Compositions and methods for inhibiting growth of SMAD4-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 ν β 6 , and the responsiveness of patient populations to α ν β 6 -active compounds and compositions (e.g., antibodies and other ligands that bind α ν 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 ν β 6 -active compounds and compositions by examining the expression of ≢ ν 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 molecule drugs, that bind to integrin α ν 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.

What is claimed is: 
     
       1. A method of inhibiting growth of a cell from a smad4-deficient tumor that is smad4 deficient, comprising:
 (a) determining the level of expression of smad4 in a cell from said cancer smad4-deficient tumor; and 
 (b) treating a cancer cell that is deficient in smad4 expression the smad4-deficient tumor with one or more an αvβ6 antagonist antibody that selectively binds to the integrin αvβ6 in said cancer cell, wherein said one or more antibody is an antibody from produced by, or a humanized version of an 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), and 3G9 (deposited under ATCC Accession No. ATCC PTA-3649), or a humanized version thereof 
 wherein said treatment results in the growth inhibition of said cancer cell smad4-deficient tumor. 
 
     
     
       2. The method of  claim 1 , wherein said tumor is a carcinoma. 
     
     
       3. The method of  claim 2 , wherein said carcinoma is an adenocarcinoma. 
     
     
       4. The method of  claim 2 , wherein said carcinoma is selected from the group consisting of a pancreatic carcinoma, a colorectal carcinoma, a cervical carcinoma, a squamous cell carcinoma, a head and neck carcinoma, a liver carcinoma, an ovarian carcinoma and a lung carcinoma. 
     
     
       5. The method of  claim 2 , wherein said carcinoma is a pancreatic carcinoma. 
     
     
       6. The method of claim  4  2, wherein said squamous cell carcinoma is an esophageal carcinoma. 
     
     
       7. The method of  claim 2 , wherein said carcinoma is a colorectal carcinoma. 
     
     
       8. The method of  claim 2 , wherein said carcinoma is a cervical carcinoma. 
     
     
       9. The method of  claim 2 , wherein said carcinoma is a head and neck carcinoma. 
     
     
       10. The method of  claim 1 , wherein said monoclonal antibody is an antibody produced by the hybridoma 3G9 (deposited under ATCC Accession No. ATCC PTA-3649) or a humanized version thereof. 
     
     
       11. The method of  claim 1 , wherein said monoclonal antibody is an antibody produced by the hybridoma 8G6 (deposited under ATCC Accession No. ATCC PTA-3645) or a humanized version thereof. 
     
     
       12. The method of  claim 1 , wherein said monoclonal antibody is a humanized monoclonal antibody. 
     
     
       13. The method of  claim 12 , wherein said humanized monoclonal antibody is hu3G9 (BG00011). 
     
     
       14. The method of  claim 12 , wherein said humanized monoclonal antibody is hu8G6. 
     
     
       15. The method of  claim 1 , wherein said antibody is conjugated with at least one detectable label. 
     
     
       16. The method of  claim 15 , 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. 
     
     
       17. The method of  claim 16 , wherein said detectable label is a chromogenic label is selected from the group consisting of diaminobenzidine and 4 hydroxyazo-benzene-2-carboxylic acid. 
     
     
       18. The method of  claim 16 , wherein said detectable label is an 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. 
     
     
       19. The method of  claim 16 , wherein said detectable label is a radioisotopic label is selected from the group consisting of 3H, 111 In, 125I, 131I, 32P, 35S, 14C, 51Cr, 57To, 58Co, 59Fe, 75Se, 152Eu, 90Y, 67Cu, 217Ci, 211At, 212Pb, 47Sc   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 109Pd   109 Pd. 
     
     
       20. The method of  claim 16 , wherein said detectable label is a non-radioactive isotopic label is selected from the group consisting of 157Gd, 55Mn, 162Dy, 52Tr, 56Fe, 99mTc  157 Gd,  55 Mn,  162 Dy,  52 Tr,  56 Fe,  99 mTc, and 112In  112 In. 
     
