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WO2011004345A1 - Polymorphismes de la protéine 1 de liaison amont et leur utilisation pour pronostiquer ou diagnostiquer une tension artérielle - Google Patents

Polymorphismes de la protéine 1 de liaison amont et leur utilisation pour pronostiquer ou diagnostiquer une tension artérielle Download PDF

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WO2011004345A1
WO2011004345A1 PCT/IB2010/053146 IB2010053146W WO2011004345A1 WO 2011004345 A1 WO2011004345 A1 WO 2011004345A1 IB 2010053146 W IB2010053146 W IB 2010053146W WO 2011004345 A1 WO2011004345 A1 WO 2011004345A1
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Prior art keywords
ubpl
polymorphic
disease
gene
blood pressure
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PCT/IB2010/053146
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English (en)
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Johan Auwerx
Rob Williams
Markku Laakso
Hana Koutnikova-Rousselin
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Ecole Polytechnique Federale De Lausanne (Epfl)
University Of Tennessee
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Publication of WO2011004345A1 publication Critical patent/WO2011004345A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Definitions

  • the present invention relates to biomarkers for diseases or disorders related to or 5 associated with the control of arterial blood pressure, such as hypertension and cardiovascular disorders biomarkers and use thereof.
  • Elevated arterial blood pressure (BP) or hypertension defined as systolic blood pressure higher or equal to 140 mmHg or diastolic blood pressure higher or equal to 90 mmHg, io is a prevalent health problem worldwide ⁇ Braunwald et al., 2008, Harrison's Principles of Internal Medicine. McGraw-Hill, New York, U.S.A.). The prevalence of hypertension is a worldwide health issue. For the US only, it is of 29% in U.S. adults 18 years and older and an additional 37% of U.S.
  • systolic BP between 120 to 139 mmHg or diastolic BP between 80 to 89 mmHg is (McDowell et al., 2008, Blood folate levels: The latest NHANES results. NCHS data briefs, no. 6, Hyattsville, MD: National Center for Health Statistics) .
  • Hypertension represents an independent risk factor for development of cardiovascular disease, including coronary heart disease, congestive heart failure, ischemic and hemorrhagic stroke, renal failure, and peripheral arterial disease.
  • BP is determined by both genetic and environmental factors, which include dietary NaCl intake, alcohol consumption, physical inactivity, stress.
  • Several genes responsible for the rare Mendelian forms of hypertension have been identified (CYPIlBl, CYP11B2, HSDIlBl, MR, SCNNlB, SCNNlG, WNKl, WNK4) and were shown to play an important role in the renal control of blood pressure (Lifton et al., 2001, Cell, 104,
  • UBP 1 gene belongs to the grainyhead family of transcription factors, has been cloned in 2002 and maps to chromosome 3p22.3.
  • UBPl is expressed in adrenal gland, placenta and in mouse heart and aorta. Its role has been described as important modulator of placental P450scc expression.
  • UBPl regulates expression of CYPIlA, the gene encoding the rate-limiting enzyme in steroidogenesis, and is expressed in placenta and adult adrenals (Huang et al, 2000, J. Biol. Chem, 275, 2852).
  • the sequence of the UBP 1 gene is known in the art.
  • the coding nucleic acid sequence of said gene can be retrieved under the number NM O 14517 at the NCBI Nucleotide Database.
  • a contiguous genomic sequence of the UBP 1 gene (SEQ ID NO: 1) can be gained from the Homo sapiens chromosome 3 genomic contig. NC 000003 at the NCBI Nucleotide database.
  • the derived protein sequence can be retrieved under the number NP 055332 at the National Center for Biotechnology Information (NCBI) Protein Database (National Center for Biotechnology Information, National Library of Medicine, Bethesda, MD 20894, USA; web address : http://www.ncbi.nlm.nih.gov/).
  • NCBI National Center for Biotechnology Information
  • the present invention is based on the finding that the type of nucleotide C/T, which is defined by SNP rsl7030583 is present at the position 33415322 and on the UBPl gene (according to NC 000003, Fig.l) is indicative for the risk of said individual to suffer from or develop cardiovascular and/or thrombotic disorders.
  • the present invention is directed towards a new and advantageous method for risk assessment and/or diagnosis and/or prognosis of diseases or disorders related to or associated with the control of arterial blood pressure, notably hypertension or a cardiovascular disorder in a subject.
  • the invention provides an in vitro method for risk assessment and/or diagnosis and/or prognosis of a disease or a disorder related to or associated with the control of arterial blood pressure in a subject, comprising the following steps:
  • step (b) Comparing the variant data obtained in step (a) to Ubpl or homologue thereof variant data from healthy and/or diseased subjects, wherein the variant data are correlated with a disease or a disorder related to or associated with the control of arterial blood pressure status in said subject.
  • the invention provides an isolated polynucleotide, a vector and oligonucleotide probes according to the invention.
  • the invention provides a diagnostic kit according to the invention.
  • FIG. 1 Schematic representation (not on scale) of exon intron organization of the UBPl and Fbxl2 genes.
  • the positions of rs2291897 (SEQ ID NO: 17), rsl7030583 (SEQ ID NO: 4) and rs2272152 (SEQ ID NO: 46) SNPs are shown by arrowheads.
  • the lower panel represents intron exon 4 border of the UBPl gene with the polymorphic nucleotide shown in majuscule.
  • Ubpl controlled protein includes but is not limited to CYPl IAl, 3 ⁇ -HSD2,
  • Ubpl homologue includes genes that are homologous to Ubpl gene and include for example the LBP-9 or Transcription factor CP2-Like-l (TFCP2L1) gene.
  • the coding nucleic acid sequence of said gene can be retrieved under the number NM_014553 at the NCBI Nucleotide Database.
  • LBP9 gene (SEQ ID NO: 72) can be gained from the Homo sapiens chromosome 2 genomic contig NC 000002.10 in the NCBI Nucleotide database.
  • the derived protein sequence can be retrieved under the number NP 055368. 1 in the NCBI Protein Database.
