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Title:
MODULATION OF GPR120 ACTIVITY IN ADIPOCYTES/FAT TISSUE
Document Type and Number:
WIPO Patent Application WO/2007/134613
Kind Code:
A1
Abstract:
The present invention relates to a G protein coupled receptor, GPR120 expressed in various mammalian tissue. The present invention further relates to modulation of the activity of these receptors as a tool in diagnosing, treating or preventing diabetes and/or obesity. The present invention also relates to screening of various compounds for targeting GPR120 receptors useful in treating, alleviating, preventing or diagnosing diabetes and/or obesity. The present invention further relates to a pharmaceutical composition useful in diagnosing, treating or preventing diabetes and/or obesity.

Inventors:
VRANG, Niels (Glerupvej 2, Rødovre, DK-2610, DK)
LARSEN, Philip Just (Glerupvej 2, Rødovre, DK-2610, DK)
LARSEN, Leif Kongskov (Glerupvej 2, Rødovre, DK-2610, DK)
Application Number:
DK2007/050060
Publication Date:
November 29, 2007
Filing Date:
May 24, 2007
Export Citation:
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Assignee:
RHEOSCIENCE A/S (Glerupvej 2, Rødovre, DK-2610, DK)
VRANG, Niels (Glerupvej 2, Rødovre, DK-2610, DK)
LARSEN, Philip Just (Glerupvej 2, Rødovre, DK-2610, DK)
LARSEN, Leif Kongskov (Glerupvej 2, Rødovre, DK-2610, DK)
International Classes:
G01N33/74
Attorney, Agent or Firm:
WINTHER, Palle (Zacco Denmark A/S, Hans Bekkevolds Allé 2, Hellerup, DK-2900, DK)
Download PDF:
Claims:

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Patent Claims:

1. A method of screening of GPR120, said method comprising: a) determining the activity of GPR120 polypeptide in the adipose tissue comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, b) determining the activity of GPR120 polypeptide in the adipose tissue in the presence and optionally in the absence of a compound known to be a regulator of a GPR120 polypeptide, c) identifying compounds capable of regulating the activity of a GPR120 polypeptide.

2. A method of detecting a compound/molecule capable of binding to GPR120, said method comprising: a) contacting a sample with GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, b) detecting binding between GPR120 and a compound and/ or molecule capable of binding GPR120.

3. The method as claimed in claim 2, wherein said method is for detecting the compound and / or molecule in adipose tissue.

4. The method as claimed in claim 2, wherein said sample is isolated from adipose tissue. 5. A method as claimed in claim 1 or claim 2, wherein the GPR120 polypeptide is expressed at the surface of the adipose tissue.

6. A method as claimed in any of the preceding claim, wherein the cell is in vitro.

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7. A method as claimed in any of the preceding claims, wherein the step of contacting is in a cell-free system.

8. A method as claimed in claim 1 or claim 2, wherein the polypeptide is coupled to a detectable label. 9. A method of treating obesity and/or diabetes comprising admistering atleast one compound to the patient in an amount sufficient to modulate GPR120 activity in adipose tissue wherein the compound is selected from: a) a polynucleotide sequence encoding a GPR120 polypeptide that is up-regulated in the mammalian adipose tissue, said polynucleotide sequence is selected from a group consisting of SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, a fragment, and variant thereof, b) a polynucleotide sequence encoding a GPR120 polypeptide that is up-regulated in the mammalian adipose tissue, wherein said polypeptide comprises the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, c) a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, and d) a polynucleotide sequence that is antisense to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof; e) a GPR120 specific antibody raised against a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof,

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f) a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID N0:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof 10. A compound that modulates the action of an expression product of

GPR120 gene sequence which is up-regulated in the adipose tissue of a diabetic and/or obese mammal, wherein the GPR120 gene sequence is selected from the group consisting of SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, a fragment, and variant thereof. 1 1. A compound as claimed in claim 10, wherein the gene sequence encodes a polypeptide selected from the group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof.

12. A compound as claimed in claimi O for treating, alleviating, or preventing obesity and/or diabetes.

13. A composition for use as a medicament, said composition comprising at least one compound selected from the group consisting of: a) a polynucleotide encoding the GPR120 polypeptide, said polynucleotide comprising the sequence selected from a group consisting of SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, a fragment, and variant thereof, b) a polynucleotide encoding the GPR120 polypeptide, wherein said polypeptide comprises the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, c) a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof,

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d) a GPR120 specific antibody raised against the GPR120 polypeptide comprising the amino acid sequence according selected from a group consisting of SEQ ID N0:15, SEQ ID N0:17, SEQ ID NO:19, a fragment, and variant thereof, e) a nucleotide sequence that is antisense to a transcript encoding a

GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, and f) a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence according to SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, or a fragment, or variant thereof.

14. A composition as claimed in claim13 for treating, alleviating, or preventing obesity and/or diabetes. 15. Use of a compound for manufacturing a pharmaceutical/medicament for modulating expression of GPR120 in adipose tissues for treating, alleviating, or preventing obesity and/or diabetes, wherein said compound is selected from the group consisting of: a) a polynucleotide sequence encoding a GPR120 polypeptide that is up-regulated in the mammalian adipose tissue, said polynucleotide sequence is selected from a group consisting of SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, a fragment, and variant thereof, b) a polynucleotide sequence encoding a GPR120 polypeptide that is up-regulated in the mammalian adipose tissue, wherein said polypeptide comprises the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof,

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c) a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, d) a polynucleotide sequence that is antisense to a transcript encoding a GPR120 polypeptide selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, e) a GPR120 specific antibody raised against a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, and f) a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof.

16. Use of a compound for diagnosis of obesity and/or diabetes, wherein said compound is selected from the group consisting of: a) a polynucleotide sequence encoding a GPR120 polypeptide that is up-regulated in the mammalian adipose tissue, said polynucleotide sequence is selected from a group consisting of SEQ ID NO:16,

SEQ ID NO:18, SEQ ID NO:20, a fragment, and variant thereof, b) a polynucleotide sequence encoding a GPR120 polypeptide that is up-regulated in the mammalian adipose tissue, wherein said polypeptide comprises the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, c) a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof,

Description:

MODULATION OF GPR120 ACTIVITY IN ADIPOCYTES/FAT TISSUE Field of the Invention

The present invention relates to a G protein coupled receptor GPR120, particularly the present invention relates to modulation of the activity of the receptor.

Background of the invention G-Protein Coupled Receptors:

G-Protein Coupled Receptors (GPCRs) comprise an important class of proteins regulating signal transduction within a cell. GPCRs along with G- Proteins are the components of a modular signalling system that connects the state of intracellular second messengers to extra-cellular inputs.

