Novel peptide involved in energy homeostasis

Abstract

The expression of a mRNA encoding a putative 76 amino acid, secreted protein (“Enho1”) was found to negatively correlate with fasting triglyceride and cholesterol levels. A recombinant adenovirus was used to increase the expression of Enho1 mRNA in two mouse models of obesity, KK-A y and Lep ob /Lep ob mice. Over-expression of Enho1 by adenovirus injection significantly, and reproducibly, reduced fasting triglyceride and cholesterol levels in both models. In addition, transgenic mice strains were made that over express Enho1 protein. Additionally, the expression of a key gene involved in lipogenesis (fatty acid synthase) and FAS protein levels were reduced by ENHO1 adenoviral treatment in Lep ob /Lep ob mice. Full-length ENHO1 peptide, or peptide derivatives, homologues, analogues, or mimetics thereof, delivered by oral intake, injection, subcutaneous patch, or intranasal routes, could be used as therapeutic or diagnostic agents for hypercholesterolemia, hypertriglyceridemia, insulin resistance, obesity, diabetes, and/or energy imbalance.

1 . A purified Enho 1 peptide, and fragment, homologs, analogs and derivatives thereof, wherein the peptide has the following characteristics:
 a. it is a substantially homogenous preparation; and   b. it has at least 70% homology to any one of the sequences set forth in SEQ ID NOS: 2 and 8-18; wherein   c. the peptide modulates energy metabolism, lipid metabolism, and/or insulin action.   
     
     
         2 . A peptide of  claim 1 , wherein the peptide is a mammalian peptide. 
     
     
         3 . A peptide of  claim 1 , wherein the peptide is a murine peptide. 
     
     
         4 . A peptide of  claim 1 , wherein the peptide is a human peptide. 
     
     
         5 . A peptide of  claim 1 , wherein the peptide is a synthetic peptide. 
     
     
         6 . A peptide of  claim 1 , wherein said homology to any one of the sequences set forth in SEQ ID NOS: 2, 10, 11, 12-18 is greater than 85%. 
     
     
         7 . A peptide of  claim 6 , wherein the homology is greater than 95%. 
     
     
         8 . A pharmaceutical composition comprising an Enho1 peptide of  claim 1 , and a pharmaceutical acceptable carrier. 
     
     
         9 . A method for treating or preventing a pathophysiology relating to homeostasis of body mass, comprising administering to a subject a therapeutically effective amount of a pharmaceutical composition comprising a Enho1 peptide according to  claim 1 , or fragments, homologs, analogs and derivatives thereof. 
     
     
         10 . A method according to  claim 9 , wherein the Enho1 peptide has at least 70% homology to any one of the sequences set forth in SEQ ID NOS: 2 and 10-18. 
     
     
         11 . A method according to  claim 9 , wherein the pathophysiology is obesity. 
     
     
         12 . A method according to  claim 9 , additionally comprising administering to a subject a compound selected from the group consisting of leptin and adiponectin. 
     
     
         13 . A method according to  claim 9 , wherein the pathophysiology is one related to obesity comprising any one of hyperglycemia, hyperinsulinemia, insulin resistance, hyperlipidemia, Type II diabetes mellitus, and non-insulin dependent diabetes mellitus. 
     
     
         14 . A method according to  claim 9 , wherein the step of administering to a subject comprises intraperitoneal administration. 
     
     
         15 . A vector comprising a least 200 nucleic acids of the open reading frame of the nucleic acid sequence of SEQ ID NO: 1. 
     
     
         16 . A cultured cell that produces a Enho1 peptide according to  claim 1 . 
     
     
         17 . The cell of  claim 16 , wherein the cell is selected from the group consisting of bacteria, yeast, mammalian cells, plant cells, and insect cells. 
     
     
         18 . A method for preparing a Enho1 peptide comprising:
 a. culturing a cell containing a nucleic acid corresponding to at least the open reading frame of SEQ ID NO: 1 under conditions that provide for expression of the peptide; and   b. recovering the expressed peptide.   
     
     
         19 . A pharmaceutical composition comprising at least the open reading frame of SEQ ID NO: 1, wherein the nucleic acid encodes a Enho1 peptide, and a pharmaceutical acceptable carrier. 
     
     
         20 . An antibody, and fragments, derivatives, homologs and analogs thereof, which immunospecifically binds to the peptide of  claim 1 . 
     