     
       21. The method of  claim 16 , wherein said detectable label is a fluorescent label is selected from the group consisting of a 152Eu  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 ophthaldehyde label and a fluorescamine label. 
     
     
       22. The method of  claim 16 , wherein said detectable label is a toxic label is selected from the group consisting of a diphtheria toxin label, a ricin label and a cholera toxin label. 
     
     
       23. The method of  claim 16 , wherein said detectable label is a 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. 
     
     
       24. The method of  claim 16 , wherein said detectable label is an X-radiographic label that is barium or cesium. 
     
     
       25. The method of  claim 16 , wherein said detectable label is a spin label that is deuterium. 
     
     
       26. The method of  claim 16 , wherein said detectable label is a nuclear magnetic resonance contrast agent label is selected from the group consisting of Gd, Mn and iron Fe. 
     
     
       27. A method of increasing the responsiveness of a smad4-deficient cancer cell to treatment with a growth-inhibiting chemotherapeutic compounds compound, comprising:
 (a) determining the level of expression of smad4 in a cell from said smad4-deficient cancer; and 
 (b) treating a the smad4-deficient cancer cell that is deficient in smad4 expression with one or more an αvβ6-antagonist antibodies antibody that selectively bind binds to the integrin αvβ6 in said cancer cell, wherein said one or more antibody is an antibody from produced by, or a humanized verison of an 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), and 3G9 (deposited under ATCC Accession No. ATCC PTA-3649), or a humanized version thereof 
 wherein said treatment with said blocking antibodies antibody results in increased responsiveness of said smad4-deficient cancer cell to one or more the growth-inhibiting chemotherapeutic compounds compound as compared to the growth inhibition of said smad4-deficient cancer cell produced by treated with said chemotherapeutic agent compound alone. 
 
     
     
       28. The method of  claim 27 , 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 Aleuritesfordii protein, a dianthin protein, a Phytolaca americana protein, momordica Momordica charantia inhibitor, curcin, crotin, sapaonaria Sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, a tricothecene, a ribonuclease, and a deoxyribonuclease. 
     
     
       29. The method of  claim 28 , wherein said growth-inhibiting chemotherapeutic compound is gemcitabine, adriamycin or paclitaxel. 
     
     
       30. The method of claim 1, wherein said antibody is an antibody produced by the hybridoma 1A8 (deposited under ATCC Accession No. ATCC PTA-3647) or a humanized version thereof. 
     
     
       31. The method of claim 27, wherein said antibody is hu3G9 (BG00011). 
     
     
       32. The method of claim 27, wherein said antibody is hu8G6. 
     
     
       33. The method of claim 27, wherein said antibody is an antibody produced by the hybridoma 3G9 (deposited under ATCC Accession No. ATCC PTA-3649) or a humanized version thereof. 
     
     
       34. The method of claim 27, wherein said antibody is an antibody produced by the hybridoma 8G6 (deposited under ATCC Accession No. ATCC PTA-3645) or a humanized version thereof. 
     
     
       35. The method of claim 27, wherein said antibody is an antibody produced by the hybridoma 1A8 (deposited under ATCC Accession No. ATCC PTA-3647) or a humanized version thereof. 
     
     
       36. The method of claim 1, wherein the antibody comprises a heavy chain and a light chain, wherein the heavy and light chains comprise complementarity determining regions (CDRs) 1, 2, and 3 as follows:
 heavy chain CDR 1 comprising an amino acid sequence set forth in SEQ ID NO.: 3;   heavy chain CDR 2 comprising an amino acid sequence set forth in SEQ ID NO.: 9;   heavy chain CDR 3 comprising an amino acid sequence set forth in SEQ ID NO.: 15;   light chain CDR 1 comprising an amino acid sequence set forth in SEQ ID NO.: 20;   light chain CDR 2 comprising an amino acid sequence set forth in SEQ ID NO.: 25; and   light chain CDR 3 comprising an amino acid sequence set forth in SEQ ID NO.: 31.   
     