GPCRs, also known as seven transmembrane, 7TM, receptors, have been characterized as including seven conserved hydrophobic stretches of about 20 to 30 amino acids, connecting at least eight divergent hydrophilic loops. The seven transmembrane regions are designated as TM1 , TM2, TM3, TM4, TM5, TM6, and TM7. TM3 is being implicated with several GPCRs as having a ligand binding site, such as the TM3 aspartat residue. TM5 serines, a TM6 asparagine, and TM6 or TM7 phenylalanines or tyrosines also are implicated in ligand binding. GPCRs upon binding to a ligand transduce a signal within the cell that results in alteration of biological or physiological property of the cell. Most GPCRs have single conserved cysteine residues in each of the first two extracellular loops, which form disulfide bonds that are believed to stabilize functional protein structure. Phosphorylation and lipidation (palmitylation or farnesylation) of cysteine residues can influence signal transduction of some GPCRs.

For some receptors, the ligand binding sites of GPCRs are believed to comprise hydrophilic sockets formed by several GPCR transmembrane domains. The hydrophilic sockets are surrounded by hydrophobic residues of

the GPCRs. The hydrophilic side of each GPCR transmembrane helix is postulated to face inward and form a polar ligand binding site.

Most GPCRs contain potential phosphorylation sites within the third cytoplasmic loop and/or the carboxy terminus. For several GPCRs, such as the beta-adrenergic receptor, phosphorylation by protein kinase A and/or specific receptor kinases mediates receptor desensitization.

The family of G-protein coupled receptors (GPCRs) includes receptors for hormones, neurotransmitters, growth factors, and viruses. Specific examples of GPCRs include receptors for such diverse agents as dopamine, calcitonin, adrenergic hormones, endotheline, cAMP, adenosine, acetylcholine, serotonin, histamine, thrombin, quinine, follicle stimulating hormone, opsins, endothelial differentiation gene-1 , rhodopsins, odorants, cytomegalovirus, G-proteins themselves, effector proteins such as phospholipase C, adenyl cyclase, and phosphodiesterase, and actuator proteins such as protein kinase A and protein kinase C. GPCRs represent a very large family of proteins that control many physiological processes and are the targets of many effective drugs.

GPCRs are coupled inside the cell by heterotrimeric G-proteins to various intracellular enzymes, ion channels, and transporters. Different G- protein α-subunits preferentially stimulate particular effectors to modulate various biological functions in a cell. Phosphorylation of cytoplasmic residues of GPCRs is an important mechanism for the regulation of some GPCRs. For example, in one form of signal transduction, the effect of hormone binding is the activation of the enzyme, adenylate cyclase, inside the cell. Enzyme activation by hormones is dependent on the presence of the nucleotide GTP. GTP also influences hormone binding. A G-protein connects the hormone receptor to adenylate cyclase. G-protein exchanges GTP for bound GDP when activated by a hormone receptor. The GTP-carrying form then binds to activated adenylate cyclase. Hydrolysis of GTP to GDP, catalyzed by the G- protein itself, returns the G-protein to its basal, inactive form. Thus, the G-

protein serves a dual role, as an intermediate that relays the signal from receptor to effector, and as a clock that controls the duration of the signal.

Over the past 15 years, nearly 350 therapeutic agents targeting 7TM receptors have been successfully introduced into the market. This indicates that these receptors have an established, proven history as therapeutic targets. Clearly, there is a need for identification and characterization of further receptors which can play a role in preventing, ameliorating, or correcting metabolic dysfunctions or diseases including, but not limited to, adiposity, insulin resistance, dyslipidemia, and arterial hypertension. Combinations of these symptoms and diseases constitute what is defined as the dysmetabolic syndrome by National Cholesterol Education Program (NCEP)'s Adult Treatment Panel III (ATP-III). Patients suffering dysmetabolic syndrome as defined by ATP-III criteria have significantly elevated risk of cardiovascular disease (myocardial infarction, ischemic heart disease, atherosclerosis, and stroke) and diabetes mellitus. Medical complications accompanying monosymptomatic obesity such as osteoarthritis, cancers, liver and gall bladder diseases, and certain sleep disorders are also in need of substantially improved therapeutic means.

As it appears to be extremely difficult to change life style and loose weight there is a massive need for an effective "obesity drug". Any effective obesity drugs having little or no side effects do not exist in the market. Much development has been focused on identifying targets for developing drugs acting on the nerve system.

GPR120: The G-protein coupled receptor GPR120 was recently identified as an additional member of the large Rhodopsin family of GPCR ' s (Fredriksson et al., 2003). GPR120 is expressed in the distal gut and it was recently demonstrated that GPR120 is found on the L-cells of the intestine (Hirasawa et al., 2005). In the same study an internalization assay was used to screen a

library containing 1000 different chemical compounds and it was found that long-chain free fatty acids, with linolenic acid as the most potent, was a ligand for this receptor (Hirasawa et al., 2005).

US6395877B1 and WO2005051373A1 both disclose the transcript for GPR120 (14273) and it is suggested that this protein is linked to cardiovascular diseases and congestive heart failure (US6395877B1 ) or diabetes or dyslipidaemia (WO2005051373A1 ).

GPR120 Nucleotide sequences:

The nucleotide sequence of Rat GPR120 (Accession number AB207868) is given in SEQ ID NO: 16. The predicted amino acid sequence of Rat GPR120 is provided in SEQ ID NO: 15. Mouse GPR120 (Accession number BC053698) nucleotide sequence is as shown in SEQ ID NO: 18, corresponding amino acid sequence of Mouse GPR120 is given in SEQ ID

NO: 17. Human GPR120 having accession number BC101175 is provided in SEQ ID NO: 20 (nucleotide sequence) and SEQ ID NO: 19 (amino acid sequence), respectively.

Diabetes

Diabetes is characterized by a decreased ability to regulate blood glucose concentrations. Diabetes can be grouped in two major classes: diabetes type I, wherein the insulin-producing cells are partly or fully destroyed by an auto-immune reaction and diabetes type II, wherein the insulin-producing cells are intact or essentially intact and wherein the decreased ability to regulate blood glucose is thought to be caused by decreased insulin-sensitivity. Diabetes type Il is often accompanying obesity. In the present invention, the term diabetes is thus equivalent to diabetes type II. A number of pathological conditions furthermore accompany obesity and/or diabetes such as hypertension, and cardiovascular diseases.