  • promoter sequence includes any in vitro transcription promoter known to the skilled person such as the promoter of the CYPl IAl gene. Its nucleic acid sequence can be retrieved under the number M60421 at the NCBI Nucleotide Database (SEQ ID NO: 2). Particularly relevant in the content of the CYPl IAl promoter is the binding site of UBP-I and LBP-9 (SEQ ID NO: 3).
  • polymorphic site refers to a region in a nucleic acid at which two or more alternative nucleotide sequences are observed in a significant number of nucleic acid samples from a population of individuals.
  • a polymorphic site may be a nucleotide sequence of two or more nucleotides, an inserted nucleotide or nucleotide sequence, a deleted nucleotide or nucleotide sequence, or a microsatellite, for example.
  • a polymorphic site that is two or more nucleotides in length may be 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15 or more, 20 or more, 30 or more, 50 or more, 75 or more, 100 or more, 500 or more, or about 1000 nucleotides in length, where all or some of the nucleotide sequences differ within the region.
  • a polymorphic site is often one nucleotide in length, which is referred to herein as a "single nucleotide polymorphism" or a "SNP.”
  • each nucleotide sequence is referred to as a "polymorphic variant” or "nucleic acid variant.”
  • polymorphic variants represented in a minority of samples from a population is sometimes referred to as a “minor allele” and the polymorphic variant that is more prevalently represented is sometimes referred to as a "major allele.”
  • minor allele the polymorphic variant represented in a minority of samples from a population
  • major allele the polymorphic variant that is more prevalently represented
  • a polymorphic variant is statistically significant and often biologically relevant if it is represented in 5% or more of a population, sometimes 10% or more, 15% or more, or 20% or more of a population, and often 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, or 50% or more of a population. Polymorphic variants are often reported in a database without determining whether the variant is represented in a significant fraction of a population.
  • a polymorphic variant according to the invention may be detected on either or both strands of a double-stranded nucleic acid. For example, a rsl7030583 having the sequence:
  • AC (SEQ ID NO: 4) at a particular position with a C or T being the polymorphic nucleotide at position 33415322 in SEQ ID NO: 1 can be reported as the following sequence:
  • polymorphic variants of UBPl include an C/T variant of the UBPl genomic sequence at positions 33415322 or the UBPl genomic sequence (SEQ ID NO: I)) also as referred as SNP rsl7030583 (SEQ ID NO: 4) and a G/A variant of within the UBPl Fbxl2 genomic sequence also as referred as SNP rs2291897 (SEQ ID NO: 17).
  • a polymorphic variant may be located within an intron or exon of a gene or within a portion of a regulatory region such as a promoter, a 5' untranslated region (UTR), a 3' UTR, and in DNA (e.g., genomic DNA (gDNA) and complementary DNA (cDNA)), RNA (e.g., mRNA, tRNA, and rRNA), or a polypeptide.
  • DNA e.g., genomic DNA (gDNA) and complementary DNA (cDNA)
  • RNA e.g., mRNA, tRNA, and rRNA
  • Polymorphic variations may or may not result in detectable differences in gene expression (mRNA and/or protein expression), polypeptide structure, polypeptide sequence, or polypeptide function.
  • Ubpl do result in detectable differences in gene expression (mRNA and/or protein expression), polypeptide structure, polypeptide sequence, or polypeptide function.
  • polymorphic variants of UBPl that are associated with a blood pressure and cardiovascular diseases or disorders.
  • SNPs SNPs
  • a total of 67 variants (SNPs) in the UBPl gene region (BP 33380933-33477747) have been genotyped in p h as e I o r I I of the HapMap project (as described on www.hapmap.org) and are shown in Table 1 below:
  • Table 2 below reports the SNPs which were genotyped as described in Example 1.
  • CHR chromosome
  • SNP Single nucleotide polymorphism
  • BP base pair position
  • Al major Allele
  • TEST ADD additive model
  • NMISS number of individuals tested
  • BETA regression coefficient for the test
  • SE standard error of the regression coefficient
  • L95 lower confidence interval limit
  • U95 upper confidence interval limit
  • STAT test statistic
  • P p-value
  • SEQ ID NO: x is for sense strand and x' is for antisense/complementary strand
  • MAF Major Allele Frequency.
  • genotyped refers to a process for determining a genotype of one or more individuals, where a "genotype” is a representation of one or more polymorphic variants in a population. Genotypes may be expressed in terms of a "haplotype,” which as used herein refers to two or more polymorphic variants occurring within genomic DNA in a group of individuals within a population. For example, two SNPs may exist within a gene where each SNP position includes a cytosine variation and an adenine variation. Certain individuals in a population may carry one allele
  • the individuals can be characterized as having a cytosine/cytosine haplotype with respect to the two SNPs in the gene.
  • phenotype refers to a trait which can be compared between individuals, such as presence or absence of a condition, a visually observable difference in appearance between individuals, metabolic variations, physiological variations, variations in the function of biological molecules, and the like.
  • An example of a phenotype is occurrence of hypertension.
  • Ubpl mediated disease or disorder includes diseases or disorders mediated by Ubpl gene or a homo log thereof.
  • Ubpl mediated disease or disorder or “diseases or disorders related to or associated with the control of arterial blood pressure” include but is not limited to hypertensive vascular disease (both essential forms and forms of hypertension within the context of the metabolic syndrome), coronary artery disease, chronic heart failure, cerebro vascular disease, diseases of the aorta and vascular diseases of the extremities, pulmonary hypertension and eclampsy.
  • suffering from a disease or condition means that a person is either presently subject to the signs and symptoms, or is more likely to develop such signs and symptoms than a normal person in the population.
  • an in vitro method for risk assessment and/or diagnosis and/or prognosis of a disease or a disorder related to or associated with the control of arterial blood pressure in a subject comprising the following steps:
  • step (a) Detecting in a nucleic acid sample from said subject the presence or absence of at least one polymorphic variant in human Ubpl gene or a homologue thereof, wherein the polymorphic variant is in a nucleic acid sequence that encodes for the UBP- 1 or homologue thereof mRNA/protein, controls expression of Ubp l gene or homologue thereof or encodes a Ubpl or homologue thereof controlled protein; and (b) Comparing the variant data obtained in step (a) to Ubpl or homologue thereof variant data from healthy and/or diseased subjects, wherein the variant data are correlated with a disease or a disorder related to or associated with the control of arterial blood pressure status in said subject.
