Vimentin Directed Diagnostics And Therapeutics For Multidrug Resistant Neoplastic Disease

Abstract

Disclosed are methods for detecting multidrug resistance in neoplastic or damaged cells or multidrug resistant (MDR) neoplastic or damaged cells by detecting an increase in the cell surface expression of vimentin protein in such cells as compared to the level of cell surface expression of vimentin protein in a normal cell or a non-MDR neoplastic cell.

1 .- 9 . (canceled) 
     
     
         10 . A method for detecting a multidrug resistant cell in a patient comprising:
 (a) administering to the patient, a vimentin binding agent operably linked to a detectable label; and   (b) detecting the label operably linked to the vimentin binding agent,
 wherein the vimentin binding agent specifically binds to cell surface-expressed vimentin present on a multidrug resistant cell in the patient. 
   
     
     
         11 . The method of  claim 10 , wherein the vimentin binding agent is an antibody or fragment thereof. 
     
     
         12 . The method of  claim 10 , wherein the vimentin binding agent is selected from the group consisting of modified LDL, NLK1 protein, vimentin, desmin, glial fibrillary acidic protein, and peripherin, fimbrin, RhoA-binding kinase alpha, and protein phosphatase 2A. 
     
     
         13 . The method of  claim 10 , wherein the vimentin binding agent is selected from the group consisting of natural ligands, synthetic small molecules, chemicals, nucleic acids, peptides, proteins, and antibodies. 
     
     
         14 . The method of  claim 10 , wherein the detectable label is selected from the group consisting of fluorophores, chemical dyes, radioactive compounds, chemoluminescent compounds, magnetic compounds, paramagnetic compounds, promagnetic compounds, enzymes that yield a colored product, enzymes that yield a chemoluminescent product, and enzymes that yield a magnetic product. 
     
     
         15 . The method of  claim 14 , wherein the multidrug resistant cell is a neoplastic cell. 
     
     
         16 . The method of  claim 15 , wherein the neoplastic cell is selected from the group consisting of a breast cancer cell, an ovarian cancer cell, a myeloma cancer cell, a lymphoma cancer cell, a melanoma cancer cell, a sarcoma cancer cell, a leukemia cancer cell, a retinoblastoma cancer cell, a hepatoma cancer cell, a glioma cancer cell, a mesothelioma cancer cell, and a carcinoma cancer cell. 
     
     
         17 . The method of  claim 15 , wherein the neoplastic cell is selected from the group consisting of a promyleocytic leukemia cell, a T lymphoblastoid cell, a breast epithelial cell, and an ovarian cell. 
     
     
         18 . The method of  claim 10 , wherein the patient is a human. 
     
     
         19 . The method of  claim 18 , wherein the patient is suffering from a disease or disorder caused by the presence of the multidrug resistant cell. 
     
     
         20 . A kit for diagnosing or detecting multidrug resistance in a test neoplastic cell comprising:
 a) a first probe for the detection of vimentin; and   b) a second probe for the detection of a multidrug resistance marker selected from the group consisting of nucleophosmin and HSC70.   
     
     
         21 . A kit for diagnosing or detecting multidrug resistance in a test neoplastic cell comprising:
 a) a first probe for the detection of vimentin; and   b) a second probe for the detection of a marker selected from the group consisting of MDR1, MDR3, MRP1, MRP5, and LRP.   
     
     
         22 . The kit of  claim 21 , wherein the probe for detecting vimentin is an anti-vimentin antibody. 
     
     
         23 . The kit of  claim 21 , wherein the probe for detecting vimentin is a vimentin ligand selected from the group consisting of LDL, NLK1 protein, vimentin, desmin, glial fibrillary acidic protein, and peripherin, fimbrin, RhoA-binding kinase alpha, and protein phosphatase 2A. 
     
     
         24 . The kit of  claim 20 , wherein the second probe is selected from the group consisting of a nucleophosmin antibody and an HSC70 antibody. 
     
     
         25 . The kit of  claim 20 , wherein the second probe is selected from the group consisting of a nucleophosmin ligand and an HSC70 ligand. 
     
     
         26 . The kit of  claim 21 , wherein the first probe detects vimentin present on the surface of the test neoplastic cell. 
     
