Human osteoporosis gene

Abstract

A role of the human BMP2 gene in osteoporosis is disclosed. Methods for diagnosis, prediction of clinical course and treatment for osteoporosis using polymorphisms in the BMP2 gene are also disclosed.

What is claimed is:  
     
         1 . An isolated nucleic acid molecule comprising: 
 a) the nucleotide sequence of SEQ ID NO: 1 and comprising at least one polymorphism as shown in Table 2 and the complement thereof;    b) a nucleic acid encoding a polypeptide having an amino acid sequence selected of SEQ ID NO: 2 which comprises at least one polymorphism as shown in Table 2 and the complement thereof;    c) a nucleic acid molecule which hybridizes under high stringency conditions to a nucleotide sequence selected from the group consisting of SEQ ID NO: 1 which comprises at least one polymorphism as shown in Table 2, and the complement thereof; and    d) a nucleic acid molecule which hybridizes under high stringency conditions to a nucleotide sequence encoding an amino acid sequence selected of SEQ ID NO: 2 which comprises at least one polymorphism as shown in Table 2 and the complement thereof.    
     
     
         2 . A method for assaying the presence of a first nucleic acid molecule in a sample, comprising contacting said sample with a second nucleic acid molecule comprising a nucleotide sequence of  claim 1 .  
     
     
         3 . A vector comprising the isolated nucleic acid molecule of  claim 1 , operatively linked to a regulatory sequence.  
     
     
         4 . A recombinant host cell comprising the vector of  claim 3 .  
     
     
         5 . A method for producing a polypeptide encoded by an isolated nucleic acid molecule, comprising culturing the recombinant host cell of  claim 4  under conditions suitable for expression of said nucleic acid molecule.  
     
     
         6 . An isolated polypeptide encoded by a BMP2 gene, or a fragment or variant of said polypeptide, comprising: 
 a) the amino acid sequence of SEQ ID NO: 2 which comprises at least one polymorphism as shown in Table 2;    b) an amino acid sequence which is greater than about 90 percent identical to an amino acid sequence of SEQ ID NO: 2; and    c) an amino acid sequence which is greater than about 95 percent identical to an amino acid sequence of SEQ ID NO: 2.    
     
     
         7 . A fusion protein comprising an isolated polypeptide of  claim 6 .  
     
     
         8 . An antibody, or an antigen-binding fragment thereof, which selectively binds to a polypeptide of  claim 6 .  
     
     
         9 . A method for assaying the presence of a polypeptide encoded by an isolated nucleic acid molecule according to  claim 1  in a sample, comprising contacting said sample with an antibody which specifically binds to the encoded polypeptide.  
     
     
         10 . A method of diagnosing a susceptibility to osteoporosis in an individual, comprising detecting a polymorphism in BMP2 gene, wherein the presence of the polymorphism in the gene is indicative of a susceptibility to osteoporosis.  
     
     
         11 . A method of diagnosing a susceptibility to osteoporosis, comprising detecting an alteration in the expression or composition of a polypeptide encoded by BMP2 gene in a test sample, in comparison with the expression or composition of a polypeptide encoded by BMP2 gene in a control sample, wherein the presence of an alteration in expression or composition of the polypeptide in the test sample is indicative of a susceptibility to osteoporosis.  
     
     
         12 . The method of  claim 11 , wherein the alteration in the expression or composition of a polypeptide encoded by BMP2 gene comprises expression of a splicing variant polypeptide in a test sample that differs from a splicing variant polypeptide expressed in a control sample.  
     
     
         13 . A method of identifying an agent which alters activity of a polypeptide of  claim 6 , comprising: 
 a) contacting the polypeptide or a derivative or fragment thereof, with an agent to be tested;    b) assessing the level of activity of the polypeptide or derivative or fragment thereof; and    c) comparing the level of activity with a level of activity of the polypeptide or active derivative or fragment thereof in the absence of the agent, wherein if the level of activity of the polypeptide or derivative or fragment thereof in the presence of the agent differs, by an amount that is statistically significant, from the level in the absence of the agent, then the agent is an agent that alters activity of the polypeptide.    
     
     
         14 . An agent which alters activity of a polypeptide encoded by BMP2 gene, wherein the agent is selected from the group consisting of: a BMP2 gene receptor; a BMP2 gene binding agent; a peptidomimetic; a fusion protein; a prodrug; an antibody; and a ribozyme.  
     
     
         15 . A method of altering activity of a polypeptide encoded by BMP2 gene, comprising contacting the polypeptide with an agent of  claim 14 .  
     
