or bovine serum albumin (BSA) as a control, with or without the antibody. Data represent the mean of triplicate measurements, and the error bar represents the standard deviation for each data point. Adhesion relative to untreated cells (NT) (set to 100%) shows cell adhesion. EF: Invasive prostate cancer cells. PC-3 cells were pre-incubated for 30 min with monoclonal anti-integrin α10 function blocking antibody mAbα10 (5 μg / ml). The control was untreated cells. Cells were then subjected to adhesion assays, allowing them to adhere to collagen I (E) and IV (F) with or without antibodies. Data represent the mean of triplicate measurements, and error bars represent standard deviation. Adhesion relative to untreated (NT) cells (set to 100%) shows cell adhesion.

[0073] Figure 9 Migration was reduced after blocking integrin α10 with monoclonal antibodies. A: Breast cancer cells: BT549 breast cancer cells were pre-incubated for 30 min with monoclonal anti-integrin α10 blocking antibody mAbα10 (mouse) or Th101 (human) or control antibody IgG2a (mouse) or Th301 (human) (all 5 μg / ml). Cells were then subjected to a migration assay using a transwell assay in which the chambers were coated with type IV collagen. Migration relative to control cells (NT = untreated) (set to 100%) was used to show cell migration. B: Prostate cancer cells: Migration of prostate cancer cells was reduced by blocking integrin α10. The figure shows the effect of anti-integrin α10 antibody mAbα10 on PC-3 cell migration in a transwell assay. PC3 cells were seeded into 24-well plates in transwell chambers coated with type I collagen and incubated with monoclonal anti-integrin mAbα10 (5 μg / ml). Cell migration was shown relative to the migration of cells incubated with the isotope control antibody IgG2a (set to 100%) (measurement duration 24 or 48 hours). C: Lung cancer cells: Lung cancer cells were seeded into transwell chambers set in 24-well plates and incubated with monoclonal anti-integrin mAbα10. Cell migration was shown relative to the migration of cells incubated with the isotope control antibody IgG2a (set to 100%).

[0074] Figure 10 Integrin α10 antibody conjugated with the drug MMAE (anti-α10-MMAE) reduced the viability of breast cancer cells. BT549 cells were incubated for four days with progressively increasing concentrations of ADC (anti-α10-MMAE) or a negative control (anti-control-MMAE). Cell viability was determined by WST-1 colorimetric assay. The percentage of cell viability was calculated by comparison with the negative control anti-MMAE set to 100%. The results showed that anti-α10-MMAE reduced the viability of BT549 cells.

[0075] Figure 11 Integrin α10 antibody reduced the proliferation of spheroidized breast cancer cells, prostate cancer cells, pancreatic cancer cells, and lung cancer cells. (A) Breast cancer cells (BT549), (B) prostate cancer cells (PC-3), and (C and D) pancreatic cancer cells (MiaPaCa-2 and PANC-1) were treated with 5 μg / ml of integrin α10 antibody mAbα10 or a control antibody (IgG2a) and seeded in spheroid culture conditions. The antibody was added again every other day for 14 days. BrdU was then added to the spheroids, incubated for 24 hours, and proliferation was determined by flow cytometry. Data are shown as mean fluorescence intensity. (E) Similarly, breast cancer cells were incubated with monoclonal anti-integrin α10 antibody (mAbα10 or Th101) or negative control antibody (Th301, IgG2a). Proliferation relative to cells not incubated with the antibody (NT = untreated) (set to 100%) was shown as determined by BrdU incorporation. (F) In lung cancer, treatment with integrin α10 antibody (mAb α10) inhibited cell proliferation by arresting the cell cycle in the G0 / G1 phase, compared to treatment with the control antibody (IgG2a). Lung cancer cells (A549) were seeded in spheroids and treated with either integrin α10 antibody (mAb 10) or the control antibody (IgG2a) at 5 μg / ml. The antibody was added again every other day for 14 days. Cells were stained with BrdU and 7-AAD, and the cell cycle was analyzed by flow cytometry. Data are presented as a percentage of the total population.

[0076] Figure 12Treatment with integrin α10 antibody inhibited breast cancer tumor growth. Total flux readings (corresponding to tumor size) were reduced in mice treated with either integrin α10 antibody mAbα10 or Th101 compared to mice treated with negative control antibodies (IgG2a and Th301). (A) Box plots show tumor growth progression based on total flux (=tumor size) readings in different experimental groups (isotype control antibody IgG2a vs. mAbα10 and negative control antibody Th301 vs. Th101) after 9 cycles (9X) of antibody treatment. Invasive breast cancer BT549 cells were infected with a lentivirus expressing luciferase and GFP (BT549Luc / GFP). Animals were injected with 2x10... 6 BT549Luc / GFP cells were injected into the right ventral region of mice. Tumor growth was measured weekly using IVIS-CT spectroscopy via bioluminescence imaging. Photon flux (photons / s) and combined mean for each mouse are shown during 9 weeks (9X) post-tumor cell injection. Data are reported as mean ± SEM of total photon flux (photons / s). (B) Mouse body weight was recorded weekly during the study. No signs of disease or reduced growth rate were observed prior to treatment initiation.

[0077] Figure 13 Monoclonal antibody mAbα10 and Th101 bind to different epitopes on integrin α10. The binding competition between antibody mAbα10 and Th101 was measured in the C2C12 cell line overexpressing integrin α10--C2C12α10(A) and the triple-negative breast cancer cell line BT549(B), and analyzed by flow cytometry. Cells were incubated with the indicated concentration (μg / ml) of antibody for 30 min, followed by incubation with secondary antibody for another 30 min. Data are expressed as mean fluorescence intensity (MFI) per 100,000 cells. Gray bars: fluorescence intensity of donkey anti-human Alexa 488, which detects the binding intensity of antibody TH101 (human anti-integrin α10); black bars: fluorescence intensity of donkey anti-mouse Alexa 647, which detects the binding intensity of antibody mAbα10 (mouse anti-integrin α10). Left side of the graph: As the concentration of antibody Th101 added to the reaction wells gradually increases (0 to 9 μg / ml) (gray bars), the signal of antibody mAbα10 (at a constant concentration of 3 μg / ml) remains stable (black bars). Right side of the graph: As the concentration of antibody mAbα10 added to the reaction wells gradually increases (0 to 9 μg / ml) (black bars), the signal of antibody Th101 (at a constant concentration of 3 μg / ml) remains stable (gray bars).

[0078] definition

[0079] As used herein, “integrin α10”, “integrin α10 subunit”, or “integrin α10 polypeptide” refers to the α10 subunit of the heterodimeric protein integrin α10β1. This indication does not exclude the presence of the β1 subunit that binds to the α10 subunit to form the integrin α10β1 heterodimer. “α(alpha)” and “α”, as well as “α10” and “α10”, are equivalent terms. As used herein, “integrin α10” can refer to the α10 subunit of the heterodimeric protein integrin α10β1, as well as polynucleotide transcripts encoding the α10 subunit of the heterodimeric protein integrin α10β1, and fragments thereof.

[0080] As used herein, “anti-integrin α10 antibody” or “anti-integrin α10 subunit antibody” refers to an antibody that recognizes and binds at least to the α10 integrin of the heterodimeric protein integrin α10β1. These antibodies may be antibodies that recognize epitopes of the heterodimeric protein integrin α10β1, wherein the epitope comprises amino acid residues of both the α10 and β1 integrin polypeptides. Regarding monoclonal antibodies, “mAba10” and “mAbα10” are equivalent terms.

[0081] As used herein, the singular forms “a,” “an,” and “the” include plural indicators unless the context clearly indicates otherwise. Thus, for example, reference to “an antibody” includes multiple such antibodies.

[0082] As used herein, "subject" refers to mammals, such as rodents, felines, canines, equines, and primates. Preferably, the subject according to the invention is a human.

[0083] As used herein, “sample” encompasses any subject and a variety of sample types obtained from any subject. Examples of samples that can be used with the disclosed methods include, but are not limited to, subjects, liquid tissue samples (such as blood) or solid tissue samples (such as biopsy material or tissue cultures) or cells derived therefrom and their progeny. For example, biological samples include cells obtained from tissue samples collected from subjects. Thus, samples encompass clinical samples, cultured cells, cell supernatants, cell lysates, and tissue samples (e.g., tissue samples from breast tissue, lung tissue, prostate tissue, pancreatic tissue, ovarian tissue, bone tissue, cartilage tissue, adipose tissue, muscle tissue, and connective tissue).

[0084] As used in this article, "invasive cancer form" refers to a tumor that proliferates rapidly and / or metastasizes quickly to distant sites and other tissues. Invasive cancer forms can be invasive tumors and / or metastatic tumors and / or high-grade tumors and / or highly proliferating tumors. Invasive cancer forms are generally associated with poorer survival prognosis. Examples of invasive cancer forms include triple-negative breast cancer, inflammatory breast cancer, squamous cell lung cancer, lung adenocarcinoma, small cell lung cancer, prostate cancer, pancreatic cancer, ovarian cancer, and sarcoma. Different cancer types can exhibit varying degrees of invasiveness.

[0085] As used herein, “cancer” refers to any malignant and / or invasive growth or tumor caused by abnormal cell growth. As used herein, “cancer” refers to a tumor named after the type of cells that form it. Cancer or tumors are composed of tumor cells or cancer cells. A portion of a cancer or tumor may be stromal cells, such as connective tissue cells, such as fibroblasts. Examples of solid tumors include, but are not limited to, sarcomas and carcinomas. The term “cancer” includes, but is not limited to, primary cancer originating in a specific site in the body, metastatic cancer that has spread from its site of origin to other parts of the body, relapse after remission of the initial primary cancer, and secondary primary cancer (i.e., a new primary cancer of a different type than the previous cancer in a person with a history of cancer).

[0086] As used herein, “detection”, “detecting”, “detection” include qualitative and / or quantitative detection (measurement level) with or without a control, and further refer to the identification of the presence, absence, or quantity of a given target (particularly targets of the integrin α10 subunit).

[0087] As used herein, “inhibition” means that the presence of the antibody of the present invention wholly or partially inhibits the binding of the ligand to the receptor and / or the inactivation of the signaling that would be triggered by the receptor upon ligand binding. This includes, for example, downstream signaling that affects cellular behavior and processes. “Inhibition” and “blocking” are used as equivalent terms herein.

[0088] As used herein, "ADCC activity" or "antibody-dependent cytotoxic activity" refers to the activity of destroying target cells (e.g., tumor cells) by activating effector cells through the binding of the Fc region of an antibody to Fc receptors present on the surface of effector cells (such as killer cells, natural killer cells, activated macrophages, etc.). The activity of the antibodies of the present invention includes ADCC activity. ADCC activity measurements and antitumor assays can be performed using any assay known in the art. Detailed Implementation

[0089] The inventors of this invention have unexpectedly discovered that the integrin α10 polypeptide (Uniprot: O75578) encoded by the gene ITGA10 is overexpressed in tissues obtained from invasive tumor biopsies (particularly from triple-negative breast cancer, inflammatory breast cancer, squamous cell lung cancer, lung adenocarcinoma, small cell lung cancer, prostate cancer, pancreatic cancer, ovarian cancer, and sarcoma). Furthermore, the inventors of this invention have also discovered that the integrin α10 polypeptide (Uniprot: O75578) encoded by the gene ITGA10 is overexpressed in metastatic tumors originating from the aforementioned cancers.

[0090] Based on this discovery, the inventors have developed methods and tools for detecting and / or diagnosing and / or treating and / or preventing cancer forms, said cancer forms being any one of breast cancer, lung cancer, prostate cancer, pancreatic cancer, and sarcoma, or said cancer forms being metastases of said cancer forms.

[0091] In one implementation, the cancer form is an invasive cancer form.

[0092] Integrin α10 polypeptide

[0093] Integrins are heterodimers composed of α and β polypeptides. Integrin α10β1 heterodimers can be detected using anti-integrin α10 specific antibodies, as well as integrin α10 binding peptides and proteins.

[0094] In one embodiment of this disclosure, the integrin α10 polypeptide is part of the integrin α10β1 heterodimer.

[0095] In one embodiment of this disclosure, the integrin α10 polypeptide is expressed on the surface of cells.

[0096] In 1998, integrin α10β1 was identified as a type II collagen-binding receptor on chondrocytes (Camper et al., 1998). Immunohistochemical analyses during development and in adult tissues have demonstrated that this marker is restricted to cartilage-containing tissues (Camper et al., 1998; Camper et al., 2001). Knockout mice lacking this marker exhibited growth plate dysplasia, reduced collagen in the matrix, and shorter long bones, further supporting its structural importance (Bengtsson et al., 2005). Amino acid sequences, variants, isotypes, and sequence annotations can be found in Uniprot accession number O75578 (ITA10_human).

[0097] Integrin α10β1 is the most abundant collagen-binding integrin in cartilage tissue, and its expression pattern differs from that of other collagen-binding integrins. In vitro and in vivo studies have identified integrin α10β1 as a unique phenotypic marker for chondrocyte differentiation and a key mediator of cell-matrix interactions required for proper cartilage development. (and Aszòdi, 2014).

[0098] Furthermore, integrin α10β1 is present on mesenchymal stem cells (MSCs), and treatment of cultured MSCs with fibroblast growth factor 2 (FGF-2) increased integrin α10β1 expression and improved in vitro chondrogenesis in aggregated cultures. Therefore, integrin α10β1 is a cell surface biomarker for MSCs with chondrogenic potential (Varas et al., 2007).

[0099] Previously, integrin α10 expression in several different mouse brain structures (including the whole brain) had been analyzed, and it had been shown that integrin α10 was not expressed or was expressed at low levels in any healthy brain tissue (WO 99 / 51639). However, recently, the protein integrin α10β1 has been found to be expressed in tissues obtained from malignant tumors of the CNS and in the subventricular zone (SVZ) (a stem cell niche in the brain) (WO 2016 / 133449).

[0100] The limited distribution of integrin α10 in healthy tissues makes it a superior biomarker for disease. Therefore, this disclosure relates to the detection of antigenic integrin α10 peptides, for example, using antibodies specifically targeting SEQ ID NO:1 (integrin α10), SEQ ID NO:2 (extracellular domain of integrin α10), or SEQ ID NO:3 (extracellular I domain of integrin α10).

[0101] In one embodiment of this disclosure, integrin α10 is a naturally occurring variant of the integrin α10 polypeptide, an isotype of the integrin α10 polypeptide, or a splice variant of the integrin α10 polypeptide.

[0102] In one embodiment, the variant of the integrin α10 antigen is at least 70% identical to SEQ ID NO:1, 2, or 3, such as a variant at least 75% identical to SEQ ID NO:1, 2, or 3, such as a variant at least 80% identical to SEQ ID NO:1, 2, or 3, such as a variant at least 85% identical to SEQ ID NO:1, 2, or 3, such as a variant at least 90% identical to SEQ ID NO:1, 2, or 3, such as a variant at least 95% identical to SEQ ID NO:1, 2, or 3, such as a variant at least 96% identical to SEQ ID NO:1, 2, or 3, such as a variant at least 97% identical to SEQ ID NO:1, 2, or 3, such as a variant at least 98% identical to SEQ ID NO:1, 2, or 3, such as a variant at least 99% identical to SEQ ID NO:1, 2, or 3, such as a variant at least 99.5% identical to SEQ ID NO:1, 2, or 3.

[0103] In one embodiment, the fragment of integrin α10 comprises at least 100 consecutive amino acids of SEQ ID NO:1, preferably at least 200 consecutive amino acids of SEQ ID NO:1, preferably at least 300 consecutive amino acids of SEQ ID NO:1, preferably at least 400 consecutive amino acids of SEQ ID NO:1, preferably at least 500 consecutive amino acids of SEQ ID NO:1, preferably at least 600 consecutive amino acids of SEQ ID NO:1, preferably at least 700 consecutive amino acids of SEQ ID NO:1, preferably at least 800 consecutive amino acids of SEQ ID NO:1, preferably at least 900 consecutive amino acids of SEQ ID NO:1, preferably at least 1000 consecutive amino acids of SEQ ID NO:1.

[0104] Integrin α10 can also be detected at the nucleotide level by analyzing the presence of, for example, mRNA transcripts in the sample, which, after translation, produce the integrin α10 antigen as defined above.

[0105] Pharmaceutical Compositions and Their Administration

[0106] In one embodiment, this disclosure relates to a composition (such as a pharmaceutical composition) comprising:

[0107] a. An antibody or fragment thereof that specifically binds to integrin α10 polypeptide, or

[0108] b. A polynucleotide or fragment or variant thereof that specifically binds to a polynucleotide transcript encoding an integrin α10 polypeptide.

[0109] The composition is used for the diagnosis and / or treatment of cancer forms as defined herein.

[0110] In one embodiment of this disclosure, a composition for diagnosing and / or treating invasive cancer forms as defined herein comprises a pharmaceutically effective amount of an antibody or a fragment thereof that specifically binds to an integrin α10 polypeptide.

[0111] In one embodiment of this disclosure, a composition for diagnosing and / or treating invasive cancer forms as defined herein comprises a pharmaceutically effective amount of a polynucleotide or a fragment or variant thereof that specifically binds to a polynucleotide transcript encoding an integrin α10 polypeptide.

[0112] As mentioned herein, a pharmaceutically effective amount is typically the amount of an anti-integrin α10 antibody or a polynucleotide that specifically binds to a polynucleotide transcript encoding an integrin α10 polypeptide, said amount inducing the desired response in an individual receiving the pharmaceutical composition.

[0113] This article describes in detail antibodies or fragments thereof that specifically bind to integrin α10 polypeptide, as well as polynucleotides, fragments and variants thereof that specifically bind to polynucleotide transcripts encoding integrin α10 polypeptide.

[0114] In one embodiment of this disclosure, a composition for diagnosing and / or treating cancer forms as defined herein comprises a pharmaceutically effective amount of an antibody or fragment thereof specifically bound to an integrin α10 polypeptide, wherein the antibody or fragment thereof is conjugated to an additional portion.

[0115] In one embodiment of this disclosure, a composition for diagnosing and / or treating an aggressive form of cancer as defined herein comprises a pharmaceutically effective amount of a polynucleotide or a fragment or variant thereof specifically bound to a polynucleotide transcript encoding an integrin α10 polypeptide, wherein said polynucleotide or fragment or variant thereof is conjugated to an additional portion.

[0116] For example, the additional portion could be a detectable portion. Antibodies that specifically bind to the integrin α10 polypeptide or a fragment thereof and are conjugated to the detectable portion can be used to detect integrin α10 expression on cells and thus determine whether the cells are malignant and / or tumor-associated cells. For example, the additional portion could be a cytotoxic portion. Antibodies that specifically bind to the integrin α10 polypeptide or a fragment thereof and are conjugated to the cytotoxic portion (such as antibody-drug conjugates (ADCs) containing an antibody specifically bound to the integrin α10 polypeptide or a fragment thereof) can be used to specifically target a cytotoxic portion and / or drug to cells expressing integrin α10 and which are malignant and / or tumor-associated cells.

[0117] For example, the additional portion may include biological response modulators. Biological response modulators are substances that regulate immune responses by enhancing or inhibiting them. Biological response modulators can be endogenous (such as those normally produced naturally in the body) or exogenous.

[0118] In one embodiment, the additional portion may include biological response modulators, such as cytokines, lymphokines, interferons, or combinations thereof.

[0119] Detectable part

[0120] The compositions for the purposes described in this disclosure may include a detectable portion, such as an antibody or fragment thereof that specifically binds to the integrin α10 polypeptide and / or a polynucleotide or fragment thereof or variant thereof that specifically binds to a polynucleotide transcript encoding the integrin α10 polypeptide may be covalently bound to the detectable portion.

[0121] In one embodiment, the detectable portion is selected from fluorophores, enzymes, or radioactive tracers or radioisotopes. In one embodiment, the detectable portion is a radioactive tracer selected from positron emitters and gamma emitters. In one embodiment, the radioactive isotope is selected from 99mTc. 111 In、 67 Ga、 68 Ga、 72 As、 89 Zr、 123 I and 201 Tl. In one embodiment, the antibody comprises a pair of detectable and cytotoxic radionuclides, such as 86 Y / 90 Y or 124 I / 211 At.

[0122] In one embodiment, the radioisotope can function simultaneously as both a detectable portion and a cytotoxic portion in a multimodal manner.

[0123] In one embodiment, the detectable portion comprises or is composed of a paramagnetic isotope, such as paramagnetic isotopes selected from the following: 157 Gd, 55 Mn, 162 Dy、 52 Cr and 56 Fe. In one embodiment, the detectable portion is detectable by imaging techniques such as SPECT, PET, MRI, optical or ultrasound imaging.

[0124] In one embodiment, the cytotoxic portion and / or the detectable portion are indirectly connected to the antibody or its antigen-binding fragment via a connecting portion.

[0125] Cytotoxicity

[0126] The compositions for the purposes described in this disclosure may include a cytotoxic portion, such as an antibody or fragment thereof that specifically binds to the integrin α10 polypeptide and / or a polynucleotide or fragment thereof or variant thereof that specifically binds to a polynucleotide transcript encoding the integrin α10 polypeptide may be covalently bound to the cytotoxic portion.

[0127] In one embodiment, the cytotoxic component is selected from toxins, chemotherapeutic agents, and radiopharmaceuticals, or combinations thereof.

[0128] In one embodiment, the cytotoxic portion is a toxin. For example, in one embodiment, the cytotoxic portion is a toxin selected from microtubule toxins, DNA toxins, and transcription toxins.

[0129] In one embodiment, the cytotoxic component is a microtubule toxin selected from the following: auristatin-based toxin, maytansinoid-based toxin, tubulosin-based toxin, and eribulin.

[0130] In one embodiment, the cytotoxic portion is selected from the following DNA toxins: DNA minor groove binders, DNA minor groove binding alkylating agents, DNA alkylating agents, and DNA cleaving agents. For example, in one embodiment, the cytotoxic portion is selected from the following DNA toxins: pyrrolobenzodiazepine (PBD), duocarmycin, duocarmycin analogs, indoline benzodiazepine, calicacin, irinotecan, and exatecan derivatives.