It thus follows that there is a long felt need in the art for novel obesity drugs and drug targets. There is also a need in the art for identifying drugs

and drug targets that may affect the fat tissue directly rather than affecting nerve tissues. Such drugs and drug targets will furthermore be useful in connection with diabetes treatment as well as treatment of conditions that accompany obesity and/or diabetes. SUMMARY OF THE INVENTION

The present invention relates to a G protein coupled receptor, GPR120. The invention further relates to screening the expression of GPR120 in various mammalian tissues and its high expression in adipose tissue. Further, the invention discloses the up-regulation of GPR120 in differentiating adipocytes. GPR120 is up-regulated in response to overfeeding /obesity. The present invention also discloses the substantially higher expression of GPR120 in the adipose tissue.

These findings together form basis for the therapeutic approaches in connection with obesity and/or diabetes for modulating GPR120 activity as well as using GPR120 as a tool for identifying obesity/diabetes drugs.

In one aspect, the invention provides a method of screening of GPR120, said method comprising: a) determining the activity of GPR120 polypeptide in the adipose tissue comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, b) determining the activity of GPR120 polypeptide in the adipose tissue in the presence and optionally in the absence of a compound known to be a regulator of a GPR120 polypeptide, c) identifying compounds capable of regulating the activity of a GPR120 polypeptide

In another aspect, the invention provides a method of detecting a compound/molecule capable of binding to GPR120, said method comprising:

a) contacting a sample with GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO: 15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, b) detecting binding between GPR120 and a compound and/ or molecule capable of binding GPR120.

Yet another aspect of the present invention relates to a method of treating obesity and/or diabetes, wherein at least one compound selected from: a) a polynucleotide sequence encoding a GPR120 polypeptide that is up- regulated in the mammalian adipose tissue, said polynucleotide sequence is selected from a group consisting of SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, a fragment, and variant thereof, b) a polynucleotide sequence encoding a GPR120 polypeptide that is up- regulated in the mammalian adipose tissue, wherein said polypeptide comprises the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, c) a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO: 19 , a fragment, and variant thereof, and d) a polynucleotide sequence that is antisense to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, e) a GPR120 specific antibody raised against a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, and

f) a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof is administered to the patient in an amount sufficient to modulate

GPR120 activity in adipose tissue.

Yet another aspect of the invention relates to a compound that modulates the action of an expression product of GPR120 gene sequence which is up-regulated in the adipose tissue of a diabetic and/or obese mammal, wherein the GPR120 gene sequence is selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, a fragment, and variant thereof.

Still another aspect of the invention is to provide a composition for use as a medicament, said composition comprising at least one compound selected from the group consisting of: a) a polynucleotide encoding the GPR120 polypeptide, said polynucleotide comprising the sequence selected from a group consisting of SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, a fragment, and variant thereof, b) a polynucleotide encoding the GPR120 polypeptide, wherein said polypeptide comprises the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, c) a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID

NO: 19, a fragment, and variant thereof, d) a GPR120 specific antibody raised against the GPR120 polypeptide comprising the amino acid sequence according selected from a group

consisting of SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, a fragment, and variant thereof, e) a nucleotide sequence that is antisense to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID

NO: 19, a fragment, and variant thereof, and f) a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence according to SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, or a fragment, or variant thereof.

Another embodiment of the invention relates to use of one of the compounds according to the invention for manufacturing a pharmaceutical for modulating expression of GPR120 in fat or adipose tissues for treating, alleviating, or preventing obesity and/or diabetes. Still another embodiment of the invention relates to use of a compound for manufacturing a pharmaceutical for modulating expression of GPR120 in fat or adipose tissues for treating, alleviating, or preventing obesity and/or diabetes, wherein said compound is selected from the group consisting of; a) a polynucleotide sequence encoding a GPR120 polypeptide that is up-regulated in the mammalian adipose tissue, said polynucleotide sequence is selected from a group consisting of SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, a fragment, and variant thereof, b) a polynucleotide sequence encoding a GPR120 polypeptide that is up-regulated in the mammalian adipose tissue, wherein said polypeptide comprises the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof,

c) a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, d) a polynucleotide sequence that is antisense to a transcript encoding a GPR120 polypeptide selected from a group consisting of SEQ ID

NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, e) a GPR120 specific antibody raised against a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, and f) a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof.

Another embodiment of the invention further discloses the use of a compound for diagnosis of obesity and/or diabetes, wherein said compound is selected from the group consisting of: a) a polynucleotide sequence encoding a GPR120 polypeptide that is up- regulated in the mammalian adipose tissue, said polynucleotide sequence is selected from a group consisting of SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, a fragment, and variant thereof, b) a polynucleotide sequence encoding a GPR120 polypeptide that is up- regulated in the mammalian adipose tissue, wherein said polypeptide comprises the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof,

c) a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO: 19, a fragment, and variant thereof, d) a polynucleotide sequence that is antisense to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, e) a GPR120 specific antibody raised against a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, and f) a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof.

Still another embodiment of the invention discloses the use of a compound that modulates the action of an expression product of GPR120 gene sequence which is up-regulated in the adipose tissue of a diabetic and/or obese mammal, for manufacturing a pharmaceutical for treating, alleviating, or preventing obesity and/or diabetes.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure: 1 shows results from a quantitative multiplex RT-PCR experiment. RNA was isolated from various rat and human tissues and RT-PCR was performed with specific primers to TATA box binding protein (TBP) and GPR120. The signals were quantified and the GPR120 signal was normalized to the TBP signal.

Figure: 2 shows a multiplex quantitative RT-PCR experiment. RNA was isolated from 3T3-L1 cells at different days during the differentiation to adipocytes and RT-PCR was performed with two sets of primers specific for

TATA box binding protein (TBP) and GPR120 or for TBP and PPARgamma2. The signals were quantified and the GPR120 and PPARgamma2 signals, respectively, were normalized to the TBP signals.

Figure: 3 shows GPR120 shows the physiological regulation of a GPR120 receptor polynucleotide in inguinal fat tissue from Zucker obese (fa/fa) and

Zucker lean (Fa/?) rats exposed to different feeding regimes. One group of rats of each genotype was given chow ad libitum ("Ad Lib") and one group of rats of each genotype was fasted for 48 hours ("48h Fast) before the termination. Figure 3 shows results from multiplex quantitative RT-PCR experiment. Inguinal white adipose tissue was isolated from the rats and the expression of GPR120 and TBP was determined by Multiplex quantitative

RT-PCR. Error bars show Standard Error of the Mean (SEM).

DETAILED DESCRIPTION OF THE INVENTION Definitions An "oligonucleotide" is a strand of nucleotide residues which has a sufficient number of bases to be used as an oligomer, amplimer or probe in a polymerase chain reaction (PCR). Oligonucleotides are prepared from genomic or cDNA sequence and are used to amplify, reveal, or confirm the presence of a similar DNA or RNA in a particular cell or tissue. Oligonucleotides or oligomers comprise portions of a DNA sequence having at least about 10 nucleotides and as many as about 35 nucleotides, preferably about 25 nucleotides.

