Biomarkers for Diagnosing Schizophrenia and Bipolar Disorder

Abstract

The invention relates to the identification and selection of novel biomarkers and the identification and selection of novel biomarker combinations which are differentially expressed in blood and useful in diagnosing schizophrenia and/or bipolar disorder as well as monitoring therapeutic efficacy of treatment for schizophrenia or bipolar disorder. The measurement of expression levels of the products of the biomarkers and combinations of biomarkers of the invention can be used to diagnose schizophrenia and/or bipolar disorder. Measurement of the expression level of products of biomarkers of the invention using polynucleotides and proteins which specifically and/or selectively hybridize to the products of the biomarkers of the invention are also encompassed within the scope of the invention as are compositions and kits containing said polynucleotides and proteins. Further encompassed by the invention is the use of the polynucleotides and proteins to monitor the efficacy of therapeutic regimens.

What is claimed is: 
     
         1 . A composition comprising a collection of two or more isolated polynucleotides, said polynucleotides which selectively hybridize to at least two biomarkers of the invention, wherein the biomarkers are selected from the group consisting of the genes: adenylosuccinate synthase (ADSS); apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3B (APOBEC3B); ataxia telangiectasia mutated (includes complementation groups A, C and D) (ATM); Charcot-Leyden crystal protein (CLC); C-terminal binding protein 1 (CTBP1); chemokine (C—X—C motif) ligand 1 (melanoma growth stimulating activity, alpha) (CXCL1); death associated transcription factor 1 (DATF1); 5100 calcium binding protein A9 (calgranulin B) (S100A9), and as set out in Table 1, and wherein the composition is used to measure the level of expression of said biomarker. 
     
     
         2 . A composition comprising a collection of two or more isolated polynucleotides which bind selectively to the RNA products of at least two biomarkers, wherein the biomarkers are selected from the group consisting of the genes: adenylosuccinate synthase (ADSS); apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3B (APOBEC3B); ataxia telangiectasia mutated (includes complementation groups A, C and D) (ATM); Charcot-Leyden crystal protein (CLC); C-terminal binding protein 1 (CTBP1); chemokine (C—X—C motif) ligand 1 (melanoma growth stimulating activity, alpha) (CXCL1); death associated transcription factor 1 (DATF1); 5100 calcium binding protein A9 (calgranulin B) (S100A9), as set out in Table 1. 
     
     
         3 . A composition comprising a collection of two or more isolated proteins which bind selectively to the protein products of at least two biomarkers, wherein the biomarkers are selected from the group consisting of the genes: adenylosuccinate synthase (ADSS); apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3B (APOBEC3B); ataxia telangiectasia mutated (includes complementation groups A, C and D) (ATM); Charcot-Leyden crystal protein (CLC); C-terminal binding protein 1 (CTBP1); chemokine (C—X—C motif) ligand 1 (melanoma growth stimulating activity, alpha) (CXCL1); death associated transcription factor 1 (DATF1); 5100 calcium binding protein A9 (calgranulin B) (S100A9), as set out in Table 1 
     
     
         4 . A composition comprising a collection of two or more isolated polynucleotides which bind selectively to the RNA products of one or more biomarkers, wherein the biomarkers are selected from the group consisting of the genes: adenylosuccinate synthase (ADSS); death associated transcription factor 1 (DATF1); as listed in Table 3. 
     
     
         5 . A composition comprising a collection of one or more isolated proteins, which bind selectively to the protein products of one or more biomarkers, wherein the biomarkers are selected from the group consisting of the genes: adenylosuccinate synthase (ADSS); death associated transcription factor 1 (DATF1); as set out in Table 3. 
     
     
         6 . A composition comprising a collection of isolated polynucleotides which bind selectively to the RNA products of biomarkers, wherein the biomarkers are selected from the group of genes: adenylosuccinate synthase (ADSS); death associated transcription factor 1 (DATF1); as set out in Table 3. 
     
     
         7 . A composition comprising a collection of two or more isolated polynucleotides which bind selectively to the RNA products of at least one biomarker, wherein the biomarkers are selected from the group as set out in Table 4. 
     
