Proteoglycan Splice Variants as Therapeutics and Diagnostics for Amyloid Diseases

Abstract

The identification of novel Syndecan-2 splice variants and their use in the diagnosis and therapeutic intervention of Alzheimer's disease and other amyloid diseases. In addition the use of new animal models expressing or devoid of syndecan-2 splice variants to effectively screen and identify potential therapeutic compounds for Alzheimer's disease.

1 . An isolated polynucleotide consisting of SEQ ID NO: 6. 
     
     
         2 . An isolated polynucleotide consisting of SEQ ID NO: 8. 
     
     
         3 . A polynucleotide encoding a protein according to SEQ ID NO:7. 
     
     
         4 . A polynucleotide encoding a protein according to SEQ ID NO:9. 
     
     
         5 . A vector comprising SEQ ID NO: 6. 
     
     
         6 . A vector comprising at least a portion of nucleotides 124-279 of SEQ ID NO: 6. 
     
     
         7 . A vector comprising SEQ ID NO: 8 or a fragment thereof. 
     
     
         8 . A splice variant of syndecan-2 having the sequence of SEQ ID NO: 7. 
     
     
         9 . A splice variant of syndecan-2 having the sequence of SEQ ID NO: 9 or a fragment thereof. 
     
     
         10 . A polypeptide obtained from the translation of the polynucleotide of  claim 1 . 
     
     
         11 . A polypeptide comprising at least a portion of amino acids 21-72 of SEQ ID NO: 7. 
     
     
         12 . A polypeptide as set forth in SEQ ID NO:9 or a fragment thereof. 
     
     
         13 . A method for detection and/or quantitation of a splice variant of syndecan-2 in a biological sample, the method comprising; synthesizing cDNA from mRNA in the sample, amplifying portions of the cDNA corresponding to the splice variant or fragments thereof and detecting/quantitating the amplified cDNA. 
     
     
         14 . The method of  claim 13 , wherein the biological sample is derived from one of tissues, cells or biological fluids. 
     
     
         15 . The method of  claim 14  wherein said tissues, cells or biological fluids are derived from humans. 
     
     
         16 . The method of  claim 13  wherein said biological fluids include blood, plasma, serum, cerebrospinal fluid, sputum, saliva, urine and stool. 
     
     
         17 . The method of  claim 13  wherein said biological fluid is cerebrospinal fluid. 
     
     
         18 . The method of  claim 13  wherein the step of amplifying portions of the cDNA is performed by RT-PCR. 
     
     
         19 . The method of  claim 18  wherein primers utilized for RT-PCR are selected from the group consisting of SEQ ID NO:1, SEQ. ID NO:2, SEQ ID NO:3, SEQ ID NO:4 and SEQ ID NO:5. 
     
     
         20 . The method of  claim 13 , whereby said method utilizes quantitative RT-PCR to determine relative levels of the splice variant in the sample. 
     
     
         21 . The method of  claim 13  whereby said method is utilized to diagnose Alzheimer's disease or determine susceptibility or progression of Alzheimer's disease related to the levels of the splice variant, wherein elevated or diminished levels of a particular splice variant are indicative of the presence of, susceptibility to, or progression of Alzheimer's disease. 
     
     
         22 . A method for the treatment of Alzheimer's disease comprising: administering to a patient a therapeutically effective amount of an oligonucleotide having a sequence complementary to the polynucleotide of  claim 1  or  2 . 
     
     
         23 . The method of  claim 22  where the oligonucleotide is antisense DNA or RNA and is complementary to a sequence selected from the group consisting of SEQ ID NO:6, SEQ ID NO: 8, and fragments thereof. 
     
     
         24 . An antibody that binds to an epitope comprising at least several of amino acids 21-72 of SEQ ID NO: 7. 
     
     
         25 . The antibody of  claim 24  where the epitope consisted of amino acids 50-65 of SEQ ID NO: 7. 
     
     
         26 . A method for the treatment of Alzheimer's disease comprising administering to a patient the antibody of  claim 24  or  25 . 
     
     
         27 . A method for the treatment of Alzheimer's disease comprising administering to a patient a immunogenic amount of the splice variant of  claim 9 . 
     
     
         28 . Use of the splice variant of  claim 8  or  9  to modulate APP processing. 
     
     
         29 . Use of the splice variant of  claim 8  or  9  to modulate activity of APP secretases. 
     
     
         30 . The use according to  claim 29  where the APP secretase is beta-secretase.