     
         21 . An antibody of  claim 20  labeled with a detectable label. 
     
     
         22 . An antibody of  claim 20 , wherein said antibody is generated using any one of the peptides of SEQ ID NOS: 2 and 8-18, and fragments, derivatives, homologs, and analogs of said peptide. 
     
     
         23 . An antibody or fragment of  claim 20 , wherein the antibody is a polyclonal antibody. 
     
     
         24 . An antibody or fragment of  claim 20 , wherein the antibody is a monoclonal antibody. 
     
     
         25 . A recombinant non-human animal containing a nucleic acid, wherein the nucleic acid is an isolated nucleic acid comprising a nucleotide sequence encoding any one of the Enho1 related peptides of SEQ ID NOS:2 and 8-18, and fragments, derivatives, homologs, and analogs thereof of those peptides. 
     
     
         26 . A method to ameliorate or decrease insulin resistance in a subject, comprising administering to a subject a therapeutically effective amount of a Enho1 peptide according to  claim 1 , and fragments, derivatives, homologs, and analogs thereof of those peptides. 
     
     
         27 . A method to ameliorate or treat symptoms of diabetes in a subject, comprising administering to the subject a therapeutically effective amount of a Enho1 peptide according to  claim 1 , and fragments, derivatives, homologs, and analogs thereof of those peptides. 
     
     
         28 . A method to prevent or delay the onset of obesity in a subject, comprising administering to the subject a therapeutically effective amount of a Enho1 peptide according to  claim 1 , and fragments, derivatives, homologs, and analogs thereof of those peptides. 
     
     
         29 . A method to ameliorate or decrease hyperlipidemia in a subject, comprising administering to the subject a therapeutically effective amount of a Enho1 peptide according to  claim 1 , and fragments, derivatives, homologs, and analogs thereof of those peptides. 
     
     
         30 . A method to decrease total cholesterol in the serum in a subject, comprising administering to the subject a therapeutically effective amount of a Enho1 peptide according to  claim 1 , and fragments, derivatives, homologs, and analogs thereof of those peptides. 
     
     
         31 . A method to decrease serum triglycerides in a subject, comprising administering to the subject a therapeutically effective amount of a Enho1 peptide according to  claim 1 , and fragments, derivatives, homologs, and analogs thereof of those peptides. 
     
     
         32 . A method of any one of  claims 28 - 33 , additionally comprising administering to a subject a compound selected from the group consisting of leptin and adiponectin. 
     
     
         33 . A non-human transgenic animal having cells that carry a exogenous promoter and a cDNA corresponding to at least the open reading frame of SEQ ID NO: 1 wherein said cells constitutively over express a Enho1 peptide with a sequence as given in SEQ ID NOS:2 and 8-18, and fragments, derivatives, homologs, and analogs thereof of those peptides. 
     
     
         34 . The transgenic animal of  claim 33 , wherein said exogenous promoter is an actin promoter. 
     
     
         35 . The transgenic animal of  claim 33 , wherein said non-human animal is a mouse. 
     
     
         36 . A transformation vector comprising at least the open reading frame of a nucleic acid sequence as recited in SEQ ID NO: 1. 
     
     
         37 . A host cell comprising at least the open reading frame of the nucleic acid sequence as recited in SEQ ID NO: 1. 
     
     
         38 . A nucleic acid construct comprising at least the open reading frame of the nucleic acid sequence as recited in SEQ ID NO: 1, wherein said sequence is operably linked to a promoter that is functional in mammals. 
     
     
         39 . A nucleic acid construct as recited in  claim 38 , wherein said promoter is an actin promoter. 
     
     
         40 . An animal transformed with a nucleic acid construct as recited in  claim 39 . 
     
     
         41 . A purified Enho1 peptide, and fragment, homologs, analogs and derivatives thereof, wherein the peptide has the following characteristics:
 a. it is a substantially homogenous preparation; and   b. it has at least 70% homology to any one of the sequences set forth in SEQ ID NOS: 10-11; wherein   c. the peptide modulates energy metabolism, lipid metabolism, and/or insulin action.   
     
     
         42 . A peptide of  claim 41 , wherein said homology to any one of the sequences set forth in SEQ ID NOS: 10-11 is greater than 85%.