[0131] In one embodiment, the cytotoxic component is a transcription toxin, such as a ribosome-inactivating protein, such as an RNA polymerase II inhibitor.

[0132] In one embodiment, the cytotoxic portion is a transcription toxin selected from the following: doxorubicin, doxorubicin derivatives, and amatoxins.

[0133] In one embodiment, the cytotoxic component is selected from the following transcription toxins: Shiga toxin and Shiga-like toxins, type I ribosome inactivating proteins, type II ribosome inactivating proteins, and saponins or combinations thereof. For example, the type I ribosome inactivating protein may be trichosanthin and / or luffin. For example, the type II ribosome inactivating protein may be ricin, lectin, and / or absinthecin.

[0134] In one embodiment, the cytotoxic portion is a chemotherapeutic agent. For example, in one embodiment, the chemotherapeutic agent may be an alkylating agent, an antimetabolite, an antimicrotubule agent, a topoisomerase inhibitor, or a cytotoxic antibiotic. For example, in one embodiment, the chemotherapeutic agent may be selected from anthracyclines, taxanes, and platinum-based agents. For example, in one embodiment, the chemotherapeutic agent may be selected from cisplatin, paclitaxel, albumin-bound paclitaxel, docetaxel, cyclophosphamide, eribulin, epirubicin, doxorubicin, carboplatin, gemcitabine, bleomycin, fluorouracil, cyclophosphamide, vinorelbine, capecitabine, ixabepilone, and ixabepilone, or combinations thereof.

[0135] Preparations

[0136] The compositions for the purposes described in this disclosure may be pharmaceutical compositions suitable for parenteral administration. Such compositions preferably comprise aqueous and non-aqueous sterile injectable solutions, which may contain wetting agents or emulsifiers, antioxidants, pH buffers, antimicrobial compounds, and solutes that make the formulation isotonic with an individual's bodily fluids (preferably blood); and aqueous and non-aqueous sterile suspensions, which may include suspending agents or thickeners. The pharmaceutical compositions may be presented in single-dose or multi-dose containers (e.g., sealed ampoules and vials) and may be stored under lyophilized conditions, requiring only the addition of a sterile liquid carrier just before use.

[0137] In one embodiment, the composition of this disclosure for the said use further comprises at least one pharmaceutically acceptable diluent, carrier, or excipient.

[0138] Preferably, the compositions of the present invention comprise one or more suitable pharmaceutical excipients for use with cells, tissues, or organisms, said excipients being non-sterile or sterile, such as pharmaceutical excipients suitable for individual administration. Such excipients may include, but are not limited to, saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations of these excipients in various amounts. The formulation shall conform to the administration regimen.

[0139] Preferably, the pharmaceutical compositions of the present invention are prepared in an injectable form as a liquid solution or suspension; furthermore, solid forms suitable for dissolving or suspending in a liquid before injection are also within the scope of the present invention. The formulation may also be emulsified or encapsulated in liposomes.

[0140] The anti-integrin α10 polypeptide antibody or the polynucleotide that specifically binds to the polynucleotide transcript encoding the integrin α10 polypeptide can be administered alone or in combination with other compounds, simultaneously or sequentially in any order.

[0141] Administration can be, for example, via injection or infusion, parenteral administration. Parenteral administration can be, for example, intravenous, intrathecal, intravenous, intramuscular, intradermal, or subcutaneous injection. Preferably, the administration is via injection or infusion, parenteral administration.

[0142] Clinical symptoms

[0143] In one embodiment of this disclosure, the cancer form to be treated and / or prevented and / or detected and / or diagnosed and / or classified and / or determined for prognosis and / or prevented from metastasizing is selected from breast cancer, lung cancer, prostate cancer, pancreatic cancer, ovarian cancer, and sarcoma.

[0144] In another embodiment of this disclosure, the cancer form to be treated and / or prevented and / or detected and / or diagnosed and / or classified and / or determined for prognosis and / or prevented from metastasizing is selected from the following invasive cancer forms: invasive breast cancer, invasive lung cancer, invasive prostate cancer, invasive pancreatic cancer, ovarian cancer, and invasive sarcoma.

[0145] As used in this article, "invasive cancer form" refers to a tumor that proliferates rapidly and / or metastasizes quickly to distant sites and other tissues. Invasive cancer forms can be invasive tumors and / or metastatic tumors and / or high-grade tumors and / or highly proliferating tumors. Invasive cancer forms are generally associated with poorer survival prognosis. Examples of invasive cancer forms include triple-negative breast cancer, inflammatory breast cancer, squamous cell lung cancer, lung adenocarcinoma, small cell lung cancer, prostate cancer, pancreatic cancer, ovarian cancer, and sarcoma. Different cancer types can exhibit varying degrees of invasiveness.

[0146] In one embodiment of this disclosure, the invasive breast cancer is selected from triple-negative breast cancer and inflammatory breast cancer.

[0147] In one embodiment of this disclosure, the invasive cancer form is a triple-negative breast cancer form, and the triple-negative breast cancer is selected from basaloid 1 breast cancer, basaloid 2 breast cancer, low tight junction protein breast cancer, metaplastic breast cancer (MBC), interferon-rich breast cancer, immunomodulatory breast cancer, mesenchymal breast cancer, mesenchymal stem cell-like breast cancer, luminal androgen receptor breast cancer, and unstable breast cancer.

[0148] Those skilled in the art will understand the characteristics of invasive breast cancer, see, for example, Arpino et al. 2015; Dai et al. 2016 and Ma et al. 2018.

[0149] In one embodiment of this disclosure, the invasive form of breast cancer is a triple-negative breast cancer form that has the morphological characteristics of invasive ductal carcinoma.

[0150] In one embodiment of this disclosure, the invasive form of breast cancer is a triple-negative breast cancer form, and it has the morphological characteristics of basal-like triple-negative breast cancer.

[0151] In one embodiment of this disclosure, the invasive form of breast cancer is a basal-like breast cancer form that has the morphological characteristics of invasive ductal carcinoma.

[0152] In one embodiment of this disclosure, the invasive form of cancer is prostate cancer, and the prostate cancer is small cell (neuroendocrine) carcinoma (SCNC).

[0153] In one embodiment of this disclosure, the invasive form of cancer is lung cancer, and the lung cancer is squamous cell lung cancer, lung adenocarcinoma, small cell lung cancer, or large cell lung cancer.

[0154] In one embodiment of this disclosure, the invasive form of cancer is lung cancer, and the lung cancer is squamous cell lung cancer.

[0155] In one embodiment of this disclosure, the invasive cancer form is pancreatic cancer, and the invasive pancreatic cancer is a neuroendocrine tumor.

[0156] In one embodiment of this disclosure, the invasive form of cancer is pancreatic cancer, and the invasive pancreatic cancer is grade I, II, or III pancreatic cancer.

[0157] In one embodiment of this disclosure, the invasive sarcoma is selected from undifferentiated pleomorphic sarcoma, myxofibrosarcoma, dedifferentiated liposarcoma, atypical lipoma, myxoinflammatory fibroblastic sarcoma, low-grade fibromyxoid sarcoma, sclerosing epithelioid fibrosarcoma, pseudomyogenic angioendothelioma, and mesenchymal chondrosarcoma.

[0158] In one embodiment of this disclosure, the invasive cancer form is a metastatic tumor, for example, the invasive cancer form can be a metastatic tumor of any of the following: invasive breast cancer, invasive lung cancer, invasive prostate cancer, invasive pancreatic cancer, ovarian cancer, and invasive sarcoma.

[0159] In one embodiment of this disclosure, the invasive cancer form is a metastatic tumor, such as a metastatic tumor of either triple-negative breast cancer or inflammatory breast cancer.

[0160] In one embodiment of this disclosure, the invasive cancer form is a metastatic tumor, such as a metastatic tumor of triple-negative breast cancer, including metastatic tumors selected from the following triple-negative breast cancers: basaloid 1 breast cancer, basaloid 2 breast cancer, low tight junction protein breast cancer, metaplastic breast cancer (MBC), interferon-rich breast cancer, immunomodulatory breast cancer, mesenchymal breast cancer, mesenchymal stem cell-like breast cancer, luminal androgen receptor breast cancer, and unstable breast cancer.

[0161] In one embodiment of this disclosure, the invasive cancer form is a metastatic tumor, such as a metastatic tumor of invasive lung cancer. In another embodiment of this disclosure, the invasive cancer form is a metastatic tumor, such as a metastatic tumor of squamous cell lung cancer, lung adenocarcinoma, small cell lung cancer, or large cell lung cancer.

[0162] In one embodiment of this disclosure, the invasive cancer form is a metastatic tumor, such as a metastatic tumor of invasive pancreatic cancer.

[0163] In one embodiment of this disclosure, the invasive cancer form is a metastatic tumor, such as a metastatic tumor of invasive prostate cancer.

[0164] In one embodiment of this disclosure, the invasive cancer form is a metastatic tumor, such as a metastatic sarcoma.

[0165] Treatment of invasive cancer forms

[0166] In one aspect, the present invention relates to the use of a composition for manufacturing an agent for treating aggressive forms of cancer, said composition comprising:

[0167] a. An antibody or fragment thereof that specifically binds to integrin α10 polypeptide, or

[0168] b. A polynucleotide or fragment or variant thereof that specifically binds to a polynucleotide transcript encoding an integrin α10 polypeptide.

[0169] The invasive cancer form is selected from invasive breast cancer, invasive lung cancer, invasive prostate cancer, invasive pancreatic cancer, and invasive sarcoma, or the invasive cancer form is a metastatic tumor.

[0170] One aspect of this disclosure relates to a method of treating an aggressive form of cancer, wherein the aggressive form of cancer is selected from aggressive breast cancer, aggressive lung cancer, aggressive prostate cancer, aggressive pancreatic cancer, and aggressive sarcoma, or wherein the aggressive form of cancer is a metastatic tumor, the method comprising administering to a subject in need a pharmaceutically effective amount of a composition, the composition comprising:

[0171] a. An antibody or fragment thereof that specifically binds to integrin α10 polypeptide, or

[0172] b. A polynucleotide or a fragment or variant thereof that specifically binds to a polynucleotide transcript encoding an integrin α10 polypeptide.

[0173] One aspect of this disclosure relates to a method for inhibiting integrin α10-mediated signaling in at least one cancer cell, the method comprising contacting the at least one cancer cell with a composition comprising an effective amount of:

[0174] a. Antibodies or antigen-binding fragments that are specific to integrin α10 peptide; and / or

[0175] b. A multinucleotide transcript or fragment or variant thereof encoding the integrin α10 polypeptide.

[0176] The at least one cancer cell is selected from invasive breast cancer cells, invasive lung cancer cells, invasive prostate cancer cells, invasive pancreatic cancer cells, invasive sarcoma cells, and metastatic tumor cells.

[0177] Those skilled in the art will understand that cell signaling includes molecular mechanisms by which cells detect and respond to external stimuli. Cell signaling also includes transcriptional and translational control mechanisms, as well as signal transduction mechanisms.

[0178] One aspect of this disclosure relates to a method for inhibiting the cellular function of at least one cancer cell, the method comprising contacting the at least one cancer cell with an effective amount of a composition comprising:

[0179] a. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0180] b. A multinucleotide transcript or fragment or variant thereof encoding the integrin α10 polypeptide.

[0181] The at least one cancer cell is selected from invasive breast cancer cells, invasive lung cancer cells, invasive prostate cancer cells, invasive pancreatic cancer cells, invasive sarcoma cells, and metastatic tumor cells.

[0182] In one embodiment, the present invention relates to a method for inhibiting the growth and / or proliferation of cells expressing integrin α10, the method comprising administering a composition comprising an effective amount of:

[0183] a. An antibody or fragment thereof that specifically binds to integrin α10 polypeptide, or

[0184] b. A polynucleotide or a fragment or variant thereof that specifically binds to a polynucleotide transcript encoding an integrin α10 polypeptide.

[0185] In one embodiment of this disclosure, the inhibition of at least one cancer cell is selected from:

[0186] a. Inhibit the proliferation of at least one of the cancer cells;

[0187] b. Inhibit the self-renewal of at least one of the cancer cells;

[0188] c. Inhibit the anchorage-independent growth of the at least one cancer cell;

[0189] d. Inhibit the migration of at least one of the cancer cells;

[0190] e. Inhibit the invasion of at least one of the cancer cells;

[0191] f. Inhibit the adhesion of at least one of the cancer cells; and / or

[0192] Its combination.

[0193] In another embodiment, the anchor-dependent growth of at least one cancer cell is inhibited by i) an antibody that specifically binds to the integrin α10 polypeptide or ii) a polynucleotide or fragment or variant thereof that specifically binds to a polynucleotide transcript encoding the integrin α10 polypeptide.

[0194] In one embodiment, this disclosure relates to a method of inhibiting at least one cancer cell, wherein the cancer cells are in an invasive and / or metastatic tumor, and wherein the inhibition is at least one of the following:

[0195] a. The growth of the invasive and / or metastatic tumors;

[0196] b. The proliferation of the invasive and / or metastatic tumors;

[0197] c. The migration of the invasive and / or metastatic tumors;

[0198] d. Invasion of the aforementioned invasive and / or metastatic tumors;

[0199] e. The onset of new invasive and / or metastatic tumors;

[0200] f. Infiltration of new invasive and / or metastatic tumors; and

[0201] Its combination.

[0202] The cells may express one or more further markers as defined herein. In one embodiment, the cells are malignant cells and / or tumor-associated cells. In another embodiment, the cells are cancer-associated fibroblasts (CAFs), stromal cells, stem cells, and / or stem cell-like cells. The method may be performed in vitro or in vivo.

[0203] In one embodiment of this disclosure, an effective quantity is included:

[0204] a. An antibody or fragment thereof that specifically binds to integrin α10 polypeptide, or

[0205] b. A combination of polynucleotides or fragments or variants thereof that specifically bind to a polynucleotide transcript encoding an integrin α10 polypeptide, capable of inducing cell death and / or inhibiting cell growth and / or inhibiting cell proliferation and / or inhibiting cell migration in cells expressing integrin α10.

[0206] In one embodiment of this disclosure, the treatment is initiated after integrin α10 peptide and / or polynucleotide transcripts are detected in cancer cells in the subject's tumor.

[0207] In one embodiment of this disclosure, the composition disclosed herein for treating an aggressive form of cancer is administered to an individual in need in combination with radiotherapy and / or surgical resection of the cancer. For example, in one embodiment, the composition disclosed herein for treating an aggressive form of cancer is administered to the individual prior to radiotherapy and / or surgical resection of the cancer. Alternatively, or additionally, in one embodiment, the composition disclosed herein for treating an aggressive form of cancer is administered to the individual after radiotherapy and / or surgical resection of the cancer.

[0208] In one embodiment of this disclosure, the treatment is preventative, ameliorative, or curative.

[0209] In one embodiment of this disclosure, the method disclosed herein targets an antigen expressed on the surface of the cell that contains an integrin α10 polypeptide or a fragment thereof.

[0210] Those skilled in the art will understand the possibility of preventing metastasis originating from invasive cancer.

[0211] One aspect of this disclosure relates to a method for preventing metastasis of a primary cancer form selected from breast cancer, lung cancer, prostate cancer, pancreatic cancer, and sarcoma, the method comprising administering a therapeutically effective dose to a patient in need:

[0212] a) an antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment is specific to integrin α10 polypeptide; and / or

[0213] b) A multinucleotide transcript or a fragment or variant thereof encoding the integrin α10 polypeptide.

[0214] Those skilled in the art will understand that methods for preventing the metastasis of said primary cancer can be applied after the primary cancer is detected.

[0215] Detection and diagnosis of aggressive cancer forms

[0216] One aspect of this disclosure relates to an antibody or fragment thereof specific to the integrin α10 polypeptide, or an agent composed of said antibody or fragment thereof, for detecting cells associated with an aggressive form of cancer in mammals, said cells expressing the integrin α10 polypeptide, and said aggressive form of cancer selected from aggressive breast cancer, aggressive lung cancer, aggressive prostate cancer, aggressive pancreatic cancer, and aggressive sarcoma, or metastatic tumors of any of said aggressive forms of cancer.

[0217] One aspect of this disclosure relates to a composition comprising:

[0218] a) An antibody or its antigen-binding fragment that specifically binds to integrin α10 polypeptide, or

[0219] b) A polynucleotide or a fragment or variant thereof that specifically binds to a polynucleotide transcript encoding an integrin α10 polypeptide.

[0220] Used to diagnose metastatic tumors of any of the following cancer forms: breast cancer, lung cancer, prostate cancer, pancreatic cancer, and sarcoma, or any of the aforementioned cancer forms.

[0221] Those skilled in the art will understand that, for example, the process of detecting disease biomarkers (such as integrin α10 in this invention) or diagnosing disease by analyzing the expression of integrin α10, as described in this invention, may include comparing the tissue to be analyzed with healthy, non-malignant, or unaffected tissue. For example, a breast cancer sample may be compared with an unaffected region in the same tissue sample or with a healthy breast tissue sample. A lung cancer sample may be compared with an unaffected region in the same tissue sample or with a healthy lung tissue sample. A prostate cancer sample may be compared with an unaffected region in the same tissue sample or with a healthy prostate tissue sample. A pancreatic cancer sample may be compared with an unaffected region in the same tissue sample or with a healthy pancreatic tissue sample. A sarcoma sample may be compared with an unaffected region in the same tissue sample or with a healthy connective tissue sample.

[0222] In the process of diagnosing cancer, those skilled in the art will understand the potential usefulness of comparing the levels of integrin α10 peptides or polynucleotides in cancer cells with those in reference cells.

[0223] In one embodiment, the diagnosed cancer comprises cells exhibiting equal or higher levels of i) the integrin α10 antigen or ii) the polynucleotide transcript observed in the same type of healthy and / or benign tissue.

[0224] In another embodiment, the diagnosed cancer comprises cells exhibiting equal or higher levels of i) the integrin α10 antigen or ii) the polynucleotide transcript observed in cancer types of the same tissue type that are less aggressive compared to the diagnosed cancer type.

[0225] In another embodiment, the diagnosed cancer comprises cells exhibiting equal or higher expression levels of i) the integrin α10 antigen or ii) the polynucleotide transcript compared to a reference cell line.

[0226] Those skilled in the art will understand that reference cell lines can be used to standardize diagnostic procedures due to their genotypic and phenotypic stability compared to primary cells. Reference cell lines can be established from healthy tissue or from cancerous tissue, where the cancer can be a more or less aggressive type, depending on the growth and metastatic tendencies of that cancer type. Established cell lines require immortalization to be able to propagate in cell culture. As used herein, a non-malignant cell line is understood to be an established cell line that does not show signs of malignancy and is phenotypically similar to cells from healthy tissue.

[0227] The reference cell line can be, for example, the cell line depicted in Table 1.

[0228]

[0229] Table 1. Overview of the cell lines used in this study and their invasiveness.

[0230] In one implementation, the reference cell line used during the diagnostic procedure is derived from healthy tissue, such as a reference cell line derived from healthy breast tissue, a reference cell line derived from healthy prostate tissue, a reference cell line derived from healthy lung tissue, a reference cell line derived from healthy pancreatic tissue, and a reference cell line derived from healthy connective tissue.

[0231] In one implementation, the reference cell line used during a breast cancer diagnostic procedure is derived from healthy tissue, such as reference cell line 184A1 derived from healthy breast cancer tissue.

[0232] In another embodiment, the reference cell line used during the diagnostic procedure is derived from a less aggressive cancer type of the same tissue type compared to the diagnosed cancer type, such as a reference cell line derived from less aggressive breast cancer, a reference cell line derived from less aggressive prostate cancer, a reference cell line derived from less aggressive lung cancer, a reference cell line derived from less aggressive pancreatic cancer, and a reference cell line derived from less aggressive sarcoma.

[0233] In one embodiment, the reference cell lines used during the diagnostic procedure for invasive cancer are selected from HCC1428 (a cell line derived from less invasive breast cancer), T47D (a cell line derived from less invasive breast cancer), 22Rv1 (a cell line derived from less invasive prostate cancer), DU145 (a cell line derived from less invasive prostate cancer), BxPC-3 (a cell line derived from less invasive pancreatic cancer), and AsPC-1 (a cell line derived from less invasive pancreatic cancer).

[0234] One aspect of this disclosure relates to a method for detecting aggressive cancer cells in a subject, the method comprising the following steps:

[0235] a. Provide the subject with tissue suspected of containing cancer cells;

[0236] b. Analyze the presence of the following in the described organization:

[0237] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0238] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0239] c. Determine the expression levels of the antigen described in i) and / or the polynucleotide transcript described in ii), and

[0240] d. The expression levels determined in c. will be compared with control levels.

[0241] The control level is the average level observed in healthy and / or benign cells of the same tissue type as the isolated sample, wherein i) the expression level of the antigen and / or ii) the polynucleotide transcript being higher than the control level indicates the presence of an aggressive form of cancer in the subject, and wherein the aggressive form of cancer is selected from aggressive breast cancer, aggressive lung cancer, aggressive prostate cancer, aggressive pancreatic cancer, and aggressive sarcoma, or a metastatic tumor of any of the aggressive forms of cancer.

[0242] Another aspect of this disclosure relates to a method for diagnosing a form of cancer in a subject, the method comprising the following steps:

[0243] a. Provide the subject with tissue suspected of containing cancer cells;

[0244] b. Analyze the presence of the following in the described organization:

[0245] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0246] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0247] c. Determine the expression levels of i) integrin α10 antigen and / or ii) polynucleotide transcripts, and

[0248] d. The expression levels determined in c. will be compared with control levels, wherein the control levels are the average expression levels of i) the antigen or ii) the polynucleotide transcript observed in the same type of healthy and / or benign tissue; and

[0249] Wherein i) the expression level of the antigen and / or ii) the polynucleotide transcript is higher than the control level, indicating the presence of a cancer form in the sample, wherein the cancer form is selected from breast cancer, lung cancer, prostate cancer, pancreatic cancer and sarcoma or a metastatic tumor of any of the cancer forms, thereby diagnosing the cancer form of the subject.

