Diagnosing predisposition to fat deposition and therapeutic methods for reducing fat deposition and treatment of associated conditions

A method for diagnosing a predisposition to leanness in a subject, which comprises detecting the presence or absence of a polymorphic variation associated with leanness at a polymorphic site in a P2X4 nucleotide sequence in a nucleic acid sample from a subject, wherein the P2X4 nucleotide sequence is selected from the group consisting of:

• (a) the nucleotide sequence of SEQ ID NO:1;
• (b) a nucleotide sequence which encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• (c) a nucleotide sequence which encodes a polypeptide that is 90% identical to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• and
• (d) a fragment of a nucleotide sequence of (i), (ii), or (iii) comprising the polymorphic site;
• whereby the presence of the polymorphic variation is indicative of leanness in the subject.

A method for identifying a polymorphic variation associated with fat deposition proximal to an incident polymorphic variation associated with fat deposition, which comprises:

• identifying a polymorphic variant proximal to the incident polymorphic variant associated with fat deposition, wherein the incident polymorphic variant is in a P2X4 nucleotide sequence and the P2X4 nucleotide sequence comprises a polynucleotide sequence selected from the group consisting of:
• (a) a polynucleotide sequence set forth in SEQ ID NO:
• 1;
• (b) a polynucleotide sequence that encodes a polypeptide having an amino acid sequence encoded by a nucleotide sequence set forth as SEQ ID NO:
• 1;
• or
• (c) a polynucleotide sequence that encodes a polypeptide having an amino acid sequence that is 90% identical to an amino acid sequence encoded by a nucleotide sequence set forth in SEQ ID NO:
• 1;
• and
• determining the presence or absence of an association of the proximal polymorphic variant with fat deposition.

A method for diagnosing a predisposition to non-insulin dependent diabetes mellitus (NIDDM) in a subject, which comprises detecting the presence or absence of a polymorphic variation associated with NIDDM at a polymorphic site in a P2X4 nucleotide sequence in a nucleic acid sample from a subject, wherein the P2X4 nucleotide sequence comprises a polynucleotide sequence selected from the group consisting of:

• (a) the nucleotide sequence of SEQ ID NO:1;
• (b) a nucleotide sequence which encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• (c) a nucleotide sequence which encodes a polypeptide that is 90% identical to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• and
• (d) a fragment of a nucleotide sequence of (i), (ii), or (iii) comprising the polymorphic site;
• whereby the presence of the polymorphic variation is indicative of a predisposition to NIDDM in the subject.

A method for identifying a polymorphic variation associated with NIDDM proximal to an incident polymorphic variation associated with NIDDM, which comprises:

• identifying a polymorphic variant proximal to the incident polymorphic variant associated with NIDDM, wherein the incident polymorphic variant is in a P2X4 nucleotide sequence and the P2X4 nucleotide sequence comprises a polynucleotide sequence selected from the group consisting of:
• (a) a polynucleotide sequence set forth in SEQ ID NO:
• 1;
• (b) a polynucleotide sequence that encodes a polypeptide having an amino acid sequence encoded by a nucleotide sequence set forth as SEQ ID NO:
• 1;
• or
• (c) a polynucleotide sequence that encodes a polypeptide having an amino acid sequence that is 90% identical to an amino acid sequence encoded by a nucleotide sequence set forth in SEQ ID NO:
• 1;
• and
• determining the presence or absence of an association of the proximal polymorphic variant with NIDDM.

An isolated nucleic acid which comprises a P2X4 nucleotide sequence selected from the group consisting of:

• (a) the nucleotide sequence of SEQ ID NO:1;
• (b) a nucleotide sequence which encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• (c) a nucleotide sequence which encodes a polypeptide that is 90% identical to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• (d) a fragment of a nucleotide sequence of (a), (b), or (c) comprising a polymorphic variation;
• and
• (e) a nucleotide sequence complementary to the nucleotide sequences of (a), (b), (c), or (d);
• wherein the P2X4 nucleotide sequence comprises one or more polymorphic variations selected from the group consisting of thymine at position 14744, an adenine at position 14977, a cytosine at position 15046, a thymine at position 21538, a guanine at position 21630, and a single nucleotide deletion at position 25264 of SEQ ID NO:1.

An oligonucleotide which comprises a nucleotide sequence complementary to a portion of a P2X4 nucleotide sequence selected from the group consisting of:

• (a) the nucleotide sequence of SEQ ID NO:1;
• (b) a nucleotide sequence which encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• and
• (c) a nucleotide sequence which encodes a polypeptide that is 90% identical to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• wherein the 3′ end of the oligonucleotide is adjacent to a polymorphic variation in the P2X4 nucleotide sequence.

A method for identifying a candidate therapeutic for fat reduction, which comprises:

• (a) introducing a test molecule to a system which comprises a nucleic acid comprising a P2X4 nucleotide sequence selected from the group consisting of:
• (i) the nucleotide sequence of SEQ ID NO:1;
• (ii) a nucleotide sequence which encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• (iii) a nucleotide sequence which encodes a polypeptide that is 90% identical to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• and
• (iv) a fragment of a nucleotide sequence of (i), (ii), or (iii);
• or
• introducing a test molecule to a system which comprises a protein encoded by a nucleotide sequence of (i), (ii), (iii), or (iv);
• and
• (b) determining the presence or absence of an interaction between the test molecule and the nucleic acid or protein,
• whereby the presence of an interaction between the test molecule and the nucleic acid or protein identifies the test molecule as a candidate therapeutic for fat reduction.