"Reporter molecules" are radionuclides, enzymes, fluorescent, chemiluminescent, or chromogenic agents which associate with a particular nucleotide or amino acid sequence, thereby establishing the presence of a certain sequence, or allowing for the quantification of a certain sequence.

"Chimeric" molecules may be constructed by introducing all or part of the nucleotide sequence of this invention into a vector containing additional nucleic acid sequence which might be expected to change any one or

several of the following GPR120 characteristics: cellular location, distribution, ligand-binding affinities, interchain affinities, degradation/turnover rate, signalling, etc.

"Active", with respect to a GPR120 polypeptide, refers to those forms, fragments, or domains of a GPR120 polypeptide which retain the biological and/or antigenic activity of a GPR120 polypeptide.

"Naturally occurring GPR120 polypeptide" refers to a polypeptide produced by cells which have not been genetically engineered and specifically contemplates various polypeptides arising from post-translational modifications of the polypeptide including but not limited to acetylation, carboxylation, glycosylation, phosphorylation, lipidation and acylation.

"Conservative amino acid substitutions" result from replacing one amino acid with another having similar structural and/or chemical properties, such as the replacement of a leucine with an isoleucine or valine, an aspartate with a glutamate, or a threonine with a serine.

"Insertions" or "deletions" are typically in the range of about 1 to 5 amino acids. The variation allowed may be experimentally determined by producing the peptide synthetically while systematically making insertions, deletions, or substitutions of nucleotides in the sequence using recombinant DNA techniques.

"Inhibitor" is any substance which retards or prevents a chemical or physiological reaction or response. Common inhibitors include but are not limited to antisense molecules, antibodies, and antagonists.

"Standard expression" is a quantitative or qualitative measurement for comparison.

It is based on a statistically appropriate number of normal samples and is created to use as a basis of comparison when performing diagnostic assays, running clinical trials, or following patient treatment profiles.

"Animal" as used herein may be defined to include human, domestic (e. g., cats, dogs, etc.), agricultural (e. g., cows, horses, sheep, etc.) or test species (e. g., mouse, rat, rabbit, etc.).

A sample derived from fat tissue denotes a sample comprising a number of components from fat tissue, e.g. transcripts expressed in fat tissue and/or proteins expressed in fat tissue. The proteins may of course have been subject to post-translational modification such as signal peptide cleavage, glycosylation, acylation, etc.

A regulator of a GPR120 polypeptide denotes any compound which is known to have the ability of modulating the activity of GPR120. Examples of GPR120 regulators comprise e.g. GPR120 specific antibodies.

As used herein, the terms "specific binding" or "specifically binding" refer to the interaction between a protein or peptide and an agonist, an antibody, or an antagonist. The present invention relates to modulation of the activity of a G protein coupled receptor, GPR120 expressed in various mammalian tissue. The present invention also relates to screening of various compounds for targeting GPR120 receptors useful in diagnosing, treating, alleviating or preventing diabetes and/or obesity. The present invention further relates to a pharmaceutical composition useful in diagnosing, treating, alleviating or preventing diabetes and/or obesity.

A nucleotide sequence encoding a GPR120 polypeptide having a sequence according to SEQ ID NO: 15, accession number AB207868; or SEQ ID NO: 17, accession number BC053698; or SEQ ID NO: 19, accession number BC101175: SEQ ID NO: 16, SEQ ID NO: 18 and SEQ ID NO: 20 are examples of nucleotide sequences encoding GPR120. The skilled person knows that the nucleotide sequence of SEQ ID NO: 16 or SEQ ID NO: 18 or SEQ ID NO: 20 can be varied to a large extent, due to the alternative codon usage, while still encoding a polypeptide having an amino acid sequence as

given in SEQ ID NO: 15 or SEQ ID NO: 17 or SEQ ID NO: 19. A nucleotide sequence encoding GPR120 may furthermore be a fragment of a sequence as given in SEQ ID NO:16 or SEQ ID NO:18 or SEQ ID NO:20 or a variant thereof with a length of at least 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 300, 400, 500, 600, 700, 800, 900, 1000 bases, and even up to1419 bases. It thus follows that a nucleotide sequence encoding GPR120 encodes at least about 10, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 150, 200, 250, 300 amino acids and even up to about 361amino acids of the sequence identified as SEQ ID NO: 15 or SEQ ID NO: 17 or SEQ ID NO: 19. A fragment of a nucleotide sequence encoding GPR120 furthermore preferably comprises at least the fraction of the molecule that encodes the ligand-binding domain of the molecule.

A variant of SEQ ID NO:16 or SEQ ID NO:18 or SEQ ID NO:20 denotes all DNA sequences encoding a GPR120 polypeptide having an amino acid sequence as given in SEQ ID NO: 15 or SEQ ID NO: 17or SEQ ID NO: 19 or a fragment or variant thereof. A variant should furthermore be understood as a DNA sequence with at least 80%, preferably at least 85%, more preferably at least 90%, most preferably at least 95%, and even most preferably at least 98% identity with SEQ ID NO: 16 or SEQ ID NO: 18 or SEQ ID NO:20. It is furthermore preferred that the ligand-binding domain of the molecule is the most highly conserved part of the molecule, preferably at least 95%, more preferably at least 98%, and even most preferably at least 99% conserved compared with SEQ ID NO:16 or SEQ ID NO:18 or SEQ ID NO:20. A variant of GPR120 denotes a polypeptide with an amino acid identity of at least 80%, preferably, at least 85%, more preferably at least 90%, even more preferably at least 95%, and most preferably at least 98% identity with the sequence as given in SEQ ID NO: 15 or SEQ ID NO: 17 or SEQ ID NO: 19 or a fragment thereof. It is furthermore preferred that the ligand-binding encoding domain of the molecule is the most highly conserved part of the

molecule, preferably at least 95%, more preferably at least 98%, and even most preferably at least 99% conserved compared with SEQ ID NO: 15 or SEQ ID NO: 17 or SEQ ID NO: 19. Conservative amino acid substitutions are preferred, especially in the ligand binding domains. In one embodiment the variant has substantially the same biological function as the molecule from which it is derived.

A GPR120 specific antibody denotes an antibody or a pool of antibodies (e.g. monoclonal or polyclonal antibodies) that has been raised against a GPR120 polypeptide according to SEQ ID NO: 15 or SEQ ID NO: 17 or SEQ ID NO: 19 or a fragment thereof and/or a variant thereof. Preparation of antibodies is well known in the art, see (Howard and Bethell, 2000) for an example.