     
         8 . A composition comprising a collection of two or more isolated polynucleotides which bind selectively to the RNA products of at least one biomarker, wherein the biomarkers are selected from the group as set out in Table 5. 
     
     
         9 . A composition comprising a collection of one or more isolated proteins which bind selectively to the protein products of at least one biomarker, wherein the biomarkers are selected from the group as set out in Table 4. 
     
     
         10 . A composition comprising a collection of one or more isolated proteins which bind selectively to the protein products of at least one biomarker, wherein the biomarkers are selected from the group as set out in Table 5. 
     
     
         11 . A method of diagnosing or prognosing schizophrenia in an individual, comprising the steps of:
 a) determining the level of one or more RNA transcripts expressed in blood obtained from said individual, wherein said one or more RNA transcripts corresponds to said one or more biomarkers of Table 3, and   b) comparing the level of each of said one or more RNA transcripts in said blood according to step a) with the level of each of said one or more RNA transcripts in blood from one or more individuals not having schizophrenia,   wherein detecting differential expression of each of said one or more RNA transcripts in the comparison of step b) is indicative of schizophrenia in the individual of step a).   
     
     
         12 . A method of diagnosing or prognosing bipolar disorder in an individual, comprising the steps of:
 a) determining the level of one or more RNA transcripts expressed in blood obtained from said individual, wherein said one or more RNA transcripts corresponds to said one or more biomarkers of Table 4, and   b) comparing the level of each of said one or more RNA transcripts in said blood according to step a) with the level of each of said one or more RNA transcripts in blood from one or more individuals not having bipolar disorder,   wherein detecting differential expression of each of said one or more RNA transcripts in the comparison of step b) is indicative of bipolar disorder in the individual of step a).   
     
     
         13 . A method of diagnosing or prognosing an individual as having either bipolar disorder or schizophrenia, comprising the steps of:
 a) determining the level of one or more RNA transcripts expressed in blood obtained from said individual, wherein said one or more RNA transcripts corresponds to said one or more biomarkers of Table 5, and   b) comparing the level of each of said one or more RNA transcripts in said blood according to step a) with the level of each of said one or more RNA transcripts in blood from one or more individuals having schizophrenia,   wherein detecting differential expression of each of said one or more RNA transcripts in the comparison of step b) is indicative of bipolar disorder in the individual of step a).   
     
     
         14 . A method of diagnosing or prognosing an individual as having either bipolar disorder or schizophrenia, comprising the steps of:
 a) determining the level of one or more RNA transcripts expressed in blood obtained from said individual, wherein said one or more RNA transcripts corresponds to said one or more biomarkers of Table 5, and   b) comparing the level of each of said one or more RNA transcripts in said blood according to step a) with the level of each of said one or more RNA transcripts in blood from one or more individuals having bipolar disorder,   wherein detecting differential expression of each of said one or more RNA transcripts in the comparison of step b) is indicative of schizophrenia in the individual of step a).   
     
     
         15 . A method of diagnosing or prognosing schizophrenia in an individual, comprising the steps of:
 a) determining the level of two or more RNA transcripts expressed in blood obtained from said individual, wherein said two or more RNA transcripts corresponds to said one or more biomarkers of Table 1 and   b) comparing the level of each of said two or more RNA transcripts in said blood according to step a) with the level of each of said two or more RNA transcripts in blood from one or more individuals having schizophrenia,   c) comparing the level of each of said two or more RNA transcripts in said blood according to step a) with the level of each of said two or more RNA transcripts in blood from one or more individuals not having schizophrenia,   d) determining whether the level of said two or more RNA transcripts of step a) classify with the levels of said transcripts in step b) as compared with levels of said transcripts in step c),   wherein said determination is indicative of said individual of step a) having schizophrenia.   
     
     
         16 . A method of diagnosing or prognosing bipolar disorder in an individual, comprising the steps of:
 a) determining the level of two or more RNA transcripts expressed in blood obtained from said individual, wherein said two or more RNA transcripts corresponds to said two or more biomarkers of Table 1 and   b) comparing the level of each of said two or more RNA transcripts in said blood according to step a) with the level of each of said two or more RNA transcripts in blood from one or more individuals having bipolar disorder,   c) comparing the level of each of said two or more RNA transcripts in said blood according to step a) with the level of each of said two or more RNA transcripts in blood from one or more individuals not having bipolar disorder,   d) determining whether the level of said two or more RNA transcripts of step a) classify with the levels of said transcripts in step b) as compared with levels of said transcripts in step c),   wherein said determination is indicative of said individual of step a) having bipolar disorder.   
     
