Wnt Signaling Inhibitor Comprising Insulin-Like Growth Factor-Binding Protein

Abstract

Provided is a novel soluble factor that modulates morphogenesis and cell proliferation, such as cardiac development and/or cardiomyocyte differentiation. Specifically provided are: an inhibitor of Wnt signalling, comprising an insulin-like growth-factor-binding protein (IGFBP), the protein being binding to a Wnt receptor, and/or a polynucleotide encoding the protein; a medicament for prevention and/or treatment of a disease due to enhanced Wnt signalling, comprising the inhibitor of Wnt signalling, and a medicament for induction of cardiomyocyte differentiation; and a method for prevention and/or treatment of a disease due to enhanced Wnt signalling and a method of inducing cardiomyocyte differentiation, the methods each comprising using the inhibitor of Wnt signalling, and a cardiomyocyte, which is obtained by the method of inducing cardiomyocyte differentiation, and a use thereof.

1 . An inhibitor of Wnt signalling, comprising at least one kind of insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor. 
     
     
         2 . An inhibitor of Wnt signalling, comprising as an active ingredient an effective amount of at least one kind of insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor, wherein the at least one kind of IGFBP is selected from the following group of proteins:
 (1) a protein that is represented by an amino acid sequence set forth in any one of SEQ ID NOS: 2, 4, 6, and 8 of Sequence Listing;   (2) a protein that has 70% or more homology to the protein according to the item (1) and is capable of binding to a Wnt receptor; and   (3) a protein that is represented by an amino acid sequence having 1 to 10 amino acid mutations in the amino acid sequence of the protein according to the item (1) and is capable of binding to a Wnt receptor.   
     
     
         3 . The inhibitor of Wnt signalling according to  claim 1 , wherein the at least one kind of insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor, is a protein represented by an amino acid sequence set forth in any one of SEQ ID NOS: 2, 4, 6, and 8 of Sequence Listing. 
     
     
         4 . The inhibitor of Wnt signalling according to  claim 1 , wherein the at least one kind of insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor, is a protein represented by an amino acid sequence set forth in SEQ ID NO: 2 of Sequence Listing. 
     
     
         5 . The inhibitor of Wnt signalling according to  claim 1 , wherein the Wnt receptor is low-density lipoprotein receptor-related protein 6 (LRP6) and Frizzled 8 (Frz8). 
     
     
         6 . An inhibitor of Wnt signalling, comprising at least one kind of polynucleotide encoding an insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor. 
     
     
         7 . An inhibitor of Wnt signalling, comprising as an active ingredient an effective amount of at least one kind of polynucleotide encoding an insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor, wherein the at least one kind of polynucleotide is selected from the following group of polynucleotides:
 (1) a polynucleotide that is represented by a base sequence set forth in any one of SEQ ID NOS: 1, 3, 5, and 7 of Sequence Listing;   (2) a polynucleotide that has 70% or more homology to the polynucleotide according to the item (1) and encodes a protein capable of binding to a Wnt receptor;   (3) a polynucleotide that is represented by a base sequence having 1 to 30 nucleotide mutations in the base sequence of the polynucleotide according to the item (1) and encodes a protein capable of binding to a Wnt receptor; and   (4) a polynucleotide that includes the polynucleotide according to any one of the items (1) to (3).   
     
     
         8 . The inhibitor of Wnt signalling according to  claim 6 , wherein the at least one kind of polynucleotide encoding an insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor, is a polynucleotide represented by a base sequence set forth in any one of SEQ ID NOS: 1, 3, 5, and 7 of Sequence Listing. 
     
     
         9 . The inhibitor of Wnt signalling according to  claim 6 , wherein the at least one kind of polynucleotide encoding an insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor, is a polynucleotide represented by a base sequence set forth in SEQ ID NO: 1 of Sequence Listing. 
     
     
         10 . The inhibitor of Wnt signalling according to  claim 6 , wherein the Wnt receptor is low-density lipoprotein receptor-related protein 6 (LRP6) and Frizzled 8 (Frz8). 
     
     
         11 . A method of inhibiting Wnt signalling, comprising using at least one kind of insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor. 
     
     
         12 . The method of inhibiting Wnt signalling according to  claim 11 , wherein the at least one kind of insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor, is at least one kind of protein selected from the following group of proteins:
 (1) a protein that is represented by an amino acid sequence set forth in any one of SEQ ID NOS: 2, 4, 6, and 8 of Sequence Listing;   (2) a protein that has 70% or more homology to the protein according to the item (1) and is capable of binding to a Wnt receptor; and   (3) a protein that is represented by an amino acid sequence having 1 to 10 amino acid mutations in the amino acid sequence of the protein according to the item (1) and is capable of binding to a Wnt receptor.   
     
