Methods of identifying adipocyte specific genes, the genes identified, and their uses

Abstract

Disclosed is a method of identifying genes that are over-expressed in adipose tissue as compared to pre-adipocyte tissue or other tissues comprising performing differential gene expression analysis between the white adipose tissue (WAT) or stromal vascular tissue (SVT) from any two different mice selected from the group consisting of wild-type, HMGI-C−/−, ob/ob, and HMGI-C−/− ob/ob genotype mice. Based on this method a number of nucleotide sequences are identified whose expression is adipocyte specific. The identified nucleotide sequences and their corresponding polypeptides are then used to prevent adipogenesis, to treat diabetes, and to screen for small-molecules that can modulate or prevent adipogenesis and to treat diabetes.

1 - 46 . (canceled) 
     
     
         47 . An isolated polynucleotide comprising
 a) a nucleotide sequence of SEQ ID NO:103;   b) a nucleotide sequence coding for the polypeptide of SEQ.ID.NO. 603;   c) a nucleotide sequence that has at least. 91% identity over its entire length to a nucleotide sequence encoding the sFRP-5 polypeptide of SEQ ID NO:603 said identity being over the entire region encoding SEQ ID NO:603; or   d) a nucleotide sequence complementary to the isolated nucleic acid molecule.   
     
     
         48 . The isolated polynucleotide of  claim 47  wherein expression of said gene upregulates its own expression in an autocrine fashion in adipocytes. 
     
     
         49 . The isolated polynucleotide of  claim 47  comprising a nucleotide sequence that is at least 91% identical with SEQ ID NO:103, said identity being over the entire length of SEQ ID NO:103. 
     
     
         50 . The isolated polynucleotide of  claim 47  comprising the nucleotide sequence of SEQ ID NO:103. 
     
     
         51 . An isolated polynucleotide comprising a nucleotide sequence which, by virtue of redundancy of the genetic code, encodes for the amino acid of SEQ ID NO:603. 
     
     
         52 . The polynucleotide of any one of  claim 47  which is DNA or RNA. 
     
     
         53 . The isolated polynucleotide of any one of  claim 47 , further comprising an expression system that is capable of transcribing the nucleic acid sequence and producing the corresponding amino acid sequence. 
     
     
         54 . A host cell comprising a polynucleotide sequence of as defined in any one of  claim 47 . 
     
     
         55 . The host cell of  claim 54 , wherein the polynucleotide sequence is stably incorporated into the genome of the host cell. 
     
     
         56 . A process for producing an sFRP-5 polypeptide comprising culturing a host of  claim 54  under conditions sufficient for the production of said polypeptide and recovering the polypeptide from the culture. 
     
     
         57 . A process for producing a cell which produces an sFRP-5 polypeptide comprising transforming or transfecting a host cell with the polynucleotide of  claim 53  such that the host cell, under appropriate culture conditions, produces an sFRP-5 polypeptide. 
     
     
         58 . An sFRP-5 polypeptide comprising an amino acid sequence which is at least 91% identical to the amino acid sequence of SEQ.ID.NO:603 over its entire length. 
     
     
         59 . The polypeptide of  claim 58  which comprises the amino acid sequence of SEQ ID NO:603. 
     
     
         60 . An antibody immunospecific for the sFRP-5 polypeptide of  claim 58 . 
     
     
         61 . A method for the treatment of a subject in need of enhanced activity or expression of the sFRP-5 polypeptide of  claim 58  comprising:
 (a) administering to the subject a therapeutically effective amount of an agonist to the polypeptide; and/or (b) administering to the subject an isolated polynucleotide comprising a nucleotide sequence that has at least 91% identity to a nucleotide sequence encoding the sFRP-5 polypeptide of SEQ.ID.NO.603 over its entire length, or a nucleotide sequence complementary to said nucleotide sequence in a form so as to effect production of such polypeptide in vivo; and/or (c) administering to a subject a therapeutically effective amount of the sFRP-5 polypeptide. 
 
     
     
         62 . A method for the treatment of a subject in need of decreased activity or expression of the sFRP-5 polypeptide of  claim 58  comprising:
 (a) administering to the subject a therapeutically effective amount of an antagonist to said polypeptide; and/or (b) administering to the subject a nucleic acid molecule that inhibits the expression of the nucleotide sequence encoding said polypeptide; and/or (c) administering to the subject a therapeutically effective amount of a polypeptide that competes with said polypeptide for its ligand, substrate, or receptor. 
 
     
     
         63 . A process for diagnosing a susceptibility to obesity in a subject related to expression or activity of the sFRP-5 polypeptide of  claim 58  in a subject comprising:
 (a) determining the presence or absence of a mutation in the nucleotide sequence encoding said sFRP-5 polypeptide in the genome of said subject; and/or (b) analyzing for the presence or amount of the sFRP-5 polypeptide expression in a sample derived from said subject. 
 
     
     
         64 . A method for identifying compounds which antagonize or agonize the sFRP-5 polypeptide of  claim 58  comprising:
 (a) contacting a candidate compound(s) with cells which express the sFRP-5 polypeptide (or cell membrane expressing the sFRP-5 polypeptide or the sFRP-5 polypeptide or a fragment thereof bound to a solid matrix) or respond to the sFRP-5 polypeptide; and 
 (b) observing the binding, or stimulation or inhibition of a functional response; or comparing the ability of the cells (or cell membrane) which were contacted with the candidate compound(s) with the same cells which were not contacted for sFRP-5 polypeptide activity. 
 
     
     
         65 - 66 . (canceled) 
     
     
         67 . A recombinant host cell produced by a method of  claim 57  or a membrane thereof expressing an sFRP-5 polypeptide. 
     
     
         68 . A method for screening for modulators of a target protein, wherein the target protein is sFRP-5 and comprises a sequence that has greater than 91% amino acid identity to SEQ ID NO:603 as measured using a sequence comparison algorithm, the method comprising the steps of
 a) contacting the target protein with a candidate agent at a first concentration and determining a level of activity of the target protein; and   b) contacting the target protein with a candidate agent at a second concentration and determining a level of activity of the target protein; wherein a difference between the level of activity of the target protein contacted with the first concentration of the candidate agent and the level of activity of the target protein contacted with the second concentration of the candidate agent indicates that the candidate agent modulates the activity of the target protein.   
     
     
         69 - 116 . (canceled)