[0250] Another aspect of this disclosure relates to a method for diagnosing a form of cancer in a subject, the method comprising the steps of:

[0251] a. Provide the subject with tissue suspected of containing cancer cells;

[0252] b. Analyze the presence of one or more cells with cancerous morphology in the tissue;

[0253] c. Analyze the presence of the following in the sample:

[0254] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0255] ii. A polynucleotide transcript or a fragment or variant thereof encoding an integrin α10 polypeptide, optionally determining the expression level of the i) integrin α10 antigen and / or the ii) polynucleotide transcript, wherein the presence of one or more cells with a cancer morphology, in combination with the expression of the integrin α10 i) antigen and / or the ii) polynucleotide transcript, indicates the presence of a cancer form in the sample, wherein the cancer form is selected from breast cancer, lung cancer, prostate cancer, pancreatic cancer, and sarcoma, or a metastatic tumor of any of the cancer forms, thereby diagnosing the cancer form of the subject.

[0256] In one embodiment of this disclosure, analyzing the presence of an antigen containing an integrin α10 polypeptide or a fragment thereof in step b. includes contacting tissue suspected of containing invasive cancer cells with the composition of this disclosure. For example, in one embodiment of this disclosure, tissue suspected of containing cancer cells may be contacted with a composition containing an antibody specific to or composed of an integrin α10 polypeptide.

[0257] In addition to analyzing the presence of an antigen containing an integrin α10 polypeptide or a fragment thereof in the sample; and / or the presence of a polynucleotide transcript or a fragment or variant thereof encoding an integrin α10 polypeptide; and determining whether the integrin α10 expression level is higher than the control level, the method for diagnosing an aggressive form of cancer in a subject and / or the method for detecting aggressive cancer cells in a subject may further include the step of morphologically characterizing the sample as containing cancer cells belonging to an aggressive form of cancer, wherein the aggressive form of cancer is selected from aggressive breast cancer, aggressive lung cancer, aggressive prostate cancer, aggressive pancreatic cancer, and aggressive sarcoma, or metastatic tumors of any of the aggressive forms of cancer.

[0258] Morphological characterization of cancerous or suspected cancerous tissue is known to those skilled in the art and is currently used to detect cancer cells and diagnose the presence of cancer. However, morphological characterization alone may only be used to distinguish cancer cells from healthy and / or benign cells of the same tissue type and is insufficient to determine whether cancer cells belong to an invasive form of cancer, such as for diagnosing the presence of an invasive form of cancer in a subject.

[0259] Therefore, in one aspect, this disclosure relates to a method for detecting aggressive cancer cells in a subject, the method comprising the following steps:

[0260] a. Provide the subject with tissue suspected of containing cancer cells;

[0261] b. Analyze the presence of one or more cells with cancerous morphology in the tissue.

[0262] c. Analyze the presence of the following in the described tissue:

[0263] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0264] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0265] d. Optionally determine the expression levels of the i) integrin α10 antigen and / or ii) polynucleotide transcripts, wherein the presence of one or more cells with cancerous morphology, in combination with the expression of the integrin α10 i) antigen and / or ii) polynucleotide transcripts, indicates the presence of an invasive cancer form in the subject, and wherein the invasive cancer form is selected from invasive breast cancer, invasive lung cancer, invasive prostate cancer, invasive pancreatic cancer, and invasive sarcoma, or a metastatic tumor of any of the invasive cancer forms.

[0266] In one embodiment, this disclosure relates to a method for detecting the presence of breast cancer cells in a subject or for diagnosing breast cancer in a subject, wherein the breast cancer is an aggressive form of breast cancer selected from triple-negative breast cancer and inflammatory breast cancer, the method comprising the following steps:

[0267] a. Provide the subject with breast tissue suspected of containing cancer cells;

[0268] b. Analyze the presence of the following in the described organization:

[0269] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0270] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0271] c. Determine the expression level of the antigen described in i) or the polynucleotide transcript described in ii), and

[0272] d. The expression levels determined in c. will be compared with control levels, wherein the control levels are the average expression levels of i) the antigen or ii) the polynucleotide transcripts observed in healthy and / or benign breast tissue; and

[0273] Wherein i) the expression level of the antigen and / or ii) the polynucleotide transcript is higher than the control level, indicating the presence of breast cancer, thereby detecting the presence of breast cancer cells in the subject and / or diagnosing breast cancer in the subject.

[0274] In one embodiment, this disclosure relates to a method for detecting the presence of lung cancer in a subject or for diagnosing lung cancer in a subject, the method comprising the following steps:

[0275] a. Provide the subject with lung tissue suspected of containing cancer cells;

[0276] b. Analyze the presence of the following in the described organization:

[0277] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0278] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0279] c. Determine the expression level of the antigen described in i) or the polynucleotide transcript described in ii), and

[0280] d. The expression levels determined in c. will be compared with control levels, wherein the control levels are the average expression levels of i) the antigen or ii) the polynucleotide transcripts observed in healthy and / or benign lung tissue; and

[0281] Wherein i) the expression level of the antigen and / or ii) the polynucleotide transcript is higher than the control level, indicating the presence of lung cancer, thereby detecting and / or diagnosing squamous cell carcinoma of the subject's lung.

[0282] In one embodiment, this disclosure relates to a method for detecting the presence of squamous cell carcinoma of the lung in a subject or for diagnosing squamous cell carcinoma of the lung in a subject, the method comprising the following steps:

[0283] a. Provide the subject with lung tissue suspected of containing cancer cells;

[0284] b. Analyze the presence of the following in the described organization:

[0285] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0286] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0287] c. Determine the expression level of the antigen described in i) or the polynucleotide transcript described in ii), and

[0288] d. The expression levels determined in c. will be compared with control levels, wherein the control levels are the average expression levels of i) the antigen or ii) the polynucleotide transcripts observed in healthy and / or benign lung tissue; and

[0289] Wherein i) the expression level of the antigen and / or ii) the polynucleotide transcript is higher than the control level, indicating the presence of lung squamous cell carcinoma, thereby detecting and / or diagnosing lung squamous cell carcinoma in the subject.

[0290] In one embodiment, this disclosure relates to a method for detecting the presence of lung adenocarcinoma in a subject or for diagnosing lung adenocarcinoma in a subject, the method comprising the following steps:

[0291] a. Provide the subject with lung tissue suspected of containing cancer cells;

[0292] b. Analyze the presence of the following in the described organization:

[0293] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0294] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0295] c. Determine the expression level of the antigen described in i) or the polynucleotide transcript described in ii), and

[0296] d. The expression levels determined in c. will be compared with control levels, wherein the control levels are the average expression levels of i) the antigen or ii) the polynucleotide transcripts observed in healthy and / or benign lung tissue; and

[0297] Wherein i) the expression level of the antigen and / or ii) the polynucleotide transcript is higher than the control level, indicating the presence of lung adenocarcinoma, thereby detecting and / or diagnosing lung adenocarcinoma in the subject.

[0298] In one embodiment, this disclosure relates to a method for detecting the presence of prostate cancer cells in a subject or for diagnosing prostate cancer in a subject, the method comprising the following steps:

[0299] a. Provide the subject with prostate tissue suspected of containing cancer cells;

[0300] b. Analyze the presence of the following in the sample:

[0301] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0302] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0303] c. Determine the expression level of the antigen described in i) or the polynucleotide transcript described in ii), and

[0304] d. The expression levels determined in c. will be compared with control levels, wherein the control levels are the average expression levels of i) the antigen or ii) the polynucleotide transcripts observed in healthy and / or benign prostate tissue; and

[0305] Wherein i) the expression level of the antigen and / or ii) the polynucleotide transcript is higher than the control level, indicating the presence of prostate cancer, thereby detecting and / or diagnosing prostate cancer in the subject.

[0306] In one embodiment, this disclosure relates to a method for detecting the presence of pancreatic cancer cells in a subject or for diagnosing pancreatic cancer in a subject, the method comprising the following steps:

[0307] a. Provide the subject with pancreatic tissue suspected of containing cancer;

[0308] b. Analyze the presence of the following in the sample:

[0309] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0310] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0311] c. Determine the expression level of the antigen described in i) or the polynucleotide transcript described in ii), and

[0312] d. The expression levels determined in c. will be compared with control levels, wherein the control levels are the average expression levels of i) the antigen or ii) the polynucleotide transcripts observed in healthy and / or benign pancreatic tissue; and

[0313] Wherein i) the expression level of the antigen and / or ii) the polynucleotide transcript is higher than the control level, indicating the presence of pancreatic cancer, thereby detecting and / or diagnosing pancreatic cancer in the subject.

[0314] In one embodiment, this disclosure relates to a method for detecting the presence of sarcoma cells in a subject or for diagnosing sarcoma in a subject, said method comprising the following steps:

[0315] a. Provide the subject with connective tissue suspected of containing cancer;

[0316] b. Analyze the presence of the following in the sample:

[0317] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0318] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0319] c. Determine the expression level of the antigen described in i) or the polynucleotide transcript described in ii), and

[0320] d. The expression levels determined in c. will be compared with control levels, wherein the control levels are the average expression levels of i) the antigen or ii) the polynucleotide transcripts observed in healthy and / or benign connective tissue; and

[0321] Wherein i) the expression level of the antigen and / or ii) the polynucleotide transcript is higher than the control level, indicating the presence of sarcoma, thereby detecting and / or diagnosing sarcoma in the subject.

[0322] The inventors of this invention have discovered that the expression level of integrin α10 can be used to differentiate subtypes of invasive breast cancer, such as triple-negative breast cancer.

[0323] Therefore, one aspect of this disclosure relates to a method for classifying triple-negative breast cancer tumor samples from a subject, the method comprising:

[0324] a. Provide a breast tissue sample from the subject;

[0325] b. Isolate breast cancer cells characterized as ER-negative, PR-negative, and HER2-negative;

[0326] c. Determine the expression levels of the following in the isolated cells:

[0327] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0328] ii. A multinucleotide transcript or fragment or variant thereof encoding an integrin α10 polypeptide, and

[0329] d. The expression level determined in c. will be compared with a control level, wherein the control level is the average expression level of the antigen containing the integrin α10 polypeptide observed in healthy and / or benign breast tissue;

[0330] Wherein i) the expression level of the antigen and / or ii) the polynucleotide transcript in the breast cancer cells is higher than the control level, and the expression status of ER negative, PR negative and HER2 negative indicates basal-like triple-negative breast cancer or luminal triple-negative breast cancer, thereby classifying the triple-negative breast cancer tumor sample as belonging to basal-like triple-negative breast cancer tumor or luminal triple-negative breast cancer tumor.

[0331] In an alternative embodiment, the luminal triple-negative breast cancer is luminal androgen receptor triple-negative breast cancer.

[0332] In one embodiment, the method for classifying triple-negative breast cancer tumor samples according to this disclosure further includes analyzing the expression levels of tight junction proteins in the samples, wherein i) the expression levels of the antigen and / or ii) the polynucleotide transcript in the breast cancer cells are higher than control levels, and the expression status of tight junction proteins that are ER-negative, PR-negative, HER2-negative and at low levels indicates basal-like triple-negative breast cancer, thereby classifying the triple-negative breast cancer tumor sample as belonging to basal-like triple-negative breast cancer tumors.

[0333] Low tight junction protein (LDTT) breast tumors are a subtype of triple-negative breast tumors. These tumors exhibit low expression of many tight junction protein genes, including tight junction protein 3, tight junction protein 4, and tight junction protein 7. Other important characteristics of LDTT tumors are that they almost always have strong immune cell infiltration, and they also exhibit stem cell characteristics and epithelial-mesenchymal transition (EMT) features.

[0334] In one embodiment, the method of classifying triple-negative breast cancer tumor samples according to this disclosure further includes classifying the triple-negative breast cancer tumor sample as belonging to type 2 basal-like triple-negative breast cancer tumors when i) the expression level of the integrin α10 antigen and / or ii) the expression level of the polynucleotide transcript is higher than the control level.

[0335] In one embodiment, the method for classifying triple-negative breast cancer tumor samples according to this disclosure further includes determining the expression of one or more polypeptides selected from cytokeratin 7, cytokeratin 8, cytokeratin 18 and cytokeratin 19, and wherein the expression of integrin α10 and one or more polypeptides selected from cytokeratin 7, cytokeratin 8, cytokeratin 18 and cytokeratin 19 indicates luminal triple-negative breast cancer.

[0336] In one embodiment, the method for classifying triple-negative breast cancer tumor samples according to this disclosure further includes determining the expression of one or more polypeptides selected from cytokeratin 5 / 6, cytokeratin 14, cytokeratin 17, p63, EGFR, and c-kit / CD117, and the expression of integrin α10 and one or more polypeptides selected from cytokeratin 5 / 6, cytokeratin 14, cytokeratin 17, p63, EGFR, 34BE12, and c-kit / CD117 indicates basal-like triple-negative breast cancer.

[0337] The inventors of this invention have unexpectedly discovered that high levels of integrin α10 expression in cells belonging to the invasive cancer forms as defined herein are directly associated with and indicate poor prognosis.

[0338] Therefore, one aspect of this disclosure relates to a method for determining the prognosis of a subject's invasive cancer form, said invasive cancer form being selected from invasive breast cancer, invasive lung cancer, invasive prostate cancer, invasive pancreatic cancer, and invasive sarcoma, or a metastatic tumor of any of said invasive cancer forms, said method comprising:

[0339] a. Provide the subject with cancerous tumor tissue;

[0340] b. Analyze the presence of the following in the sample:

[0341] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0342] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0343] c. Determine the expression levels of i) integrin α10 antigen and / or ii) polynucleotide transcripts.

[0344] d. The expression levels determined in c. will be compared with control levels, wherein the control levels are the average expression levels of the integrin α10i) antigen and / or ii) polynucleotide transcripts observed in healthy and / or benign tissues of the same tissue type as the sample;

[0345] e. When the expression levels of i) the antigen and / or ii) the polynucleotide transcript are higher than the control levels, a poor prognosis is determined for the invasive cancer form.

[0346] Another aspect of this disclosure relates to a method for determining the prognosis of a subject's aggressive cancer form, the method comprising:

[0347] a. Provide the subject with cancerous tumor tissue;

[0348] b. Analyze the presence of one or more cells with cancerous morphology in the tissue;

[0349] c. Analyze the presence of the following in the sample:

[0350] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0351] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0352] d. Optionally determine the expression levels of the i) integrin α10 antigen and / or ii) polynucleotide transcripts.

[0353] e. When one or more cells with a cancerous morphology are present in the tissue in combination with the expression of the integrin α10i) antigen and / or ii) polynucleotide transcript, a poor prognosis is determined for the aggressive cancer form.

[0354] The invasive cancer form is selected from invasive breast cancer, invasive lung cancer, invasive prostate cancer, invasive pancreatic cancer, and invasive sarcoma, or a metastatic tumor of any of the invasive cancer forms.

[0355] The methods used to determine the prognosis of a subject’s invasive cancer form can be applied to any of the invasive cancer forms described herein, see, for example, the “Clinical Symptoms” section.

[0356] In one embodiment, this disclosure relates to a method for determining the prognosis of a subject’s invasive cancer form, wherein the prognosis is overall survival or recurrence-free survival.

[0357] In one embodiment of this disclosure, the method disclosed herein targets an antigen expressed on the surface of the cell that contains an integrin α10 polypeptide or a fragment thereof.

[0358] The methods described in this disclosure can be performed in vivo or in vitro.

[0359] In one embodiment, the method for detecting aggressive cancer cells in a subject, as disclosed herein, is performed in vivo. In another embodiment, the method for diagnosing an aggressive form of cancer in a subject, as disclosed herein, is performed in vitro, and the tissue is a tissue sample obtained from the subject.

[0360] In one embodiment, the method for diagnosing an aggressive form of cancer in a subject, as disclosed herein, is performed in vivo. In another embodiment, the method for diagnosing an aggressive form of cancer in a subject, as disclosed herein, is performed in vitro, and the tissue is a tissue sample obtained from the subject.

[0361] In one embodiment, the method for determining the prognosis of an aggressive cancer in a subject, as disclosed herein, is performed in vivo. In another embodiment, the method for determining the prognosis of an aggressive cancer in a subject, as disclosed herein, is performed in vitro, and the tissue is a tissue sample obtained from the subject.

[0362] In one embodiment of this disclosure, the following are analyzed to indicate the presence of the sample:

[0363] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0364] ii. The step of encoding a polynucleotide transcript or a fragment or variant thereof encoding an integrin α10 polypeptide includes imaging the tissue and / or tissue sample.

[0365] In one embodiment of this disclosure, the step of determining the expression levels of i) integrin α10 antigen and / or ii) polynucleotide transcripts includes imaging the tissue and / or tissue sample.

[0366] For example, imaging can be performed by administering to the subject and / or to the tissue sample a labeled portion of an antigen that can bind to an antigen containing an integrin α10 polypeptide or a fragment thereof. For example, imaging can be performed by administering to the subject and / or to the tissue sample a labeled anti-integrin α10 antibody as defined herein.

[0367] Antibodies against integrin α10 peptide

[0368] In one embodiment, the composition of this disclosure for diagnosing and / or treating and / or preventing invasive cancer forms comprises an anti-integrin α10 specific antibody that binds to the integrin α10 polypeptide during an immune response. Preferably, the antibody binds to the extracellular domain of the integrin α10 polypeptide; however, in some embodiments, the anti-integrin α10 antibody has overlapping specificity for the entire integrin α10β1 heterodimer complex. This could, for example, mean that the antibody of the present invention binds to an epitope covering both the α10 and β1 polypeptides.

[0369] The antibodies and their functional equivalents can be produced by any suitable method known to those skilled in the art.

[0370] In one embodiment, the antibody of the present invention is produced in a hybridoma cell line (e.g., mAb 365 hybridoma cell line, deposited on February 4, 2003, at the German Collection of Microbial Cultures, accession number GmbH, DSM ACC2583) to produce an antibody that binds to the extracellular integrin α10 domain. To generate the hybridoma, a gene knockout mouse of integrin α10β1 can be used. The knockout mouse is described in WO 03 / 101497, which is incorporated herein by reference.

[0371] In one embodiment, the antibody of the present invention is

[0372] a) Monoclonal antibodies produced using hybridoma cell lines deposited at the German Culture Collection (DSM-ACC2583); or

[0373] b) Antibodies that competitively bind to the same epitope as those produced by monoclonal antibodies from hybridomas deposited at the German Collection of Microbial Cultures (accession number DSM ACC2583); or

[0374] c)a) or b) fragments, wherein the fragments are capable of specifically binding to the extracellular I domain of the integrin α10 polypeptide chain.

[0375] In another embodiment, the anti-integrin α10 antibody of the present invention has been identified by an antibody panning process.

[0376] The antibody of the present invention, defined by the amino acid sequence of SEQ ID NO:4-11, is hereby designated as Th101. The Th101 antibody is disclosed herein by way of its six CDRs (complementarity-determining regions) (SEQ ID NO:4 to 9), its heavy chain variable region (SEQ ID NO:10), and its light chain variable region (SEQ ID NO:11). The identification and production of the Th101 antibody used in the examples are described in WO 08 / 075038, which is incorporated herein by reference.

[0377] In one embodiment, the anti-integrin α10 antibody of the present invention comprises:

[0378] Heavy chain variable region, the heavy chain variable region includes

[0379] a) An amino acid sequence comprising or consisting of SEQ ID NO:4;

[0380] b) A CDR-H2 consisting of or containing the amino acid sequence of SEQ ID NO:5; and

[0381] c) An amino acid sequence comprising or consisting of SEQ ID NO:6;

[0382] and / or

[0383] Light chain variable region, the light chain variable region comprising

[0384] d) Containing the amino acid sequence of SEQ ID NO:7 or CDR-L1 composed of it;

[0385] e) CDR-L2 containing or consisting of the amino acid sequence of SEQ ID NO:8; and

[0386] f) Contains the amino acid sequence of SEQ ID NO:9 or is composed of CDR-L3.

[0387] Or a variant of any one of SEQ ID NO:4 to 9, wherein any one amino acid has been changed to another amino acid, provided that no more than three amino acids have been changed in this way, for example, two or one amino acid have been changed in this way.

[0388] In another embodiment, the anti-integrin α10 antibody of the present invention comprises a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:10.

[0389] In another embodiment, the anti-integrin α10 antibody of the present invention comprises a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:11.

[0390] The integrin α10 antibody may be administered to the mammal multiple times, such as twice, for example three times, such as three to five times, for example five to ten times, such as ten to twenty times, for example twenty to fifty times, such as more than fifty times. It is also possible to administer different integrin α10 antigens to the same mammal simultaneously or sequentially in any order.

[0391] Generally, the integrin α10 antibody will be in the form of an aqueous solution or suspension before administration. Furthermore, the integrin α10 antigen can be mixed with one or more other compounds. For example, the integrin α10 antigen can be mixed with one or more suitable adjuvants and / or with one or more carriers.

[0392] An adjuvant is any substance whose mixing with the applied antigen increases or otherwise alters the immune response to said antigen. Suitable adjuvants are well known to those skilled in the art.

[0393] A carrier is a scaffold structure, such as a polypeptide or polysaccharide, capable of associating with an antigen. The carrier can exist independently of the adjuvant. Suitable carriers are well known to those skilled in the art.

[0394] Methods for preparing monoclonal antibodies, mixtures of monoclonal antibodies, or polyclonal antibodies are known in the art and are described, for example, in Ed Harlow and David Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 1988.

[0395] In one embodiment, the anti-integrin α10 antibody of the present invention is an antibody capable of inhibiting the biological and functional activity of integrin α10 polypeptide.

[0396] In another embodiment, the anti-integrin α10 antibody is administered as part of an antibody-drug complex (ADC). In the ADC, the antibody is linked to a moiety, such as a drug or toxin that improves the disease. After specifically binding to the integrin α10 polypeptide, the ADC is internalized into the cell, thereby delivering the moiety into the cell.