     
         27 . The kit of  claim 21 , wherein the second probe detects a marker present of the surface of the test neoplastic cell. 
     
     
         28 . The kit of  claim 21 , wherein the second probe is selected from the group consisting of: an MDR1 antibody, an MDR3 antibody, an MRP1 antibody, an MRP3 antibody, and an LRP antibody. 
     
     
         29 . A cell surface vimentin in situ detection probe for the detection of cell surface vimentin in a patient, comprising a vimentin binding component and a detectable label for detection in situ. 
     
     
         30 . The cell surface vimentin in situ detection probe of  claim 29 , wherein the vimentin binding component is an antibody. 
     
     
         31 . The cell surface vimentin in situ detection probe of  claim 29 , wherein the detectable label is Technetium. 
     
     
         32 . A cell surface vimentin-targeted agent for treating or preventing a multi-drug resistant neoplasm, comprising a vimentin binding component and a therapeutic component, wherein the vimentin binding component targets the therapeutic component to the multi-drug resistant neoplasm and thereby treats the multi-drug resistant neoplasm. 
     
     
         33 . The agent of  claim 32 , wherein the vimentin binding component is an anti-vimentin antibody. 
     
     
         34 . The agent of  claim 32 , wherein the vimentin binding component is selected from the group consisting of LDL, NLK1 protein, vimentin, desmin, glial fibrillary acidic protein, peripherin, fimbrin, RhoA-binding kinase alpha, and protein phosphatase 2A. 
     
     
         35 . The agent of  claim 32 , wherein said vimentin binding component is selected from the group consisting of natural ligands, synthetic small molecules, chemicals, nucleic acids, peptides, proteins, antibodies, and vimentin binding fragments thereof. 
     
     
         36 . The agent of  claim 32 , wherein the therapeutic component is selected from the group consisting of Actinomycin, Adriamycin, Altretamine, Asparaginase, Bleomycin, Busulfan, Capecitabine, Carboplatin, Carmustine, Chlorambucil, Cisplatin, Cladribine, Cyclophosphamide, Cytarabine, Dacarbazine, Dactinomycin, Daunorubicin, Docetaxel, Doxorubicin, Epoetin, Etoposide, Fludarabine, Fluorouracil, Gemcitabine, Hydroxyurea, Idarubicin, Ifosfamide, Imatinib, Irinotecan, Lomustine, Mechlorethamine, Melphalan, Mercaptopurine, Methotrexate, Mitomycin, Mitotane, Mitoxantrone, Paclitaxel, Pentostatin, Procarbazine, Taxol, Teniposide, Topotecan, Vinblastine, Vincristine, and Vinorelbine. 
     
     
         37 . The agent of  claim 32 , wherein the therapeutic component is in a liposome formulation. 
     
     
         38 . The agent of  claim 32 , wherein the therapeutic component is a radioisotope. 
     
     
         39 . The agent of  claim 38 , wherein the radioisotope is selected from the group consisting of  90 Y,  125 I,  131 I,  211 At, and  213 Bi. 
     
     
         40 . The agent of  claim 32 , wherein the therapeutic component is a toxin capable of killing or inducing the killing of the targeted multi-drug resistant neoplastic cell. 
     
     
         41 . The agent of  claim 40 , wherein the toxin is selected from the group consisting of a Pseudomonas exotoxin, a diphtheria toxin, a plant ricin toxin, a plant abrin toxin, a plant saporin toxin, a plant gelonin toxin, and pokeweed antiviral protein. 
     
     
         42 . The agent of  claim 32 , wherein the vimentin binding component binds to the surface of the target cell and the therapeutic element is internalized and arrests growth of the cell, compromises viability of the cell or kills the cell. 
     
     
         43 . A vaccine for treating or preventing a multi-drug resistant neoplasm, comprising a vimentin polypeptide, or vimentin polypeptide subsequence thereof, and at least one pharmaceutically acceptable vaccine component. 
     
     
         44 . The vaccine of  claim 43 , wherein the vimentin polypeptide or polypeptide subsequence is a human vimentin polypeptide sequence of SEQ ID NO.: 1. 
     