     
         17 . A method of diagnosing or prognosing an individual as having bipolar disorder or schizophrenia, comprising the steps of:
 a) determining the level of two or more RNA transcripts expressed in blood obtained from said individual, wherein said two or more RNA transcripts corresponds to said two or more biomarkers of Table 1 and   b) comparing the level of each of said two or more RNA transcripts in said blood according to step a) with the level of each of said two or more RNA transcripts in blood from one or more individuals having bipolar disorder,   c) comparing the level of each of said two or more RNA transcripts in said blood according to step a) with the level of each of said two or more RNA transcripts in blood from one or more individuals having schizophrenia,   d) determining whether the level of said two or more RNA transcripts of step a) classify with the levels of said transcripts in step b) as compared with levels of said transcripts in step c),   wherein said determination is indicative of said individual of step a) having bipolar disorder.   
     
     
         18 . A method of diagnosing or prognosing schizophrenia in an individual, comprising the steps of:
 a) determining the level of one or more RNA transcripts expressed in blood obtained from said individual, wherein said one or more RNA transcripts corresponds to said one or more biomarkers of Table 1 and   b) using the results from step (a) in combination with a classifier so as to determine a diagnosis with respect to schizophrenia.   
     
     
         19 . A method of diagnosing or prognosing bipolar disorder in an individual, comprising the steps of:
 a) determining the level of one or more RNA transcripts expressed in blood obtained from said individual, wherein said one or more RNA transcripts corresponds to said one or more biomarkers of Table 1 and   b) using the results from step (a) in combination with a classifier so as to determine a diagnosis with respect to bipolar disorder.   
     
     
         20 . A kit for diagnosing or prognosing schizophrenia comprising:
 a) at least two sets of biomarker specific priming means wherein each set of biomarker specific priming means produces double stranded DNA complementary to a unique biomarker selected from Table 1; wherein each first priming means of said sets contains a sequence which can selectively hybridize to RNA, cDNA or an EST complementary to one of said biomarkers to create an extension product and each said second priming means of said sets is capable of selectively hybridizing to said extension product;   b) an enzyme with reverse transcriptase activity;   c) an enzyme with thermostable DNA polymerase activity, and   d) a labeling means;   wherein each of said primer sets is used to detect the quantitative expression levels of said biomarker in a test subject.   
     
     
         21 . A kit for diagnosing or prognosing bipolar disorder comprising:
 a) at least two sets of biomarker specific priming means wherein each set of biomarker specific priming means produces double stranded DNA complementary to a unique biomarker selected from Table 1; wherein each first priming means of said sets contains a sequence which can selectively hybridize to RNA, cDNA or an EST complementary to one of said biomarkers to create an extension product and each said second priming means of said sets is capable of selectively hybridizing to said extension product;   b) an enzyme with reverse transcriptase activity;   c) an enzyme with thermostable DNA polymerase activity, and   d) a labeling means;   
       wherein each said primer set is used to detect the quantitative expression levels of a biomarker in a test subject. 
     
     
         22 . A method of diagnosing or prognosing schizophrenia in an individual, comprising the steps of:
 a) determining the level of two or more proteins expressed in blood obtained from said individual, wherein said two or more proteins are encoded by two or more biomarkers of Table 1, and   b) comparing the level of each of said two or more proteins in said blood according to step a) with the level of each of said two or more proteins in blood from one or more individuals having schizophrenia,   c) comparing the level of each of said two or more proteins in said blood according to step a) with the level of each of said two or more proteins in blood from one or more individuals not having schizophrenia,   d) determining whether the level of said two or more proteins of step a) classify with the levels of said proteins in step b) as compared with levels of said proteins in step c),   
       wherein said determination is indicative of said individual of step a) having schizophrenia. 
     