[0397] In one embodiment, the antibody included in the composition of this disclosure for diagnosing and / or treating invasive forms of cancer has an isotype selected from IgA, IgD, IgG, IgE, and IgM. In a further embodiment, the antibody is an IgG isotype, such as IgG1, IgG2 (e.g., IgG2a), IgG3, and IgG4.

[0398] In one embodiment, the antibody that specifically binds to the integrin α10 polypeptide according to this disclosure is a single-chain antibody.

[0399] This disclosure considers monoclonal and polyclonal antibodies and fragments thereof, antigen-binding fragments and recombinant proteins capable of binding integrin α10 peptide.

[0400] In one embodiment, the antibody that specifically binds to the integrin α10 polypeptide for diagnosis and / or treatment, according to this disclosure, is a polyclonal antibody.

[0401] In one embodiment, the antibody that specifically binds to the integrin α10 polypeptide according to this disclosure is a humanized antibody or a human antibody.

[0402] In one embodiment, the antibody that specifically binds to the integrin α10 polypeptide according to this disclosure is a mouse antibody.

[0403] In one embodiment, the antibody that specifically binds to the integrin α10 polypeptide according to this disclosure is a monoclonal antibody.

[0404] In one embodiment, the antibody that specifically binds to the integrin α10 polypeptide according to this disclosure is a polyclonal antibody.

[0405] In one embodiment, the antibody included in the compositions of this disclosure for diagnosing and / or treating aggressive forms of cancer is an antibody fragment. An antigen-binding fragment of an antibody is a fragment that retains the antibody's ability to specifically bind to an antigen. Examples of antibody fragments of the present invention include antibody fragments selected from: Fab fragments, Fab' fragments, F(ab')2 fragments, and Fv fragments, such as single-chain variable fragments (scFv) and single-domain antibodies.

[0406] The antibodies included in the compositions for diagnosing and / or treating aggressive forms of cancer disclosed in this document may also be chimeric antibodies, i.e., antibodies comprising regions derived from different species. Such chimeric antibodies may, for example, comprise a variable region from one animal species and a constant region from another animal species. For example, a chimeric antibody may be an antibody having a variable region derived from a mouse monoclonal antibody and a human constant region. Such antibodies may also be referred to as humanized antibodies. For example, a chimeric humanized antibody may be entirely human.

[0407] In one embodiment, the antibody included in the compositions of this disclosure for diagnosing and / or treating invasive forms of cancer is a xenospecific antibody (such as a bispecific antibody), which is a protein or polypeptide containing two distinct antigen-binding sites with different specificities. For example, the bispecific antibody can recognize and bind to (a) one epitope on integrin α10 and (b) another epitope on integrin α10. Thus, it can recognize and bind to two different epitopes within the same antigen. The term "xenospecific antibody" is intended to include any protein or polypeptide containing more than two distinct antigen-binding sites with different specificities. Therefore, the present invention includes, but is not limited to, bispecific, trispecific, tetraspecific, and other multispecific antibodies against integrin α10 polypeptides.

[0408] In some embodiments, antibodies or fragments thereof that specifically bind to the integrin α10 polypeptide according to this disclosure may be conjugated to portions, such as additional portions. Such conjugation can improve and facilitate both the treatment and diagnosis of aggressive cancer forms.

[0409] In one embodiment, the antibody is covalently bound to a detectable portion, such as a fluorophore, enzyme, radiotracer, or radioisotope. The integrin α10 antigen can also be detected by detecting peptides, proteins, or polypeptides other than the integrin α10 polypeptide, wherein these other peptides, proteins, or polypeptides are specifically bound to the integrin α10 antigen. In one embodiment, the peptide, protein, or polypeptide is linked to an enzyme, fluorophore, or radiotracer. The radiotracer may be, for example, selected from positron emitters or gamma emitters. The conjugation of the antibody to the detectable portion facilitates and improves the detection of the antibody, which in turn facilitates the detection of cells expressing integrin α10 in a sample, and thus facilitates the diagnosis of invasive cancer forms.

[0410] Those skilled in the art can select standard laboratory equipment for detecting anti-integrin α10 antibodies based on the condition and physical state of the sample.

[0411] In one implementation, those skilled in the art will use flow cytometry (such as fluorescence activated cell sorting (FACS)) for the detection step.

[0412] Typical immunological methods well known in the art include, but are not limited to, Western blotting, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunohistochemistry (IHC), immunofluorescence assay (IF), and fluorescence in situ hybridization (FISH).

[0413] Integrin α10 can be detected using methods well-known in the fields of detection and imaging (such as clinical imaging), such as conventional fluorescence microscopy, confocal microscopy, two-photon microscopy, stimulated emission loss (STED), etc.

[0414] In one embodiment, the anti-integrin α10 antibody is a functional blocking antibody, such as an antibody capable of inhibiting and / or blocking the migration and / or proliferation of cells expressing integrin α10.

[0415] Molecular probes for detecting integrin α10

[0416] The presence of integrin α10 antigen or polynucleotide encoding integrin α10 in the biological sample can also be analyzed by using a molecular probe (protein or polynucleotide) capable of binding or hybridizing to integrin α10 mRNA, cDNA or protein to detect its expression in the biological sample, or by using PCR, preferably Q-PCR.

[0417] In one embodiment, the integrin α10-specific polynucleotide probe is attached to a detectable portion optionally capable of emitting photons. Using this embodiment, a light source capable of exciting the detectable portion (e.g., a fluorophore) can be used to illuminate a subject for diagnosis or research. Methods for detecting photons include, but are not limited to, PET scans and SPECT scans.

[0418] In some embodiments, the detectable portion is selected from fluorophores, enzymes, or radiotracers.

[0419] Polynucleotide transcripts can also be detected using PCR, preferably Q-PCR.

[0420] In a further embodiment, the presence of integrin α10 in a biological sample is detected by using an integrin α10 nucleic acid probe that binds to integrin α10 RNA or cDNA in a hybridization reaction.

[0421] Exemplary nucleic acid integrin-α10 specific targeting components include DNA probes, antisense RNA, or RNAi, such as microRNA, short interfering RNA (siRNA), and short hairpin RNA (shRNA).

[0422] Typical methods known in the art for detecting nucleic acids include, but are not limited to, RNA blotting, DNA blotting, polymerase chain reaction (PCR), microarrays, and in situ hybridization.

[0423] In a further embodiment, the presence of integrin α10 in a biological sample is detected by using an integrin α10-binding peptide or protein. Such peptides or proteins can be recombinantly manufactured, chemically synthesized, or purified from natural sources.

[0424] In a further embodiment, the presence of integrin α10 in a biological sample is detected in vivo by using an integrin α10-specific antibody, or an integrin α10-binding peptide or protein, or an integrin α10 nucleic acid probe that binds to integrin α10 RNA or cDNA in a hybridization reaction.

[0425] Typical methods for detecting cell surface antigens and polynucleotides in vivo are well known in the art, including but not limited to positron emission tomography (PET), x-ray computed tomography (CT), magnetic resonance imaging (MRI) and functional magnetic resonance imaging (fMRI), ultrasound, and single-photon emission computed tomography (SPECT). In particular, cell surface antigens can be imaged in vivo using immunolabeling with radiotracers that bind to antibodies or other specific binding proteins.

[0426] Preferably, the antibodies used for in vivo imaging are antibody fragments (such as Fab fragments) and single-chain antibodies because they are small in size and do not have effector functions.

[0427] project

[0428] 1. A composition comprising:

[0429] a. An antibody or its antigen-binding fragment that specifically binds to integrin α10 polypeptide, or

[0430] b. A polynucleotide or fragment or variant thereof that specifically binds to a polynucleotide transcript encoding an integrin α10 polypeptide.

[0431] Used for the treatment and / or prevention of cancers selected from the following forms: breast cancer, lung cancer, prostate cancer, pancreatic cancer, and sarcoma, or metastatic tumors of any of these cancer forms.

[0432] 2. The composition for the purpose according to any one of the foregoing items, wherein the breast cancer is selected from triple-negative breast cancer and inflammatory breast cancer.

[0433] 3. The composition for the purpose according to any one of the preceding items, wherein the triple-negative breast cancer is selected from basal-like 1 breast cancer, basal-like 2 breast cancer, low tight junction protein breast cancer, metaplastic breast cancer (MBC), interferon-rich breast cancer, immunomodulatory breast cancer, mesenchymal breast cancer, mesenchymal stem cell-like breast cancer, luminal androgen receptor breast cancer, and unstable breast cancer.

[0434] 4. The composition for the purpose according to any one of the preceding items, wherein the lung cancer is selected from squamous cell lung cancer, lung adenocarcinoma, large cell lung cancer and small cell lung cancer.

[0435] 5. The composition for the use according to any one of the preceding items, wherein the prostate cancer is small cell neuroendocrine carcinoma (SCNC).

[0436] 6. The composition for the purpose according to any one of the preceding items, wherein the pancreatic cancer is a neuroendocrine tumor.

[0437] 7. The composition for the purpose according to any one of the preceding items, wherein the pancreatic cancer and / or neuroendocrine tumor is grade I, II or III pancreatic cancer.

[0438] 8. The composition for the use according to any one of the preceding items, wherein the sarcoma is selected from undifferentiated pleomorphic sarcoma, myxofibrosarcoma, dedifferentiated liposarcoma, atypical lipoma, myxoinflammatory fibroblastic sarcoma, low-grade fibromyxoid sarcoma, sclerosing epithelioid fibrosarcoma, pseudomyogenic angioendothelioma, and mesenchymal chondrosarcoma.

[0439] 9. The composition for the purpose according to any one of the preceding items, wherein the integrin α10 polypeptide is expressed on the surface of malignant cells and / or tumor-associated cells.

[0440] 10. The composition for the purpose according to any one of the preceding items, wherein the antibody is a monoclonal antibody, a polyclonal antibody, a chimeric antibody, a single-chain antibody, or a fragment thereof.

[0441] 11. The composition for the purpose according to any one of the preceding items, wherein the antibody is a non-human antibody, a chimeric antibody, a bispecific antibody, a humanized antibody, or a human antibody.

[0442] 12. The composition for the purpose according to any one of the preceding items, wherein the antibody is a mouse monoclonal antibody.

[0443] 13. The composition for the said use according to any one of the preceding items, wherein the antibody is a human monoclonal antibody.

[0444] 14. The composition for the purpose according to any one of the preceding items, wherein the antibody has an isotype selected from IgA, IgD, IgG, IgE and IgM.

[0445] 15. The composition for the said use according to any one of the preceding items, wherein said antibody is:

[0446] a) Monoclonal antibodies produced using hybridoma cell lines deposited at the German Culture Collection (DSM-ACC2583); or

[0447] b) Antibodies that competitively bind to the same epitope as those produced by monoclonal antibodies from hybridomas deposited at the German Collection of Microbial Cultures (accession number DSM ACC2583); or

[0448] c)a) or b) fragments, wherein the fragments are capable of specifically binding to the extracellular I domain of the integrin α10 polypeptide chain.

[0449] 16. A composition for the said use according to any one of the preceding items, wherein the antibody or antigen-binding fragment thereof comprises:

[0450] Heavy chain variable region, the heavy chain variable region includes

[0451] a) An amino acid sequence comprising or consisting of SEQ ID NO:4;

[0452] b) A CDR-H2 consisting of or containing the amino acid sequence of SEQ ID NO:5; and

[0453] c) An amino acid sequence comprising or consisting of SEQ ID NO:6;

[0454] and / or

[0455] Light chain variable region, the light chain variable region comprising

[0456] d) Containing the amino acid sequence of SEQ ID NO:7 or CDR-L1 composed of it;

[0457] e) CDR-L2 containing or consisting of the amino acid sequence of SEQ ID NO:8; and

[0458] f) Contains the amino acid sequence of SEQ ID NO:9 or is composed of CDR-L3.

[0459] Or a variant of any one of SEQ ID NO:4 to 9, wherein any one amino acid has been changed to another amino acid, provided that no more than three amino acids have been changed in this way, for example, two or one amino acid have been changed in this way.

[0460] 17. A composition for the purpose according to any one of the preceding items, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:10.

[0461] 18. A composition for the purpose according to any one of the preceding items, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:11.

[0462] 19. A composition for the purpose according to any one of the preceding items, wherein the antibody or its antigen-binding fragment and / or the polynucleotide or its fragment or variant is conjugated to an additional portion.

[0463] 20. A composition for the purpose according to any one of the preceding items, wherein the additional portion comprises a detectable portion, such as a detectable portion selected from: fluorophores, enzymes, and radiotracers or radioisotopes.

[0464] 21. The composition for the stated use according to any one of the preceding items, wherein the additional portion comprises a cytotoxic portion.

[0465] 22. The composition for the stated use according to any one of the preceding items, wherein the cytotoxic portion is selected from toxins, chemotherapeutic agents, and radiopharmaceuticals or combinations thereof.

[0466] 23. The composition for the stated use according to any one of the preceding items, wherein the cytotoxic portion is a toxin.

[0467] 24. The composition for the stated use according to any one of the preceding items, wherein the toxin is selected from microtubule toxins, DNA toxins, and transcription toxins.

[0468] 25. The composition for the stated use according to any one of the preceding items, wherein the microtubule toxin is selected from auristatin-based toxins, maytansine-based toxins, microtubule-lysin-based toxins, and eribulin.

[0469] 26. The composition for the stated use according to any one of the preceding items, wherein the DNA toxin is selected from DNA major groove binding agents, DNA minor groove binding alkylating agents, DNA alkylating agents, and DNA cleaving agents.

[0470] 27. The composition for the stated use according to any one of the preceding items, wherein the DNA toxin is selected from pyrrolobenzodiazepine (PBD), pyroxine, pyroxine analogues, indoline benzodiazepine, cazidromycin, irinotecan, and ethanotecan derivatives.

[0471] 28. The composition for the stated use according to any one of the preceding items, wherein the transcription toxin is an RNA polymerase II inhibitor.

[0472] 29. The composition for the stated use according to any one of the preceding items, wherein the transcription toxin is selected from doxorubicin, doxorubicin derivatives and amatoxins.

[0473] 30. The composition for the stated use according to any one of the preceding items, wherein the transcription toxin is selected from Shiga toxin and Shiga-like toxin, type I ribosome inactivating protein, type II ribosome inactivating protein and saponin or a combination thereof.

[0474] 31. The composition for the purpose according to any one of the preceding items, wherein the type I ribosome inactivating protein is trichosanthes pollen protein and / or loofah seed protein.

[0475] 32. The composition for the stated use according to any one of the preceding items, wherein the type II ribosome inactivating protein is ricin, lectin, and / or absinthecin.

[0476] 33. The composition for the stated use according to any one of the preceding items, wherein the additional portion comprises a biological response modifier.

[0477] 34. The composition for the stated use according to any one of the preceding items, wherein the biological response modifier is a cytokine, lymphokine, interferon, or a combination thereof.

[0478] 35. The composition for the stated use according to any one of the preceding items, wherein the chemotherapeutic agent is an alkylating agent, an antimetabolite, an antimicrotubule agent, a topoisomerase inhibitor, or a cytotoxic antibiotic.

[0479] 36. The composition for the stated use according to any one of the preceding items, wherein the chemotherapeutic agent is selected from anthracyclines, taxanes, and platinum agents.

[0480] 37. The composition for the stated use according to any one of the preceding items, wherein the chemotherapeutic agent is selected from cisplatin, paclitaxel, albumin-bound paclitaxel, docetaxel, cyclophosphamide, eribulin, epirubicin, doxorubicin, carboplatin, gemcitabine, bleomycin, fluorouracil, cyclophosphamide, vinorelbine, capecitabine, ixaprilone, and ixaprilone, or combinations thereof.

[0481] 38. A composition for the stated use according to any one of the preceding items, wherein the composition further comprises at least one pharmaceutically acceptable diluent, carrier, or excipient.

[0482] 39. The composition for the use according to any one of the preceding items, wherein the integrin α10 polypeptide is a naturally occurring variant of the integrin α10 polypeptide, an isotype of the integrin α10 polypeptide, or a splice variant of the integrin α10 polypeptide.

[0483] 40. A composition for the purpose according to any one of the preceding items, wherein the antibody and / or the polynucleotide is capable of inducing cell death and / or inhibiting cell growth and / or inhibiting cell proliferation and / or inhibiting cell migration in cells expressing integrin α10 polypeptide and / or polynucleotide transcript.

[0484] 41. The composition for the stated use according to any one of the preceding items, wherein the cells are malignant cells and / or tumor-associated cells.

[0485] 42. The composition for the purpose according to any one of the preceding items, wherein the malignant cell or tumor-associated cell is a cancer-associated fibroblast (CAF), stromal cell, stem cell, and / or stem cell-like cell.

[0486] 43. The composition for the purpose according to any one of the preceding items, wherein the integrin α10 polypeptide is part of the integrin α10β1 heterodimer.

[0487] 44. The composition for the stated purpose according to any one of the preceding items, wherein the treatment is preventative, ameliorative, or curative.

[0488] 45. The composition for the use according to any one of the preceding items, wherein the treatment is initiated after integrin α10 polypeptide and / or polynucleotide transcripts are detected in cancer cells in the tumor of the subject.

[0489] 46. ​​The composition for the stated purpose according to any one of the preceding items, wherein the composition is administered to an individual in need in combination with radiotherapy and / or surgical resection of cancer.

[0490] 47. The composition for the stated purpose according to any one of the preceding items, wherein the composition is administered to an individual in need prior to radiotherapy and / or surgical removal of cancer.

[0491] 48. The composition for the stated purpose according to any one of the preceding items, wherein the composition is administered to an individual in need after radiotherapy and / or surgical resection of cancer.

[0492] 49. A composition comprising:

[0493] a) An antibody or its antigen-binding fragment that specifically binds to integrin α10 polypeptide, or

[0494] b) A polynucleotide or a fragment or variant thereof that specifically binds to a polynucleotide transcript encoding an integrin α10 polypeptide.

[0495] Used to diagnose metastatic tumors of any of the following cancer forms: breast cancer, lung cancer, prostate cancer, pancreatic cancer, and sarcoma, or any of the aforementioned cancer forms.

[0496] 50. The composition according to item 49, wherein the cancer comprises cells exhibiting equal or higher levels of i) the integrin α10 antigen or ii) the polynucleotide transcript observed in the same type of healthy and / or benign tissue.

[0497] 51. The composition according to item 49, wherein the cancer comprises cells exhibiting equal or higher levels of i) the integrin α10 antigen or ii) the polynucleotide transcript observed in cancer types of the same tissue type that are less aggressive compared to the diagnosed cancer type.

[0498] 52. The composition according to item 49, wherein the cancer comprises cells exhibiting equal or higher expression levels of i) the integrin α10 antigen or ii) the polynucleotide transcript compared to a reference cell line.

[0499] 53. The composition according to item 52, wherein the reference cell line is derived from healthy tissue.

[0500] 54. The composition according to any one of items 52 to 53, wherein the reference cell line is selected from reference cell lines derived from healthy breast tissue, reference cell lines derived from healthy prostate tissue, reference cell lines derived from healthy lung tissue, reference cell lines derived from healthy pancreatic tissue, and reference cell lines derived from healthy connective tissue.

[0501] 55. The composition according to any one of items 52 to 54, wherein the reference cell line is cell line 184A1 derived from healthy breast cancer tissue.

[0502] 56. The composition according to item 52, wherein the reference cell line is derived from a cancer type of the same tissue type that is less aggressive than the diagnosed cancer type.

[0503] 57. The composition according to item 56, wherein the reference cell line is selected from reference cell lines derived from less aggressive breast cancer, reference cell lines derived from less aggressive prostate cancer, reference cell lines derived from less aggressive lung cancer, reference cell lines derived from less aggressive pancreatic cancer, and reference cell lines derived from less aggressive sarcoma.

[0504] 58. The composition according to any one of items 56 to 57, wherein the reference cell line is selected from the following: HCC1428, a cell line derived from less aggressive breast cancer; T47D, a cell line derived from less aggressive breast cancer; 22Rv1, a cell line derived from less aggressive prostate cancer; DU145, a cell line derived from less aggressive prostate cancer; BxPC-3, a cell line derived from less aggressive pancreatic cancer; and AsPC-1, a cell line derived from less aggressive pancreatic cancer.

[0505] 59. A method of treating a form of cancer, wherein the form of cancer is selected from breast cancer, lung cancer, prostate cancer, pancreatic cancer, and sarcoma, or wherein the form of cancer is a metastatic tumor, the method comprising administering to a subject in need a pharmaceutically effective amount of a composition, the composition comprising:

[0506] a. An antibody or fragment thereof that specifically binds to integrin α10 polypeptide, or

[0507] b. A polynucleotide or a fragment or variant thereof that specifically binds to a polynucleotide transcript encoding an integrin α10 polypeptide.

[0508] 60. A method for detecting cancer cells in a subject, the method comprising the following steps:

[0509] a. Provide the subject with tissue suspected of containing cancer cells;

[0510] b. Analyze the presence of the following in the described organization:

[0511] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0512] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0513] c. Determine the expression levels of i) integrin α10 antigen and / or ii) polynucleotide transcripts, and

[0514] d. The expression levels determined in c. will be compared with control levels.

[0515] The control level is the average expression level of i) the antigen or ii) the polynucleotide transcript observed in healthy and / or benign cells of the same tissue type as the isolated sample.

[0516] Wherein i) the expression level of the antigen and / or ii) the polynucleotide transcript being higher than the control level indicates the presence of a form of cancer in the subject, and wherein the form of cancer is selected from breast cancer, lung cancer, prostate cancer, pancreatic cancer and sarcoma or a metastatic tumor of any of the forms of cancer.

[0517] 61. A method for detecting cancer cells in a subject, the method comprising the following steps:

[0518] a. Provide the subject with tissue suspected of containing cancer cells;

[0519] b. Analyze the presence of one or more cells with cancerous morphology in the tissue.

[0520] c. Analyze the presence of the following in the described tissue:

[0521] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0522] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0523] d. Optionally determine the expression levels of the i) integrin α10 antigen and / or ii) polynucleotide transcripts.