     
         45 . The vaccine of  claim 43 , wherein the vimentin polypeptide subsequence is at least eight amino acids long. 
     
     
         46 . The vaccine of  claim 45 , wherein the vimentin polypeptide subsequence comprises a hapten. 
     
     
         47 . The vaccine of  claim 43 , wherein the pharmaceutically acceptable vaccine component is an adjuvant. 
     
     
         48 . The vaccine of  claim 47 , wherein the adjuvant is selected from the group consisting of aluminum hydroxide, aluminum phosphate, calcium phosphate, oil emulsion, a bacterial product, whole inactivated bacteria, an endotoxins, cholesterol, a fatty acid, an aliphatic amine, a paraffinic compound, a vegetable oil, monophosphoryl lipid A, a saponin, and squalene. 
     
     
         49 . A method of treating or preventing a multidrug resistant neoplasm in a subject comprising administering a cell surface vimentin-targeted therapeutic agent of  claim 32 . 
     
     
         50 . The method of  claim 49 , wherein the neoplasm is selected from the group consisting of a breast cancer, an ovarian cancer, a myeloma, a lymphoma, a melanoma, a sarcoma, a leukemia, a retinoblastoma, a hepatoma, a glioma, a mesothelioma, and a carcinoma. 
     
     
         51 . The method of  claim 49 , wherein the subject is a human patient. 
     
     
         52 . The method of  claim 51 , wherein the human patient is suffering from a disease or disorder caused by the presence of the multi-drug resistant cell. 
     
     
         53 . The method of  claim 49 , wherein the neoplasm is from a tissue selected from the group consisting of blood, bone marrow, spleen, lymph node, liver, thymus, kidney, brain, skin, gastrointestinal tract, eye, breast, prostate, and ovary. 
     
     
         54 . A method of treating or preventing a multidrug resistant neoplasm in a subject comprising administering a vimentin vaccine of  claim 43 . 
     
     
         55 . The method of  claim 54 , wherein the neoplasm is selected from the group consisting of a breast cancer, an ovarian cancer, a myeloma, a lymphoma, a melanoma, a sarcoma, a leukemia, a retinoblastoma, a hepatoma, a glioma, a mesothelioma, and a carcinoma. 
     
     
         56 . The method of  claim 54 , wherein the subject is a human patient. 
     
     
         57 . The method of  claim 56 , wherein the human patient is suffering from a disease or disorder caused by the presence of the multi-drug resistant cell. 
     
     
         58 . The method of  claim 54 , wherein the neoplasm is from a tissue selected from the group consisting of blood, bone marrow, spleen, lymph node, liver, thymus, kidney, brain, skin, gastrointestinal tract, eye, breast, prostate, and ovary. 
     
     
         59 - 65 . (canceled) 
     
     
         66 . A method for detecting a neoplastic cell in a patient comprising:
 (a) administering to the patient, a vimentin binding agent operably linked to a detectable label; and   (b) detecting the label operably linked to the vimentin binding agent,
 wherein the vimentin binding agent specifically binds to cell surface-expressed vimentin present on a neoplastic cell in the patient. 
   
     
     
         67 . The method of  claim 66 , wherein the vimentin binding agent is an antibody or fragment thereof. 
     
     
         68 . The method of  claim 66 , wherein the vimentin binding agent is selected from the group consisting of modified LDL, NLK1 protein, vimentin, desmin, glial fibrillary acidic protein, and peripherin, fimbrin, RhoA-binding kinase alpha, and protein phosphatase 2A. 
     
     
         69 . The method of  claim 66 , wherein the vimentin binding agent is selected from the group consisting of natural ligands, synthetic small molecules, chemicals, nucleic acids, peptides, proteins, antibodies, and fragments thereof. 
     
     
         70 . The method of  claim 66 , wherein the detectable label is selected from the group consisting of fluorophores, chemical dyes, radioactive compounds, chemoluminescent compounds, magnetic compounds, paramagnetic compounds, promagnetic compounds, enzymes that yield a colored product, enzymes that yield a chemoluminescent product, and enzymes that yield a magnetic product. 
     
