Tumor Suppressor Killin

Abstract

The present invention relates to a new tumor suppressor, designated Killin. Also described are diagnostic and therapeutic uses of the Killin protein and the killin gene, alone or in combination with traditional cancer therapies.

1 - 6 . (canceled) 
     
     
         7 . A nucleic acid of about 15 to about 5000 base pairs comprising from about 15 contiguous base pairs of SEQ ID NO:2, or the complement thereof. 
     
     
         8 . The nucleic acid of  claim 7 , comprising from about 20 contiguous base pairs of SEQ ID NO:2, or the complement thereof. 
     
     
         9 . The nucleic acid of  claim 7 , comprising from about 30 contiguous base pairs of SEQ ID NO:2, or the complement thereof. 
     
     
         10 . The nucleic acid of  claim 7 , comprising from about 50 contiguous base pairs of SEQ ID NO:2, or the complement thereof. 
     
     
         11 . The nucleic acid of  claim 7 , comprising about 100 contiguous base pairs of SEQ ID NO:2, or the complement thereof. 
     
     
         12 . The nucleic acid of  claim 7 , comprising about 150 contiguous base pairs of SEQ ID NO:2. 
     
     
         13 . The nucleic acid of  claim 7 , comprising about 250 contiguous base pairs of SEQ ID NO:2, or the complement thereof. 
     
     
         14 . The nucleic acid of  claim 7 , comprising about 500 contiguous base pairs of SEQ ID NO:2, or the complement thereof. 
     
     
         15 . The nucleic acid of  claim 7 , comprising about 1000 contiguous base pairs of SEQ ID NO:2, or the complement thereof. 
     
     
         16 . The nucleic acid of  claim 7 , comprising about 2500 contiguous base pairs of SEQ ID NO:2, or the complement thereof. 
     
     
         17 . The nucleic acid of  claim 7 , comprising about 4000 contiguous base pairs of SEQ ID NO:2, or the complement thereof. 
     
     
         18 . A peptide comprising about 10-50 contiguous amino acids of SEQ ID NO:1. 
     
     
         19 . The peptide of  claim 18 , comprising about 15 contiguous amino acids of SEQ ID NO:1. 
     
     
         20 . The peptide of  claim 18 , comprising about 25 contiguous amino acids of SEQ ID NO:1. 
     
     
         21 . The peptide of  claim 18 , comprising about 50 contiguous amino acids of SEQ ID NO:1. 
     
     
         22 . The peptide of  claim 18 , comprising residues 8-49 of of SEQ ID NO:1. 
     
     
         23 . A polypeptide comprising the sequence of SEQ ID NO:1. 
     
     
         24 . An expression cassette comprising a polynucleotide encoding a polypeptide having the sequence of SEQ ID NO:1 or a fragment thereof, wherein said polynucleotide is under the control of a promoter operable in eukaryotic cells. 
     
     
         25 - 33 . (canceled) 
     
     
         34 . A method for suppressing growth of a cancer cell comprising contacting said cells with an expression cassette comprising a polynucleotide encoding a polypeptide having the sequence of SEQ ID NO:1 or a fragment thereof, wherein said polynucleotide is under the control of a promoter operable in eukaryotic cells. 
     
     
         35 . The method of  claim 34 , wherein said promoter is heterologous to the polynucleotide sequence. 
     
     
         36 . The method of  claim 35 , wherein said promoter is selected from the group consisting of hsp68, SV40, CMV, MKC, GAL4 UAS , HSV and β-actin. 
     
     
         37 . The method of  claim 35 , wherein said promoter is a tissue specific promoter. 
     
     
         38 . The method of  claim 35 , wherein said promoter is an inducible promoter. 
     
     
         39 . The method of  claim 35 , wherein said expression cassette is contained in a viral vector. 
     
     
         40 . The method of  claim 35 , wherein said viral vector is selected from the group consisting of a retroviral vector, an adenoviral vector, and adeno-associated viral vector, a vaccinia viral vector, and a herpesviral vector. 
     
