Compositions and Methods for Inhibiting Growth of SMAD-4 Deficient Cancers

Abstract

The present invention is in the fields of cell biology, immunology and oncology. The invention relates to the discovery that there is a relationship between the expression levels of the tumor suppressor gene smad4 (also known as dpc4) and integrin α v β 6 , and the responsiveness of patient populations to α v β 6 -active compounds and compositions (e.g., antibodies and other ligands that bind α v β 6 ), particularly in cancer cells from such patient populations, more particularly on carcinomas such as pancreatic carcinomas. The invention thus provides methods for determining the responsiveness of tumor cells (particularly those from pancreatic tumors) to such α v β 6 -active compounds and compositions by examining the expression of α v β 6 and smad4 by the tumor cells, as well as methods of diagnosis and treatment/prevention of tumor progression using ligands, including antibodies and small molecule drugs, that bind to integrin α v β 6 on the surfaces of tumor cells and/or that block one or more components of the TGF-β pathway, particularly in smad4-deficient tumor cells.

1 - 31 . (canceled) 
     
     
         32 . A method of inhibiting TGF-β signalling pathway in a cell from a tumor that is smad4 deficient, comprising:
 (a) determining the level of expression of smad4 in a cell from said tumor; and 
 (b) treating a tumor cell that is deficient in smad4 expression with one or more with one or more agents that inhibits the TGF-β signalling pathway in said tumor cell, 
 wherein said treatment blocks TGF-B activation in said smad4 deficient tumor cell and causes growth inhibition or cell death of said tumor cell wherein said agent that inhibits the TGF-β signalling pathway in said tumor cell is selected from the group consisting of: 
 one or more antibody produced by a hybridoma selected from the group consisting of 2A1 (deposited under ATCC Accession No. ATCC PTA-3896), 2E5 (deposited under ATCC Accession No. ATCC PTA-3897), 1A8 (deposited under ATCC Accession No. ATCC PTA-3647), 2B10 deposited under ATCC Accession No. ATCC PTA-3648), 2B1 deposited under ATCC Accession No. ATCC PTA-3646), 1G10 (deposited under ATCC Accession No. ATCC PTA-3898), 7G5 (deposited under ATCC Accession No. ATCC PTA-3899), 8G6 (deposited under ATCC Accession No. ATCC PTA-3645), 3G9 (deposited under ATCC Accession No. ATCC PTA-3649), or a humanized version thereof. 
 
     
     
         33 - 34 . (canceled) 
     
     
         35 . The method of  claim 32 , wherein said tumor is a carcinoma. 
     
     
         36 . The method of  claim 35 , wherein said carcinoma is an adenocarcinoma. 
     
     
         37 . The method of  claim 35 , wherein said carcinoma is selected from the group consisting of a pancreatic carcinoma, a colorectal carcinoma, a cervical carcinoma, squamous cell carcinoma, a head and neck carcinoma, a liver carcinoma, an ovarian carcinoma and a lung carcinoma. 
     
     
         38 . The method of  claim 35 , wherein said carcinoma is a pancreatic carcinoma. 
     
     
         39 . The method of  claim 37 , wherein said squamous cell carcinoma is an esophageal carcinoma. 
     
     
         40 . The method of  claim 35 , wherein said carcinoma is a colorectal carcinoma. 
     
     
         41 . The method of  claim 35 , wherein said carcinoma is a cervical carcinoma. 
     
     
         42 . The method of  claim 35 , wherein said carcinoma is a head and neck carcinoma. 
     
     
         43 - 46 . (canceled) 
     
     
         47 . The method of  claim 32 , wherein said monoclonal antibody is 3G9. 
     
     
         48 . The method of  claim 32 , wherein said monoclonal antibody is 8G6. 
     
     
         49 . (canceled) 
     
     
         50 . The method of  claim 32 , wherein said humanized monoclonal antibody is hu3G9 (BG00011). 
     
     
         51 . The method of  claim 32 , wherein said humanized monoclonal antibody is hu8G6. 
     
     
         52 . The method of  claim 32 , wherein said antibody is conjugated with at least one detectable label or to one or more cytotoxic compounds or agents which cause the death of the cell or tissue upon binding of the αvβ6-binding antibody-toxic compound conjugate to one or more αvβ6 integrins on the cell or tissue. 
     