     
         71 . The method of  claim 66 , wherein the neoplastic cell is selected from the group consisting of a breast cancer cell, an ovarian cancer cell, a myeloma cancer cell, a lymphoma cancer cell, a melanoma cancer cell, a sarcoma cancer cell, a leukemia cancer cell, a retinoblastoma cancer cell, a hepatoma cancer cell, a glioma cancer cell, a mesothelioma cancer cell, and a carcinoma cancer cell. 
     
     
         72 . The method of  claim 66 , wherein the neoplastic cell is selected from the group consisting of a promyleocytic leukemia cell, a T lymphoblastoid cell, a breast epithelial cell, and an ovarian cell. 
     
     
         73 . The method of  claim 66 , wherein the patient is a human. 
     
     
         74 . The method of  claim 73 , wherein the patient is suffering from a disease or disorder caused by the presence of the neoplastic cell. 
     
     
         75 . A kit for diagnosing or detecting neoplasia, comprising:
 a) a first probe for the detection of vimentin; and   b) a second probe for the detection of a neoplasia marker selected from the group consisting of nucleophosmin and HSC70.   
     
     
         76 . The kit of  claim 75 , wherein the probe for detecting vimentin is an anti-vimentin antibody or binding fragment thereof. 
     
     
         77 . The kit of  claim 75 , wherein the probe for detecting vimentin is a vimentin ligand selected from the group consisting of LDL, NLK1 protein, vimentin, desmin, glial fibrillary acidic protein, and peripherin, fimbrin, RhoA-binding kinase alpha, and protein phosphatase 2A. 
     
     
         78 . The kit of  claim 75 , wherein the second probe is selected from the group consisting of a nucleophosmin antibody and an HSC70 antibody. 
     
     
         79 . The kit of  claim 75 , wherein the second probe is selected from the group consisting of a nucleophosmin ligand and an HSC70 ligand. 
     
     
         80 . The kit of  claim 75 , wherein the first probe detects vimentin present on the surface of the test cell if it is neoplastic. 
     
     
         81 . The kit of  claim 75 , wherein the second probe detects a marker present of the surface of the test cell if it is neoplastic. 
     
     
         82 . A cell surface vimentin-targeted agent for treating a cancerous neoplastic cell growth comprising a vimentin binding component and a therapeutic component, wherein the vimentin binding component targets the therapeutic component to the neoplastic cell growth and thereby treats the cancer. 
     
     
         83 . The agent of  claim 82 , wherein the vimentin binding component is an anti-vimentin antibody. 
     
     
         84 . The agent of  claim 82 , wherein the vimentin binding component is selected from the group consisting of LDL, NLK1 protein, vimentin, desmin, glial fibrillary acidic protein, and peripherin, fimbrin, RhoA-binding kinase alpha, and protein phosphatase 2A. 
     
     
         85 . The agent of  claim 82 , wherein said vimentin binding component is selected from the group consisting of natural ligands, synthetic small molecules, chemicals, nucleic acids, peptides, proteins, antibodies, and vimentin binding fragments thereof. 
     
     
         86 . The agent of  claim 82 , wherein the therapeutic component is selected from the group consisting of Actinomycin, Adriamycin, Altretamine, Asparaginase, Bleomycin, Busulfan, Capecitabine, Carboplatin, Carmustine, Chlorambucil, Cisplatin, Cladribine, Cyclophosphamide, Cytarabine, Dacarbazine, Dactinomycin, Daunorubicin, Docetaxel, Doxorubicin, Epoetin, Etoposide, Fludarabine, Fluorouracil, Gemcitabine, Hydroxyurea, Idarubicin, Ifosfamide, Imatinib, Irinotecan, Lomustine, Mechlorethamine, Melphalan, Mercaptopurine, Methotrexate, Mitomycin, Mitotane, Mitoxantrone, Paclitaxel, Pentostatin, Procarbazine, Taxol, Teniposide, Topotecan, Vinblastine, Vincristine, and Vinorelbine and combinations thereof. 
     
     
         87 . The agent of  claim 82 , wherein the therapeutic component is in a liposome formulation. 
     
     
         88 . The agent of  claim 82 , wherein the therapeutic component is a radioisotope. 
     