  • a method according to the invention wherein at least one of the variants is a polymorphic site associated with at least one single nucleotide polymorphism (SNP) listed in Table 2, in particular wherein said at least one SNP is selected from rs2291897, rs2272152 and rsl7030583.
  • SNP single nucleotide polymorphism
  • a method according to the invention comprising detecting an at-risk allele of a SNP associated with a disease or disorder related to or associated with the control of arterial blood pressure status in said subject, wherein the SNP is located within a sequence selected from the Table 1 consisting of sequences identified by SEQ ID NOs: 4 to 71 and the complements of sequences identified by SEQ ID NOs: 4' to 71 '.
  • a method according to the invention comprising assaying for the presence of a first polynucleotide having a SNP according to the invention in a sample, comprising contacting said sample with a second polynucleotide, wherein said second polynucleotide comprises a nucleotide sequence selected from the group consisting of sequences identified by SEQ ID NOs: 4 to 71 and the complements of sequences identified by SEQ ID NOs: 4' to 71 ', wherein said second polynucleotide hybridizes to said first polynucleotide under stringent conditions.
  • a method according to the invention wherein said method is for monitoring the effect of a therapy administered to a subject having a disease or a disorder related to or associated with the control of arterial blood.
  • a method according to the invention wherein an increase in the level of Ubpl (or homologue thereof) variants compared to healthy control indicates that said subject is suffering from or has a predisposition to develop a disease or a disorder related to or associated with the control of arterial blood pressure.
  • a method according to the invention wherein the said subject's age is lower or of about 55 years.
  • a method according to the invention wherein the disease or disorder is hypertension or a cardiovascular disorder.
  • an isolated polynucleotide comprising a SNP located within a sequence selected from the group consisting of sequences identified by SEQ ID NOs: 4 to 71 and the complements of sequences identified by SEQ ID NOs: 4' to 71 '.
  • an oligonucleotide probe an oligonucleotide probe according to the invention.
  • an oligonucleotide probe according to the invention which is capable of detecting a polymorphism in the Ubpl gene (SEQ ID NO: 1).
  • an oligonucleotide probe according to the invention which is capable of detecting a polymorphism in the Ubpl gene homologue, LBP-9 (SEQ ID NO: 72).
  • a vector comprising an isolated polynucleotide containing a SNP located within a sequence selected from the group consisting of sequences identified by SEQ ID NOs: 4 to 71 and the complements of sequences identified by SEQ ID NOs: 4' to 71 ', wherein said isolated polynucleotide is operably linked to a regulatory sequence.
  • a diagnostic kit comprising one or more probe or primer which is capable of hybridizing to a polymorphic variant of an Ubpl gene or homo log thereof thereby determining whether the Ubpl gene or homolog thereof contains a polymorphic variant.
  • diagnostic kit wherein the said probe or primer that binds to SEQ ID NO: 1 , or a sequence complementary thereto.
  • a diagnostic kit comprising one or more oligonucleotide probes according to the invention.
  • a diagnostic kit for the detection of SNP haplotypes associated with a disease or a disorder related to or associated with the control of arterial blood pressure.
  • a diagnostic kit comprising at least one primer selected from the group consisting of SEQ ID NOs: 4 to 71 and the complements of sequences identified by SEQ ID NOs: 4' to 71 '.
  • Methods for identifying a polymorphic variation associated with a disease according to the invention that is proximal to an incident polymorphic variation associated with a Ubpl mediated disease or disorder comprises identifying a polymorphic variant proximal to the incident polymorphic variant associated with a Ubpl mediated disease or disorder, where the incident polymorphic variant is in a Ubpl gene or homolog thereof or regulatory sequence.
  • the presence or absence of an association of the proximal polymorphic variant with the disease then is determined using a known association method, such as a method described herein.
  • the incident polymorphic variant is present in a Ubpl gene or regulatory sequence.
  • the polymorphic variant is identified using a known method, including, but not limited to, sequencing a region surrounding the incident polymorphic variant in a group of nucleic acid samples.
  • a proximal polymorphic variant often is identified in a region surrounding the incident polymorphic variant.
  • this surrounding region is about 50 kb flanking the first polymorphic variant (e.g., about 50 kb 5' of the first polymorphic variant and about 50 kb 3' of the first polymorphic variant), and the region sometimes is composed of shorter flanking sequences, such as flanking sequences of about 40 kb, about 30 kb, about 25 kb, about 20 kb, about 15 kb, about 10 kb, about 7 kb, about 5 kb, or about 2 kb 5' and 3' of the incident polymorphic variant.
  • the region is composed of longer flanking sequences, such as flanking sequences of about 55 kb, about 60 kb, about 65 kb, about 70 kb, about 75 kb, about 80 kb, about 85 kb, about 90 kb, about 95 kb, or about 100 kb 5' and 3' of the incident polymorphic variant.
  • flanking sequences such as flanking sequences of about 55 kb, about 60 kb, about 65 kb, about 70 kb, about 75 kb, about 80 kb, about 85 kb, about 90 kb, about 95 kb, or about 100 kb 5' and 3' of the incident polymorphic variant.
  • polymorphic variants associated with an Ubpl mediated disease or disorder are identified iteratively.
  • a first proximal polymorphic variant is associated with a Ubpl mediated disease or disorder using the methods described herein and then another polymorphic variant proximal to the first proximal polymorphic variant is identified and the presence or absence of an association of one or more other polymorphic variants proximal to the first proximal polymorphic variant with a Ubpl mediated disease or disorder is determined.
  • allelotyping or genotyping data from the additional polymorphic variants may be used to identify a functional mutation or a region of linkage disequilibrium.
  • polymorphic variants identified or discovered within a region comprising the first polymorphic variant associated with a Ubpl mediated disease or disorder are genotyped using the genetic methods and sample selection techniques described herein, and it can be determined whether those polymorphic variants are in linkage disequilibrium with the first polymorphic variant. The size of the region in linkage disequilibrium with the first polymorphic variant also can be assessed using these genotyping methods.
