Vector utilizing ribonuclease reductase UTR and use for inhibiting neoplastic cells

Abstract

The present invention relates to oligonucleotides having the sequence of an untranslated region (UTR) of mRNA from a housekeeping gene and which can modulate the tumorigenicity of neoplastic cells. The present invention further provides pharmaceutical compositions and methods of inhibiting the growth of neoplastic/malignant cells in a mammal with oligonucleotides having sequences of untranslated regions of the mRNA of housekeeping genes. In a preferred embodiment the UTR is the 3′-UTR and the housekeeping gene is ribonucleotide reductase and its regulatory sequences.

1 - 35 . (canceled)  
     
     
         36 . A vector comprising a nucleic acid sequence corresponding to a sequence segment of at least 7 consecutive nucleotides of a 3′ untnanslated region of a mammalian ribonucleotide reductase mRNA, said nucleic acid sequence operatively liked to an expression control sequence, wherein said nucleic acid sequence is substantially free of coding sequence of said mRNA, and wherein said nucleic acid sequence inhibits neoplastic cell growth or metastasis.  
     
     
         37 . The vector according to claim  1 , wherein said nucleic acid sequence is from the 3′ untranslated region of the ribonucleotide reductase R1 mRNA as set forth in SEQ ID NO:1.  
     
     
         38 . The vector according to claim  1 , wherein said nucleic acid sequence is from the 3′untranslated region of the ribonucleotide reductase R2 mRNA as set forth in SEQ ID NO:2.  
     
     
         39 . The vector according to claim  1 , wherein said nucleic acid sequence comprises a sequence as set forth in any one of SEQ ID NOs:6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 or 49.  
     
     
         40 . The vector according to claim  1 , wherein said nucleic acid sequence comprises a sequence as set forth in any one of SEQ ID NOs:6, 7, 8, 9, 10, 11, or 12.  
     
     
         41 . The vector according to claim  1 , wherein said nucleic acid sequence comprises a sequence as set forth in any one of SEQ ID NOs:44, 45, 46, 47, 48, or 49.  
     
     
         42 . The vector according to claim  1 , wherein said vector is targeted to a specific cell type.  
     
     
         43 . The vector according to claim  1 , wherein said vector is a viral vector.  
     
     
         44 . The vector according to claim  1 , wherein said vector is an adenoviral vector or a retroviral vector.  
     
     
         45 . A vector comprising a transcription control sequence operatively linked to two nucleic acid sequences linked together, wherein a first of said nucleic acid sequences has a sequence as set forth in any one of SEQ ID NOs: 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or 49.  
     
     
         46 . The vector according to claim  10 , wherein a second of said nucleic acid sequences has a sequence as set forth in any one of SEQ ID NOs: 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or 49.  
     
     
         47 . A pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent, and a vector comprising a nucleic acid sequence corresponding to a sequence segment of at least 7 consecutive nucleotides of a 3′ untranslated region of a mammalian ribonucleotide reductase mRNA, said nucleic acid sequence operatively linked to an expression control sequence, wherein said nucleic acid sequence is substantially free of coding sequence of said mRNA, and wherein said nucleic acid sequence inhibits neoplastic cell growth or metastasis.  
     
     
         48 . The pharmaceutical composition according to claim  12 , wherein said nucleic acid sequence is from the 3′ untranslated region of the ribonucleotide reductase R1 mRNA as set forth in SEQ ID NO:1.  
     
     
         49 . The pharmaceutical composition according to claim  12 , wherein said nucleic acid sequence is from the 3′untranslated region of the ribonucleotide reductase R2 mRNA as set forth in SEQ ID NO:2.  
     
     
         50 . The pharmaceutical composition according to claim  12 , wherein said nucleic acid sequence comprises a sequence as set forth in any one of SEQ ID NOs:6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or 49.  
     
     
         51 . The pharmaceutical composition according to claim  12 , wherein said nucleic acid sequence comprises a sequence as set forth in any one of SEQ ID NOs:6, 7, 8, 9, 10, 11, or 12.  
     
     
         52 . The pharmaceutical composition according to claim  12 , wherein said nucleic acid sequence comprises a sequence as set forth in any one of SEQ ID NOs:44, 45, 46, 47, 48, or 49.  
     
     
         53 . A pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent, and a vector comprising an expression control sequence operatively linked to two nucleic acid sequences linked together, wherein a first of said nucleic acid sequences has a sequence as set forth in any one of SEQ ID NOs: 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or 49.  
     