     
         23 . A method of diagnosing or prognosing bipolar disorder in an individual, comprising the steps of:
 a) determining the level of two or more proteins expressed in blood obtained from said individual, wherein said two or more proteins are encoded by two or more biomarkers of Table 1, and   b) comparing the level of each of said two or more proteins in said blood according to step a) with the level of each of said two or more proteins in blood from one or more individuals having bipolar disorder   c) comparing the level of each of said two or more proteins in said blood according to step a) with the level of each of said two or more proteins in blood from one or more individuals not having bipolar disorder,   d) determining whether the level of said two or more proteins of step a) classify with the levels of said proteins in step b) as compared with levels of said proteins in step c),   
       wherein said determination is indicative of said individual of step a) having bipolar disorder. 
     
     
         24 . A method of diagnosing or prognosing an individual with bipolar disorder as compared with schizophrenia, comprising the steps of:
 a) determining the level of two or more proteins expressed in blood obtained from said individual, wherein said two or more proteins are encoded by two or more biomarkers of Table 1, and   b) comparing the level of each of said two or more proteins in said blood according to step a) with the level of each of said two or more proteins in blood from one or more individuals having bipolar disorder   c) comparing the level of each of said two or more proteins in said blood according to step a) with the level of each of said two or more proteins in blood from one or more individuals having schizophrenia,   d) determining whether the level of said two or more proteins of step a) classify with the levels of said proteins in step b) as compared with levels of said proteins in step c),   
       wherein said determination is indicative of said individual of step a) having bipolar disorder. 
     
     
         25 . A method of diagnosing or prognosing schizophrenia in an individual, comprising the steps of:
 a) determining the level of two or more protein products expressed in blood obtained from said individual, wherein said two or more protein products corresponds to two or more biomarkers of Table 1 and   b) using the results from step (a) in combination with a classifier designed to differentiate schizophrenia from non schizophrenia so as to determine a diagnosis with respect to schizophrenia.   
     
     
         26 . A method of diagnosing or prognosing bipolar disorder in an individual, comprising the steps of:
 a) determining the level of two or more protein products expressed in blood obtained from said individual, wherein said two or more protein products corresponds to two or more biomarkers of Table 1 and   b) using the results from step (a) in combination with a classifier designed to differentiate bipolar disorder from non bipolar disorder so as to determine a diagnosis with respect to bipolar disorder.   
     
     
         27 . A method of diagnosing or prognosing and individual as having bipolar disorder or schizophrenia, comprising the steps of:
 a) determining the level of two or more protein products expressed in blood obtained from said individual, wherein said two or more protein products corresponds to two or more biomarkers of Table 1 and   b) using the results from step (a) in combination with a classifier designed to differentiate bipolar disorder from schizophrenia so as to determine a diagnosis with respect to bipolar disorder or schizophrenia.   
     
     
         28 . A method of developing a classifier useful for diagnosing schizophrenia, said method comprising:
 (a) measuring the level of expression of the products of the biomarkers identified in Table 1 in a training population wherein said training population is comprised of two subgroups, a first subgroup diagnosed as having schizophrenia and said second subgroup diagnosed as not having schizophrenia.   (b) apply one or more mathematical models to the levels of expression of step (a) to develop one or more classifiers which differentiate between said first subgroup and said second subgroup.   
     
     
         29 . A method of developing a classifier useful for diagnosing bipolar disorder, said method comprising:
 (a) measuring the level of expression of the products of the biomarkers identified in Table 1 in a training population wherein said training population is comprised of two subgroups, a first subgroup diagnosed as having bipolar disorder and said second subgroup diagnosed as not having bipolar disorder.   (b) apply one or more mathematical models to the levels of expression of step (a) to develop one or more classifiers which differentiate between said first subgroup and said second subgroup.   
     
     
         30 . A method of developing a classifier useful for diagnosing bipolar disorder or schizophrenia, said method comprising:
 (a) measuring the level of expression of the products of the biomarkers identified in Table 1 in a training population wherein said training population is comprised of two subgroups, a first subgroup diagnosed as having bipolar disorder and said second subgroup diagnosed as having schizophrenia.   (b) apply one or more mathematical models to the levels of expression of step (a) to develop one or more classifiers which differentiate between said first subgroup and said second subgroup.