  • Methods for determining the presence or absence of a polymorphic variant include, for example, detection of a polymorphic variant in a nucleic acid sequence such as genomic DNA, cDNA, mRNA, tRNA, rRNA, etc.
  • Variants may be located in any region of a nucleic acid sequence including coding regions, exons, introns, intron/exon borders and regulatory regions, such as promoters, enhancers, termination sequences, etc.
  • Certain polymorphic variants may be associated with differences in gene expression (mRNA and/or protein), post-transcriptional regulation and/or protein activity.
  • determining the presence or absence of the polymorphic variant may involve determining the level of transcription, mRNA maturation, splicing, translation, protein level, protein stability, and/or protein activity.
  • Polymorphic variants that lead to a change in protein sequence may also be determined by identifying a change in protein sequence and/or structure.
  • the methods described herein may be used to determine the genotype of a subject with respect to both copies of the polymorphic site present in the genome.
  • the complete genotype may be characterized as -/-, as -/+, or as +/+, where a minus sign indicates the presence of the reference sequence at the polymorphic site, and the plus sign indicates the presence of a polymorphic variant other than the reference sequence. If multiple polymorphic variants exist at a site, this can be appropriately indicated by specifying which ones are present in the subject. Any of the detection means described herein may be used to determine the genotype of a subject with respect to one or both copies of the polymorphism present in the subject's genome.
  • oligonucleotide arrays represent one suitable means for doing so.
  • Other methods including methods in which reactions (e.g., amplification, hybridization) are performed in individual vessels, e.g., within individual wells of a multi-well plate or other vessel may also be performed so as to detect the presence of multiple polymorphic variants (e.g., polymorphic variants at a plurality of polymorphic sites) in parallel or substantially simultaneously according to certain embodiments of the invention.
  • Diagnostic procedures may also be performed in situ directly upon tissue sections (fixed and/or frozen) obtained from a patient such that no nucleic acid purification is necessary.
  • Nucleic acids may be used as probes and/or primers for such in situ procedures (e.g., Nuovo 1992, PCR in situ hybridization: protocols and applications, Raven Press, New York).
  • Polymorphic variants may be detected in a sample from a subject using a biological sample from said patient such as, for example, samples of blood, serum, urine, saliva, cells (including cell lysates), tissue, hair, etc..
  • Biological samples suitable for use in accordance with the methods described herein will comprise an Ubpl (or homolog thereof) nucleic acid or polypeptide sequence.
  • Biological samples may be obtained using known techniques such as venipuncture to obtain blood samples or biopsies to obtain cell or tissue samples.
  • Cells, tissue or clinical samples can be from heart, kidney, brain, liver, bone marrow, colon, breast, prostate, thyroid, gall bladder, lung, adrenals, muscle, fat, nerve fibers, pancreas, skin, etc.
  • Preferred samples include blood, white blood cells, liver, muscle, kidney, fat and other tissues.
  • the cells or tissues can be pretreated with a therapeutic substance useful for the control of arterial blood pressure or other pharmacological agents either in vivo or following isolation.
  • haplotypes, SNPs or alleles of the Ubpl gene or homolog thereof as described herein could be done on the samples collected above. It is of course understood that in general the genetic analysis need not be done on the same sample used for subsequent biochemical analysis. Any sample, tissue or biopsy obtained from the given patient should be sufficient to determine the genetic haplotype of the Ubpl gene or homolog thereof as well as genetic analysis of any other gene.
  • the haplotype is schematically represented as +/+, +/- or -/- for the Ubpl or homolog thereof allele of interest.
  • the sample can then be subjected to a number of other biochemical and/or biological studies. These include quantitative measurement of mRNA or protein by methods known in the art and/or described herein, such as the measurement of Ubpl or homolog thereof mRNA or protein and the determination of mRNAs, protein and enzymes that are controlled by Ubpl or homolog thereof such as CYPl IAl , and hormones and signaling factors that are produced by CYPl IAl . In addition to the measurement of protein and mRNA of specific gene products as described above, the measurement of endogenous Ubpl or homolog thereof activity in various samples may be performed. Detection methods
  • Examples of techniques for detecting differences of at least one nucleotide between two nucleic acids include, but are not limited to, selective oligonucleotide hybridization, selective amplification, or selective primer extension.
  • a preferred detection method is allele specific hybridization using probes overlapping the polymorphic site and having about 5, 10, 20, 25, or 30 nucleotides around the polymorphic site.
  • oligonucleotide probes may be prepared in which the known polymorphic nucleotide is placed centrally (allele-specific probes) and then hybridized to target DNA under conditions which permit hybridization only if a perfect match is found (Saiki et al., 1989, Proc. Natl. Acad. Sci. USA, 86:6230).
  • Such allele specific oligonucleotide hybridization techniques may be used for the simultaneous detection of several nucleotide changes in different polymorphic regions of gene.
  • probes for detecting specific polymorphic variants of the polymorphic site located in the Ubpl gene or homo log thereof are probes comprising about 5, 10, 20, 25, 30, 50, 75 or 100 nucleotides of SEQ ID NO: 1 or about 5, 10, 20, 25, 30, 50, 75 or 100 nucleotides of a sequence complementary to SEQ ID NO: 1.
  • oligonucleotides having nucleotide sequences of specific polymorphic variants are attached to a hybridizing membrane and this membrane is then hybridized with labeled sample nucleic acid. Analysis of the hybridization signal will then reveal the identity of the polymorphic variants of the sample nucleic acid.
  • a solid phase support e.g., a "chip”.
  • Oligonucleotides can be bound to a solid support by a variety of processes, including lithography.
  • a chip can hold up to 250,000 oligonucleotides (GeneChip, Affymetrix). Mutation detection analysis using these chips comprising oligonucleotides, also termed "DNA probe arrays" is described e.g., in Cronin et al, 1996, Human Mutation, 7:244 and in Kozal et al., 1996, Nature Medicine, 2: 753.
  • a chip comprises all the polymorphic variants of at least one polymorphic region of a gene.
  • the solid phase support is then contacted with a test nucleic acid and hybridization to the specific probes is detected.
  • the identity of numerous polymorphic variants of one or more genes can be identified in a simple hybridization experiment.
