Viral vector targeting cancer stem cells

Abstract

[Problem] To provide a novel and effective method for treating cancer, method for preventing cancer, and method for suppressing metastasis by targeting cancer stem cells, the methods being capable of labeling and damaging cancer stem cells; and to provide a method for damaging and a method for identifying cancer stem cells. [Solution] The present invention provides a viral vector having a promoter that is specifically expressed in cancer stem cells and that can be used for treatment and diagnosis. Furthermore, the present invention provides a method for treating cancer, a method for preventing cancer, and a method for suppressing metastasis using this viral vector, and further provides a method for damaging and a method for identifying cancer stem cells. Furthermore, the present invention provides a labeling agent and a toxic agent for cancer stem cells containing the vector as the active ingredient, and further provides a diagnostic drug, a therapeutic drug, and a metastasis suppressant for cancer.

1 - 51 . (canceled) 
     
     
         52 . A virus vector comprising a CD133 promoter operably linked to a desired gene, wherein said virus vector can express said desired gene specifically in a cancer stem cell. 
     
     
         53 . The virus vector of  claim 52 , wherein the desired gene is a gene encoding a protein which is essential for replication of virus and the virus vector can replicate specifically in a cancer stem cell. 
     
     
         54 . The virus vector of  claim 53 , wherein the virus vector is oncolytic. 
     
     
         55 . The virus vector of  claim 53 , wherein the virus vector is adenovirus, herpes simplex virus, myxoma virus, reovirus, vesicular stomatitis virus, Newcastle disease virus, vaccinia virus, RS virus, Sendai virus, measles virus, Coxsackie virus, or Seneca Valley virus. 
     
     
         56 . The virus vector of  claim 53 , wherein the virus vector is adenovirus. 
     
     
         57 . The adenovirus vector of  claim 56 , wherein the desired gene is an adenovirus E1A or E1B gene. 
     
     
         58 . The adenovirus vector of  claim 57 , wherein the E1A is an E1A lacking the Rb-binding region (E1AΔ24) or the E1B is an E1B lacking the p53-binding region (E1BΔ55K). 
     
     
         59 . The virus vector of  claim 53 , further comprising a marker gene or a cytotoxic gene. 
     
     
         60 . The virus vector of  claim 53 , further comprising an exogenous cancer-specific promoter. 
     
     
         61 . The adenovirus vector of  claim 57 , wherein the adenovirus vector is selected from the following (a) to (c):
 (a) an adenovirus vector having the following transcription units:
 (a1) a CD133 transcription unit consisting of a CD133 promoter and an E1A gene or E1AΔ24 gene operably linked downstream of said promoter, 
 (a2) a transcription unit consisting of a eukaryotic cell promoter, a cancer-specific promoter or CD133 promoter, and an E1B gene or E1BΔ55K gene operably linked downstream of said promoter, or a transcription unit consisting of an E1BΔ19K gene operably linked downstream of a cancer-specific promoter, and 
 (a3) optionally, an additional transcription unit consisting of a eukaryotic cell promoter, a cancer-specific promoter or CD133 promoter, and a marker gene or cytotoxic gene operably linked downstream of said promoter; 
   (b) an adenovirus vector having the following transcription units:
 (b1) a transcription unit consisting of a eukaryotic cell promoter, a cancer-specific promoter or CD promoter, and an E1A gene or E1AΔ24 gene operably linked downstream of said promoter, 
 (b2) a CD133 transcription unit consisting of a CD133 promoter and a E1B gene or E1BΔ55K gene operably linked downstream of said CD133 promoter, and 
 (b3) optionally, an additional transcription unit consisting of a eukaryotic cell promoter, a cancer-specific promoter or CD133 promoter, and a marker gene or cytotoxic gene operably linked downstream of said promoter; and 
   (c) an adenovirus vector having the following transcription units:
 (c1) a transcription unit consisting of a eukaryotic cell promoter, a cancer-specific promoter or CD 133 promoter, and an E1A gene or E1AΔ24 gene operably linked downstream of said promoter, 
 (c2) a transcription unit consisting of a eukaryotic cell promoter, a cancer-specific promoter or CD133 promoter, and an E1B gene or E1BΔ55K gene operably linked downstream of said promoter, and 
 (c3) a CD133 transcription unit consisting of a CD133 promoter and a marker gene or cytotoxic gene operably linked downstream of said CD 133 promoter. 
   
     
     
         62 . The virus vector of  claim 52 , wherein the desired gene is a marker gene. 
     