A method for reducing fat deposition in a subject, which comprises contacting a P2X4 nucleic acid with one or more cells of a subject in need thereof, wherein the P2X4 nucleic acid comprises a nucleotide sequence selected from the group consisting of:

• (a) the nucleotide sequence of SEQ ID NO:1;
• (b) a nucleotide sequence which encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• (c) a nucleotide sequence which encodes a polypeptide that is 90% identical to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• and
• (d) a fragment of a nucleotide sequence of (i), (ii), or (iii);
• whereby contacting the one or more cells of the subject with the P2X4 nucleic acid reduces fat deposition.

A method for reducing fat deposition in a subject, which comprises contacting a P2X4 protein with one or more cells of a subject in need thereof, wherein the P2X4 protein is encoded by a P2X4 nucleotide sequence which comprises a polynucleotide sequence selected from the group consisting of:

• (a) the polynucleotide sequence of SEQ ID NO:1;
• (b) a polynucleotide sequence which encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• (c) a polynucleotide sequence which encodes a polypeptide that is 90% identical to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• and
• (d) a fragment of a polynucleotide sequence of (i), (ii), or (iii);
• whereby contacting the one or more cells of the subject with the P2X4 protein reduces fat deposition.

A method for reducing fat deposition in a subject, which comprises:

• detecting the presence or absence of a polymorphic variant associated with fat deposition in a P2X4 nucleotide sequence in a nucleic acid sample from a subject, wherein the P2X4 nucleotide sequence comprises a polynucleotide sequence selected from the group consisting of:
• (a) the polynucleotide sequence of SEQ ID NO:1;
• (b) a polynucleotide sequence which encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• (c) a polynucleotide sequence which encodes a polypeptide that is 90% identical to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• and
• (d) a fragment of a polynucleotide sequence of (i), (ii), or (iii);
• and
• administering a treatment that reduces fat deposition to a subject from whom the sample originated where the presence of a polymorphic variation associated with fat reduction is detected in the P2X4 nucleotide sequence.

A method for identifying a candidate therapeutic for alleviating NIDDM, which comprises:

• (a) introducing a test molecule to a system which comprises a nucleic acid comprising a P2X4 nucleotide sequence selected from the group consisting of:
• (i) the nucleotide sequence of SEQ ID NO:
• 1;
• (ii) a nucleotide sequence which encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• (iii) a nucleotide sequence which encodes a polypeptide that is 90% identical to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• and
• (iv) a fragment of a nucleotide sequence of (i), (ii), or (iii);
• or
• introducing a test molecule to a system which comprises a protein encoded by a nucleotide sequence of (i), (ii), (iii), or (iv);
• and
• (b) determining the presence or absence of an interaction between the test molecule and the nucleic acid or protein,
• whereby the presence of an interaction between the test molecule and the nucleic acid or protein identifies the test molecule as a candidate therapeutic for treating NIDDM.

A method for alleviating NIDDM in a subject, which comprises contacting a P2X4 nucleic acid with one or more cells of a subject in need thereof, wherein the P2X4 nucleic acid comprises a nucleotide sequence selected from the group consisting of:

• (a) the nucleotide sequence of SEQ ID NO:1;
• (b) a nucleotide sequence which encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• (c) a nucleotide sequence which encodes a polypeptide that is 90% identical to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• and
• (d) a fragment of a nucleotide sequence of (i), (ii), or (iii);
• whereby contacting the one or more cells of the subject with the P2X4 nucleic acid alleviates NIDDM.

A method for alleviating NIDDM in a subject, which comprises contacting a P2X4 protein with one or more cells of a subject in need thereof, wherein the P2X4 protein is encoded by a P2X4 nucleotide sequence which comprises a polynucleotide sequence selected from the group consisting of:

• (a) the polynucleotide sequence of SEQ ID NO:1;
• (b) a polynucleotide sequence which encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• (c) a polynucleotide sequence which encodes a polypeptide that is 90% identical to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• and
• (d) a fragment of a polynucleotide sequence of (i), (ii), or (iii);
• whereby contacting the one or more cells of the subject with the P2X4 protein alleviates NIDDM.

A method for alleviating NIDDM in a subject, which comprises:

• detecting the presence or absence of a polymorphic variant associated with NIDDM in a P2X4 nucleotide sequence in a nucleic acid sample from a subject, wherein the P2X4 nucleotide sequence comprises a polynucleotide sequence selected from the group consisting of:
• (a) the polynucleotide sequence of SEQ ID NO:1;
• (b) a polynucleotide sequence which encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• (c) a polynucleotide sequence which encodes a polypeptide that is 90% identical to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:3;
• and
• (d) a fragment of a polynucleotide sequence of (i), (ii), or (iii);
• and
• administering a treatment that alleviates NIDDM to a subject from whom the sample originated where the presence of a polymorphic variation associated with NIDDM is detected in the P2X4 nucleotide sequence.