  • the identity of the polymorphic variant at any of the polymorphic sites described herein can be determined in a single hybridization experiment.
  • the identity of the polymorphic variant at the SNPs from Table 1 (SEQ ID NOs: 4 to 71) and the complements of sequences identified by SEQ ID NOs: 4' to 71 ' may be determined in a single hybridization experiment.
  • Oligonucleotides used as primers for specific amplification may carry the polymorphic variant of interest in the center of the molecule (so that amplification depends on differential hybridization) (Gibbs et al., 1989, Nucleic Acids Res., 17:2437-2448) or at the extreme 3' end of one primer where, under appropriate conditions, a mismatch can prevent or reduce polymerase extension ⁇ Prossner, 1993, Tibtech, 11:238; Newton et al, 1989, Nucl. Acids Res., 17:2503). This technique is also termed "PROBE” for Probe Oligo Base Extension.
  • Various detection methods described herein involve first amplifying at least a portion of a gene prior to identifying the polymorphic variant. Amplification can be performed, e.g., by PCR and/or LCR, according to methods known in the art. In one embodiment, genomic DNA of a cell is exposed to two PCR primers and amplification is carried out for a number of cycles that is sufficient to produce the required amount of amplified DNA.
  • the primers may be about 5-50, about 10-50, about 10-40, about 10-30, about 10-25, about 15-50, about 15-40, about 15-30, about 15-25, or about 25-50 nucleotides in length and may be designed to hybridize to sites about 40-500 base pairs apart (e.g., to amplify a nucleotide sequence of about 40-500 base pairs in length).
  • Additional amplification methods include, for example, self-sustained sequence replication (Guatelli et al., 1990, Proc. Natl. Acad. Sci. U.S.A. 87:1874-1878), transcriptional amplification system (Kwoh et al., 1989, Proc. Natl. Acad. Sci. U.S.A. 86:1173-1177), Q-Beta Replicase (Lizardi et al, 1988, Bio/Technology, 6:1197), or any other nucleic acid amplification method, followed by the detection of the amplified molecules using techniques well known to those of skill in the art. These detection schemes are especially useful for the detection of nucleic acid molecules that may be present in very low numbers.
  • sequencing reactions Any of a variety of sequencing reactions known in the art can be used to directly sequence at least a portion of a gene and detect polymorphic variants by comparing the sequence of the sample sequence with the corresponding control sequence.
  • Exemplary sequencing reactions include those based on techniques developed by Maxam and Gilbert, 1977, Proc. Natl. Acad. Sci. USA, 74:560 or Sanger et al, 1977, Proc. Nat.
  • a specific polymorphic variant in a DNA sample from a subject can be shown by restriction enzyme analysis.
  • a specific polymorphic variant can result in a nucleotide sequence comprising a restriction site which is absent from a nucleotide sequence of another polymorphic variant.
  • alterations in electrophoretic mobility may be used to identify the polymorphic variant.
  • SSCP single strand conformation polymorphism
  • Single-stranded DNA fragments of sample and control nucleic acids are denatured and allowed to renature.
  • the secondary structure of single-stranded nucleic acids varies according to sequence and the resulting alteration in electrophoretic mobility enables the detection of even a single base change.
  • the DNA fragments may be labeled or detected with labeled probes.
  • RNA rather than DNA
  • the subject method utilizes heteroduplex analysis to separate double stranded heteroduplex molecules on the basis of changes in electrophoretic mobility (Keen et al., 1991, Trends Genet, 7:5).
  • the identity of a polymorphic variant of a may be obtained by analyzing the movement of a nucleic acid comprising the polymorphic variant in polyacrylamide gels containing a gradient of denaturant, e.g., denaturing gradient gel electrophoresis (DGGE).
  • DGGE denaturing gradient gel electrophoresis
  • DNA will be modified to insure that it does not completely denature, for example by adding a GC clamp of approximately 40 bp of high-melting GC-rich DNA by PCR.
  • a temperature gradient may be used in place of a denaturing agent gradient to identify differences in the mobility of control and sample DNA (Rosenbaum and Reissner, 1987, Biophys. Chem., 265:1275).
  • identification of the polymorphic variant is carried out using an oligonucleotide ligation assay (OLA), as described, e.g., in Landegren et al, 1988, Science, 241:1077-1080.
  • OLA oligonucleotide ligation assay
  • the OLA protocol uses two oligonucleotides which are designed to be capable of hybridizing to abutting sequences of a single strand of a target.
  • One of the oligonucleotides is linked to a separation marker, e.g., biotinylated, and the other is detectably labeled. If the precise complementary sequence is found in a target molecule, the oligonucleotides will hybridize such that their termini abut, and create a ligation substrate.
  • Ligation then permits the labeled oligonucleotide to be recovered using a biotin ligand, such as avidin.
  • a biotin ligand such as avidin.
  • some nucleic acid detection assays that combine attributes of PCR and OLA to achieve the exponential amplification of target DNA which is then detected using OLA.
  • Polymorphic variants may also be identified using methods for detecting single nucleotide polymorphisms. Because single nucleotide polymorphisms constitute sites of variation flanked by regions of invariant sequence, their analysis requires no more than the determination of the identity of the single nucleotide present at the site of variation and it is unnecessary to determine a complete gene sequence for each patient. Several methods have been developed to facilitate the analysis of such single nucleotide polymorphisms.
  • a single base polymorphism can be detected by using a specialized exonuclease-resistant nucleotide, as disclosed, e.g., in U.S. Patent No. 4,656,127.
  • a primer complementary to the allelic sequence immediately 3' to the polymorphic site is permitted to hybridize to a target molecule obtained from a subject. If the polymorphic site on the target molecule contains a nucleotide that is complementary to the particular exonuclease-resistant nucleotide derivative present, then that derivative will be incorporated onto the end of the hybridized primer. Such incorporation renders the primer resistant to exonuclease, and thereby permits its detection.
  • a solution-based method is used for determining the identity of a polymorphic variant (WO 91/02087).
  • a primer is employed that is complementary to allelic sequences immediately 3' to a polymorphic site. The method determines the identity of the nucleotide at that site using labeled dideoxynucleotide derivatives, which, if complementary to the nucleotide of the polymorphic site will become incorporated onto the terminus of the primer.