[0524] The presence of one or more cells with a cancerous morphology, combined with the expression of integrin α10i) antigen and / or ii) polynucleotide transcripts, indicates the presence of a cancerous morphology in the subject, wherein the cancerous morphology is selected from breast cancer, lung cancer, prostate cancer, pancreatic cancer, and sarcoma, or a metastatic tumor of any of the cancerous morphologies.

[0525] 62. A method for diagnosing a form of cancer in a subject, the method comprising the steps of:

[0526] a. Provide the subject with tissue suspected of containing cancer cells;

[0527] b. Analyze the presence of the following in the described organization:

[0528] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0529] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0530] c. Determine the expression levels of i) integrin α10 antigen and / or ii) polynucleotide transcripts, and

[0531] d. The expression levels determined in c. will be compared with control levels, wherein the control levels are the average expression levels of i) the antigen or ii) the polynucleotide transcript observed in the same type of healthy and / or benign tissue; and

[0532] Wherein i) the expression level of the antigen and / or ii) the polynucleotide transcript is higher than the control level, indicating the presence of a cancer form in the sample, wherein the cancer form is selected from breast cancer, lung cancer, prostate cancer, pancreatic cancer and sarcoma or a metastatic tumor of any of the cancer forms, thereby diagnosing the cancer form of the subject.

[0533] 63. A method for diagnosing a form of cancer in a subject, the method comprising the steps of:

[0534] a. Provide the subject with tissue suspected of containing cancer cells;

[0535] b. Analyze the presence of one or more cells with cancerous morphology in the tissue;

[0536] c. Analyze the presence of the following in the sample:

[0537] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0538] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0539] d. Optionally determine the expression levels of the i) integrin α10 antigen and / or ii) polynucleotide transcripts, and

[0540] The presence of one or more cells with a cancerous morphology, in combination with the expression of integrin α10i) antigen and / or ii) polynucleotide transcripts, indicates the presence of a cancerous form in the sample, wherein the cancerous form is selected from breast cancer, lung cancer, prostate cancer, pancreatic cancer, and sarcoma, or a metastatic tumor of any of the cancerous forms, thereby diagnosing the cancerous form of the subject.

[0541] 64. The method according to any one of items 60 to 63, wherein step b. further comprises administering to the subject the composition or a fragment thereof according to any one of items 1 to 58.

[0542] 65. The method according to any one of items 60 to 64, the method further comprising morphologically characterizing the sample as containing cancer cells belonging to a cancerous form, wherein the cancerous form is selected from breast cancer, lung cancer, prostate cancer, pancreatic cancer, and sarcoma, or a metastatic tumor of any one of said cancerous forms.

[0543] 66. The method according to any one of items 60 to 65, wherein the method is used to detect the presence of breast cancer in a subject or to diagnose breast cancer in a subject, wherein the breast cancer is a form of breast cancer selected from triple-negative breast cancer and inflammatory breast cancer, wherein the method includes comparing the expression level of the integrin α10 polypeptide determined in b. with a control level, wherein the control level is the average expression level of i) the antigen or ii) the polynucleotide transcript observed in healthy and / or benign breast tissue cells, and a higher level of the antigen containing the integrin α10 polypeptide than the control level indicates the presence of breast cancer in the subject.

[0544] 67. The method according to any one of items 60 to 65, wherein the method is used to detect the presence of lung cancer in a subject or to diagnose lung cancer in a subject, wherein the method includes comparing the expression level of the integrin α10 polypeptide determined in b. with a control level, wherein the control level is the average expression level of i) the antigen or ii) the polynucleotide transcript observed in non-cancerous lung cells, and a higher level of the antigen containing the integrin α10 polypeptide than the control level indicates the presence of lung cancer in the subject.

[0545] 68. The method according to any one of items 60 to 65, wherein the method is used to detect the presence of squamous cell carcinoma of the lung in a subject or to diagnose squamous cell carcinoma of the lung in a subject, wherein the method includes comparing the expression level of the integrin α10 polypeptide determined in b. with a control level, wherein the control level is the average expression level of i) the antigen or ii) the polynucleotide transcript observed in non-cancerous lung cells, and a higher level of the antigen containing the integrin α10 polypeptide than the control level indicates the presence of squamous cell carcinoma of the lung in the subject.

[0546] 69. The method according to any one of items 60 to 65, wherein the method is used to detect the presence of lung adenocarcinoma in a subject or to diagnose lung adenocarcinoma in a subject, wherein the method includes comparing the expression level of the integrin α10 polypeptide determined in b. with a control level, wherein the control level is the average expression level of i) the antigen or ii) the polynucleotide transcript observed in non-cancerous lung cells, and a higher level of the antigen containing the integrin α10 polypeptide than the control level indicates the presence of lung adenocarcinoma in the subject.

[0547] 70. The method according to any one of items 60 to 65, wherein the method is used to detect the presence of prostate cancer cells in a subject or to diagnose prostate cancer in a subject, wherein the method includes comparing the expression level of the integrin α10 polypeptide determined in b. with a control level, wherein the control level is the average expression level of i) the antigen or ii) the polynucleotide transcript observed in non-cancerous prostate cells, and a higher level of the antigen containing the integrin α10 polypeptide than the control level indicates the presence of a form of prostate cancer in the subject.

[0548] 71. The method according to any one of items 60 to 65, wherein the method is used to detect the presence of pancreatic cancer cells in a subject or to diagnose pancreatic cancer in a subject, wherein the method includes comparing the expression level of the integrin α10 peptide determined in b. with a control level, wherein the control level is the average expression level of i) the antigen or ii) the polynucleotide transcript observed in non-cancerous pancreatic cells, and wherein a higher expression level of the integrin α10 peptide than the control level indicates the presence of a form of pancreatic cancer in the subject.

[0549] 72. The method according to any one of items 60 to 65, wherein the method is used to detect the presence of sarcoma in a subject or to diagnose sarcoma in a subject, wherein the method includes comparing the expression level of the integrin α10 polypeptide determined in b. with a control level, wherein the control level is the average expression level of i) the antigen or ii) the polynucleotide transcript observed in non-cancerous sarcoma cells, and a higher level of the antigen containing the integrin α10 polypeptide than the control level indicates the presence of sarcoma in the subject.

[0550] 73. A method for classifying triple-negative breast cancer tumor samples from a subject, the method comprising:

[0551] a. Provide the subject with breast tissue suspected of containing cancer cells;

[0552] b. Isolate breast cancer cells characterized as ER-negative, PR-negative, and HER2-negative;

[0553] c. Determine the expression levels of the following in the isolated cells:

[0554] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0555] ii. A multinucleotide transcript or fragment or variant thereof encoding an integrin α10 polypeptide, and

[0556] d. The expression levels determined in c. will be compared with control levels, wherein the control levels are the average expression levels of i) the antigen or ii) the polynucleotide transcripts observed in healthy and / or benign breast tissue;

[0557] Wherein i) the expression level of the antigen and / or ii) the polynucleotide transcript in the breast cancer cells is higher than the control level, and the expression status of ER negative, PR negative and HER2 negative indicates basal-like triple-negative breast cancer or luminal triple-negative breast cancer, thereby classifying the triple-negative breast cancer tumor sample as belonging to basal-like triple-negative breast cancer tumor or luminal triple-negative breast cancer tumor.

[0558] 74. The method according to Item 73, wherein the expression of integrin α10 and one or more polypeptides selected from cytokeratin 7, cytokeratin 8, cytokeratin 18 and cytokeratin 19 indicates luminal triple-negative breast cancer.

[0559] 75. The method according to Item 73, wherein the expression of integrin α10 and one or more polypeptides selected from cytokeratin 5 / 6, cytokeratin 14, cytokeratin 17, p63, EGFR, 34BE12 and c-kit / CD117 indicates basal-like triple-negative breast cancer.

[0560] 76. A method for determining the prognosis of a subject's cancer form, the method comprising:

[0561] a. Provide the subject with cancerous tumor tissue;

[0562] b. Analyze the presence of the following in the sample:

[0563] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0564] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0565] c. Determine the expression levels of i) integrin α10 antigen and / or ii) polynucleotide transcripts.

[0566] d. The expression levels determined in c. will be compared with control levels, wherein the control levels are the average expression levels of i) the antigen and / or ii) the polynucleotide transcripts observed in healthy and / or benign tissues of the same tissue type as the sample;

[0567] e. When the expression levels of the integrin α10i) antigen and / or ii) polynucleotide transcript are higher than the control levels, a poor prognosis for the cancer form is determined.

[0568] The cancer form is selected from breast cancer, lung cancer, prostate cancer, pancreatic cancer, and sarcoma, or a metastatic tumor of any of the cancer forms.

[0569] 77. A method for determining the prognosis of a subject's cancer form, the method comprising:

[0570] a. Provide the subject with cancerous tumor tissue;

[0571] b. Optionally, analyze the presence of one or more cells with cancerous morphology in the tissue;

[0572] c. Analyze the presence of the following in the sample:

[0573] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0574] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide,

[0575] d. Optionally determine the expression levels of the i) integrin α10 antigen and / or ii) polynucleotide transcripts, and compare the determined expression levels with control levels, wherein the control levels are the average expression levels of the i) antigen and / or ii) polynucleotide transcripts observed in healthy and / or benign tissues of the same tissue type as the sample;

[0576] e. When one or more cells with cancerous morphology are present in the tissue in combination with the expression of the integrin α10i) antigen and / or ii) polynucleotide transcript, and / or

[0577] When the expression levels of the integrin α10i) antigen and / or ii) polynucleotide transcript are higher than the control levels,

[0578] Determine the poor prognosis of the aforementioned cancer form.

[0579] The cancer form is selected from breast cancer, lung cancer, prostate cancer, pancreatic cancer, and sarcoma, or a metastatic tumor of any of the cancer forms.

[0580] 78. The method according to any one of items 76 to 77, wherein the prognosis is overall survival or recurrence-free survival.

[0581] 79. The method according to any one of items 60 to 78, wherein the presence of the following is analyzed in the sample:

[0582] i. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0583] ii. A multinucleotide transcript or fragment or variant thereof encoding integrin α10 polypeptide

[0584] The steps include imaging the tissue and / or tissue sample.

[0585] 80. The method according to any one of items 60 to 79, wherein the step of determining the expression level of the i) integrin α10 antigen and / or ii) polynucleotide transcript includes imaging the tissue and / or tissue sample.

[0586] 81. A method for preventing metastasis of a primary cancer form selected from breast cancer, lung cancer, prostate cancer, pancreatic cancer, and sarcoma, the method comprising administering a therapeutically effective dose to a patient in need:

[0587] a) an antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment is specific to integrin α10 polypeptide; and / or

[0588] b) A multinucleotide transcript or a fragment or variant thereof encoding the integrin α10 polypeptide.

[0589] 82. The method according to item 81, wherein the a) antibody or antigen-binding fragment and / or b) polynucleotide transcript are administered after the primary cancer is detected.

[0590] 83. A method for inhibiting integrin α10-mediated signal transduction in at least one cancer cell, the method comprising contacting the at least one cancer cell with a composition comprising an effective amount of:

[0591] a. Antibodies or antigen-binding fragments that are specific to integrin α10 peptide; and / or

[0592] b. A multinucleotide transcript or fragment or variant thereof encoding the integrin α10 polypeptide.

[0593] The at least one cancer cell is selected from breast cancer cells, lung cancer cells, prostate cancer cells, pancreatic cancer cells, sarcoma cells, and metastatic tumor cells.

[0594] 84. A method for inhibiting the cellular function of at least one cancer cell, the method comprising contacting the at least one cancer cell with a composition comprising an effective amount of:

[0595] a. Antibodies or antigen-binding fragments that are specific to integrin α10 peptide; and / or

[0596] b. A multinucleotide transcript or fragment or variant thereof encoding the integrin α10 polypeptide.

[0597] The at least one cancer cell is selected from breast cancer cells, lung cancer cells, prostate cancer cells, pancreatic cancer cells, sarcoma cells, and metastatic tumor cells.

[0598] 85. The method according to any one of items 60 to 84, wherein the antigen comprising integrin α10 polypeptide or a fragment thereof is expressed on the surface of the cell.

[0599] 86. A method for inhibiting the cellular function of at least one cancer cell according to any one of items 84 to 85, wherein the at least one cancer cell is in a tumor and / or metastatic tumor, and wherein the inhibition is selected from:

[0600] a. Inhibit the proliferation of at least one of the cancer cells;

[0601] b. Inhibit the self-renewal of at least one of the cancer cells;

[0602] c. Inhibit the anchorage-independent growth of the at least one cancer cell;

[0603] d. Inhibit the migration of at least one of the cancer cells;

[0604] e. Inhibit the invasion of at least one of the cancer cells;

[0605] f. Inhibit the activity of at least one of the cancer cells;

[0606] g. Inhibiting the adhesion of at least one of the cancer cells; and / or

[0607] Its combination.

[0608] 87. A method for inhibiting the cellular function of at least one cancer cell according to any one of items 84 to 86, wherein the at least one cancer cell is in a tumor and / or metastatic tumor, and the inhibition of the cellular function of the at least one cancer cell inhibits at least one of the following:

[0609] a. The growth of the tumor and / or metastatic tumor;

[0610] b. The proliferation of the tumor and / or metastatic tumors;

[0611] c. The migration of the tumor and / or metastatic tumors;

[0612] d. Invasion of the tumor and / or metastatic tumors;

[0613] e. The spread of new tumors and / or metastatic tumors;

[0614] f. The onset of new tumors and / or metastatic tumors;

[0615] g. Infiltration of new tumors and / or metastatic tumors; and / or

[0616] Its combination.

[0617] 88. The method according to any one of items 83 to 87, wherein the at least one cancer cell is characterized by the following expression levels:

[0618] a. An antigen containing an integrin α10 polypeptide or a fragment thereof; and / or

[0619] b. A multinucleotide transcript or fragment thereof or variant encoding the integrin α10 polypeptide

[0620] Higher than the control level, wherein the control level is the average expression level of a. the antigen and / or b. the polynucleotide observed in healthy and / or benign cells of the same tissue type as the cancer cells.

[0621] 89. The method and / or composition according to any one of items 49 to 88, wherein the composition is according to any one of items 1 to 48.

[0622] 90. The method according to any one of items 59 to 89, wherein the method further comprises administering to the subject a composition or a fragment thereof according to any one of items 1 to 48.

[0623] 91. The method according to any one of items 60 to 90, wherein the method is an in vitro method, and wherein the tissue is a tissue sample obtained from the subject.

[0624] 92. The composition of claim 49, wherein the diagnosis is an in vitro diagnostic.

[0625] 93. Use of the composition in the manufacture of an agent for treating and / or preventing forms of cancer, said composition comprising:

[0626] a. An antibody or fragment thereof that specifically binds to integrin α10 polypeptide, or

[0627] b. A polynucleotide or fragment or variant thereof that specifically binds to a polynucleotide transcript encoding an integrin α10 polypeptide.

[0628] The cancer form is selected from breast cancer, lung cancer, prostate cancer, and pancreatic cancer, or a metastatic tumor of any of the cancer forms.

[0629] 94. The composition, use, or method according to any one of items 1 to 93 above, wherein the cancer form is selected from the following invasive cancer forms: invasive breast cancer, invasive lung cancer, invasive prostate cancer, invasive pancreatic cancer, and invasive sarcoma.

[0630] Example

[0631] Example 1: Protein expression of integrin α10 in tissues derived from invasive tumors (including breast, lung, pancreatic tumors, and sarcomas), as visualized by immunohistochemistry, compared to expression in unaffected tissue regions.

[0632] Materials and methods:

[0633] Human material consisting of breast, lung, pancreas, and sarcoma tissue specimens was used. A standard protocol for immunohistochemistry was optimized using an HRP polymer-conjugated secondary antibody (DAKO Envision anti-rabbit, Denmark) (see Renshaw 2007, ISBN 10:1904842038, Scion Publishing Ltd, UK) (see below), and then reacted with diaminobenzidine and hydrogen peroxide.

[0634] For breast and pancreatic tissue: Use paraffin sections (4 or 8 μm).

[0635] According to the standard protocol, slides are dewaxed and rehydrated by immersing them in xylene, followed by immersion in a series of ethanol solutions and water. Optimization of the labeling protocol involves treating paraffin sections for antigen retrieval by immersing slides in an acidic buffer solution (citrate buffer: 10 mM sodium citrate, 0.05% Tween 20, pH 6.0) followed by heat treatment (at 92°C–95°C).

[0636] For lung and sarcoma tissue: Frozen sections (8 μm) were used, following the same protocol as described above. Optimization of the labeling protocol involved post-fixation of the frozen sections with acetone (100%) at -20°C, followed by quenching in 0.3% hydrogen peroxide, and then blocking (PBS containing 1% BSA and 0.05% Triton-X100). A 3x3 min rinse in PBS was performed between the pretreatment and incubation steps.

[0637] Add Triton-X100 (0.05%) to the primary antibody dilution buffer. Rinse in PBS 3 x 3 min between pretreatment and incubation steps. Counterstain sections with Mayer's hematoxylin to visualize cell nuclei.

[0638] The primary antibody used was a polyclonal rabbit anti-integrin α10 antibody at a concentration of 3 μg / ml.

[0639] result:

[0640] Integrin α10 in aggressive cancer forms, including invasive ductal carcinoma ( Figure 1 A) and triple-negative breast cancer tissue ( Figure 2 Integrin α10 is highly specifically expressed in breast tissue, while its expression is negligible in morphologically unaffected breast tissue. Figure 1 B). As expected, integrin α10 is located on the cell membrane (arrow).

[0641] In addition, integrin α10 was significantly expressed in squamous cell lung cancer cells (data not shown) and pancreatic cancer cells and proliferative islet cells in pancreatic ductal adenocarcinoma tissue (data not shown), but not in surrounding morphologically healthy tissues. Integrin α10 was significantly expressed in undifferentiated pleomorphic sarcoma cells. Figure 3 ).

[0642] in conclusion:

[0643] The results showed that integrin α10 was significantly expressed in patient tissues obtained from several different cancer forms considered to be the most aggressive.

[0644] Example 2: Integrin α10 expression visualized by immunofluorescence in breast and prostate cancer cell lines Materials and methods:

[0645] Cells of the triple-negative breast cancer cell line BT549 and the luminal A breast cancer cell line T47D (originally obtained from the American Type Culture Collection (ATCC)) were grown in monolayers on Ibidi μ-slide 8-well microscopy slides (Ibidi Labware, Germany), and cells of the prostate cancer cell line PC-3 were grown in spheroids. The immunofluorescence protocol described in Example 2 of PCT / EP2017 / 070838 was followed. The primary antibody used was an alternative mouse monoclonal antibody against integrin α10, used at 1.7 μg / ml. The secondary antibodies were fluorophore-conjugated antibodies (anti-mouse AF488 or AF647 conjugated and / or anti-rabbit AF488 or RodRX, all manufactured in donkeys, from Jackson Immunoresearch, USA). All secondary antibodies were diluted 1:200 in PBS containing 1% BSA and incubated for 30 min.

[0646] result:

[0647] Immunofluorescence, followed by confocal microscopy, showed that integrin α10 was present in both different breast cancer cell lines ( Figure 4 A- Figure 4 B) and prostate cancer cell lines ( Figure 4 C) Both are specifically and strongly expressed on the cell membrane.

[0648] in conclusion:

[0649] The results showed that integrin α10 was significantly expressed in cell lines from different aggressive cancers.

[0650] Example 3: Monolayer culture of human non-malignant cell lines and cancer cell lines and flow cytometry analysis of integrin α10 protein expression

[0651] Materials and methods:

[0652] Breast cancer cell lines: 184A1, HCC1428, T47D, MDA-MB-231, and BT549; lung cancer cell lines: A549 (adenocarcinoma) and U-1752 (squamous cell lung cancer); prostate cancer cell lines: 22Rv1, Du145, and PC-3; pancreatic cancer cell lines: BxPC-3, AsPC-1, PANC-1, and MiaPaCa-2 were originally obtained from the American Type Culture Collection (ATCC).

[0653] The 184A1 cell line was established from healthy breast tissue obtained from breast reduction surgery. Cells derived from said tissue were exposed to benzo(a)pyrene, and a transformation line was established. This cell line appears to be immortalized, but not malignant. All other cell lines were derived from malignant tumors and are malignant cells (see table below).

[0654] Immunostaining of cells was performed by incubating cells in the dark at 4°C for 30 min with an alternative mouse monoclonal anti-integrin 10 antibody at a concentration of 1.0 μg / ml (Alexa Fluor 647 conjugate). After incubation with the primary antibody for 30 min, cells were washed twice with DPBS (Hyclone, SH3002802) containing 1% FBS and 0.1% sodium azide, and then analyzed by flow cytometry using a BD Accuri C6 flow cytometer.

[0655]

[0656] Table 1. Overview of the cell lines used in this study and their invasiveness.

[0657] result:

[0658] Results from breast cancer studies showed that integrin α10 was moderately expressed in T47D cells cultured in monolayers and highly expressed in the triple-negative cell line BT549 cultured in monolayers. Figure 5 A and Figure 5 B). When cultured in a monolayer, invasive prostate cancer cells PC-3 showed the highest expression of integrin α10 (B). Figure 5 E and Figure 5 F). Additionally, the most invasive and high-grade pancreatic cancer cell lines (Grade III), MiaPaCa-2 and PANC-1, also showed the highest integrin α10 expression. Figure 5 I). In contrast, less aggressive pancreatic cancer cells BxPC-3 and AsPC-1 showed little or no expression of integrin α10, particularly in spheroid cultures (see Example 4). Figure 5 J). Lung cancer cells cultured in monolayers showed very low expression (J). Figure 5 K), however, expression increases when cells are cultured in spheroids. Figure 5 L and Experiment 4).