A nucleotide sequence that is antisense to a GPR120 sequence denotes a sequence that has the capability of base pairing specifically with a GPR120 transcript encoding a GPR120 polypeptide having an amino acid sequence according to SEQ ID NO: 15 or SEQ ID NO: 17 or SEQ ID NO: 19 or a variant, or a fragment thereof. The antisense sequence may be in the form of a single stranded DNA, RNA, PNA, or LNA molecule. The antisense sequence has a length of at least about 15, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 125, 150, 175, 200, 300, 400, 500, 600, or 700 bases. Upon base pairing of the antisense sequence with the GPR120 transcript within a cell, expression of GPR120 is significantly down regulated in said cell due to rapid degradation of double stranded mRNA/antisense complex.

Small interfering RNAs (siRNAs) were recently discovered in plants and in animals. Since their discovery in the nematode Caenorhabditis elegans (Fire et al., 1998), these 21-23-nucleotide double-stranded RNAs bearing 3 ' overhanging ends have shown a tremendous potential for the silencing of genes in experimental as well as in therapeutical settings. Vast amounts of literature has been published on siRNA structure and mechanism of action as well as on the use of siRNA in gene silencing, see the following

references for recent reviews: (Alisky and Davidson, 2004; Gilmore et al., 2004; Karagiannis and El-Osta, 2004; Wadhwa et al., 2004; Zhang and Hua, 2004).

A sample derived from fat tissue denotes a sample comprising a number of components from fat tissue, e.g. transcripts expressed in fat tissue and/or proteins expressed in fat tissue. The proteins may of course have been subject to post-translational modification such as signal peptide cleavage, glycosylation, acylation, etc.

A regulator of a GPR120 polypeptide denotes any compound which is known to have the ability of modulating the activity of GPR120. Examples of GPR120 regulators comprise e.g. GPR120 specific antibodies.

As used herein, the terms "specific binding" or "specifically binding" refer to the interaction between a protein or peptide and an agonist, an antibody, or an antagonist. In one embodiment the present invention provides a method of screening the expression of GPR120 in adipose tissue, wherein the activity of GPR120 polypeptide in the adipose tissue was determined in the presence and optionally in the absence of a compound known to be a regulator of a GPR120 polypeptide. In another embodiment the invention teaches differential expression of

GPR120 in response to adipocyte differentiation of 3T3-L1 cell (a mouse fibroblast/preadipocyte cell line).

Another embodiment of the present invention discloses the substantially higher expression of GPR120 polypeptide in the adipose tissue of diabetic and obese rat.

In another aspect, the present invention relates to a method of screening a GPR120 polypeptide for an interaction partner said method comprising of:

a) contacting a GPR120 polypeptide comprising the amino acid sequence according to SEQ ID NO 15 or SEQ ID NO 17 or SEQ ID NO 19, or a fragment, or variant thereof with a compound or an array of test compounds; and b) detecting binding between GPR120 and a compound capable of binding GPR120.

In a preferred embodiment, said method is a method for screening fat or adipose tissue for an interaction partner of a GPR120 polypeptide, said method comprising of: a) contacting a sample derived from fat tissue with a GPR120 polypeptide comprising the amino acid sequence according to SEQ ID NO 15 or SEQ ID NO 17 or SEQ ID NO 19, or a fragment, or variant thereof, and b) detecting binding between GPR120 and a compound capable of binding GPR120.

Another aspect of the present invention relates to a method of screening for therapeutic agents useful in the treatment of obesity and/or diabetes and/or cardiovascular diseases, said method comprising the following steps: a) determining the activity of a GPR120 polypeptide comprising the amino acid sequence according to SEQ ID NO 15 or SEQ ID NO 17 or SEQ ID NO 19, or a fragment or variant thereof, in the presence and optionally also in the absence of a test compound, b) determining the activity of a GPR120 polypeptide in the presence and optionally also in the absence of a compound known to be a regulator of a GPR120 polypeptide. c) identifying compounds capable of modulating GPR120 activity.

In a preferred embodiment according to the present invention, the GPR120 polypeptide is expressed at the surface of a cell. The cell is preferably grown in vitro.

In another preferred embodiment, the test compounds and the GPR120 polypeptide are contacted in a cell-free system.

In another preferred embodiment, the polypeptide is coupled to a detectable label.

In another preferred embodiment, the test compound is coupled to a detectable label. In another preferred embodiment, the test compound or test sample displaces a ligand which is first bound to the polypeptide.

In another preferred embodiment, the polypeptide is attached to a solid support.

Yet another embodiment provides a method of treating obesity and/or diabetes, wherein at least one compound selected from: a) a polynucleotide sequence encoding a GPR120 polypeptide that is up- regulated in the mammalian adipose tissue, said polynucleotide sequence is selected from a group consisting of SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, a fragment, and variant thereof, b) a polynucleotide sequence encoding a GPR120 polypeptide that is up- regulated in the mammalian adipose tissue, wherein said polypeptide comprises the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, c) a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO: 19, a fragment, and variant thereof, and

d) a polynucleotide sequence that is antisense to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof; e) a GPR120 specific antibody raised against a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, f) a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof is administered to the patient in an amount sufficient to modulate GPR120 activity in adipose tissue. In one embodiment the present invention provides a method of treating obesity and/or diabetes, wherein at least one compound selected from: a) a polynucleotide sequence that is antisense to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID

NO:19, a fragment, and variant thereof; b) a GPR120 specific antibody raised against a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, c) a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof

is administered to the patient in an amount sufficient to modulate GPR120 activity in adipose tissue.

Another aspect of the invention provides use of a compound for manufacturing a pharmaceutical for modulating expression of GPR120 in adipose tissues for treating, alleviating, or preventing obesity and/or diabetes, wherein said compound is selected from the group consisting of: a) a polynucleotide sequence encoding a GPR120 polypeptide that is up-regulated in the mammalian adipose tissue, said polynucleotide sequence is selected from a group consisting of SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, a fragment, and variant thereof, b) a polynucleotide sequence encoding a GPR120 polypeptide that is up-regulated in the mammalian adipose tissue, wherein said polypeptide comprises the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, c) a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, d) a polynucleotide sequence that is antisense to a transcript encoding a GPR120 polypeptide selected from a group consisting of SEQ ID

NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, e) a GPR120 specific antibody raised against a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, and f) a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected

from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO: 19, a fragment, and variant thereof.

Yet another aspect of the present invention provides use of a compound for diagnosis of obesity and/or diabetes, wherein said compound is selected from the group consisting of: a) a polynucleotide sequence encoding a GPR120 polypeptide that is up-regulated in the mammalian adipose tissue, said polynucleotide sequence is selected from a group consisting of SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, a fragment, and variant thereof, b) a polynucleotide sequence encoding a GPR120 polypeptide that is up-regulated in the mammalian adipose tissue, wherein said polypeptide comprises the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, c) a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, d) a polynucleotide sequence that is antisense to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17,

SEQ ID NO:19, a fragment, and variant thereof, e) a GPR120 specific antibody raised against a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, and f) a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO: 19, a fragment, and variant thereof.