  • GBATM Genetic Bit Analysis
  • Goelet et al. WO 92/15712
  • the method uses mixtures of labeled terminators and a primer that is complementary to the sequence 3' to a polymorphic site.
  • the labeled terminator that is incorporated is thus determined by, and complementary to, the nucleotide present in the polymorphic site of the target molecule being evaluated.
  • the method of Goelet et al. is preferably a heterogeneous phase assay, in which the primer or the target molecule is immobilized to a solid phase.
  • a polymorphic variant is located in an exon (either a coding or non-coding exon)
  • the identity of the polymorphic variant can be determined by analyzing the molecular structure of the mRNA, pre-mRNA, or cDNA.
  • the molecular structure can be determined using any of the above described methods for determining the molecular structure of the genomic DNA, e.g., sequencing and SSCP.
  • profiles may also be assessed in such detection schemes. Fingerprint profiles may be generated, for example, by utilizing a differential display procedure, Northern analysis and/or RT- PCR.
  • Additional methods may be used for determining the identity of a polymorphic variant located in the coding region of a gene. For example, identification of a polymorphic variant which encodes a protein having a sequence variation can be performed using an antibody that specifically recognizes the protein variant, for example, using immunohisto chemistry, immunoprecipitation or immunoblotting techniques. Antibodies to protein variants may be prepared according to methods known in the art and as described herein.
  • polymorphic variants may be detected by determining variations in Ubpl protein expression and/or activity.
  • the expression level i.e., abundance
  • expression pattern e.g., temporal or spatial expression pattern, which includes subcellular localization, cell type specificity
  • size e.g., size, sequence, association with other cellular constituents (e.g., in a complex such as a UBPl complex), etc.
  • a control e.g., the expression level or expression pattern that would be expected in a sample obtained from a normal subject.
  • detection and/or comparison may be performed using any of a number of suitable methods known in the art including, but not limited to, immunoblotting (Western blotting), immunohistochemistry, ELISA, radioimmunoassay, protein chips (e.g., comprising antibodies to the relevant proteins), etc.
  • Historical data e.g., the known expression level, activity, expression pattern, or size in the normal population
  • Such methods may utilize UBPl antibodies that can distinguish between UBPl variants that differ at sites encoded by polymorphic variants.
  • Antibodies that specifically bind to antigenic determinants that comprise a region encoded by a polymorphic site of UBPl are useful in accordance with the methods described herein. According to certain embodiments, such antibodies are able to distinguish between UBPl polypeptides that differ by a single amino acid. Any of the antibodies described herein may be labeled. The methods described herein may also utilize panels of antibodies able to specifically bind to a variety of polymorphic variants of Ubp 1.
  • preferred antibodies will possess high affinity, e.g., a IQ of ⁇ 200 nM, and preferably, of ⁇ 100 nM for a specific polymorphic variant of UBPl.
  • Exemplary antibodies do not show significant reactivity (e.g., less than about 50%, 25%, 10%, 5%, 1%, or less, cross reactivity) with a different Ubpl polymorphic variant.
  • polymorphic variants may be determined by determining a change in level of activity of a UBPl controlled protein. Such activity may be measured in a biological sample obtained from a subject. Methods for measuring UBPl controlled protein activity could include determination of the mRNA, proteins and enzyme activities that are controlled by UBPl . Methods of the invention
  • a disease or a disorder related to or associated with the control of arterial blood pressure notably hypertension or a cardiovascular disorder in a subject, using one or more polymorphic variants of Ubpl or a homolog thereof.
  • the methods disclosed herein may be used, for example, to identify a subject suffering from or susceptible to develop hypertension or a cardiovascular disorder, e.g., to identify a subject that would benefit from a treatment with an agent useful for preventing or treating hypertension or a cardiovascular disorder or from a specific life style change (e.g., diet, exercise program) useful for preventing hypertension or a cardiovascular disorder to develop, based on the presence or absence of one or more polymorphic variants in a subject.
  • a panel of polymorphic variants may be defined that predict the risk of an Ubpl mediated disease or disorder and/or predict drug response to a therapeutic agent.
  • This predictive panel is then used for genotyping of patients on a platform that can genotype multiple polymorphic variants, such as SNPs, at the same time (Multiplexing).
  • Preferred platforms include, for example, gene chips (Affymetrix) or the Luminex LabMAP reader. The subsequent identification and evaluation of a patient's haplotype can then help to guide specific and individualized therapy.
  • kits that may be used to determine the presence or absence of one or more polymorphic variants of Ubpl or a homolog thereof. Such kits may be used to diagnose, or predict a subject's susceptibility to, an Ubpl mediated disease or disorder. This information could then be used, for example, to optimize treatment with an agent useful for treating a disease or a disorder related to or associated with the control of arterial blood pressure such as hypertension or a cardiovascular disorder for subjects having one or more polymorphic variants.
  • the kit comprises a probe or primer which is capable of hybridizing to a polymorphic variant of an Ubpl gene or a homolog thereof thereby determining whether the Ubpl gene or homolog thereof contains a polymorphic variant that is associated with a risk of having or developing an Ubpl mediated disease or disorder.
  • the kit may further comprise instructions for use in diagnosing a subject as having, or having a predisposition, towards developing an Ubpl mediated disease or disorder.
  • the probe or primers of the kit can be a probe or primer that binds to SEQ ID NO: 1, or a sequence complementary thereto. Such probe or primers may bind, for example, at and/or flanking a polymorphic site of Ubpl or a homolog thereof, such as the sites set forth in Table 1 or as described herein above.
  • probes according to the invention will be apparent to the molecular biologist of ordinary skill.
  • Such probes are of any convenient length such as up to 50 bases, up to 40 bases, more conveniently up to 30 bases in length, such as for example 8-25 or 8-15 bases in length.
  • such probes will comprise base sequences entirely complementary to the corresponding wild type or variant locus in the gene.
  • one or more mismatches may be introduced, provided that the discriminatory power of the oligonucleotide probe is not unduly affected.
  • the probes of the invention may carry one or more labels to facilitate detection.