[0659] in conclusion:

[0660] The results showed that integrin α10 expression was highest in the most aggressive cancer types. This suggests a correlation between integrin α10 and the invasiveness of different cancers, and thus potentially with migration and invasion. This observation was readily observed in a single culture system, and even more pronounced in a three-dimensional (3D) culture system (see Experiment 4). Due to improved cell-cell interactions, cell-ECM interactions, and cell populations and structures similar to those in vivo, the 3D cell culture model is a superior model compared to the traditional 2D monolayer culture model.

[0661] Example 4: Detection and analysis of integrin α10 protein in spheroidization assay

[0662] Materials and methods:

[0663] Breast cancer cell lines 184A1, HCC1428, T47D, MDA-MB-231, and BT549, or lung cancer cell lines A549 and U-1752, or prostate cancer cell lines 22Rv1, DU145, and PC-3, or pancreatic cancer cell lines BxPC-3, AsPC-1, PANC-1, and MiaPACa-2, or sarcoma cell line MFH152, were seeded in serum-free, ultra-low adsorption plates (CLS3471, Corning) to form non-adherent spheroids with self-renewal capabilities (breast cancer: called breast globules; prostate cancer: called prostate globules). The non-scaffold-based cultures formed aggregates of cells commonly referred to as spheroids or spheres. The plating medium used for spheroid formation consisted of DMEM / F12 (1:1) w / Glutamax (31331-08, Gibco) supplemented with B27 (12587-010, Gibco), 20 ng / ml human basic fibroblast growth factor (Miltenyi Biotec), 20 ng / ml human epidermal growth factor (Miltenyi Biotec), and 100 U / ml penicillin and 100 U / ml streptomycin. Cells were incubated in an incubator for approximately 10 days without movement (especially the first 5 days). Immunostaining and flow cytometry analysis were performed using an alternative mouse monoclonal anti-integrin α10 antibody as described in Example 3. The 3D culture system provides a superior in vitro model, allowing the study of cellular responses in an environment similar to that in vivo.

[0664] result:

[0665] Results from breast cancer experiments demonstrated that when invasive triple-negative breast cancer cells MDA-MB-231 and BT549 were cultured in 3D conditions (spheres) simulating in vitro tumor growth, the expression of integrin α10 was significantly increased. Figure 5 C- Figure 5D). Non-malignant or less invasive breast cell lines 184A1 and HCC1428 did not increase integrin α10 in spheroid cultures. The invasive prostate cancer cell line PC-3 also upregulated integrin α10 protein expression when cells grew in spheroidal forms. Figure 5 G- Figure 5 H)(used with monolayer cultures: Figure 5 E- Figure 5 (Comparison with F). Consistently, compared with monolayer cultures, invasive pancreatic cancer cells PANC-1 and MiaPaCa-2 readily formed spheroids and significantly increased integrin α10 protein expression (F). Figure 5 I- Figure 5 In contrast, cell lines BxPC-3 and AsPC-1 did not increase integrin α10 expression, and furthermore, AsPC-1 did not form spheroids in culture. Lung cancer cells A549 and U-1752, which grew as spheroids, significantly increased integrin α10 protein levels. Figure 5 K- Figure 5 L). Similarly, compared to monolayer cultures, the invasive sarcoma cell line MFH152 readily formed spheroids and exhibited significantly increased expression of integrin α10 at the protein level (L). Figure 5 M).

[0666] in conclusion:

[0667] The results showed that integrin α10 expression was highest in the most aggressive cancer types. Spheroid culture (3D cell culture) (a superior method for mimicking in vivo tumor growth) further increased integrin α10 expression in aggressive cancer cells. This provides additional support for the correlation between integrin α10 and aggressive cancer.

[0668] Example 5: Detection and analysis of integrin α10 mRNA (ITGA10) expression in human breast cancer, prostate cancer, and pancreatic cancer cell lines cultured in monolayers or spheres.

[0669] Materials and methods:

[0670] RNA extraction and quantitative PCR were performed on different cell lines cultured in monolayers or spheres (to simulate tumor growth). Total RNA was extracted from cells using the RNeasy Plus Mini Kit (Qiagen) and reverse transcribed into cDNA using the SuperScript cDNA Synthesis Kit (Life Technologies). For quantitative PCR, TaqMan gene expression premix (Life Technologies) and TaqMan probes (Thermo Fisher Scientific) were used: GAPDH (Mm99999915_g1) and ITGA10 (Mm01265767_m1). The cycle threshold of the target gene was normalized to the geometric mean of the housekeeping gene GAPDH to obtain ΔCt. 2^(-ΔCt) was calculated. -ΔCt (to conduct the final analysis.) result:

[0671] Compared to cell lines with lower invasiveness and less aggressiveness (i.e., non-malignant breast cells 184A1 and breast cancer cells HCC1428 and T47D, prostate cancer cells 22RV1 and DU145, and pancreatic cancer cells BxPC-3 and AsPC-1), more invasive and aggressive cancer cells (i.e., triple-negative breast cancer cells MDA-MB-231 and BT549) showed greater invasiveness and aggressiveness. Figure 6 A- Figure 6 B) Prostate cancer cells PC-3 ( Figure 6 C- Figure 6 D), and pancreatic cancer cells PANC-1 and MiaPaCa-2 ( Figure 6 E- Figure 6 In F), the level of integrin α10 mRNA (ITGA10) was higher, as shown in the corresponding figure. Furthermore, compared to monolayer conditions, the level of integrin α10 (ITGA10) mRNA was significantly increased when cells were cultured in spherical conditions (see [reference]). Figure 6 A and Figure 6 B. Figure 6 C and Figure 6 D, and Figure 6 E and Figure 6 F).

[0672] in conclusion:

[0673] The results demonstrated that integrin α10 mRNA expression was highest in the most aggressive cancer types. Spheroid culture (3D cell culture) (a superior method for mimicking in vivo tumor growth) further increased integrin α10 expression in aggressive cancer cells. This provides additional support for the correlation between integrin α10 and aggressive cancer.

[0674] Example 6: Overall survival curves for ITGA10 gene expression in different cancer indications.

[0675] Materials and methods:

[0676] Overall survival curves for ITGA10 gene expression in 255 patients with triple-negative breast cancer were analyzed (Gyorffy B et al., 2010) (A). Patients were divided into low ITGA10 expression and high ITGA10 expression groups based on the median cutoff value. Kaplan-Meier plots were constructed, and the log-rank test was used to determine the difference in recurrence-free survival. The cutoff value used in the analysis was 224.

[0677] Overall survival curves for squamous cell lung cancer (GSE4573 database, 88 samples), prostate adenocarcinoma (TCGA-prostate adenocarcinoma database, 413 samples), localized pancreatic ductal adenocarcinoma (GSE211501 database, 102 samples), and sarcoma (TCGA-sarcoma database, 234 samples) were analyzed using the PROGgeneV2 tool, targeting high and low ITGA10 expression groups based on median values. This analysis allowed for the investigation of the prognostic significance of ITGA10-related gene expression in the corresponding microarray datasets. (Goswami CP et al., 2013)

[0678] result:

[0679] Survival curves show the difference in overall survival between patients with high (solid) or low (dashed) ITGA10 expression. Figure 7 Patients were divided into high and low ITGA10 expression groups based on survival analysis and the median cutoff value of the log-rank test. The p-value represents the log-rank test for the difference in overall survival. For all indicated cancer forms, the high ITGA10 expression group had lower survival rates compared to the low ITGA10 expression group.

[0680] in conclusion:

[0681] High expression of ITGA10 was associated with lower overall survival in the indicated cancer types.

[0682] Example 7: Inhibition of cell adhesion after blocking integrin α10 with antibody

[0683] Materials and methods:

[0684] One day prior to the adhesion assay, 48-well plates were coated with type I collagen (Sigma, C7661-5MG), collagen IV (Sigma, C5533-5MG), or bovine serum albumin (BSA). On the day of the experiment, the plates were incubated with 0.25% BSA at 37°C for 30 min to block nonspecific binding. Simultaneously, cancer cells (breast cancer cells BT549 and prostate cancer cells PC-3) were harvested and suspended in HBSS to form a single-cell suspension. Cells were pre-incubated for 30 min with or without a mouse monoclonal antibody mAbα targeting integrin α10 peptide at a concentration of 10 μg / ml. As a negative control, a mouse monoclonal isotope control antibody IgG2a was used. Cells were then allowed to adhere for 60 min, and unadhered cells were removed by washing with HBSS. Adhered cells were fixed with 96% ethanol and stained with 0.1% crystal violet. Absorbance was measured at 590 nm using a plate reader.

[0685] result:

[0686] Treatment of breast BT549 cells with a functional blocking monoclonal antibody targeting integrin α10 significantly reduced BT549 cell adhesion to collagen I and IV. Figure 8 When cells are in a monolayer ( Figure 8 A- Figure 8 B) and mammary glomeruli ( Figure 8 C- Figure 8 This effect was observed when cells were cultured in D) and compared with untreated or IgG2a-treated cells. Similarly, treatment with the functionally blocking monoclonal integrin α10 antibody significantly reduced cell adhesion of PC-3 cells cultured in spheroids to collagen I and IV compared with untreated cells as a control. Figure 8 E- Figure 8 F). BSA-coated wells were used as negative controls because cell adhesion to the extracellular matrix (ECM) is mediated by integrin receptors, and cells do not adhere to the BSA-coated wells.

[0687] in conclusion:

[0688] The results showed that the integrin α10 function blocking antibody could block adhesion, thereby potentially affecting key tumor cell functions such as proliferation, migration and growth.

[0689] Example 8: Reduced cell migration after blocking integrin α10 with monoclonal antibodies

[0690] Materials and methods:

[0691] Migration assays of BT549 breast cancer cells, A549 lung cancer cells, and PC-3 prostate cancer cells were performed using Boyden Chambers (Corning) containing polycarbonate filters with an 8 μm pore size. For BT549 breast cancer cells, the filters were coated with collagen IV (Sigma, C5533-5MG); for PC-3 prostate cancer cells, the filters were coated with type I collagen (Sigma, C7661-5MG); or for A549 lung cancer cells, the filters were not coated. For migration assays, a working solution of collagen (0.01 mg / ml) was prepared from a stock solution (1 mg / ml) containing PBS. The lower compartment was filled with 10% FBS medium as a chemical inducer and also contained a monoclonal antibody against integrin α10. Cancer cells were incubated with 5 μg / ml of the antibody for 30 min, and then the cells were added to the upper compartment of the Boyden Chamber. Cells in the lower compartment were fixed and stained with crystal violet after 24 h or 48 h. OD 590nm was measured in a reader (SpectraMax ABS, MolecularDevices).

[0692] Breast cancer cell migration assay: Cells were incubated with mouse anti-integrin α10 monoclonal antibodies (mAbα10) or human anti-integrin α10Th101 (see Example 12) that bind to different epitopes of integrin α10. As a negative control, cells were incubated with the negative control antibody Th301 human IgG1VH / λ or the negative / isotope control antibody IgG2a (mouse monoclonal IgG2a, κ), or any antibody treatment was omitted (NT = untreated).

[0693] Prostate cancer cell migration assay: Cells were incubated with mouse anti-integrin α10 monoclonal antibody (mAbα10) and allowed to migrate for 24 or 48 hours. As a negative control, cells were incubated with isotope control antibody IgG2a.

[0694] Lung cancer migration assay: Cells were incubated with mouse anti-integrin α10 monoclonal antibody (mAbα10) for 24 or 48 hours. As a negative control, cells were incubated with isotope control antibody IgG2a.

[0695] result:

[0696] Breast cancer cells: Compared with untreated cells or cells incubated with control antibodies, incubation with monoclonal anti-integrin α10 antibody mAbα10 or Th101 reduced cell migration. Figure 9 A). The two anti-integrin α10 antibodies have similar effects.

[0697] Prostate cancer cells: Compared with cells incubated with the control antibody, incubation with anti-integrin α10 antibody mAb α10 reduced cell migration. Figure 9 B). This effect is amplified when cells are measured at 48 hours, compared to 24 hours.

[0698] Lung cancer cells: Similarly, compared with cells incubated with the control antibody, incubation with monoclonal anti-integrin α10 antibody mAbα10 reduced cell migration. Figure 9 C).

[0699] in conclusion:

[0700] The results demonstrated that the monoclonal integrin α10 antibody could block integrin α10 and inhibit breast cancer cells and prostate cancer cells (respectively). Figure 9 A and Figure 9 B) and lung cancer cells ( Figure 9 C) migration.

[0701] Example 9: Reduced viability of breast cancer cells treated with integrin α10 antibody-drug conjugate

[0702] Materials and methods:

[0703] BT549 breast cancer cells were cultured as a monolayer in 96-well plates and treated with anti-integrin α10-MMAE ADC (antibody-drug conjugate or anti-control-MMAE ADC). The ADC is an alternative conjugate of mouse monoclonal integrin α10 antibody (IgG1(κ)) or isotype control IgG1 antibody (anti-control) with the microtubule inhibitor methylaurestatin E (MMAE). Cells were incubated at 37°C for 4 days with 23, 69, and 207 nM ADC. Cell viability was determined using the WST-1 assay (Roche, Mannheim, Germany) according to the manufacturer's recommendations.

[0704] result:

[0705] Antibody-drug conjugates (ADCs) consisting of an integrin α10 antibody (anti-α10-MMAE) conjugated to the potent cytotoxic MMAE induced cell death in breast cancer cells. Figure 10 In contrast, no cell death was observed when cells were treated with an isotype control IgG1 antibody conjugated to MMAE (anti-control-MMAE). Higher concentrations of the antibody-drug conjugate were associated with lower viability of breast cancer cells.

[0706] in conclusion:

[0707] The results demonstrated the specific cytotoxic effects of the integrin α10 antibody conjugate.

[0708] Example 10: Monoclonal α10 antibody treatment inhibits the in vitro cell proliferation of invasive breast cancer, pancreatic cancer, and lung cancer cells. Materials and methods:

[0709] Sphere measurement ( Figure 11 A- Figure 11 D and Figure 11 F): Breast cancer cells (BT549), prostate cancer cells (PC-3), pancreatic cancer cells (MiaPaCa-2 and PANC-1), and lung cancer cells (A549) were seeded in serum-free medium in 6-well ultra-low adsorption plates (CLS3471, Corning) to form non-adherent spheroids. Simultaneously with seeding, cells were treated with 10 μg / ml of monoclonal anti-integrin α10 antibody mAbα10 or a control antibody (IgG2a). Antibody was added every other day for 14 days. After 14 days of treatment, BrdU (final concentration 10 μM) was added to the spheroids, and the cells were incubated for 24 hours. Cells were then collected for BrdU analysis by flow cytometry according to the instructions from the BD Pharmigen APC BrdU Flow Kit (catalog number 552598). The mean fluorescence intensity of BrdU staining was calculated. For lung cancer cells (A549), in addition to BrdU staining, 7-aminoactinomycin D (7-AAD) was added to stain total DNA. Cell cycle analysis was based on both BrdU and 7-AAD staining.

[0710] Monolayer determination ( Figure 11 E): Adherent BT549 breast cancer cells were seeded in 96-well plates coated with type IV collagen and treated with monoclonal anti-integrin α10 antibody (mAbα10 or Th101) to obtain a final concentration of 5 μg / ml directly after cell seeding. After 24 hours of antibody treatment, BrdU was added for 2.5 hours, and proliferation was measured using the BrdU cell proliferation ELISA kit (Roche Diagnostics GmbH) according to the manufacturer's instructions.

[0711] Cell cycle analysis ( Figure 11 F):Immunofluorescence staining and flow cytometry analysis using incorporated bromodeoxyuridine (BrdU) are used to determine the frequency and nature of individual cells that have synthesized DNA. In this method, BrdU (an analogue of the DNA precursor thymidine) is incorporated into newly synthesized DNA via cell entry and processing during the S phase of the cell cycle (DNA synthesis). The incorporated BrdU is stained with a specific anti-BrdU fluorescent antibody. The level of cell-associated BrdU is then measured by flow cytometry. Typically, staining with a dye that binds to total DNA (such as 7-aminoactinomycin D (7-AAD)) is combined with immunofluorescence BrdU staining. With this combination, two-color flow cytometry analysis allows for the enumeration and characterization of cells actively synthesizing DNA (BrdU-incorporated) in terms of their cell cycle position (i.e., G0 / G1, S, or G2+M phases defined by 7-AAD staining intensity).

[0712] result:

[0713] Sphere measurement: The results demonstrated that the functional blocking monoclonal α10 antibody reduced the number of invasive breast cancer cells ( Figure 11 A) Prostate cancer cells ( Figure 11 B) and pancreatic cancer cells ( Figure 11 C- Figure 11 The proliferation of D) was compared with that after treatment with the control antibody IgG2a, which showed no reduction in proliferation. Cell cycle analysis showed that in the lung cancer cell line A549, treatment with monoclonal α10 antibody inhibited cell proliferation by arresting cells in the G0 / G1 phase compared with treatment with the control antibody IgG2a. Figure 11 F).

[0714] Single-layer measurement: Figure 11 E showed that, compared with incubation with or without the control antibody, anti-integrin α10 antibodies (mAbα10 and Th101) blocked the cell proliferation of breast cancer cells in monolayer culture.

[0715] in conclusion:

[0716] These results confirm that integrin α10 is involved in the cell proliferation process, thus indicating that anti-integrin α10 antibodies have the potential as a treatment for anti-proliferative cancers.

[0717] Example 11: Treatment with integrin α10 antibody inhibited the progression of breast tumor growth in vivo.

[0718] Materials and methods:

[0719] All in vivo experiments were conducted in 6-8 week old female NMRI-nu immunodeficient mice (n=5 / group) purchased from Janvier Labs (France). Animal welfare and experimental procedures were performed in accordance with international standards, and animals were maintained under specific pathogen-free (SPF) conditions. All experimental procedures were approved by the Malmö and Lund Animal Ethics Committees (…). Approved by the Swedish Medical Association and the LundAnimal Ethics Committee (Sweden). For tumor induction, mice were inoculated with 2x10 MMR matrix gel via subcutaneous injection into the right ventral region. 6 Invasive BT549 breast cancer cells were injected. Tumor growth was monitored by non-invasive 2D bioluminescence (BLI) imaging using IVIS-CT spectroscopy (PerkinElmer, Massachusetts, USA) two to three weeks post-injection. Mice showing tumor growth signals were randomly assigned to two groups (control and treatment) based on their mean BLI signal intensity and mean total flux (photons / second) values ​​recorded in the defined target region (ROI). Different antibodies against integrin α10 were used in this study: mouse monoclonal mAbα10 and human monoclonal antibody Th101. The concentration of each antibody injected into mice was calculated to be 5 mg / kg, and animals received the antibodies via intraperitoneal injection until the endpoint. Tumor growth was monitored using bioluminescence 2D and 3D micro-CT imaging. Briefly, mice were anesthetized with 3% isoflurane gas prior to imaging and intraperitoneally injected with 150 mg D-luciferin / kg body weight in PBS. 2D images were acquired sequentially at five intervals between different exposure segments (emission: open filter, f / stop: 1, binding: 8). The BLI signal intensity was quantified in total flux (photons / second) after subtracting the average background signal (Bkg) from the target area of ​​measurement (ROI) using real-time image analysis software (PerkinElmer, Massachusetts, USA). Mouse weight was recorded weekly before antibody treatment. No signs of disease or reduced growth rate were observed.

[0720] result:

[0721] The results showed that, compared with mice treated with negative control antibodies (IgG2a and Th301), mice treated with integrin α10 antibody mAbα10 or Th101 had lower total flux readings. Figure 12 A). This indicates a reduction in tumor growth. Animal weight was monitored from the start of treatment, and no signs of disease or weight loss were observed in animals treated with the appropriate antibodies. Figure 12 B).

[0722] in conclusion:

[0723] The results showed that different antibodies against integrin α10 (here mAbα10 (=Ab365) or Th101) could reduce tumor growth in vivo, thus demonstrating the usefulness of anti-integrin α10 antibodies as a treatment for proliferative cancers.

[0724] Example 12: Different epitopes of monoclonal antibody mAbα10 and Th101 binding to integrin α10.

[0725] Materials and methods:

[0726] The binding competition assay for mouse monoclonal antibody mAbα10 and human monoclonal antibody Th101 was performed in C2C12α10 cells overexpressing integrin α10 and triple-negative breast cancer BT549 cells. Cells were incubated with single or mixed antibodies at indicated concentrations (μg / ml) for 30 min, washed twice, and stained with secondary antibody for 30 min. Donkey anti-human Alexa 488 was used as the secondary antibody for human monoclonal antibody Th101. Donkey anti-mouse Alexa 647 was used as the secondary antibody for mouse monoclonal antibody mAbα10. Antibody binding was analyzed by flow cytometry, in which the fluorescence signal of the secondary antibody was detected.

[0727] Data are expressed as average fluorescence intensity per 100,000 cells.

[0728] result:

[0729] Flow cytometry data showed that the binding strength of antibody mAbα10 did not change significantly with the increase of Th101 antibody concentration. Figure 13 A- Figure 13 B). As the concentration of antibody Th101 added to the reaction wells gradually increased (0 to 9 μg / ml) (grey bar), the signal representing antibody mAbα10 (at a constant concentration of 3 μg / ml) remained stable (black bar) (left side of the figure).

[0730] Conversely, the binding strength of Th101 is not affected by the increase in mAbα10 concentration. Figure 13 A- Figure 13 B). As the concentration of antibody mAbα10 added to the reaction wells gradually increased (0 to 9 μg / ml) (black bar), the signal representing antibody Th101 (at a constant concentration of 3 μg / ml) remained stable (gray bar) (right side of the figure).

[0731] Two cell lines, C2C12α10 cells ( Figure 13 A) and BT549 cells ( Figure 13 B) yielded similar results.

[0732] in conclusion:

[0733] If antibodies bind to the same epitope, increasing the concentration of one antibody would lead to a decrease in the binding of the other antibody due to competition for the same binding site. Since this was not the case, the results indicate that the two monoclonal integrin α10 antibodies, mAbα10 and Th101, bind to different epitopes of the integrin α10 antigen.

[0734] Example 13: Sequence

[0735] Sequence information of human integrin α10 polypeptide

[0736] SEQ ID NO:1

[0737] Human integrin α10 full-length polypeptide

[0738] See the sequence list.