     
         16 . A method of identifying an agent which alters interaction of the polypeptide of  claim 6  with a BMP2 gene binding agent, comprising: 
 a) contacting the polypeptide or a derivative or fragment thereof, the binding agent and with an agent to be tested;  
 b) assessing the interaction of the polypeptide or derivative or fragment thereof with the binding agent; and  
 c) comparing the level of interaction with a level of interaction of the polypeptide or derivative or fragment thereof with the binding agent in the absence of the agent, wherein if the level of interaction of the polypeptide or derivative or fragment thereof in the presence of the agent differs, by an amount that is statistically significant, from the level of interaction in the absence of the agent, then the agent is an agent that alters interaction of the polypeptide with the binding agent.  
 
     
     
         17 . An agent which alters interaction of a BMP2 gene polypeptide with a first BMP2 gene binding agent, selected from the group consisting of: a BMP2 gene receptor; a second BMP2 gene binding agent; a peptidomimetic; a fusion protein; a prodrug; an antibody; and a ribozyme.  
     
     
         18 . A method of altering interaction of a BMP2 gene polypeptide with a BMP2 gene binding agent, comprising contacting the BMP2 gene polypeptide and/or the BMP2 gene binding agent with an agent of  claim 17 .  
     
     
         19 . A method of identifying an agent which alters expression of BMP2 gene, comprising the steps of: 
 a) contacting a solution containing a nucleic acid of  claim 1  or a derivative or fragment thereof with an agent to be tested;    b) assessing the level of expression of the nucleic acid, derivative or fragment; and    c) comparing the level of expression with a level of expression of the nucleic acid, derivative or fragment in the absence of the agent, wherein if the level of expression of the nucleotide, derivative or fragment in the presence of the agent differs, by an amount that is statistically significant, from the expression in the absence of the agent, then the agent is an agent that alters expression of BMP2 gene.    
     
     
         20 . A method of identifying an agent which alters expression of BMP2 gene, comprising the steps of: 
 a) contacting a solution containing a nucleic acid comprising the promoter region of BMP2 gene operably linked to a reporter gene, with an agent to be tested;    b) assessing the level of expression of the reporter gene; and    c) comparing the level of expression with a level of expression of the reporter gene in the absence of the agent, wherein if the level of expression of the reporter gene in the presence of the agent differs, by an amount that is statistically significant, from the level of expression in the absence of the agent, then the agent is an agent that alters expression of BMP2 gene.    
     
     
         21 . A method of identifying an agent which alters expression of BMP2 gene, comprising the steps of: 
 a) contacting a solution containing a nucleic acid of  claim 1  or a derivative or fragment thereof with an agent to be tested;    b) assessing expression of the nucleic acid, derivative or fragment; and    c) comparing expression with expression of the nucleic acid, derivative or fragment in the absence of the agent, wherein if expression of the nucleotide, derivative or fragment in the presence of the agent differs, by an amount that is statistically significant, from the expression in the absence of the agent, then the agent is an agent that alters expression of BMP2 gene.    
     
     
         22 . The method of  claim 21 , wherein the expression of the nucleotide, derivative or fragment in the presence of the agent comprises expression of one or more splicing variant(s) that differ in kind or in quantity from the expression of one or more splicing variant(s) the absence of the agent.  
     
     
         23 . An agent which alters expression of BMP2 gene, selected from the group consisting of: antisense nucleic acid to BMP2 gene; a BMP2 gene polypeptide; a BMP2 gene receptor; a BMP2 gene binding agent; a peptidomimetic; a fusion protein; a prodrug thereof; an antibody; and a ribozyme.  
     
     
         24 . A method of altering expression of BMP2 gene, comprising contacting a cell containing BMP2 gene with an agent of  claim 23 .  
     
     
         25 . A method of identifying a polypeptide which interacts with a BMP2 gene polypeptide, comprising employing a two yeast hybrid system using a first vector which comprises a nucleic acid encoding a DNA binding domain and a BMP2 gene polypeptide, splicing variant, or fragment or derivative thereof, and a second vector which comprises a nucleic acid encoding a transcription activation domain and a nucleic acid encoding a test polypeptide, wherein if transcriptional activation occurs in the two yeast hybrid system, the test polypeptide is a polypeptide which interacts with a BMP2 polypeptide.  
     
     
         26 . A BMP2 gene therapeutic agent selected from the group consisting of: a BMP2 gene or fragment or derivative thereof; a polypeptide encoded by BMP2 gene; a BMP2 gene receptor; a BMP2 gene binding agent; a peptidomimetic; a fusion protein; a prodrug; an antibody; an agent that alters BMP2 gene expression; an agent that alters activity of a polypeptide encoded by BMP2 gene; an agent that alters posttranscriptional processing of a polypeptide encoded by BMP2 gene; an agent that alters interaction of a BMP2 gene with a BMP2 gene binding agent; an agent that alters transcription of splicing variants encoded by BMP2 gene; and a ribozyme.  
     