     
         53 . The method of  claim 52 , wherein said detectable label is selected from the group consisting of a chromogenic label, an enzyme label, a radioisotopic label, a non-radioactive isotopic label, a fluorescent label, a toxic label, a chemiluminescent label, an X-radiographic label, a spin label and a nuclear magnetic resonance contrast agent label. 
     
     
         54 . The method of  claim 53 , wherein said chromogenic label is selected from the group consisting of diaminobenzidine and 4 hydroxyazo-benzene-2-carboxylic acid. 
     
     
         55 . The method of  claim 53 , wherein said enzyme label is selected from the group consisting of malate dehydrogenase, staphylococcal nuclease, delta 5 steroid isomerase, yeast alcohol dehydrogenase, alpha glycerol phosphate dehydrogenase, triose phosphate isomerase, peroxidase, alkaline phosphatase, asparaginase, glucose oxidase, β galactosidase, ribonuclease, urease, catalase, glucose 6 phosphate dehydrogenase, glucoamylase and acetylcholine esterase. 
     
     
         56 . The method of  claim 53 , wherein said radioisotopic label is selected from the group consisting of  3 H,  111 In,  125 I,  131 I,  32 P,  35 S,  14 C,  51 Cr,  57 To,  58 Co,  59  Fe,  75 Se,  152 Eu,  90 Y,  67 Cu,  217 Ci,  211 At,  212 Pb,  47 Sc, and  109 Pd. 
     
     
         57 . The method of  claim 53 , wherein said non-radioactive isotopic label is selected from the group consisting of  157 Gd,  55 Mn,  162 Dy,  52 Tr,  56 Fe,  99m Tc and  112 In. 
     
     
         58 . The method of  claim 53 , wherein said fluorescent label is selected from the group consisting of a  152 Eu label, a fluorescein label, an isothiocyanate label, a rhodamine label, a phycoerythrin label, a phycocyanin label, an allophycocyanin label, a Green Fluorescent Protein (GFP) label, an o phthaldehyde label and a fluorescamine label. 
     
     
         59 . The method of  claim 53 , wherein said toxic label is selected from the group consisting of a diphtheria toxin label, a ricin label and a cholera toxin label. 
     
     
         60 . The method of  claim 53 , wherein said chemiluminescent label is selected from the group consisting of a luminol label, an isoluminol label, an aromatic acridinium ester label, an imidazole label, an acridinium salt label, an oxalate ester label, a luciferin label, a luciferase label and an aequorin label. 
     
     
         61 . The method of  claim 53 , wherein said X-radiographic label is barium or cesium. 
     
     
         62 . The method of  claim 53 , wherein said spin label is deuterium. 
     
     
         63 . The method of  claim 53 , wherein said nuclear magnetic resonance contrast agent label is selected from the group consisting of Gd, Mn and iron. 
     
     
         64 . (canceled) 
     
     
         65 . The method of  claim 32 , wherein said antibody is administered in combination with a growth-inhibiting chemotherapeutic compound. 
     
     
         66 . The method of  claim 65 , wherein said growth-inhibiting chemotherapeutic compound is selected from the group consisting of cisplatin, carboplatin, oxaliplatin, paclitaxel, gemcitabine, adriamycin, melphalan, methotrexate, 5-fluorouracil, etoposide, mechlorethamine, cyclophosphamide, bleomycin, a calicheamicin, a maytansine, a trichothene, CC1065, diphtheria A chain,  Pseudomonas aeruginosa  exotoxin A chain, ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, an  Aleurites fordii  protein, a dianthin protein, a  Phytolaca americana  protein,  momordica charantia  inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, a tricothecene, a ribonuclease, and a deoxyribonuclease. 
     
     
         67 . The method of  claim 52 , wherein said cytotoxic compound is one or more compounds selected from the group consisting of maytansine, cisplatin, carboplatin, oxaliplatin, paclitaxel, gemcitabine, adriamycin, melphalan, methotrexate, 5-fluorouracil, etoposide, mechlorethamine, cyclophosphamide, bleomycin, a calicheamicin, a maytansine, a trichothene, CC1065, diphtheria A chain,  Pseudomonas aeruginosa  exotoxin A chain, ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, an  Aleurites fordii  protein, a dianthin protein, a  Phytolaca americana  protein,  momordica charantia  inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, a tricothecene, a ribonuclease, and a deoxyribonuclease. 
     
     
         68 . The method of  claim 32  wherein said antibody is produced by the hydridoma 8G6 (deposited under ATCC Accession No. ATCC PTA-3645) or a humanized version thereof and said antibody is conjugated to a cytotoxic agent.