     
         63 . The virus vector of  claim 62 , further comprising an exogenous cancer-specific promoter. 
     
     
         64 . The virus vector of  claim 62 , further comprising a cytotoxic gene. 
     
     
         65 . The virus vector of  claim 52 , wherein the desired gene is a cytotoxic gene. 
     
     
         66 . The virus vector of  claim 65 , further comprising an exogenous cancer-specific promoter. 
     
     
         67 . The adenovirus vector of  claim 65 , further comprising a marker gene. 
     
     
         68 . The virus vector of  claim 62 , wherein the virus vector is oncolytic virus. 
     
     
         69 . The virus vector of  claim 62 , wherein the virus vector is adenovirus, herpes simplex virus, myxoma virus, reovirus, vesicular stomatitis virus, Newcastle disease virus, vaccinia virus, RS virus, Sendai virus, measles virus, Coxsackie virus, or Seneca Valley virus. 
     
     
         70 . The virus vector of  claim 52 , wherein the CD133 promoter is any one of nucleic acid molecule selected from the following (i) to (iv):
 (i) a nucleic acid molecule comprising the nucleotide sequence of any one of SEQ ID NOS: 1 to 5,   (ii) a nucleic acid molecule comprising a nucleotide sequence having 85% homology to any one of SEQ ID NOS: 1 to 5,   (iii) a nucleic acid molecule which can hybridize under stringent conditions with the nucleic acid molecule having the nucleotide sequence of any one of SEQ ID NOS: 1 to 5 or having complement of said sequence, and   (iv) a nucleic acid molecule having a nucleotide sequence of any one of SEQ ID NOS: 1 to 5, wherein a part of nucleotide is substituted or deleted, or additional nucleotide is added or inserted.   
     
     
         71 . The virus vector of  claim 52 , wherein the CD133 promoter is promoter 5 of CD133 promoter. 
     
     
         72 . A composition comprising the virus vector of  claim 52 , which give damage specifically to cancer stem cells. 
     
     
         73 . A labeling agent for labeling specifically cancer stem cells, which comprises the virus vector of  claim 52 . 
     
     
         74 . The composition of  claim 72 , further comprising one or more agents selected from the following (i) and (ii):
 (i) a drug for decreasing antiviral action, and   (ii) an anticancer agent;   
       wherein said agents are administered simultaneously, continuously, or separately at certain intervals. 
     
     
         75 . A method for labeling cancer stem cells, comprising a step of administering the virus vector of  claim 52  to a patient in need thereof. 
     
     
         76 . A method for diagnosing cancer, comprising a step of administering the virus vector of  claim 52  to a patient in need thereof. 
     
     
         77 . A prophylactic method, metastasis-suppressing method or therapeutic method for cancer, comprising a step of administering the virus vector of  claim 52  to a patient in need thereof. 
     
     
         78 . The prophylactic method, metastasis-suppressing method or therapeutic method of  claim 77 , further comprising a step of infecting the cancer stem cells of the patient with the virus vector. 
     
     
         79 . The virus vector of  claim 52 , wherein said cancer stem cells is brain tumor stem cells or glioblastoma stem cells. 
     
     
         80 . A composition for expressing a desired gene in brain tumor stem cells, which comprises a virus vector comprising a CD133 promoter operably linked to said desired gene, wherein said virus vector can express said desired gene specifically in a cancer stem cell. 
     
     
         81 . The composition of  claim 80 , wherein the desired gene is a gene encoding a protein which is essential for replication of virus and the virus vector can replicate specifically in a cancer stem cell. 
     
     
         82 . The composition of  claim 81 , wherein the virus vector is oncolytic. 
     
     
         83 . The composition of  claim 81 , wherein the virus vector is adenovirus, herpes simplex virus, myxoma virus, reovirus, vesicular stomatitis virus, Newcastle disease virus, vaccinia virus, RS virus, Sendai virus, measles virus, Coxsackie virus, or Seneca Valley virus. 
     
     
         84 . The composition of  claim 81 , wherein the virus vector is adenovirus. 
     
     
         85 . The composition of  claim 84 , wherein the desired gene is an adenovirus E1A or E1B gene. 
     
     
         86 . The composition of  claim 84 , wherein the E1A is an E1A lacking the Rb-binding region (E1AΔ24) or the E1B is an E1B lacking the p53-binding region (E1BΔ55K). 
     
     
         87 . The composition of  claim 81 , further comprising a marker gene or a cytotoxic gene. 
     
     
         88 . The virus vector of  claim 81 , further comprising an exogenous cancer-specific promoter. 
     