     
         41 . The method of  claim 34 , wherein said expression cassette further comprises a polyadenylation signal. 
     
     
         42 . The method of  claim 34 , wherein said expression cassette comprises a second polynucleotide encoding a second polypeptide. 
     
     
         43 . The method of  claim 42 , wherein said second polynucleotide is under the control of a second promoter. 
     
     
         44 . A cell comprising an expression cassette comprising a polynucleotide encoding a polypeptide having the sequence of SEQ ID NO:1 or a fragment thereof, wherein said polynucleotide is under the control of a promoter operable in eukaryotic cells. 
     
     
         45 . A monoclonal antibody that binds immunologically to a polypeptide having the sequence of SEQ ID NO:1, or an immunologic fragment thereof. 
     
     
         46 - 49 . (canceled) 
     
     
         50 . A method of diagnosing a cancer comprising the steps of:
 (i) obtaining a tissue sample from a subject; and   (ii) assessing the expression or structure of Killin in cells of said sample.   
     
     
         51 . The method of  claim 50 , wherein said cancer is selected from the group consisting of brain, lung, liver, spleen, kidney, lymph node, small intestine, pancreas, blood cells, colon, stomach, breast, endometrium, prostate, testicle, ovary, skin, head and neck, esophagus, bone marrow and blood cancer. 
     
     
         52 . The method of  claim 50 , wherein step (ii) comprises assessing Killin expression. 
     
     
         53 . The method of  claim 50 , wherein step (ii) comprises assessing Killin structure. 
     
     
         54 . The method of  claim 50 , wherein said cancer is a colon cancer or lung cancer. 
     
     
         55 . The method of  claim 50 , wherein said sample is a tissue or fluid sample. 
     
     
         56 . The method of  claim 50 , wherein said assessing comprises assaying for a Killin-encoding nucleic acid from said sample. 
     
     
         57 . The method of  claim 56 , further comprising subjecting said sample to conditions suitable to amplify said nucleic acid. 
     
     
         58 . The method of  claim 50 , wherein said assessing comprises contacting said sample with an antibody that binds immunologically to a Killin polypeptide. 
     
     
         59 . The method of  claim 58 , further comprising subjecting proteins of said sample to ELISA. 
     
     
         60 . The method of  claim 50 , wherein assessing involves evaluating the level of Killin expression. 
     
     
         61 . The method of  claim 50 , further comprising the step of comparing the expression of Killin with the expression of Killin in non-cancer samples. 
     
     
         62 . The method of  claim 50 , wherein assessing involves evaluating the structure of the Killin gene or transcript. 
     
     
         63 . The method of  claim 62 , wherein said evaluating comprises an assay selected from the group consisting of sequencing, wild-type oligonucleotide hybridization, mutant oligonucleotide hybridization, SSCP, PCR and RNase protection. 
     
     
         64 . The method of  claim 63 , wherein a said evaluating is wild-type or mutant oligonucleotide hybridization and said oligonucleotide is configured in an array on a chip or wafer. 
     
     
         65 . A method for altering the phenotype of a tumor cell comprising the step of administering to a cell a tumor suppressor designated Killin or a fragment thereof under conditions permitting the uptake of said tumor suppressor by said tumor cell. 
     
     
         66 . The method of  claim 65 , wherein said tumor cell is derived from a tissue selected from the group consisting of brain, lung, liver, spleen, kidney, lymph node, small intestine, blood cells, pancreas, colon, stomach, breast, endometrium, prostate, testicle, ovary, skin, head and neck, esophagus, bone marrow and blood tissue. 
     
     
         67 - 68 . (canceled) 
     
     
         69 . A method for altering the phenotype of a tumor cell comprising the step of contacting the cell with a nucleic acid (i) encoding a tumor suppressor designated Killin or a fragment thereof and (ii) a promoter active in said tumor cell, wherein said promoter is operably linked to the region encoding said tumor suppressor, under conditions permitting the uptake of said nucleic acid by said tumor cell. 
     