     
         13 . The method of inhibiting Wnt signalling according to  claim 11 , wherein the at least one kind of insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor, is a protein represented by an amino acid sequence set forth in any one of SEQ ID NOS: 2, 4, 6, and 8 of Sequence Listing. 
     
     
         14 . The method of inhibiting Wnt signalling according to  claim 11 , wherein the at least one kind of insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor, is a protein represented by an amino acid sequence set forth in SEQ ID NO: 2 of Sequence Listing. 
     
     
         15 . The method of inhibiting Wnt signalling according to  claim 11 , wherein the Wnt receptor is low-density lipoprotein receptor-related protein 6 (LRP6) and Frizzled 8 (Frz8). 
     
     
         16 . A method of inhibiting Wnt signalling, comprising using at least one kind of polynucleotide encoding an insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor. 
     
     
         17 . The method of inhibiting Wnt signalling according to  claim 16 , wherein the at least one kind of polynucleotide encoding an insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor, is at least one kind of polynucleotide selected from the following group of polynucleotides:
 (1) a polynucleotide that is represented by a base sequence set forth in any one of SEQ ID NOS: 1, 3, 5, and 7 of Sequence Listing;   (2) a polynucleotide that has 70% or more homology to the polynucleotide according to the item (1) and encodes a protein capable of binding to a Wnt receptor;   (3) a polynucleotide that is represented by a base sequence having 1 to 30 nucleotide mutations in the base sequence of the polynucleotide according to the item (1) and encodes a protein capable of binding to a Wnt receptor; and   (4) a polynucleotide that includes the polynucleotide according to any one of the items (1) to (3).   
     
     
         18 . The method of inhibiting Wnt signalling according to  claim 16 , wherein the at least one kind of polynucleotide encoding an insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor, is a polynucleotide represented by a base sequence set forth in any one of SEQ ID NOS: 1, 3, 5, and 7 of Sequence Listing. 
     
     
         19 . The method of inhibiting Wnt signalling according to  claim 16 , wherein the at least one kind of polynucleotide encoding an insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor, is a polynucleotide represented by a base sequence set forth in SEQ ID NO: 1 of Sequence Listing. 
     
     
         20 . The method of inhibiting Wnt signalling according to  claim 16 , wherein the Wnt receptor is low-density lipoprotein receptor-related protein 6 (LRP6) and Frizzled 8 (Frz8). 
     
     
         21 . A use of an insulin-like growth-factor-binding protein (IGFBP), the protein being capable of binding to a Wnt receptor, and/or a polynucleotide encoding the protein, in manufacture of an inhibitor of Wnt signalling. 
     
     
         22 . A medicament for prevention and/or treatment of a disease due to enhanced Wnt signalling, comprising as an active ingredient an effective amount of the inhibitor of Wnt signalling according to  claim 1 . 
     
     
         23 . A method for prevention and/or treatment of a disease due to enhanced Wnt signalling, comprising administering to a subject an effective amount of the inhibitor of Wnt signalling according to  claim 1 . 
     
     
         24 . A use of the inhibitor of Wnt signalling according to  claim 1  in prevention and/or treatment of a disease due to enhanced Wnt signalling. 
     
     
         25 . A medicament for induction of cardiomyocyte differentiation, comprising as an active ingredient an effective amount of the inhibitor of Wnt signalling according to  claim 1 . 
     
     
         26 . A method of inducing cardiomyocyte differentiation, comprising bringing an effective amount of the inhibitor of Wnt signalling according to  claim 1  into contact with a cell capable of differentiating into a cardiomyocyte. 
     
     
         27 . The method of inducing cardiomyocyte differentiation according to  claim 26 , wherein the cell capable of differentiating into a cardiomyocyte is a pluripotent stem cell. 
     
     
         28 . The method of inducing cardiomyocyte differentiation according to  claim 26 , wherein the cell capable of differentiating into a cardiomyocyte is an embryonic stem cell. 
     
     
         29 . A method of inducing cardiomyocyte differentiation, comprising administering to a subject an effective amount of the inhibitor of Wnt signalling according to  claim 1 . 
     
     
         30 . A cardiomyocyte, which is obtained by the method of inducing cardiomyocyte differentiation according to  claim 26 . 
     
     
         31 . A use of a cardiomyocyte, which is obtained by the method of inducing cardiomyocyte differentiation according to  claim 26 .