     
         27 . A pharmaceutical composition comprising a BMP2 gene therapeutic agent of  claim 26 .  
     
     
         28 . The pharmaceutical composition of  claim 27 , wherein the BMP2 gene therapeutic agent is an isolated nucleic acid molecule comprising a BMP2 gene or fragment or derivative thereof.  
     
     
         29 . The pharmaceutical composition of  claim 27 , wherein the BMP2 gene therapeutic agent is a polypeptide encoded by the BMP2 gene.  
     
     
         30 . The pharmaceutical composition of  claim 27  wherein the receptor is BMP2R2, BMPR1A or BMPR1B.  
     
     
         31 . A method of treating osteoporosis in an individual, comprising administering a BMP2 gene therapeutic agent to the individual, in a therapeutically effective amount.  
     
     
         32 . The method of  claim 31 , wherein the BMP2 gene therapeutic agent is a BMP2 gene agonist or a BMP2 gene antagonist.  
     
     
         33 . A transgenic animal comprising a nucleic acid selected from the group consisting of: an exogenous BMP2 gene and a nucleic acid encoding a BMP2 gene polypeptide.  
     
     
         34 . A method for assaying a sample for the presence of a BMP2 gene nucleic acid, comprising: 
 a) contacting said sample with a nucleic acid comprising a contiguous nucleotide sequence which is at least partially complementary to a part of the sequence of said BMP2 gene nucleic acid under conditions appropriate for hybridization, and    b) assessing whether hybridization has occurred between a BMP2 gene nucleic acid and said nucleic acid comprising a contiguous nucleotide sequence which is at least partially complementary to a part of the sequence of said BMP2 gene nucleic acid.    
     
     
         35 . The method of  claim 34 , wherein said nucleic acid comprising a contiguous nucleotide sequence is completely complementary to a part of the sequence of said BMP2 gene nucleic acid.  
     
     
         36 . The method of  claim 34 , comprising amplification of at least part of said BMP2 gene nucleic acid.  
     
     
         37 . The method of  claim 34 , wherein said contiguous nucleotide sequence is 100 or fewer nucleotides in length and is either: a) at least 80% identical to a contiguous sequence of nucleotides in SEQ ID NO: 1 which comprises at least one polymorphism as shown in Table 2; b) at least 80% identical to the complement of a contiguous sequence of nucleotides in SEQ ID NO: 1 which comprises at least one polymorphism as shown in Table 2; or c) capable of selectively hybridizing to said BMP2 gene nucleic acid.  
     
     
         38 . A reagent for assaying a sample for the presence of a BMP2 gene nucleic acid, said reagent comprising a nucleic acid comprising a contiguous nucleotide sequence which is at least partially complementary to a part of the nucleotide sequence of said BMP2 gene nucleic acid.  
     
     
         39 . The reagent of  claim 38 , wherein the nucleic acid comprises a contiguous nucleotide sequence which is completely complementary to a part of the nucleotide sequence of said BMP2 gene nucleic acid.  
     
     
         40 . A reagent kit for assaying a sample for the presence of a BMP2 gene nucleic acid, comprising in separate containers: 
 a) one or more labeled nucleic acids comprising a contiguous nucleotide sequence which is at least partially complementary to a part of the nucleotide sequence of said BMP2 gene nucleic acid, and    b) reagents for detection of said label.    
     
     
         41 . The reagent kit of claim  40 , wherein the labeled nucleic acid comprises a contiguous nucleotide sequences which is completely complementary to a part of the nucleotide sequence of said BMP2 gene nucleic acid.  
     
     
         42 . A reagent kit for assaying a sample for the presence of a BMP2 gene nucleic acid, comprising one or more nucleic acids comprising a contiguous nucleotide sequence which is at least partially complementary to a part of the nucleotide sequence of said BMP2 gene nucleic acid, and which is capable of acting as a primer for said BMP2 gene nucleic acid when maintained under conditions for primer extension.  
     
     
         43 . A kit comprising: 
 a) at least one antibody selected from the group consisting of: an antibody specific for the BMP2 protein comprising a serine at amino acid position 189 and an alanine at amino acid position 37; and an antibody specific for the BMP2 protein comprising an arginine at amino acid position 189 and a serine at amino acid position 37; and    b) reference BMP2 protein sample.