Still another aspect of the invention relates to use of a compound that modulates the action of an expression product of GPR120 gene sequence which is up-regulated in the adipose tissue of a diabetic and/or obese mammal, for manufacturing a pharmaceutical for treating, alleviating, or preventing obesity and/or diabetes.

The present invention also relates to use of a compound that modulates the action of an expression product of GPR120 gene sequence which is up-regulated in the adipose tissue of a diabetic and/or obese mammal, for diagnosis of obesity and/or diabetes. Further the present invention relates to a compound that modulates the action of an expression product of GPR120 gene sequence which is up- regulated in the adipose tissue of a diabetic and/or obese mammal, wherein the GPR 120 gene sequence is selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, a fragment, and variant thereof. In another embodiment the invention discloses a composition for use as a medicament, said composition comprising at least one compound selected from the group consisting of: a) a polynucleotide encoding the GPR120 polypeptide, said polynucleotide comprising the sequence selected from a group consisting of SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, a fragment, and variant thereof, b) a polynucleotide encoding the GPR120 polypeptide, wherein said polypeptide comprises the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, c) a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof,

d) a GPR120 specific antibody raised against the GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, e) a nucleotide sequence that is antisense to a transcript encoding a

GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, and f) a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence according to SEQ ID

NO:15, SEQ ID NO:17, SEQ ID NO:19, or a fragment, or variant thereof.

In one embodiment the present invention thus relates to a composition for use as a medicament, said composition comprising at least one compound selected from the group consisting of: a) a GPR120 specific antibody raised against the GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, b) a nucleotide sequence that is antisense to a transcript encoding a

GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, and c) a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence according to SEQ ID

NO:15, SEQ ID NO:17, SEQ ID NO:19, or a fragment, or variant thereof.

In another embodiment the composition for use as a medicament comprises a polynucleotide encoding a GPR120 polypeptide, said

polynucleotide comprising the sequence of SEQ ID NO 16, SEQ ID NO 18, or SEQ ID NO 20, a fragment, or variant thereof.

In another embodiment the composition comprises a polynucleotide encoding a GPR120 polypeptide, wherein said polypeptide comprises the amino acid sequence according to SEQ ID NO 15, or SEQ ID NO 17, or SEQ ID NO 19, or a fragment, or variant thereof.

In another embodiment the composition comprises a GPR120 polypeptide comprising the amino acid sequence according to SEQ ID NO 15 or SEQ ID NO 17 or SEQ ID NO 19, or a fragment, or variant thereof In another embodiment the composition comprises a GPR120 specific antibody raised against a GPR120 polypeptide comprising the amino acid sequence according to SEQ ID NO 15 or SEQ ID NO 17 or SEQ ID NO 19, or a fragment, or variant thereof.

In another embodiment the composition comprises a nucleotide sequence that is antisense to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence according to SEQ ID NO 15 or SEQ ID NO 17 or SEQ ID NO 19, or a fragment, or variant thereof

In another embodiment the composition comprises a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence according to SEQ ID NO 15 or SEQ ID NO 17 or SEQ ID NO 19, or a fragment, or variant thereof.

In an embodiment the invention also discloses the use of a compound that modulates the action of an expression product of GPR120 gene sequence which is up-regulated in the adipose tissue of a diabetic and/or obese mammal, for diagnosis of obesity and/or diabetes.

Expression profiling of GPR120 polypeptide (SEQ ID NO: 15, and SEQ ID NO: 19) encoded by polynucleotide as shown in SEQ ID NO: 16 and SEQ ID NO: 20 in various tissue of rat and human was carried out. For this total RNA was extracted by using the methods well known in the art.

For the rat expression profile, fresh tissue was sampled from Sprague

Dawley rats from the following anatomically defined areas: Cortex, prefrontral cortex, brain, striatum, colliculus superior, hippocampus, amygdale, cerebellum, thalamus, hypothalamus, raphe, nucleus tractus solitarius (NTS), Brain stem, medulla spinalis, epididymal white adipose tissue, perirenal white adipose tissue, mesenteric white adipose tissue, subcutaneneous white adipose tissue, inguinal white adipose tissue, interscapular brown adipose tissue, antrum of stomach, fundus of stomach, corpus of stomach, duodenum, ileum, jejunum, colon, thymus, adrenal, pancreas, pituitary, tongue, muscle, heart, kidney, liver, spleen, and lung.

For the human expression profile, following tissues were used: Cerebral cortex, parietal lobe, total brain, pons, olfactory, medulla oblongata, temporal lobe, cerebellum (left), occipital lobe, frontal lobe, diencephalon, hypothalamus, thalamus, hippocampus, spinal cord, adipose, breast, stomach, pylorus of stomach, fundus of stomach, cardia of stomach, corpus of stomach, duodenum, ileum, jejunum, colon, colon from type 2 diabetic patient, thymus, adrenal, pancreas, prostate, uterus, ovary, skeletal muscle, heart, peripheral blood leukocyte, bladder, placenta, kidney, liver, fetal liver, spleen, lung. It was found that GPR 120 gene is expressed in the intestine in accordance with the results from Hirasawa et al (Hirasawa et al., 2005) in relatively high levels, but surprisingly it was found that the gene also has high expression in adipose tissue in both rats and humans (Figure 1 ). The detailed procedure of expression analysis of GPR120 is provided in Example 1. In another embodiment the invention discloses the up-regulation of

GPR120 polypeptide (SEQ ID NO: 17) encoded by polynucleotide sequence (SEQ ID NO: 18) in differentiating adipocytes of 3T3-L1 cell (a mouse fibroblast/preadipocyte cell line). For this, RNA was isolated from differentiating adipocytes and multiplex PCR was performed. The adipocyte marker PPARγ2 is upregulated during the adipocyte differentiation, showing

that the adipocytes are indeed differentiating, and, importantly, that the expression of GPR120 is increased during the differentiation to adipocytes (Figure 2). Detail procedure is provided in Example 2.

Expression of GPR120 in adipose tissue of lean rat (Fa/?) and obese and/or diabetic rat (Zucker fa/fa) was analyzed and it was found that the expression of GPR120 is substantially higher in the adipose tissue from the

Zucker fa/fa rats than in the adipose tissue from the Fa/? rats, probably because the adipose tissue in the Zucker fa/fa rats is more differentiated than the adipose tissue in the Fa/? rats (Figure 3). Details are provided in Example 3.

EXAMPLES

The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and the description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as there invention nor are they intended to represent that the experiments below are all and only experiments performed.