  • Kits for amplifying a region of a gene comprising a polymorphic variant of Ubpl of interest may comprise one, two or more primers.
  • a kit may comprise a microarray suitable for detection of a variety of Ubp l or homolog thereof polymorphic variants. Examples of such microarrays are described further herein above.
  • kits provided herein may comprise one or more antibodies that are capable of specifically recognizing a polypeptide variant of Ubpl or a homolog thereof arising from a polymorphic variant of an Ubpl nucleic acid sequence.
  • the kit may include a panel of antibodies able to specifically bind to a variety of polypeptide variants of Ubpl encoded by polymorphic variants of Ubpl nucleic acid sequences.
  • the kits may further comprise additional components such as substrates for an enzymatic reaction.
  • the antibodies may be used for research, diagnostic, and/or therapeutic purposes.
  • kits may comprise appropriate packaging and instructions for use in the methods of the invention. Such kits may further comprise appropriate buffer(s) and polymerase(s) such as thermostable polymerases, for example taq polymerase. Such kits may also comprise accompanying primers and/or control primers or probes.
  • An accompanying primer is one that is part of the pair of primers used to perform PCR. Such primer usually complements the template strand precisely.
  • GWA Gene Wide Association analysis
  • BMI body mass index
  • BP body mass index
  • T2DM type-2 diabetes mellitus
  • polymorphisms may be responsible for disease or phenotypic variation by, for example, causing a functional alteration in an encoded protein
  • many polymorphisms appear to be silent in that no known detectable difference in phenotype exists between individuals having different alleles.
  • polymorphisms may be physically and/or genetically linked to genes or DNA sequences in which mutations or variations confer susceptibility to and/or play a causative role in disease (i.e., they are located within a contiguous piece of DNA).
  • polymorphisms that are physically linked to such mutations or variations will generally be inherited together with the mutation or alteration.
  • Polymorphisms are thus useful for genetic mapping and identification of candidate genes, in which mutations or variations may play a causative role in disease.
  • detection of particular polymorphic variants (alleles) is useful for diagnosis of disease or susceptibility to disease as described herein.
  • Studies provided herein have linked increased systolic and diastolic blood pressure with polymorphic variants of UBPl.
  • EXAMPLE 1 GENETIC ANALYSIS OF UBPl
  • the following example describes a clinical genetic study showing the association of genetic variations in the human UBPl gene is associated with blood pressure variation.
  • the probands were randomly selected among type 2 diabetic patients living in the regions of five centers in Europe. They were recruited over a 4-year period through advertisements in public media and in the hospitals. The acceptance rate of volunteers was at least 70% in the different centers.
  • BP was measured with a mercury sphygmomanometer in the sitting position after a 5 min rest. The average of two measurements was used in statistical analyses. BP of individuals who were on BP medication (8.6% of study subjects) was adjusted by adding 15 mmHg to systolic and 10 mmHg to diastolic BP. Height and weight were measured to the nearest 0.5 cm and 0.1 kg, respectively and BMI (kg/m 2 ) calculated. Because the primary aim of the study was to investigate the genes associated with insulin secretion and insulin sensitivity, every participant underwent detailed metabolic studies, as described ⁇ Laakso et al.,
  • Gender calls from X chromosome genotype data was verified to be in concordance with the reported gender of each individual.
  • pairwise identity-by-descent (IBD) analysis was performed. Individuals with genotyping call rates less than 95% were excluded, resulting in total of 903 out of 970 samples passing these criteria. Of these individuals, 869 had the required phenotype data and these individuals advanced to the actual association analysis. Total genotyping call rate in remaining individuals was 99.6%.
  • Markers advanced to the actual association analysis if they passed the following quality control criteria 1) had a 95% genotype call rate (4007 markers excluded), 2) had a minor allelic frequency (MAF) >1% (23107 markers excluded) and 3) demonstrated Hardy- Weinberg Equilibrium (HWE) with a P > le-05 (890 markers excluded). Total of 534 287 markers passed these quality control criteria. In addition, for each marker, tests for MAF, HWE and missingness were performed, and this information was used when evaluating marker quality for replication.
  • Phase 2 HapMap (release 23) CEU (Utah residents with Northern and Western European ancestry from the CEPH collection) founder population data (60 individuals with MAF > 0.01 and genotyping rate > 0.95) were used to impute markers that were not directly genotyped.
  • METSIM METabolic Syndrome In Men
  • METSIM METabolic Syndrome In Men
  • BP Blood pressure
  • BP Blood pressure
  • the average of 3 measurements was used to calculate systolic and diastolic BPs. Height and weight were measured to the nearest 0.5 cm and 0.1 kg, respectively.
  • Body mass index was calculated as weight (kg) divided by height (m) squared.
  • the study protocol was accepted by the Ethics Committee of the University of Kuopio and Kuopio University Hospital. Genotyping was performed using the TaqMan Allelic Discrimination Assays (Applied Biosystems) or using the iPLEX Sequenom MassARRAY platform.
  • the pooled effect size of this SNP was 1.5 (0.4) for systolic and 0.8 (0.3) mmHg with diastolic BP.
  • rs2291897, rs2272152, rsl7030583 belong to the FBXL2/UBP1 gene locus (Fig. 1).
  • This locus belongs to a haploblock including also other genes (CLASP2, PDC61P, SUSD5). Therefore, the SNPs genotyped in the METSIM cohort are those giving the most significant P value from CLASP2 (rs9841066), PDC61P (rs9311032, rs9858195), and SUSD5 (rs4678778, rs9836433, rsl0222597) loci (SEQ ID NO:73 to 78 and their reverse complement 73' to 78') to exclude the possibility that these genes could be responsible for the association signal.

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Abstract

La présente invention concerne des méthodes permettant d’effectuer un diagnostic et un pronostic à l'aide de variants polymorphes de l’Ubp 1 et de sondes et kits associés.