     
       37. The method of claim 1, wherein the antibody comprises a heavy chain and a light chain, wherein the heavy and light chains comprise CDRs 1, 2, and 3 as follows:
 heavy chain CDR 1 comprising an amino acid sequence set forth in SEQ ID NO.: 1;   heavy chain CDR 2 comprising an amino acid sequence set forth in SEQ ID NO.: 6;   heavy chain CDR 3 comprising an amino acid sequence set forth in SEQ ID NO.: 12;   light chain CDR 1 comprising an amino acid sequence set forth in SEQ ID NO.: 18;   light chain CDR 2 comprising an amino acid sequence set forth in SEQ ID NO.: 24; and   light chain CDR 3 comprising an amino acid sequence set forth in SEQ ID NO.: 28.   
     
     
       38. The method of claim 1, wherein the antibody comprises a heavy chain and a light chain, wherein the heavy and light chains comprise CDRs 1, 2, and 3 as follows:
 heavy chain CDR 1 comprising an amino acid sequence set forth in SEQ ID NO.: 2;   heavy chain CDR 2 comprising an amino acid sequence set forth in SEQ ID NO.: 7;   heavy chain CDR 3 comprising an amino acid sequence set forth in SEQ ID NO.: 13;   light chain CDR 1 comprising an amino acid sequence set forth in SEQ ID NO.: 19;   light chain CDR 2 comprising an amino acid sequence set forth in SEQ ID NO.: 24; and   light chain CDR 3 comprising an amino acid sequence set forth in SEQ ID NO.: 29.   
     
     
       39. The method of claim 27, wherein the antibody comprises a heavy chain and a light chain, wherein the heavy and light chains comprise CDRs 1, 2, and 3 as follows:
 heavy chain CDR 1 comprising an amino acid sequence set forth in SEQ ID NO.: 3;   heavy chain CDR 2 comprising an amino acid sequence set forth in SEQ ID NO.: 9;   heavy chain CDR 3 comprising an amino acid sequence set forth in SEQ ID NO.: 15;   light chain CDR 1 comprising an amino acid sequence set forth in SEQ ID NO.: 20;   light chain CDR 2 comprising an amino acid sequence set forth in SEQ ID NO.: 25; and   light chain CDR 3 comprising an amino acid sequence set forth in SEQ ID NO.: 31.   
     
     
       40. The method of claim 27, wherein the antibody comprises a heavy chain and a light chain, wherein the heavy and light chains comprise CDRs 1, 2, and 3 as follows:
 heavy chain CDR 1 comprising an amino acid sequence set forth in SEQ ID NO.: 1;   heavy chain CDR 2 comprising an amino acid sequence set forth in SEQ ID NO.: 6;   heavy chain CDR 3 comprising an amino acid sequence set forth in SEQ ID NO.: 12;   light chain CDR 1 comprising an amino acid sequence set forth in SEQ ID NO.: 18;   light chain CDR 2 comprising an amino acid sequence set forth in SEQ ID NO.: 24; and   light chain CDR 3 comprising an amino acid sequence set forth in SEQ ID NO.: 28.   
     
     
       41. The method of claim 27, wherein the antibody comprises a heavy chain and a light chain, wherein the heavy and light chains comprise CDRs 1, 2, and 3 as follows:
 heavy chain CDR 1 comprising an amino acid sequence set forth in SEQ ID NO.: 2;   heavy chain CDR 2 comprising an amino acid sequence set forth in SEQ ID NO.: 7;   heavy chain CDR 3 comprising an amino acid sequence set forth in SEQ ID NO.: 13;   light chain CDR 1 comprising an amino acid sequence set forth in SEQ ID NO.: 19;   light chain CDR 2 comprising an amino acid sequence set forth in SEQ ID NO.: 24; and   light chain CDR 3 comprising an amino acid sequence set forth in SEQ ID NO.: 29.