     
         89 . The composition of  claim 85 , wherein The composition is selected from the following (a) to (c):
 (a) an adenovirus vector having the following transcription units:
 (a1) a CD133 transcription unit consisting of a CD133 promoter and an E1A gene or E1AΔ24 gene operably linked downstream of said promoter, 
 (a2) a transcription unit consisting of a eukaryotic cell promoter, a cancer-specific promoter or CD 133 promoter, and an E1B gene or E1BΔ55K gene operably linked downstream of said promoter, or a transcription unit consisting of an E1BΔ19K gene operably linked downstream of a cancer-specific promoter, and 
 (a3) optionally, an additional transcription unit consisting of a eukaryotic cell promoter, a cancer-specific promoter or CD133 promoter, and a marker gene or cytotoxic gene operably linked downstream of said promoter; 
   (b) an adenovirus vector having the following transcription units:
 (b1) a transcription unit consisting of a eukaryotic cell promoter, a cancer-specific promoter or CD133 promoter, and an E1A gene or E1AΔ24 gene operably linked downstream of said promoter, 
 (b2) a CD133 transcription unit consisting of a CD133 promoter and a E1B gene or E1BΔ55K gene operably linked downstream of said CD133 promoter, and 
 (b3) optionally, an additional transcription unit consisting of a eukaryotic cell promoter, a cancer-specific promoter or CD133 promoter, and a marker gene or cytotoxic gene operably linked downstream of said promoter; and 
   (c) an adenovirus vector having the following transcription units:
 (c1) a transcription unit consisting of a eukaryotic cell promoter, a cancer-specific promoter or CD 133 promoter, and an E1A gene or E1AΔ24 gene operably linked downstream of said promoter, 
 (c2) a transcription unit consisting of a eukaryotic cell promoter, a cancer-specific promoter or CD133 promoter, and an E1B gene or E1BΔ55K gene operably linked downstream of said promoter, and 
 (c3) a CD133 transcription unit consisting of a CD133 promoter and a marker gene or cytotoxic gene operably linked downstream of said CD133 promoter. 
   
     
     
         90 . The composition of  claim 80 , wherein the desired gene is a marker gene. 
     
     
         91 . The composition of  claim 90 , further comprising an exogenous cancer-specific promoter. 
     
     
         92 . The composition of  claim 90 , further comprising a cytotoxic gene. 
     
     
         93 . The composition of  claim 80 , wherein the desired gene is a cytotoxic gene. 
     
     
         94 . The composition of  claim 93 , further comprising an exogenous cancer-specific promoter. 
     
     
         95 . The composition of  claim 93 , further comprising a marker gene. 
     
     
         96 . The composition of  claim 90 , wherein the virus is oncolytic virus. 
     
     
         97 . The composition of  claim 90 , wherein the virus is adenovirus, herpes simplex virus, myxoma virus, reovirus, vesicular stomatitis virus, Newcastle disease virus, vaccinia virus, RS virus, Sendai virus, measles virus, Coxsackie virus, or Seneca Valley virus. 
     
     
         98 . The composition of  claim 80 , wherein the CD133 promoter is any one of nucleic acid molecule selected from the following (i) to (iv):
 (i) a nucleic acid molecule comprising the nucleotide sequence of any one of SEQ ID NOS: 1 to 5,   (ii) a nucleic acid molecule comprising a nucleotide sequence having 85% homology to any one of SEQ ID NOS: 1 to 5,   (iii) a nucleic acid molecule which can hybridize under stringent conditions with the nucleic acid molecule having the nucleotide sequence of any one of SEQ ID NOS: 1 to 5 or having complement of said sequence, and   (iv) a nucleic acid molecule having a nucleotide sequence of any one of SEQ ID NOS: 1 to 5, wherein a part of nucleotide is substituted or deleted, or additional nucleotide is added or inserted.   
     
     
         99 . The composition of  claim 80 , wherein the CD133 promoter is promoter 5 of CD133 promoter. 
     
     
         100 . A therapeutic agent, metastasis-suppressing agent or prophylactic agent or diagnostic agent for cancer, comprising the composition of  claim 80 . 
     
     
         101 . A labeling agent comprising the composition of  claim 80 . 
     
     
         102 . The therapeutic agent or prophylactic agent or diagnostic agent of  claim 100 , further comprising one or more agents selected from the following (i) and (ii):
 (i) a drug for decreasing antiviral action, and   (ii) an anticancer agent;   
       wherein said agents are administered simultaneously, continuously, or separately at certain intervals.