PCT/IB2010/053146 2009-07-09 2010-07-09 Polymorphismes de la protéine 1 de liaison amont et leur utilisation pour pronostiquer ou diagnostiquer une tension artérielle WO2011004345A1 (fr)

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4656127A (en) 1983-04-22 1987-04-07 Amersham International Plc. Method of detecting mutations in DNA and RNA
WO1991002087A1 (fr) 1989-08-11 1991-02-21 Bertin & Cie Procede rapide de detection et/ou d'identification d'une seule base sur une sequence d'acide nucleique, et ses applications
WO1992015712A1 (fr) 1991-03-05 1992-09-17 Molecular Tool, Inc. Determination d'acides nucleiques par extension de la polymerase d'oligonucleotides a l'aide de melanges terminateurs
WO1994016101A2 (fr) 1993-01-07 1994-07-21 Koester Hubert Sequençage d'adn par spectrometrie de masse
US5571676A (en) 1995-06-07 1996-11-05 Ig Laboratories, Inc. Method for mismatch-directed in vitro DNA sequencing
US5580732A (en) 1992-04-03 1996-12-03 The Perkin Elmer Corporation Method of DNA sequencing employing a mixed DNA-polymer chain probe

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4656127A (en) 1983-04-22 1987-04-07 Amersham International Plc. Method of detecting mutations in DNA and RNA
WO1991002087A1 (fr) 1989-08-11 1991-02-21 Bertin & Cie Procede rapide de detection et/ou d'identification d'une seule base sur une sequence d'acide nucleique, et ses applications
WO1992015712A1 (fr) 1991-03-05 1992-09-17 Molecular Tool, Inc. Determination d'acides nucleiques par extension de la polymerase d'oligonucleotides a l'aide de melanges terminateurs
US5580732A (en) 1992-04-03 1996-12-03 The Perkin Elmer Corporation Method of DNA sequencing employing a mixed DNA-polymer chain probe
WO1994016101A2 (fr) 1993-01-07 1994-07-21 Koester Hubert Sequençage d'adn par spectrometrie de masse
US5547835A (en) 1993-01-07 1996-08-20 Sequenom, Inc. DNA sequencing by mass spectrometry
US5571676A (en) 1995-06-07 1996-11-05 Ig Laboratories, Inc. Method for mismatch-directed in vitro DNA sequencing

Non-Patent Citations (30)

* Cited by examiner, † Cited by third party
Title
COHEN ET AL., ADV. CHROMATOGR., vol. 36, 1996, pages 127 - 162
COTTON, MUTAT. RES., vol. 285, 1993, pages 125 - 144
COWLEY, NAT. REV. GENET., vol. 7, 2006, pages 11
CRONIN ET AL., HUMAN MUTATION, vol. 7, 1996, pages 244
DATABASE DBsnp [online] NCBI; 24 October 2001 (2001-10-24), XP002604630, retrieved from http://www.ncbi.nlm.nih.gov/projects/SNP/snp_ref.cgi?rs=2291897 Database accession no. rs2291897 *
DATABASE DBsnp [online] NCBI; 24 October 2001 (2001-10-24), XP002604631, retrieved from http://www.ncbi.nlm.nih.gov/projects/SNP/snp_ref.cgi?rs=2272152 Database accession no. rs2272152 *
DATABASE DBsnp [online] NCBI; 28 October 2004 (2004-10-28), XP002604629, retrieved from http://www.ncbi.nlm.nih.gov/projects/SNP/snp_ref.cgi?rs=17030583 Database accession no. rs17030583 *
GASPARINI ET AL., MOL. CELL PROBES, vol. 6, 1992, pages 1
GIBBS ET AL., NUCLEIC ACIDS RES., vol. 17, 1989, pages 2437 - 2448
GRIFFIN ET AL., APPL. BIOCHEM. BIOTECHNOL., vol. 38, 1993, pages 147 - 159
GUATELLI ET AL., PROC. NATL. ACAD. SCI. U.S.A., vol. 87, 1990, pages 1874 - 1878
HUANG ET AL., J. BIOL. CHEM, vol. 275, 2000, pages 2852
JI ET AL., NAT. GENET., vol. 40, 2008, pages 592
KEEN ET AL., TRENDS GENET., vol. 7, 1991, pages 5
KELLY TANIKA N ET AL: "Novel genetic variants in the alpha-adducin and guanine nucleotide binding protein beta-polypeptide 3 genes and salt sensitivity of blood pressure.", AMERICAN JOURNAL OF HYPERTENSION, July 2009 (2009-07-01), XP002604627, Retrieved from the Internet <URL:http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2882159/pdf/nihms202640.pdf?tool=pmcentrez> [retrieved on 20101014] *
KOZAL ET AL., NATURE MEDICINE, vol. 2, 1996, pages 753
KUPPUSWAMY ET AL., PROC. NATL. ACAD. SCI. (U.S.A.), vol. 88, 1991, pages 1143 - 1147
KWOH ET AL., PROC. NATL. ACAD. SCI. U.S.A., vol. 86, 1989, pages 1173 - 1177
LAAKSO ET AL., DIABETOLOGIA, vol. 51, 2008, pages 502
LANDEGREN ET AL., SCIENCE, vol. 241, 1988, pages 1077 - 1080
LEVY DANIEL ET AL: "Framingham Heart Study 100K Project: genome-wide associations for blood pressure and arterial stiffness", BMC MEDICAL GENETICS, vol. 8, no. Suppl. 1, 2007, XP002604628, ISSN: 1471-2350 *
LIFTON ET AL., CELL, vol. 104, 2001, pages 545
NEWTON ET AL., NUCL. ACIDS RES., vol. 17, 1989, pages 2503
NYREN ET AL., ANAL. BIOCHEM., vol. 208, 1993, pages 171 - 175
PREZANT ET AL., HUM. MUTAT., vol. 1, 1992, pages 159 - 164
PROSSNER, TIBTECH, vol. 11, 1993, pages 238
ROSENBAUM; REISSNER, BIOPHYS. CHEM., vol. 265, 1987, pages 12 75
SAIKI ET AL., PROC. NATL. ACAD. SCI. USA, vol. 86, 1989, pages 6230
SYVANEN ET AL., AMER. J. HUM. GENET., vol. 52, 1993, pages 46 - 59
TEMELKOVA-KURKTSCHIEV ET AL., J. CLIN. ENDOCRINOL. METAB., vol. 89, 2004, pages 4238

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