Isolated nucleic acids and polypeptides associated with glucose homeostasis disorders and method of detecting the same
AssigneeDuke University
InventorsY.T. Chen, Alison J. McVie-Wylie
Life of this patent
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Abstract
An isolated and substantially pure nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13, the nucleic acid sequence including: a nucleic acid sequence coding for a glucose transporting protein and having the sequence shown in SEQ ID NO: 1; or a nucleic acid sequence having at least 70% sequence identity with the nucleic acid sequence shown in SEQ ID NO: 1. The disclosed nucleic acid sequences map to a locus associated with human Type II diabetes mellitus and, therefore, therapeutic and diagnostic screening methods, which accommodate naturally and artificially occurring polymorphisms, are also disclosed.
Claims66
An isolated and substantially pure nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13, the nucleic acid sequence comprising:
- (a) a nucleic acid sequence coding for a glucose transporting protein and having the sequence shown in SEQ ID NO:
- 1;
- or (b) a nucleic acid sequence having at least 70% sequence identity with the nucleic acid sequence shown in SEQ ID NO:
- 1.
An antisense oligonucleotide complementary to an mRNA transcribed from a nucleic acid sequence of SEQ ID NO:
- 1, and comprising from 10 to 35 bases, said antisense oligonucleotide being complementary to at least 10 consecutive bases of SEQ ID NO:
- 1.
A method of facilitating a diagnosis of susceptibility to a glucose homeostasis disorder, the method comprising:
- (a) obtaining a biological sample from a subject;
- (b) isolating a target nucleic acid sequence from the biological sample, the target nucleic acid sequence encoding a glucose transporter polypeptide located on chromosome 20q13 between D20S119 and D20S178;
- (c) sequencing the target nucleic acid sequence;
- and
- (d) determining a sequence variation between a wild type nucleic acid sequence and the isolated target nucleic acid sequence, the presence of a variation between the wild type nucleic acid sequence and the isolated target nucleic acid sequence indicating susceptibility of the subject to a glucose homeostasis-related disorder.
A method of identifying a subject having a polymorphism of the nucleic acid sequence shown in SEQ ID NO:
- 1, the method comprising:
- (a) sequencing a target nucleic acid of a sample from a subject by dideoxy sequencing;
- and
- b) identifying a polymorphism by comparing the nucleic acid sequence of the target nucleic acid sequence to the DNA sequence shown in SEQ ID NO:
- 1.
A method of identifying a subject having a polymorphism of the nucleic acid sequence shown in SEQ ID NO:
- 1, the method comprising:
- (a) contacting a target nucleic acid sample from a subject with a reagent that detects the presence of a polymorphism in the nucleic acid sequence shown in SEQ ID NO:
- 1;
- and
- (b) detecting the reagent.
A substantially pure preparation of a polypeptide having at least 70% sequence identity with the amino acid sequence shown in SEQ ID NO:
- 2.
A substantially pure preparation of a polypeptide having the amino acid sequence shown in SEQ ID NO:
- 2.
A method of screening a biological sample for the presence of a glucose transporter polypeptide encoded by a nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13, the method comprising:
- (a) obtaining a biological sample;
- (b) contacting the biological sample with an antibody immunoreactive with the glucose transporter polypeptide to form an antibody-polypeptide conjugate;
- and
- (c) detecting the conjugate.
A method of screening a biological sample for the presence of antibodies immunoreactive with a glucose transporter polypeptide encoded by a nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13, the method comprising
- (a) exposing a biological sample is exposed to a glucose transporter polypeptide under biological conditions and for a period of time sufficient for antibody-polypeptide conjugate formation;
- and
- (b) detecting the conjugate.
A method of a facilitating a diagnosis of a disorder affecting glucose homeostasis, the method comprising:
- (a) obtaining a biological sample from a subject;
- (b) determining an amount of a glucose transporter polypeptide encoded by a nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13 that is present in the biological sample, wherein the presence of a reduced amount of the glucose transporter polypeptide as compared to a standard facilitates a diagnosis of a disorder affecting glucose homeostasis.
A method of a facilitating a diagnosis of a disorder affecting glucose homeostasis, the method comprising:
- (a) obtaining a glucose transporter polypeptide encoded by a nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13 from a subject;
- (b) determining an activity level of a glucose transporter polypeptide from the subject;
- and
- (c) detecting a variation in glucose transport activity between a wild type glucose transporter polypeptide and the glucose transporter polypeptide from the subject, the presence of a glucose transport activity variation between the wild type glucose transporter polypeptide and the glucose transporter polypeptide from the subject facilitating a diagnosis of a disorder affecting glucose homeostasis.