[0739] SEQ ID NO:2

[0740] Extracellular domain of human integrin α10

[0741] See the sequence list.

[0742] SEQ ID NO:3

[0743] I domain of human integrin α10

[0744] See the sequence list.

[0745] Sequence information of anti-integrin α10 antibody Th101

[0746] SEQ ID NO:4:

[0747] Variable heavy chain complementarity-determining region 1 (CDR-H1)

[0748] FTFSDYGMN

[0749] SEQ ID NO:5:

[0750] Variable heavy chain complementarity-determining region 2 (CDR-H2)

[0751] VISYDGSNKYYADSVKG

[0752] SEQ ID NO:6:

[0753] Variable heavy chain complementarity-determining region 3 (CDR-H3)

[0754] GGNWGVFDY

[0755] SEQ ID NO:7:

[0756] Variable light chain complementarity-determining region 4 (CDR-L1)

[0757] SGSSSNIGSNPVHSEQ ID NO.8:

[0758] Variable light chain complementarity-determining region 5 (CDR-L2)

[0759] ENNKRPS

[0760] SEQ ID NO.9:

[0761] Variable light chain complementarity-determining region 6 (CDR-L3)

[0762] AAWDDSLSGQGV

[0763] SEQ ID NO.10:

[0764] Heavy chain variable region

[0765] EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYGMNWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGGNWGVFDYWGQGTLVTVSS

[0766] SEQ ID NO.11:

[0767] Light chain variable region

[0768] QSVLTQPPSASGTPGQRVTISSCSGSSSNIGSNPVHWYQQLPGTAPKLLIYENNKRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGQGVFGGGTKLTVLG

[0769] References

[0770] Arpino G,Milano M,De Placido S.Features of aggressive breast cancer,The Breast.2015;24(5):594-600.

[0771] Bengtsson T,Aszodi A,Nicolae C,Hunziker EB,Lundgren-Akerlund E, R.Loss of alpha10beta1integrin expression leads to moderate dysfunction ofgrowth plate chondrocytes,J Cell Sci.2005;118(Pt 5):929-36.

[0772] Bianchini G,Balko JM,Mayer IA,et al.Triple-negative breast cancer:challenges and opportunities of a heterogeneous disease.Nat Rev Clin Oncol2016;13:674–90.

[0773] Bombardelli L and Berns A.The steady progress of targeted therapies,promising advances for lung cancer,Ecancermedicalscience 2016;10:638.

[0774] Camper L,Hellman U,Lundgren-Akerlund.Isolation,Cloning,and SequenceAnalysis of the Integrin Subunitα10,aβ1-associated Collagen Binding Integrin,Expressed on Chondrocytes,J Biol Chem 1998;273(32):20383-9.

[0775] Camper L,Holmvall K, C,AszódiA,Lundgren-AkerlundE.Distribution of the collagen-binding integrin alpha10beta1during mousedevelopment,Cell Tissue Res.2001;306(1):107-16.

[0776] Dai X,Li T,Bai Z,Yang Y,Liu X,Zhan J,Shi B.Breast cancer intrinsicsubtype classification,clinical use and future trends,Am J Cancer Res.2015;5(10):2929-2943.

[0777] Dai X,Xiang L,Li T,Bai Z.Cancer Hallmarks,Biomarkers and BreastCancer Molecular Subtypes,Journal of Cancer.2016;7(10):1281-94.

[0778] Einstein DJ,Arai S,Balk SP.Targeting the androgen receptor andovercoming resistance in prostate cancer,Curr Opin Oncol.2019;31(3):175-18.

[0779] Goswami CP and Nakshatri H.PROGgene:gene expression based survivalanalysis web application for multiple cancers.J Clin Bioinforma.2013 Oct 28;3(1):22.

[0780] Gyorffy B,Lanczky A,Eklund AC,Denkert C,Budczies J,Li Q,Szallasi Z.Anonline survival analysis tool to rapidly assess the effect of 22,277 genes onbreast cancer prognosis using microarray data of 1809 patients,Breast CancerRes Treatment,2010 Oct;123(3):725-31.

[0781] Hariri N,Hasteh F,Walavalkar V,Roma AA,and Fadare,O.Estrogenreceptor,progesterone receptor,and human epidermal growth factor receptor-2testing in breast cancer:assessing the value of repeated centralized testingin excision specimens,Appl Immunohistochem Mol Morphol.2019;27(1):1-7.

[0782] Ilic M,Ilic I.Epidemiology of pancreatic cancer.World Journal ofGastroenterology 2016;44:9694-9705.

[0783] Lundgren- E,AszòdiA.α10β1:a collagen receptor critical inskeletal development,Adv Exp Med Biol.2014;819:61-71.

[0784] Ma L,Liang Z,Zhou H,Qu L.Applications of RNA Indexes for PrecisionOncology in Breast Cancer,Genomics Proteomics Bioinformatics.2018;16(2):108–119.

[0785] Malorni L,Shetty PB,De Angelis C,et al.Clinical and biologic featuresof triple-negative breast cancers in a large cohort of patients with long-term follow-up.Breast Cancer Res Treat 2012;136:795–804.

[0786] Moschos SJ,Drogowski LM,Reppert SL,Kirkwood JM.Integrins and Cancer,Oncology.2007 Aug;21(9 Suppl 3):13-20.

[0787] Rakha EA,El-Sayed ME,Green AR,Lee AH,Robertson JF,Ellis IO Prognosticmarkers in triple-negative breast cancer.Cancer.2007 1;109(1):25-32

[0788] Saad F,Shore N,Zhang T,Sharma S,Cho HK,Jacobs IA.Emerging therapeutictargets for patients with advanced prostate cancer.Cancer Treat Rev.2019;19;76:1-9.

[0789] U.S.National Institute of Health,National Cancer Institute.SEERCancer Statistics Review,1975–2015.

[0790] Varas L,Ohlsson LB,Honeth G,Olsson A,Bengtsson T,Wiberg C,BockermannR, S,Richter J,Pennington D,Johnstone B,Lundgren-Akerlund E,KjellmanC.Alpha10 integrin expression is up-regulated on fibroblast growth factor-2-treated mesenchymal stem cells with improved chondrogenic differentiationpotential,Stem Cells Dev.2007;16(6):965-78.WHO classification of tumors ofsoft tissue and bone(2013).

[0791] sequence list <110> Tarkinta JSC <120> Integrin α10 and invasive cancer forms <130> P4894PC00 <160> 11 <170> PatentIn version 3.5 <210> 1 <211> 1167 <212> PRT <213> Homo sapiens <220> <221> Unclassified features <222> (1)..(1167) <223> Human integrin α10 full-length polypeptide <400> 1 Met Glu Leu Pro Phe Val Thr His Leu Phe Leu Pro Leu Val Phe Leu 1 5 10 15 Thr Gly Leu Cys Ser Pro Phe Asn Leu Asp Glu His His Pro Arg Leu 20 25 30 Phe Pro Gly Pro Pro Glu Ala Glu Phe Gly Tyr Ser Val Leu Gln His 35 40 45 Val Gly Gly Gly Gln Arg Trp Met Leu Val Gly Ala Pro Trp Asp Gly 50 55 60 Pro Ser Gly Asp Arg Arg Gly Asp Val Tyr Arg Cys Pro Val Gly Gly 65 70 75 80 Ala His Asn Ala Pro Cys Ala Lys Gly His Leu Gly Asp Tyr Gln Leu 85 90 95 Gly Asn Ser Ser His Pro Ala Val Asn Met His Leu Gly Met Ser Leu 100 105 110 Leu Glu Thr Asp Gly Asp Gly Gly Phe Met Ala Cys Ala Pro Leu Trp 115 120 125 Ser Arg Ala Cys Gly Ser Ser Val Phe Ser Ser Gly Ile Cys Ala Arg 130 135 140 Val Asp Ala Ser Phe Gln Pro Gln Gly Ser Leu Ala Pro Thr Ala Gln 145 150 155 160 Arg Cys Pro Thr Tyr Met Asp Val Val Ile Val Leu Asp Gly Ser Asn 165 170 175 Ser Ile Tyr Pro Trp Ser Glu Val Gln Thr Phe Leu Arg Arg Leu Val 180 185 190 Gly Lys Leu Phe Ile Asp Pro Glu Gln Ile Gln Val Gly Leu Val Gln 195 200 205 Tyr Gly Glu Ser Pro Val His Glu Trp Ser Leu Gly Asp Phe Arg Thr 210 215 220 Lys Glu Glu Val Val Arg Ala Ala Lys Asn Leu Ser Arg Arg Glu Gly 225 230 235 240 Arg Glu Thr Lys Thr Ala Gln Ala Ile Met Val Ala Cys Thr Glu Gly 245 250 255 Phe Ser Gln Ser His Gly Gly Arg Pro Glu Ala Ala Arg Leu Leu Val 260 265 270 Val Val Thr Asp Gly Glu Ser His Asp Gly Glu Glu Leu Pro Ala Ala 275 280 285 Leu Lys Ala Cys Glu Ala Gly Arg Val Thr Arg Tyr Gly Ile Ala Val 290 295 300 Leu Gly His Tyr Leu Arg Arg Gln Arg Asp Pro Ser Ser Phe Leu Arg 305 310 315 320 Glu Ile Arg Thr Ile Ala Ser Asp Pro Asp Glu Arg Phe Phe Phe Asn 325 330 335 Val Thr Asp Glu Ala Ala Leu Thr Asp Ile Val Asp Ala Leu Gly Asp 340 345 350 Arg Ile Phe Gly Leu Glu Gly Ser His Ala Glu Asn Glu Ser Ser Phe 355 360 365 Gly Leu Glu Met Ser Gln Ile Gly Phe Ser Thr His Arg Leu Lys Asp 370 375 380 Gly Ile Leu Phe Gly Met Val Gly Ala Tyr Asp Trp Gly Gly Ser Val 385 390 395 400 Leu Trp Leu Glu Gly Gly His Arg Leu Phe Pro Pro Arg Met Ala Leu 405 410 415 Glu Asp Glu Phe Pro Pro Ala Leu Gln Asn His Ala Ala Tyr Leu Gly 420 425 430 Tyr Ser Val Ser Ser Met Leu Leu Arg Gly Gly Arg Arg Leu Phe Leu 435 440 445 Ser Gly Ala Pro Arg Phe Arg His Arg Gly Lys Val Ile Ala Phe Gln 450 455 460 Leu Lys Lys Asp Gly Ala Val Arg Val Ala Gln Ser Leu Gln Gly Glu 465 470 475 480 Gln Ile Gly Ser Tyr Phe Gly Ser Glu Leu Cys Pro Leu Asp Thr Asp 485 490 495 Arg Asp Gly Thr Thr Asp Val Leu Leu Val Ala Ala Pro Met Phe Leu 500 505 510 Gly Pro Gln Asn Lys Glu Thr Gly Arg Val Tyr Val Tyr Leu Val Gly 515 520 525 Gln Gln Ser Leu Leu Thr Leu Gln Gly Thr Leu Gln Pro Glu Pro Pro 530 535 540 Gln Asp Ala Arg Phe Gly Phe Ala Met Gly Ala Leu Pro Asp Leu Asn 545 550 555 560 Gln Asp Gly Phe Ala Asp Val Ala Val Gly Ala Pro Leu Glu Asp Gly 565 570 575 His Gln Gly Ala Leu Tyr Leu Tyr His Gly Thr Gln Ser Gly Val Arg 580 585 590 Pro His Pro Ala Gln Arg Ile Ala Ala Ala Ser Met Pro His Ala Leu 595 600 605 Ser Tyr Phe Gly Arg Ser Val Asp Gly Arg Leu Asp Leu Asp Gly Asp 610 615 620 Asp Leu Val Asp Val Ala Val Gly Ala Gln Gly Ala Ala Ile Leu Leu 625 630 635 640 Ser Ser Arg Pro Ile Val His Leu Thr Pro Ser Leu Glu Val Thr Pro 645 650 655 Gln Ala Ile Ser Val Val Gln Arg Asp Cys Arg Arg Arg Gly Gln Glu 660 665 670 Ala Val Cys Leu Thr Ala Ala Leu Cys Phe Gln Val Thr Ser Arg Thr 675 680 685 Pro Gly Arg Trp Asp His Gln Phe Tyr Met Arg Phe Thr Ala Ser Leu 690 695 700 Asp Glu Trp Thr Ala Gly Ala Arg Ala Ala Phe Asp Gly Ser Gly Gln 705 710 715 720 Arg Leu Ser Pro Arg Arg Leu Arg Leu Ser Val Gly Asn Val Thr Cys 725 730 735 Glu Gln Leu His Phe His Val Leu Asp Thr Ser Asp Tyr Leu Arg Pro 740 745 750 Val Ala Leu Thr Val Thr Phe Ala Leu Asp Asn Thr Thr Lys Pro Gly 755 760 765 Pro Val Leu Asn Glu Gly Ser Pro Thr Ser Ile Gln Lys Leu Val Pro 770 775 780 Phe Ser Lys Asp Cys Gly Pro Asp Asn Glu Cys Val Thr Asp Leu Val 785 790 795 800 Leu Gln Val Asn Met Asp Ile Arg Gly Ser Arg Lys Ala Pro Phe Val 805 810 815 Val Arg Gly Gly Arg Arg Lys Val Leu Val Ser Thr Thr Leu Glu Asn 820 825 830 Arg Lys Glu Asn Ala Tyr Asn Thr Ser Leu Ser Ile Ile Phe Ser Arg 835 840 845 Asn Leu His Leu Ala Ser Leu Thr Pro Gln Arg Glu Ser Pro Ile Lys 850 855 860 Val Glu Cys Ala Ala Pro Ser Ala His Ala Arg Leu Cys Ser Val Gly 865 870 875 880 His Pro Val Phe Gln Thr Gly Ala Lys Val Thr Phe Leu Leu Glu Phe 885 890 895 Glu Phe Ser Cys Ser Ser Leu Leu Ser Gln Val Phe Gly Lys Leu Thr 900 905 910 Ala Ser Ser Asp Ser Leu Glu Arg Asn Gly Thr Leu Gln Glu Asn Thr 915 920 925 Ala Gln Thr Ser Ala Tyr Ile Gln Tyr Glu Pro His Leu Leu Phe Ser 930 935 940 Ser Glu Ser Thr Leu His Arg Tyr Glu Val His Pro Tyr Gly Thr Leu 945 950 955 960 Pro Val Gly Pro Gly Pro Glu Phe Lys Thr Thr Leu Arg Val Gln Asn 965 970 975 Leu Gly Cys Tyr Val Val Ser Gly Leu Ile Ile Ser Ala Leu Leu Pro 980 985 990 Ala Val Ala His Gly Gly Asn Tyr Phe Leu Ser Leu Ser Gln Val Ile 995 1000 1005 Thr Asn Asn Ala Ser Cys Ile Val Gln Asn Leu Thr Glu Pro Pro 1010 1015 1020 Gly Pro Pro Val His Pro Glu Glu Leu Gln His Thr Asn Arg Leu 1025 1030 1035 Asn Gly Ser Asn Thr Gln Cys Gln Val Val Arg Cys His Leu Gly 1040 1045 1050 Gln Leu Ala Lys Gly Thr Glu Val Ser Val Gly Leu Leu Arg Leu 1055 1060 1065 Val His Asn Glu Phe Phe Arg Arg Ala Lys Phe Lys Ser Leu Thr 1070 1075 1080 Val Val Ser Thr Phe Glu Leu Gly Thr Glu Glu Gly Ser Val Leu 1085 1090 1095 Gln Leu Thr Glu Ala Ser Arg Trp Ser Glu Ser Leu Leu Glu Val 1100 1105 1110 Val Gln Thr Arg Pro Ile Leu Ile Ser Leu Trp Ile Leu Ile Gly 1115 1120 1125 Ser Val Leu Gly Gly Leu Leu Leu Leu Ala Leu Leu Val Phe Cys 1130 1135 1140 Leu Trp Lys Leu Gly Phe Phe Ala His Lys Lys Ile Pro Glu Glu 1145 1150 1155 Glu Lys Arg Glu Glu Lys Leu Glu Gln 1160 1165 <210> 2 <211> 1100 <212> PRT <213> Homo sapiens <220> <221> Unclassified features <222> (1)..(1100) <223> Extracellular domain of human integrin α10 <400> 2 Phe Asn Leu Asp Glu His His Pro Arg Leu Phe Pro Gly Pro Pro Glu 1 5 10 15 Ala Glu Phe Gly Tyr Ser Val Leu Gln His Val Gly Gly Gly Gln Arg 20 25 30 Trp Met Leu Val Gly Ala Pro Trp Asp Gly Pro Ser Gly Asp Arg Arg 35 40 45 Gly Asp Val Tyr Arg Cys Pro Val Gly Gly Ala His Asn Ala Pro Cys 50 55 60 Ala Lys Gly His Leu Gly Asp Tyr Gln Leu Gly Asn Ser Ser His Pro 65 70 75 80 Ala Val Asn Met His Leu Gly Met Ser Leu Leu Glu Thr Asp Gly Asp 85 90 95 Gly Gly Phe Met Ala Cys Ala Pro Leu Trp Ser Arg Ala Cys Gly Ser 100 105 110 Ser Val Phe Ser Ser Gly Ile Cys Ala Arg Val Asp Ala Ser Phe Gln 115 120 125 Pro Gln Gly Ser Leu Ala Pro Thr Ala Gln Arg Cys Pro Thr Tyr Met 130 135 140 Asp Val Val Ile Val Leu Asp Gly Ser Asn Ser Ile Tyr Pro Trp Ser 145 150 155 160 Glu Val Gln Thr Phe Leu Arg Arg Leu Val Gly Lys Leu Phe Ile Asp 165 170 175 Pro Glu Gln Ile Gln Val Gly Leu Val Gln Tyr Gly Glu Ser Pro Val 180 185 190 His Glu Trp Ser Leu Gly Asp Phe Arg Thr Lys Glu Glu Val Val Arg 195 200 205 Ala Ala Lys Asn Leu Ser Arg Arg Glu Gly Arg Glu Thr Lys Thr Ala 210 215 220 Gln Ala Ile Met Val Ala Cys Thr Glu Gly Phe Ser Gln Ser His Gly 225 230 235 240 Gly Arg Pro Glu Ala Ala Arg Leu Leu Val Val Val Thr Asp Gly Glu 245 250 255 Ser His Asp Gly Glu Glu Leu Pro Ala Ala Leu Lys Ala Cys Glu Ala 260 265 270 Gly Arg Val Thr Arg Tyr Gly Ile Ala Val Leu Gly His Tyr Leu Arg 275 280 285 Arg Gln Arg Asp Pro Ser Ser Phe Leu Arg Glu Ile Arg Thr Ile Ala 290 295 300 Ser Asp Pro Asp Glu Arg Phe Phe Phe Asn Val Thr Asp Glu Ala Ala 305 310 315 320 Leu Thr Asp Ile Val Asp Ala Leu Gly Asp Arg Ile Phe Gly Leu Glu 325 330 335 Gly Ser His Ala Glu Asn Glu Ser Ser Phe Gly Leu Glu Met Ser Gln 340 345 350 Ile Gly Phe Ser Thr His Arg Leu Lys Asp Gly Ile Leu Phe Gly Met 355 360 365 Val Gly Ala Tyr Asp Trp Gly Gly Ser Val Leu Trp Leu Glu Gly Gly 370 375 380 His Arg Leu Phe Pro Pro Arg Met Ala Leu Glu Asp Glu Phe Pro Pro 385 390 395 400 Ala Leu Gln Asn His Ala Ala Tyr Leu Gly Tyr Ser Val Ser Ser Met 405 410 415 Leu Leu Arg Gly Gly Arg Arg Leu Phe Leu Ser Gly Ala Pro Arg Phe 420 425 430 Arg His Arg Gly Lys Val Ile Ala Phe Gln Leu Lys Lys Asp Gly Ala 435 440 445 Val Arg Val Ala Gln Ser Leu Gln Gly Glu Gln Ile Gly Ser Tyr Phe 450 455 460 Gly Ser Glu Leu Cys Pro Leu Asp Thr Asp Arg Asp Gly Thr Thr Asp 465 470 475 480 Val Leu Leu Val Ala Ala Pro Met Phe Leu Gly Pro Gln Asn Lys Glu 485 490 495 Thr Gly Arg Val Tyr Val Tyr Leu Val Gly Gln Gln Ser Leu Leu Thr 500 505 510 Leu Gln Gly Thr Leu Gln Pro Glu Pro Pro Gln Asp Ala Arg Phe Gly 515 520 525 Phe Ala Met Gly Ala Leu Pro Asp Leu Asn Gln Asp Gly Phe Ala Asp 530 535 540 Val Ala Val Gly Ala Pro Leu Glu Asp Gly His Gln Gly Ala Leu Tyr 545 550 555 560 Leu Tyr His Gly Thr Gln Ser Gly Val Arg Pro His Pro Ala Gln Arg 565 570 575 Ile Ala Ala Ala Ser Met Pro His Ala Leu Ser Tyr Phe Gly Arg Ser 580 585 590 Val Asp Gly Arg Leu Asp Leu Asp Gly Asp Asp Leu Val Asp Val Ala 595 600 605 Val Gly Ala Gln Gly Ala Ala Ile Leu Leu Ser Ser Arg Pro Ile Val 610 615 620 His Leu Thr Pro Ser Leu Glu Val Thr Pro Gln Ala Ile Ser Val Val 625 630 635 640 Gln Arg Asp Cys Arg Arg Arg Gly Gln Glu Ala Val Cys Leu Thr Ala 645 650 655 Ala Leu Cys Phe Gln Val Thr Ser Arg Thr Pro Gly Arg Trp Asp His 660 665 670 Gln Phe Tyr Met Arg Phe Thr Ala Ser Leu Asp Glu Trp Thr Ala Gly 675 680 685 Ala Arg Ala Ala Phe Asp Gly Ser Gly Gln Arg Leu Ser Pro Arg Arg 690 695 700 Leu Arg Leu Ser Val Gly Asn Val Thr Cys Glu Gln Leu His Phe His 705 710 715 720 Val Leu Asp Thr Ser Asp Tyr Leu Arg Pro Val Ala Leu Thr Val Thr 725 730 735 Phe Ala Leu Asp Asn Thr Thr Lys Pro Gly Pro Val Leu Asn Glu Gly 740 745 750 Ser Pro Thr Ser Ile Gln Lys Leu Val Pro Phe Ser Lys Asp Cys Gly 755 760 765 Pro Asp Asn Glu Cys Val Thr Asp Leu Val Leu Gln Val Asn Met Asp 770 775 780 Ile Arg Gly Ser Arg Lys Ala Pro Phe Val Val Arg Gly Gly Arg Arg 785 790 795 800 Lys Val Leu Val Ser Thr Thr Leu Glu Asn Arg Lys Glu Asn Ala Tyr 805 810 815 Asn Thr Ser Leu Ser Leu Ile Phe Ser Arg Asn Leu His Leu Ala Ser 820 825 830 Leu Thr Pro Gln Arg Glu Ser Pro Ile Lys Val Glu Cys Ala Ala Pro 835 840 845 Ser Ala His Ala Arg Leu Cys Ser Val Gly His Pro Val Phe Gln Thr 850 855 860 Gly Ala Lys Val Thr Phe Leu Leu Glu Phe Glu Phe Ser Cys Ser Ser 865 870 875 880 Leu Leu Ser Gln Val Phe Val Lys Leu Thr Ala Ser Ser Asp Ser Leu 885 890 895 Glu Arg Asn Gly Thr Leu Gln Asp Asn Thr Ala Gln Thr Ser Ala Tyr 900 905 910 Ile Gln Tyr Glu Pro His Leu Leu Phe Ser Ser Glu Ser Thr Leu His 915 920 925 Arg Tyr Glu Val His Pro Tyr Gly Thr Leu Pro Val Gly Pro Gly Pro 930 935 940 Glu Phe Lys Thr Thr Leu Arg Val Gln Asn Leu Gly Cys Tyr Val Val 945 950 955 960 Ser Gly Leu Ile Ile Ser Ala Leu Leu Pro Ala Val Ala His Gly Gly 965 970 975 Asn Tyr Phe Leu Ser Leu Ser Gln Val Ile Thr Asn Asn Ala Ser Cys 980 985 990 Ile Val Gln Asn Leu Thr Glu Pro Pro Gly Pro Pro Val His Pro Glu 995 1000 1005 Glu Leu Gln His Thr Asn Arg Leu Asn Gly Ser Asn Thr Gln Cys 1010 1015 1020 Gln Val Val Arg Cys His Leu Gly Gln Leu Ala Lys Gly Thr Glu 1025 1030 1035 Val Ser Val Gly Leu Leu Arg Leu Val His Asn Glu Phe Phe Arg 1040 1045 1050 Arg Ala Lys Phe Lys Ser Leu Thr Val Val Ser Thr Phe Glu Leu 1055 1060 1065 Gly Thr Glu Glu Gly Ser Val Leu Gln Leu Thr Glu Ala Ser Arg 1070 1075 1080 Trp Ser Glu Ser Leu Leu Glu Val Val Gln Thr Arg Pro Ile Leu 1085 1090 1095 Ile Ser 1100 <210> 3 <211> 198 <212> PRT <213> Homo sapiens <220> <221> Unclassified features <222> (1)..(198) <223> I domain of human integrin α10 <400> 3 Cys Pro Thr Tyr Met Asp Val Val Ile Val Leu Asp Gly Ser Asn Ser 1 5 10 15 Ile Tyr Pro Trp Ser Glu Val Gln Thr Phe Leu Arg Arg Leu Val Gly 20 25 30 Lys Leu Phe Ile Asp Pro Glu Gln Ile Gln Val Gly Leu Val Gln Tyr 35 40 45 Gly Glu Ser Pro Val His Glu Trp Ser Leu Gly Asp Phe Arg Thr Lys 50 55 60 Glu Glu Val Val Arg Ala Ala Lys Asn Leu Ser Arg Arg Glu Gly Arg 65 70 75 80 Glu Thr Lys Thr Ala Gln Ala Ile Met Val Ala Cys Thr Glu Gly Phe 85 90 95 Ser Gln Ser His Gly Gly Arg Pro Glu Ala Ala Arg Leu Leu Val Val 100 105 110 Val Thr Asp Gly Glu Ser His Asp Gly Glu Glu Leu Pro Ala Ala Leu 115 120 125 Lys Ala Cys Glu Ala Gly Arg Val Thr Arg Tyr Gly Ile Ala Val Leu 130 135 140 Gly His Tyr Leu Arg Arg Gln Arg Asp Pro Ser Ser Phe Leu Arg Glu 145 150 155 160 Ile Arg Thr Ile Ala Ser Asp Pro Asp Glu Arg Phe Phe Phe Asn Val 165 170 175 Thr Asp Glu Ala Ala Leu Thr Asp Ile Val Asp Ala Leu Gly Asp Arg 180 185 190 Ile Phe Gly Leu Glu Gly 195 <210> 4 <211> 9 <212> PRT <213> Homo sapiens <220> <221> Combination <222> (1)..(9) <223> Variable heavy chain complementarity-determining region 1 (CDR-H1) <400> 4 Phe Thr Phe Ser Asp Tyr Gly Met Asn 1 5 <210> 5 <211> 17 <212> PRT <213> Homo sapiens <220> <221> Combination <222> (1)..(17) <223> Variable heavy chain complementarity-determining region 2 (CDR-H2) <400> 5 Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 6 <211> 9 <212> PRT <213> Homo sapiens <220> <221> Combination <222> (1)..(9) <223> Variable heavy chain complementarity-determining region 3 (CDR-H3) <400> 6 Gly Gly Asn Trp Gly Val Phe Asp Tyr 1 5 <210> 7 <211> 13 <212> PRT <213> Homo sapiens <220> <221> Combination <222> (1)..(13) <223> Variable light chain complementarity-determining region 4 (CDR-L1) <400> 7 Ser Gly Ser Ser Ser Ser Asn Ile Gly Ser Asn Pro Val His 1 5 10 <210> 8 <211> 7 <212> PRT <213> Homo sapiens <220> <221> Combination <222> (1)..(7) <223> Variable light chain complementarity-determining region 5 (CDR-L2) <400> 8 Glu Asn Asn Lys Arg Pro Ser 1 5 <210> 9 <211> 12 <212> PRT <213> Homo sapiens <220> <221> Combination <222> (1)..(12) <223> Variable light chain complementarity-determining region 6 (CDR-L3) <400> 9 Ala Ala Trp Asp Asp Ser Leu Ser Gly Gln Gly Val 1 5 10 <210> 10 <211> 118 <212> PRT <213> Homo sapiens <220> <221> chain <222> (1)..(118) <223> Heavy chain variable region <400> 10 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Asn Trp Gly Val Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 11 <211> 112 <212> PRT <213> Homo sapiens <220> <221> chain <222> (1)..(112) <223> light chain variable region <400> 11 Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30 Pro Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Glu Asn Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95 Ser Gly Gln Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110