Example 1

Expression profiling in rat and human tissues

The procedure is based on method described by Jensen et al. (Jensen et al., 1996). Expression profiling for rat GPR120 polypeptide (SEQ ID NO: 15) encoded by polynucleotide as shown in SEQ ID NO: 16 was carried out. For this fresh tissue was sampled from Sprague Dawley rats from the following anatomically defined areas: Cortex, prefrontral cortex, brain, striatum, colliculus superior, hippocampus, amygdale, cerebellum, thalamus, hypothalamus, raphe, nucleus tractus solitarius (NTS), Brain stem, medulla spinalis, epididymal white adipose tissue, perirenal white adipose tissue, mesenteric white adipose tissue, subcutaneneous white adipose tissue, inguinal white adipose tissue, interscapular brown adipose tissue, antrum of stomach, fundus of stomach, corpus of stomach, duodenum, ileum, jejunum,

colon, thymus, adrenal, pancreas, pituitary, tongue, muscle, heart, kidney, liver, spleen, and lung.

Total RNA was extracted from the tissue samples using TRI reagent, following the manufacturer's instructions. For the expression profiling of human GPR120 polypeptide (SEQ ID

NO: 19) encoded by polynucleotide sequence as shown in SEQ ID NO: 20, total RNA from the following tissues was purchased from Biochain Inc.: Cerebral cortex, parietal lobe, total brain, pons, olfactory, medulla oblongata, temporal lobe, cerebellum (left), occipital lobe, frontal lobe, diencephalon, hypothalamus, thalamus, hippocampus, spinal cord, adipose, breast, stomach, pylorus of stomach, fundus of stomach, cardia of stomach, corpus of stomach, duodenum, ileum, jejunum, colon, colon from type 2 diabetic patient, thymus, adrenal, pancreas, prostate, uterus, ovary, skeletal muscle, heart, peripheral blood leukocyte, bladder, placenta, kidney, liver, fetal liver, spleen, lung.

First-strand cDNA was prepared using 1 μg total RNA, the Superscript Il RT kit, and random hexamer primers. The cDNA was diluted 1 :6 in distilled water. A PCR mixture was prepared. For 13.5 μl, 1.35 μl 1Ox polymerase buffer with MgCI 2 , 0.20 μl dNTP (4 mM, 2 mM dCTP), 0,25 μl of each primer (10 mM), 0.125 μl Taq polymerase, 0.0625 μl 33P-α-dCTP (10 mCi/ml), 1.5 μl cDNA solution, and finally distilled water to 13.5 μl was used .

Two primer sets were included in each reaction, one set specific for GPR120, the second set specific for TBP:

Rat: GPR120 (product size 208 bp)

5'- GACCAGGAAATTCCGATTTG-3' SEQ ID NO: 1

5'- CTGGTGGCTCTCGGAGTATG-3' SEQ ID NO: 2

TBP (product size 186 bp)

5'- ACCCTTCACCAATGACTCCTATG-3' SEQ ID NO: 3 δ'-TGACTGCAGCAAATCGCTTGG-S' SEQ ID NO: 4

Human: GPR120 (product size 210 bp)

5'- CGCTCATCTGGGGCTATTC-3' SEQ ID NO: 5

5'- TTTTGGAGTAACTGATCACAATGAC -3' SEQ ID NO: 6

TBP (product size 187 bp)

5'- TGGCTCTCATGTACCCTTGC-3' SEQ ID NO: 7 5'- TGCACAAATAATGCCCCTTC-3' SEQ ID NO: 8

All samples were subjected to 25 cycles of amplification by PCR: PCR conditions comprises: an initial denaturation at 94 0 C for 2 min, 25 cycles of denaturation at 94 0 C for 30 sec, annealing at 55 0 C for 30 sec and extension at 72 0 C for 30 sec, and final extension at 72 0 C for 5 min. The number of cycles was chosen in the range where the limiting factor for the amount of product is the amount of input template cDNA. The final PCR reactions were mixed with 98% formamide denaturing loading buffer and loaded in duplicate and separated on a 6% (w/v) polyacrylamide gel, containing 7M urea. The gel was subsequently dried, exposed to a phosphorimager screen, and the resulting image was analyzed. Finally, the GPR120 expression was normalized to the TBP expression.

The analysis showed (Figure 1) in accordance with the results from (Hirasawa et al., 2005) that the GPR120 gene is expressed in the intestine in relatively high levels, but surprisingly, that the gene also has a high expression in adipose tissue in both rats and humans.

Example 2

Differential expression of GPR120 in response to adipocyte differentiation

3T3-L1 cells (mouse fibroblast/preadipocyte cell line) were grown to confluence and induced to differentiate as described by Student et al.

(Student et al., 1980). Briefly, preadipocytes were maintained in Dulbecco's modified Eagle's medium (DMEM) with 10 % calf serum and 2 days after reaching confluence (day 0), differentiation was induced by culturing the cells in DMEM with 10% fetal calf serum (FCS), 167 nM insulin (INS), 0.5 mM methylisobutylxanthine (MIX), and 0.25 μM dexamethasone (DEX) for two days, DMEM with FCS and INS for two days and DMEM with FCS until RNA was isolated.

RNA was isolated on different days during the differentiation process and reverse transcribed, followed by multiplex PCR performed essentially as described above (Example 1 ) except that the mouse-specific GPR120 primers SEQ ID NO: 9 and SEQ ID NO: 10 (product length 214 bp) were used with the TBP primers (SEQ ID NO: 11 and SEQ ID NO: 12) (product length 186 bp).

5'- CGCATAGGAGAAATCTCATGG -3' SEQ ID NO: 9 5'- AAACCATGAGCAGGAAGAGC-S' SEQ ID NO: 10

5'- ACCCTTCACCAATGACTCCTATG-3' SEQ ID NO: 11 δ'-TGACTGCAGCAAATCGCTTGG-S' SEQ ID NO: 12

In parallel, separate multiplex PCR reactions were performed with primers (SEQ ID NO: 13 and SEQ ID NO: 14) specific for the adipocyte differentiation marker PPARγ2, again using TBP as a reference. δ'-AGTGTGAATTACAGCAAATCTC-S' SEQ ID NO: 13 δ'-ATGGTAATTTCTTGTGAAGTGC-S', SEQ ID NO: 14

The adipocyte marker PPARγ2 is upregulated during the adipocyte differentiation, showing that the adipocytes are indeed differentiating, and, importantly, that the expression of GPR120 is increased during the differentiation to adipocytes (Figure 2). It is presently not known, whether stimulation or inhibition of the

GPR120 receptor activity can influence the differentiation process, but it is believed that stimulation or inhibition of the GPR120 receptor can affect preadipocyte and/or adipocyte function (e.g. insulin sensitivity) as well as the differentiation process leading from preadipocytes to adipocytes. Example 3

The adipose expression of GPR120 is affected in Zucker fa/fa rats

Sixteen male Zucker fa/fa rats and sixteen male Zucker lean FA/? rats of 8 weeks old were obtained from Charles River (Charles River Laboratories, USA). The rats were group-housed (2 per cage) for a week, then singly housed for a week. The rats were housed in a 12/12h light-dark cycle (light from 0600-1800 h) with controlled temperature conditions (22- 24°C). From the time of arrival and throughout the experiment standard rodent chow (Altromin standard #1324 chow; C. Petersen, Ringsted, Denmark) and water was available ad libitum unless otherwise stated. Treatment Groups

On day 0 of the experiment, animals of each genotype were randomised into two groups according to weight.