A method of screening for a modulator of a glucose transporter polypeptide encoded by a nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13, the method comprising:
- (a) establishing replicate test and control samples that comprise the glucose transporter polypeptide;
- (b) administering a candidate substance to the test sample;
- (c) measuring the biological activity of the polypeptide in the test and the control samples;
- and
- (d) determining whether the candidate substance modulates biological activity based on a comparison of the test and control samples.
A method of designing a modulator of a glucose transporter polypeptide encoded by a nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13, the method comprising:
- (a) designing a potential modulator of the glucose transporter polypeptide that will form non-covalent bonds with amino acids in a substrate binding site based upon the structure the glucose transporter polypeptide;
- (b) synthesizing the modulator;
- and
- (c) determining whether the potential modulator modulates the activity of the glucose transporter polypeptide.
A method for identifying a candidate compound as a modulator of a glucose transporter polypeptide encoded by a nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13, the method comprising:
- (a) contacting a cell sample with a candidate compound to be tested, the cell sample containing at least one cell containing a DNA construct comprising a modulatable transcriptional regulatory sequence of a gene that encodes the glucose transporter polypeptide and a reporter gene which is capable of producing a detectable signal;
- (b) evaluating an amount of signal produced in relation to a control sample;
- and
- (c) identifying a candidate compound as a modulator of glucose transporter polypeptide biological activity based on the amount of signal produced in relation to a control sample.
The method of 37, wherein the reporter gene encodes the glucose transporter polypeptide.
A method of modulating the biological activity in a cell of a glucose transporter polypeptide encoded by a nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13 or biological equivalent thereof, the method comprising administering to the cell an effective amount of a substance capable of modulating activity of the glucose transporter polypeptide in the cell to thereby modulate glucose transporter polypeptide biological activity in the cell.
A pharmaceutical composition comprising a therapeutically effective amount of a modulator of a biological activity of a glucose transporter polypeptide encoded by a nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13 or biological equivalent thereof;
- and a pharmaceutically acceptable diluent or vehicle.
A method for modulation of a glucose transporter level in a cell, the method comprising:
- (a) delivering to a target cell an effective amount of a nucleic acid sequence that encodes a glucose transporter polypeptide encoded by a nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13 or biological equivalent thereof;
- and
- (b) maintaining the cell under conditions sufficient for expression of said polypeptide.
A nucleic acid sequence which modulates the transcription of a glucose transporter polypeptide encoded by a nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13 comprising the nucleic sequence shown in SEQ ID NO:
- 4 or a DNA fragment thereof.
A method of screening for a modulator of a glucose transport polypeptide encoded by a nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13, the method comprising:
- (a) providing cells from a cDNA expression library on a substrate, the cells comprising cDNA coding for the glucose transporter polypeptide;
- (b) causing protein synthesis by the cells;
- (c) subjecting the cells to a library of test samples;
- (d) detecting an interaction between a test sample and a cell expressing the glucose transporter polypeptide;
- (e) identifying a test sample that interacts with a cell expressing the glucose transporter polypeptide;
- and
- (f) isolating the test sample that interacts with the glucose transporter polypeptide.
A method of screening for a modulator of a glucose transport polypeptide encoded by a nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13, the method comprising:
- (a) affixing distinct colonies of cells from a cDNA expression library on a substrate, the cells comprising cDNA coding for the glucose transporter polypeptide;
- (b) causing protein synthesis by said colonies on the substrate;
- (c) subjecting the colonies of cells to a library of test samples;
- (d) detecting an interaction between a test sample and a cell expressing the glucose transporter polypeptide;
- (e) identifying a test sample that interacts with a cell expressing a glucose transporter polypeptide;
- and
- (f) isolating a test sample that interacts with a glucose transporter polypeptide.
A method of screening for a modulator of a glucose transport polypeptide encoded by a nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13, the method comprising:
- (a) providing a library of test samples;
- (b) contacting a glucose transporter polypeptide with each test sample;
- (c) detecting an interaction between a test sample and a glucose transporter polypeptide
- (d) identifying a test sample that interacts with a glucose transporter polypeptide;
- and
- (e) isolating a test sample that interacts with a glucose transporter polypeptide.
Dependencies are inferred from the claim text.