     
         89 . The agent of  claim 88 , wherein the radioisotope is selected from the group consisting of  90 Y,  111 In,  125 I,  131 I,  211 At, and  213 Bi. 
     
     
         90 . The agent of  claim 82 , wherein the therapeutic component is a toxin capable of killing or inducing the killing of the targeted neoplastic cell. 
     
     
         91 . The agent of  claim 90 , wherein the toxin is selected from the group consisting of a Pseudomonas exotoxin, a diphtheria toxin, a plant ricin toxin, a plant abrin toxin, a plant saporin toxin, a plant gelonin toxin, and pokeweed antiviral protein. 
     
     
         92 . The agent of  claim 82 , wherein the vimentin binding component binds to the surface of the target cell and the therapeutic element is internalized and arrests growth of the cell, compromises viability of the cell, or kills the cell. 
     
     
         93 . A vaccine for treating or preventing a neoplasm comprising a vimentin polypeptide, or vimentin polypeptide subsequence thereof, and at least one pharmaceutically acceptable vaccine component. 
     
     
         94 . The vaccine of  claim 93 , wherein the vimentin polypeptide or polypeptide subsequence is a human vimentin polypeptide sequence set forth in SEQ ID NO.: 1. 
     
     
         95 . The vaccine of  claim 93 , wherein the vimentin polypeptide subsequence is at least eight amino acids long. 
     
     
         96 . The vaccine of  claim 95 , wherein the vimentin polypeptide subsequence comprises a hapten. 
     
     
         97 . The vaccine of  claim 93 , wherein the pharmaceutically acceptable vaccine component is an adjuvant. 
     
     
         98 . The vaccine of  claim 97 , wherein the adjuvant is selected from the group consisting of aluminum hydroxide, aluminum phosphate, calcium phosphate, oil emulsion, a bacterial product, whole inactivated bacteria, an endotoxins, cholesterol, a fatty acid, an aliphatic amine, a paraffinic compound, a vegetable oil, monophosphoryl lipid A, a saponin, and squalene. 
     
     
         99 . A method of treating or preventing a neoplasm in a subject comprising administering a cell surface vimentin-targeted therapeutic agent of  claim 82 . 
     
     
         100 . The method of  claim 99 , wherein the neoplasm is selected from the group consisting of a breast cancer, an ovarian cancer, a myeloma, a lymphoma, a melanoma, a sarcoma, a leukemia, a retinoblastoma, a hepatoma, a glioma, a mesothelioma, and a carcinoma. 
     
     
         101 . The method of  claim 99 , wherein the subject is a human patient. 
     
     
         102 . The method of  claim 101 , wherein said human patient is suffering from a disease or disorder caused by the presence of the multi-drug resistant cell. 
     
     
         103 . The method of  claim 99 , wherein the neoplasm is from a tissue selected from the group consisting of blood, bone marrow, spleen, lymph node, liver, thymus, kidney, brain, skin, gastrointestinal tract, eye, breast, prostate and ovary. 
     
     
         104 . A method of treating or preventing a neoplasm in a subject comprising administering a vimentin vaccine of  claim 93 . 
     
     
         105 . The method of  claim 104 , wherein the neoplasm is selected from the group consisting of a breast cancer, an ovarian cancer, a myeloma, a lymphoma, a melanoma, a sarcoma, a leukemia, a retinoblastoma, a hepatoma, a glioma, a mesothelioma, and a carcinoma. 
     
     
         106 . The method of  claim 104 , wherein said subject is a human patient. 
     
     
         107 . The method of  claim 106 , wherein said human patient is suffering from a disease or disorder caused by the presence of the neoplastic cell. 
     
     
         108 . The method of  claim 104 , wherein the neoplasm is from a tissue selected from the group consisting of blood, bone marrow, spleen, lymph node, liver, thymus, kidney, brain, skin, gastrointestinal tract, eye, breast, prostate, and ovary. 
     
     
         109 . The method of  claim 10 , wherein the vimentin binding agent is an anti-vimentin antibody. 
     
     
         110 . The method of  claim 109 , wherein the anti-vimentin antibody is operably linked to a radiolabel.