Thus, there were four groups in the experiment A: fa/fa allowed free access to standard rodent chow. B: Fa/? allowed free access to standard rodent chow. C: fa/fa fasted for 48 hours (food removed at 9:00 AM) D: Fa/? Fasted for 48 hours (food removed at 9:00 AM)

All animals were placed in cages with wire mesh bottom upon start of the starvation period. All animals were killed by CO2 anaesthesia and decapitation 48-h after the beginning of the starvation period. Epididymal adipose tissue was removed and homogenized in Trizol, and the homogenate stored at -8O 0 C until RNA purification was performed. The RNA purification, cDNA synthesis, and multiplex were performed as in example 1 above, using the rat-specific primer sets.

The expression of GPR120 is substantially higher in the adipose tissue from the Zucker fa/fa rats than in the adipose tissue from the Fa/? rats, probably because the adipose tissue in the fa/fa Zucker rats is more differentiated than the adipose tissue in the Fa/? rats.

Example 4:

Knock-down of GPR120 in differentiating adipocytes using si RNA

3T3-L1 cells were grown in DMEM supplemented with 10% foetal bovine serum (FBS), 100U/ml penicillin and 100μg/ml streptomycin in a humidified atmosphere of 5% CO2 at 37°C. 3T3-L1 cells were differentiated with DMEM, 10% serum, 0.5mM 3-lsobutyl-1 methylxanthine (IBMX), 10μg/ml insulin, 1 μM dexamethasone and 10OnM pioglitazone for 2 days. From the day 3 on, cells were incubated with DMEM, 10% serum, 10μg/ml insulin, and 10OnM pioglitazone.

Transfections with siRNA molecules targeting the GPR120 mRNA were performed using the Jet PEI reagent on day 1 and day 3.

Next the cells were subjected to the following procedures: The fatty acid used to activate GPR120 was; α-linolenic acid (18:3n-3). Other fatty acids such as linoleic acid (18:2n-6), γ-linolenic acid (18:3n-3), oleic acid (18:1n-9) and stearic acid (18:0) were used. The fatty acids were dissolved in DMSO and added to the differentiated cells under serum free conditions at doses of 10-, 30 and 100μM.

After 1 , 2, 4h RNA and protein was harvested and the following biochemical parameters measured:

1. Protein expression of P-Erk1/2

2. RNA expression of enzymes involved in β-oxidation (CPT1 , CPT2, ACO) and lipolysis (LPL)

3. β-oxidation (biochemically)

4. Lipolysis (biochemically)

Example 5:

GPR120 activation and fat cell metabolism 3T3-L1 cells were induced to differentiate by standard MDI-treatment.

At day 10 the adipocytes were considered to be mature and fatty acids was added.

The fatty acid used to activate GPR120 was α-linolenic acid (18:3n-3).

Other fatty acids such as linoleic acid (18:2n-6), γ-linolenic acid (18:3n-3), oleic acid (18:1n-9) and stearic acid (18:0) were used. The fatty acids were dissolved in DMSO and added to the differentiated cells under serum free conditions at doses of 10-, 30 and 100μM.

After 1 , 2, 4h RNA and protein was harvested and the following biochemical parameters measured: 1. Protein expression of P-Erk1/2

2. RNA expression of enzymes involved in β-oxidation (CPT1 , CPT2, ACO) and lipolysis (LPL)

3. β-oxidation (biochemically)

4. Lipolysis (biochemically) Example 6:

Forced overexpression of GPR120

3T3-L1 cells were grown in DMEM supplemented with 10% foetal bovine serum (FBS), 100U/ml penicillin and 100μg/ml streptomycin in a humidified atmosphere of 5% CO2 at 37°C. 3T3-L1 cells were differentiated with DMEM, 10% serum, 0.5mM 3-lsobutyl-1 methylxanthine (IBMX), 10μg/ml

insulin, 1 μM dexamethasone and 10OnM pioglitazone for 2 days. From the day 3 on, cells were incubated with DMEM, 10% serum, 10μg/ml insulin, and 10OnM pioglitazone.

Transfections were performed using the Jet PEI reagent. For stable transfection of plasmids containing full length GPR120 constructs Jet PEI was mixed with plasmid DNA, according to the manufacturer's instructions, and the mixtures were left on the cells in the incubator for 4h. Twenty-four h following transfection, G418 or puromycin was added at a final concentration of 1 mg/ml or 2.5 μg/ml respectively. The medium plus G418 or puromycin was replaced 3 times/week until no surviving cells were observed.

Next, the cells were subjected to the following procedures: The fatty acid used to activate GPR120 was α-linolenic acid (18:3n-3). Other fatty acids such as linoleic acid (18:2n-6), γ-linolenic acid (18:3n-3), oleic acid (18:1n-9) and stearic acid (18:0) were used. The fatty acids were dissolved in DMSO and added to the differentiated cells under serum free conditions at doses of 10-, 30 and 100μM.

After 1 , 2, 4h RNA and protein was harvested and the following biochemical parameters measured: 1. Protein expression of P-Erk1/2 2. RNA expression of enzymes involved in β-oxidation (CPT1 , CPT2, ACO) and lipolysis (LPL)

3. β-oxidation (biochemically)

4. Lipolysis (biochemically)

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d) a polynucleotide sequence that is antisense to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, e) a GPR120 specific antibody raised against a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof, and f) a siRNA molecule that is specific to a transcript encoding a GPR120 polypeptide comprising the amino acid sequence selected from a group consisting of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, a fragment, and variant thereof.

17. Use of a compound that modulates the action of an expression product of GPR120 gene sequence which is up-regulated in the adipose tissue of a diabetic and/or obese mammal, for manufacturing a pharmaceutical for treating, alleviating, or preventing obesity and/or diabetes.

18. Use of a compound that modulates the action of an expression product of GPR120 gene sequence which is up-regulated in the adipose tissue of a diabetic and/or obese mammal, for diagnosis of obesity and/or diabetes.