Novel mRNA splice variant of the doublecortin-like kinase gene and its use in cancer diagnosis and therapy

Abstract

Methods for treating neuroblastoma in a subject are provided. The methods include administering to the subject nucleic acid that is capable of causing a significant reduction of the amount of doublecortin like protein (DCL) in the subject. The nucleic acids include antisense oligonucleotides and small interfering RNA.

1 . A method for treating neuroblastoma comprising administering to a subject in need thereof a composition comprising a therapeutically effective amount of a nucleic acid fragment of SEQ ID NO: 1 or SEQ ID NO: 2, or of a variant of SEQ ID NO: 1 or SEQ ID NO: 2, wherein said nucleic acid fragment is capable of causing a significant reduction of the amount of doublecortin like protein (DCL) in the subject. 
     
     
         2 . The method of  claim 1 , wherein the DCL-protein has the amino acid sequence as set forth in SEQ ID NO: 3 or SEQ ID NO: 4. 
     
     
         3 . The method of  claim 1 , wherein the nucleic acid fragment is one or more selected from the group consisting of an antisense RNA oligonucleotide, an antisense DNA oligonucleotide and a small interfering RNA (siRNA). 
     
     
         4 . The method of  claim 1 , wherein the nucleic acid fragment is one or more selected from the group consisting of SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 19 and SEQ ID NO: 20. 
     
     
         5 . The method of  claim 1 , wherein the composition comprises one or more targeting compounds capable of targeting neuroblastoma cells in vivo or in vitro. 
     
     
         6 . The method of  claim 5 , wherein the targeting compound is an immunoliposome, a monoclonal antibody or a ligand or ligand mimetic. 
     
     
         7 . The method of  claim 3 , wherein the one or more antisense RNA oligonucleotide comprises 2′-O-methyl RNA or 2′-O-allyl RNA. 
     
     
         8 . The method of  claim 3 , wherein the one or more antisense RNA oligonucleotide or antisense DNA oligonucleotide comprises peptide nucleic acid (PNA). 
     
     
         9 . The method of  claim 3 , wherein the one or more antisense RNA oligonucleotide or antisense DNA oligonucleotide comprises locked nucleic acid (LNA). 
     
     
         10 . The method of  claim 1 , wherein the concentration of the nucleic acid in the composition is in a range of 0.05-5.0 μmol/ml and the composition is infused at a rate of 1-100 ml/kg body weight of the subject. 
     
     
         11 . The method of  claim 1 , wherein the nucleic acid fragment is one or more antisense RNA oligonucleotide or antisense DNA oligonucleotide of 17-23 nucleotides, comprising one or more sequences selected from the group consisting of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, and sequences derived thereof in which 1, 2 or 3 nucleotides has been added, replaced or deleted. 
     
     
         12 . The method of  claim 3 , wherein the siRNA comprises one or more selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12.