     
         54 . The pharmaceutical composition according to claim  18 , wherein a second of said nucleic acid sequences has a sequence as set forth in any one of SEQ ID NOs: 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or 49.  
     
     
         55 . A method of inhibiting growth of neoplastic cells in a human or: mouse comprising the step of contacting said neoplastic cells with an effective amount of a vector comprising a nucleic acid sequence corresponding to a sequence segment of at least 7 consecutive nucleotides of a 3′ untranslated region of a mammalian ribonucleotide reductase mRNA, said nucleic acid sequence operatively linked to an expression control sequence, wherein said nucleic acid sequence is substantially free of coding sequence of said mRNA, and wherein said nucleic acid sequence inhibits neoplastic cell growth.  
     
     
         56 . The method according to claim  20 , wherein said nucleic acid sequence is from the 3′ untranslated region of the ribonucleotide reductase R1 mRNA as set forth in SEQ ID NO:1.  
     
     
         57 . The method according to claim  20 , wherein said nucleic acid sequence is from the 3′untranslated region of the ribonucleotide reductase R2 mRNA as set forth in SEQ ID NO:2.  
     
     
         58 . The method according to claim  20 , wherein said nucleic acid sequence comprises a sequence as set forth in any one of SEQ ID NOs:6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or 49.  
     
     
         59 . The method according to claim  20 , wherein said nucleic acid sequence comprises a sequence as set forth in any one of SEQ ID NOs:6, 7, 8, 9, 10, 11, or 12.  
     
     
         60 . The method according to claim  20 , wherein said nucleic acid sequence comprises a sequence as set forth in any one of SEQ ID NOs:44, 45, 46, 47, 48, or 49.  
     
     
         61 . The method according to claim  20 , wherein said vector is targeted to a specific cell type.  
     
     
         62 . The method according to claim  20 , wherein said vector is a viral vector.  
     
     
         63 . The method according to claim  20 , wherein said vector is an adenoviral vector or a retroviral vector.  
     
     
         64 . The method according to claim  20 , wherein said neoplastic cells are selected from the group consisting of bladder, colon, lung, breast, and pancreatic cancer cells.  
     
     
         65 . The method according to claim  20 , wherein said neoplastic cells are resistant to a chemotherapeutic compound.  
     
     
         66 . The method according to claim  30 , wherein said chemotherapeutic compound is hydroxyurea.  
     
     
         67 . The method according to claim  20 , wherein said method further comprises administering one or more anti-cancer drugs to said human or mouse.  
     
     
         68 . A method of inhibiting metastasis of neoplastic cells in a human or mouse comprising the step of contacting said neoplastic cells with an effective amount of a vector comprising a nucleic acid sequence corresponding to a sequence segment of at least 7 consecutive nucleotides of a 3′ untranslated region of a mammalian ribonucleotide reductase R2 mRNA, said nucleic acid-sequence operatively linked to an expression control sequence, wherein said nucleic acid sequence is substantially free of coding sequence of said mRNA, and wherein said nucleic acid sequence inhibits metastasis of neoplastic cells.  
     
     
         69 . The method according to claim  33 , wherein said nucleic acid sequence is from the 3′untranslated region of the ribonucleotide reductase R2 mRNA as set forth in SEQ ID NO:2.  
     
     
         70 . The method according to claim  33 , wherein said nucleic acid sequence comprises a sequence as set forth in any one of SEQ ID NOs:6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, or 43.  
     
     
         71 . The method according to claim  33 , wherein said nucleic acid sequence comprises a sequence as set forth in any one of SEQ ID NOs:6, 7, 8,9, 10, 11, or 12.  
     
     
         72 . The method according to claim  33  wherein said vector is targeted to a specific cell type.  
     
     
         73 . The method according to claim  33 , wherein said vector is a viral vector.  
     
     
         74 . The method according to claim  33 , wherein said vector is an adenoviral vector or a retroviral vector.  
     
     
         75 . The method according to claim  33 , wherein said neoplastic cells are selected from the group consisting of bladder, colon, lung, breast, and pancreatic cancer cells.  
     
     
         76 . The method according to claim  33 , wherein said neoplastic cells are resistant to a chemotherapeutic compound.  
     
     
         77 . The method according to  claim 41 , wherein said chemotherapeutic compound is hydroxyurea.  
     
     
         78 . The method according to claim  33 , wherein said method further comprises administering one or more anti cancer drugs to said human or mouse.