Claims

1. Use of an anti-integrin α10 antibody or an antigen-binding fragment thereof in the preparation of a medicament for treating and / or preventing invasive cancer forms, wherein said antibody or antigen-binding fragment specifically binds to an integrin α10 polypeptide, said invasive cancer forms being selected from: triple-negative breast cancer, inflammatory breast cancer, squamous cell lung cancer, lung adenocarcinoma, large cell lung cancer, small cell lung cancer, small cell neuroendocrine carcinoma (SCNC), and invasive pancreatic cancer. The antibody or its antigen-binding fragment is: a) Monoclonal antibodies produced using hybridoma cell lines deposited at the German Culture Collection Center (DSM-ACC2583); or b) A monoclonal antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 10 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:

11.

2. The use according to claim 1, wherein the triple-negative breast cancer is selected from basaloid 1 breast cancer, basaloid 2 breast cancer, low tight junction protein breast cancer, metaplastic breast cancer (MBC), interferon-rich breast cancer, immunomodulatory breast cancer, mesenchymal breast cancer, mesenchymal stem cell-like breast cancer, luminal androgen receptor breast cancer, and unstable breast cancer.

3. The use according to claim 1 or 2, wherein the invasive pancreatic cancer is a neuroendocrine tumor.

4. The use according to claim 1 or 2, wherein the invasive pancreatic cancer and / or neuroendocrine tumor is grade I, II or III pancreatic cancer.

5. The use according to claim 1 or 2, wherein the integrin α10 polypeptide is expressed on the surface of malignant cells and / or tumor-associated cells.

6. The use according to claim 1 or 2, wherein the antibody has an isotype selected from IgA, IgD, IgG, IgE and IgM.

7. The use according to claim 1 or 2, wherein the antibody or its antigen-binding fragment is conjugated to an additional portion.

8. The use according to claim 1 or 2, wherein the additional portion comprises a detectable portion.

9. The use according to claim 8, wherein the detectable portion is selected from fluorophores, enzymes, and radiotracers.

10. The use according to claim 8, wherein the detectable portion is selected from radioactive isotopes.

11. The use according to claim 1 or 2, wherein the additional portion comprises a cytotoxic portion.

12. The use according to claim 1 or 2, wherein the cytotoxic portion is selected from toxins, chemotherapeutic agents, and radiopharmaceuticals or combinations thereof.

13. The use according to claim 1 or 2, wherein the cytotoxic portion is a toxin.

14. The use according to claim 1 or 2, wherein the toxin is selected from microtubule toxins, DNA toxins, and transcription toxins.

15. The use according to claim 1 or 2, wherein the microtubule toxin is selected from auristatin-based toxins, maytansine-based toxins, microtubule-lysin-based toxins, and eribulin.

16. The use according to claim 1 or 2, wherein the transcription toxin is an RNA polymerase II inhibitor.

17. The use according to claim 1 or 2, wherein the transcription toxin is selected from doxorubicin, doxorubicin derivatives and amatoxins.

18. The use according to claim 1 or 2, wherein the transcription toxin is selected from Shiga toxin and Shiga-like toxin, type I ribosome inactivating protein, type II ribosome inactivating protein and saponin or a combination thereof.

19. The use according to claim 1 or 2, wherein the type I ribosome inactivating protein is trichosanthes pollen protein and / or loofah seed protein.

20. The use according to claim 1 or 2, wherein the type II ribosome inactivating protein is ricin, lectin, and / or absinthecin.

21. The use according to claim 1 or 2, wherein the integrin α10 polypeptide is a naturally occurring variant of the integrin α10 polypeptide or an isotype of the integrin α10 polypeptide.

22. The use according to claim 1 or 2, wherein the antibody is capable of inducing cell death in cells expressing integrin α10 polypeptide and / or inhibiting cell growth and / or inhibiting cell proliferation and / or inhibiting cell migration.

23. The use according to claim 1 or 2, wherein the cells are malignant cells and / or tumor-associated cells.

24. The use according to claim 1 or 2, wherein the malignant cell or tumor-associated cell is a cancer-associated fibroblast (CAF), stromal cell, stem cell, and / or stem cell-like cell.

25. The use according to claim 1 or 2, wherein the integrin α10 polypeptide is part of the integrin α10 β1 heterodimer.

26. The use according to claim 1 or 2, wherein the treatment is preventative, ameliorative, or curative.

27. Use of an antibody or an antigen-binding fragment thereof in the preparation of a medicament for the in vitro diagnosis of invasive cancer, wherein the antibody or antigen-binding fragment thereof specifically binds to an integrin α10 polypeptide, the invasive cancer being selected from: triple-negative breast cancer, inflammatory breast cancer, squamous cell lung cancer, lung adenocarcinoma, large cell lung cancer, small cell lung cancer, small cell neuroendocrine carcinoma (SCNC), and invasive pancreatic cancer. The antibody or its antigen-binding fragment is: a) Monoclonal antibodies produced using hybridoma cell lines deposited at the German Culture Collection Center (DSM-ACC2583); or b) A monoclonal antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 10 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:

11.

28. Use of an antibody or an antigen-binding fragment thereof in the preparation of a pharmaceutical agent for use in a method for detecting invasive cancer cells in a subject, wherein the antibody or antigen-binding fragment thereof specifically binds to an integrin α10 polypeptide, the method comprising the steps of: a. Provide the subject with tissue suspected of containing cancer cells; b. Analyze the presence of antigens containing integrin α10 polypeptide or fragments thereof in the tissue by specifically binding the antibody or its antigen-binding fragment to the integrin α10 polypeptide. c. Determine the expression level of the integrin α10 antigen, and d. The expression levels determined in c. will be compared with control levels. The control level is the average expression level of the antigen observed in healthy and / or benign cells of the same tissue type as the isolated sample. The higher expression level of the antigen compared to the control level indicates the presence of an invasive form of cancer in the subject, and the invasive form of cancer is selected from triple-negative breast cancer, inflammatory breast cancer, squamous cell lung cancer, lung adenocarcinoma, large cell lung cancer, small cell lung cancer, small cell neuroendocrine carcinoma (SCNC), and invasive pancreatic cancer. The antibody or its antigen-binding fragment is: a) Monoclonal antibodies produced using hybridoma cell lines deposited at the German Culture Collection Center (DSM-ACC2583); or b) A monoclonal antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 10 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:

11.

29. Use of an antibody or an antigen-binding fragment thereof in the preparation of a pharmaceutical agent for use in a method for detecting invasive cancer cells in a subject, wherein the antibody or antigen-binding fragment thereof specifically binds to an integrin α10 polypeptide, the method comprising the steps of: a. Provide the subject with tissue suspected of containing cancer cells; b. Optionally, analyze the presence of one or more cells with cancerous morphologies in the tissue. c. The presence of antigens containing integrin α10 polypeptide or fragments thereof in the tissue is analyzed by the specific binding of the antibody or its antigen-binding fragment to the integrin α10 polypeptide. d. Optionally determine the expression level of the integrin α10 antigen. The presence of one or more cells with cancerous morphology, combined with the expression of integrin α10 antigen, indicates the presence of an invasive cancer form in the subject, wherein the invasive cancer form is selected from triple-negative breast cancer, inflammatory breast cancer, squamous cell lung cancer, lung adenocarcinoma, large cell lung cancer, small cell lung cancer, small cell neuroendocrine carcinoma (SCNC), and invasive pancreatic cancer. The antibody or its antigen-binding fragment is: a) Monoclonal antibodies produced using hybridoma cell lines deposited at the German Culture Collection Center (DSM-ACC2583); or b) A monoclonal antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 10 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:

11.

30. Use of an antibody or an antigen-binding fragment thereof in the preparation of a pharmaceutical agent for use in a method for diagnosing an invasive form of cancer in a subject, wherein the antibody or antigen-binding fragment thereof specifically binds to an integrin α10 polypeptide, the method comprising the steps of: a. Provide the subject with tissue suspected of containing cancer cells; b. Analyze the presence of antigens containing integrin α10 polypeptide or fragments thereof in the tissue by specifically binding the antibody or its antigen-binding fragment to the integrin α10 polypeptide. c. Determine the expression level of the integrin α10 antigen, and d. The expression level determined in c. will be compared with a control level, wherein the control level is the average expression level of the antigen; and The expression level of the antigen being higher than the control level indicates the presence of an invasive cancer form in the sample, wherein the invasive cancer form is selected from triple-negative breast cancer, inflammatory breast cancer, squamous cell lung cancer, lung adenocarcinoma, large cell lung cancer, small cell lung cancer, small cell neuroendocrine carcinoma (SCNC), and invasive pancreatic cancer. This allows for the diagnosis of the subject's aggressive cancer form. The antibody or its antigen-binding fragment is: a) Monoclonal antibodies produced using hybridoma cell lines deposited at the German Culture Collection Center (DSM-ACC2583); or b) A monoclonal antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 10 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:

11.

31. Use of an antibody or an antigen-binding fragment thereof in the preparation of a pharmaceutical agent for use in a method for diagnosing an invasive form of cancer in a subject, wherein the antibody or antigen-binding fragment thereof specifically binds to an integrin α10 polypeptide, the method comprising the steps of: a. Provide the subject with tissue suspected of containing cancer cells; b. Optionally, analyze the presence of one or more cells with cancerous morphology in the tissue; c. The presence of antigens containing integrin α10 peptide or its fragments in the sample is analyzed by the specific binding of the antibody or its antigen-binding fragment to the integrin α10 peptide. d. Optionally determine the expression level of the integrin α10 antigen, and The presence of one or more cells exhibiting cancerous morphology, combined with the expression of integrin α10 antigen, indicates the presence of an invasive cancer form in the sample, wherein the invasive cancer form is selected from triple-negative breast cancer, inflammatory breast cancer, squamous cell lung cancer, lung adenocarcinoma, large cell lung cancer, small cell lung cancer, small cell neuroendocrine carcinoma (SCNC), and invasive pancreatic cancer. This allows for the diagnosis of the subject's aggressive cancer form. The antibody or its antigen-binding fragment is: a) Monoclonal antibodies produced using hybridoma cell lines deposited at the German Culture Collection Center (DSM-ACC2583); or b) A monoclonal antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 10 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:

11.

32. Use of an antibody or an antigen-binding fragment thereof in the preparation of a pharmaceutical agent for use in a method for classifying triple-negative breast cancer tumor samples of a subject, wherein the antibody or antigen-binding fragment thereof specifically binds to an integrin α10 polypeptide, the method comprising: a. Provide the subject with breast tissue suspected of containing cancer cells; b. Isolate breast cancer cells characterized as ER-negative, PR-negative, and HER2-negative; c. Determine the expression level of the antigen containing the integrin α10 peptide or its fragment in the isolated cells by the specific binding of the antibody or its antigen-binding fragment to the integrin α10 peptide, and d. The expression level determined in c. will be compared with a control level, wherein the control level is the average expression level of the antigen observed in healthy and / or benign breast tissue; The expression level of the antigen in the breast cancer cells was higher than the control level, and the expression status of ER-negative, PR-negative, and HER2-negative indicated basal-like triple-negative breast cancer or luminal triple-negative breast cancer. This allows the triple-negative breast cancer tumor samples to be classified as either basal-like triple-negative breast cancer tumors or luminal triple-negative breast cancer tumors. The antibody or its antigen-binding fragment is: a) Monoclonal antibodies produced using hybridoma cell lines deposited at the German Culture Collection Center (DSM-ACC2583); or b) A monoclonal antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 10 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:

11.

33. The use according to any one of claims 28 to 32, wherein the step of analyzing the sample contains an antigen comprising integrin α10 polypeptide or a fragment thereof. This includes imaging the tissue and / or tissue samples.

34. The use according to any one of claims 28 to 32, wherein the step of determining the expression level of the integrin α10 antigen includes imaging the tissue and / or tissue sample.

35. The use according to claim 28, wherein the antibody or antigen-binding fragment is administered after the primary cancer is detected.

36. Use of an antibody or an antigen-binding fragment thereof in the preparation of a medicament for inhibiting integrin α10-mediated signaling in at least one cancer cell, wherein the antibody or antigen-binding fragment thereof specifically binds to an integrin α10 polypeptide, wherein the at least one cancer cell is contacted with an effective amount of an antibody or antigen-binding fragment thereof that is specific to the integrin α10 polypeptide, wherein the at least one cancer cell is selected from invasive breast cancer cells, invasive lung cancer cells, invasive prostate cancer cells, invasive pancreatic cancer cells, and metastatic tumor cells. The antibody or its antigen-binding fragment is: a) Monoclonal antibodies produced using hybridoma cell lines deposited at the German Culture Collection Center (DSM-ACC2583); or b) A monoclonal antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 10 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:

11.

37. Use of an antibody or an antigen-binding fragment thereof in the preparation of a medicament for inhibiting the cellular function of at least one cancer cell, wherein the antibody or antigen-binding fragment thereof specifically binds to an integrin α10 polypeptide, wherein the at least one cancer cell is contacted with an effective amount of an antibody or antigen-binding fragment specifically targeting the integrin α10 polypeptide, wherein the at least one cancer cell is selected from invasive breast cancer cells, invasive lung cancer cells, invasive prostate cancer cells, invasive pancreatic cancer cells, and metastatic tumor cells. The antibody or its antigen-binding fragment is: a) Monoclonal antibodies produced using hybridoma cell lines deposited at the German Culture Collection Center (DSM-ACC2583); or b) A monoclonal antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 10 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:

11.

38. The use according to any one of claims 28 to 32 and 36-37, wherein the antigen comprising the integrin α10 polypeptide or a fragment thereof is expressed on the surface of the cell.

39. The use according to claim 37, wherein the at least one cancer cell is in an invasive and / or metastatic tumor, and wherein the inhibition is selected from: a. Inhibit the proliferation of at least one of the cancer cells; b. Inhibit the self-renewal of at least one of the cancer cells; c. Inhibit the anchorage-independent growth of the at least one cancer cell; d. Inhibit the migration of at least one of the cancer cells; e. Inhibit the invasion of at least one of the cancer cells; f. Inhibit the activity of at least one of the cancer cells; g. Inhibiting the adhesion of at least one of the cancer cells; and / or Its combination.

40. The use according to claim 37, wherein the at least one cancer cell is in an invasive and / or metastatic tumor, and the inhibition of the cellular function of the at least one cancer cell inhibits at least one of the following: a. The growth of the invasive and / or metastatic tumors; b. The proliferation of the invasive and / or metastatic tumors; c. The migration of the invasive and / or metastatic tumors; d. Invasion of the aforementioned invasive and / or metastatic tumors; e. Spread of new invasive and / or metastatic tumors; f. The onset of new invasive and / or metastatic tumors; g. Infiltration of new invasive and / or metastatic tumors; and / or Its combination.

41. The use according to claim 36 or 37, wherein the at least one cancer cell is characterized in that the expression level of an antigen comprising an integrin α10 polypeptide or a fragment thereof is higher than that of a control, wherein the control level is the average expression level of the antigen observed in healthy and / or benign cells of the same tissue type as the cancer cell.

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