     
         70 . The method of  claim 69 , wherein said tumor cell is derived from a tissue selected from the group consisting of brain, lung, liver, spleen, kidney, lymph node, small intestine, blood cells, pancreas, colon, stomach, breast, endometrium, prostate, testicle, ovary, skin, head and neck, esophagus, bone marrow and blood tissue. 
     
     
         71 . The method of  claim 70 , wherein the a phenotype is selected from the group consisting of apoptosis, angiogenesis, proliferation, migration, contact inhibition, soft agar growth or cell cycling. 
     
     
         72 . The method of  claim 70 , wherein said nucleic acid is encapsulated in a liposome. 
     
     
         73 . The method of  claim 70 , wherein said nucleic acid is a viral vector selected from the group consisting of retrovirus, adenovirus, adeno-associated virus, vaccinia virus and herpesvirus. 
     
     
         74 . The method of  claim 73 , wherein said nucleic acid is encapsulated in a viral particle. 
     
     
         75 . A method for treating subject with cancer comprising the step of administering to said subject a tumor suppressor designated Killin or a fragment thereof. 
     
     
         76 . The method of  claim 75 , wherein said tumor cell is derived from a tissue selected from the group consisting of brain, lung, liver, spleen, kidney, lymph node, small intestine, blood cells, pancreas, colon, stomach, breast, endometrium, prostate, testicle, ovary, skin, head and neck, esophagus, bone marrow and blood tissue. 
     
     
         77 . (canceled) 
     
     
         78 . A method for treating a subject with cancer comprising the step of administering to said subject a nucleic acid (i) encoding a tumor suppressor designated Killin or a fragment thereof and (ii) a promoter active in eukaryotic cells, wherein said promoter is operably linked to the region encoding said tumor suppressor. 
     
     
         79 . The method of  claim 78 , wherein said tumor cell is derived from a tissue selected from the group consisting of brain, lung, liver, spleen, kidney, lymph node, small intestine, blood cells, pancreas, colon, stomach, breast, endometrium, prostate, testicle, ovary, skin, head and neck, esophagus, bone marrow and blood tissue. 
     
     
         80 . (canceled) 
     
     
         81 . A non-human transgenic eukaryote lacking a functional Killin gene. 
     
     
         82 . (canceled) 
     
     
         83 . A non-human transgenic eukaryote that overexpresses Killin as compared to a similar non-transgenic eukaryote. 
     
     
         84 . (canceled) 
     
     
         85 . A method of screening a candidate substance for anti-tumor activity comprising the steps of:
 (i) providing a cell lacking functional Killin polypeptide;   (ii) contacting said cell with said candidate substance; and   (iii) determining the effect of said candidate substance on said cell.   
     
     
         86 - 94 . (canceled) 
     
     
         95 . An isolated and purified nucleic acid that hybridizes, under high stringency conditions, to a DNA segment comprising SEQ ID NO:2. 
     
     
         96 . The method of  claim 75 , further comprising treating said subject with a second anti-cancer therapy. 
     
     
         97 - 98 . (canceled) 
     
     
         99 . The method of  claim 78 , further comprising treating said subject with a second anti-cancer therapy. 
     
     
         100 - 101 . (canceled) 
     
     
         102 . A method of screening a candidate substance for anti-tumor activity comprising the steps of:
 (i) providing a cell expression a functional Killin peptide or polypeptide;   (ii) contacting said cell with said candidate substance; and   (iii) determining Killin DNA binding or nuclear localization,   wherein an increase in Killin DNA binding or nuclear localization, as compared to a similar cell not treated with said candidate substance, indicates that said candidate substance has anti-tumor activity.   
     
     
         103 . A nucleic acid segment comprising SEQ ID NO:3. 
     
     
         104 . A method of screening for an activator of Killin expression comprising:
 (i) providing a cell comprising a Killin promoter operably linked to a nucleic acid segment encoding expressible marker;   (ii) contacting said cell with a candidate substance; and   (iii) assessing the expression of said marker,   wherein an increase in expression of said marker, as compared to expression in a cell not contacted with said candidate substance, identifies said candidate substance as an activator of Killin expression.   
     
     
         105 - 107 . (canceled)