[go: up one dir, main page]

WO2008109110A2 - Système de régulation de la pression sanguine - Google Patents

Système de régulation de la pression sanguine Download PDF

Info

Publication number
WO2008109110A2
WO2008109110A2 PCT/US2008/002938 US2008002938W WO2008109110A2 WO 2008109110 A2 WO2008109110 A2 WO 2008109110A2 US 2008002938 W US2008002938 W US 2008002938W WO 2008109110 A2 WO2008109110 A2 WO 2008109110A2
Authority
WO
WIPO (PCT)
Prior art keywords
gene
nucleic acid
adamtslό
animal
hypertension
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2008/002938
Other languages
English (en)
Other versions
WO2008109110A3 (fr
Inventor
Bina Joe
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Toledo
Original Assignee
University of Toledo
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University of Toledo filed Critical University of Toledo
Publication of WO2008109110A2 publication Critical patent/WO2008109110A2/fr
Publication of WO2008109110A3 publication Critical patent/WO2008109110A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2267/00—Animals characterised by purpose
    • A01K2267/03—Animal model, e.g. for test or diseases
    • A01K2267/035—Animal model for multifactorial diseases
    • 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
    • C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
    • 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
    • C12Q2600/136—Screening for pharmacological compounds
    • 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
    • C12Q2600/158—Expression markers
    • 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
    • C12Q2600/172—Haplotypes

Definitions

  • Essential hypertension which is the most common cardiovascular abnormality, is characterized by sustained blood pressure (BP) with no identifiable cause l .
  • BP blood pressure
  • Genetic susceptibility factors account for approximately 27% of an individual's risk of developing essential hypertension ⁇ . Mapping experiments in humans is by far the only way by which one could attempt to identify the genetic role players in elevated BP. Such studies have successfully identified genetic determinants of monogenic forms of hypertension J ⁇ * , but few have emerged successful in defining the underlying susceptibility factors for essential hypertension, which represents 95% of all forms of hypertension. This void has a bearing on current clinical management practices for essential hypertension, which are limited to symptomatic treatments with select hypotensive agents.
  • the present invention provides an improved system for defining underlying susceptibility factors for essential hypertension.
  • a method for selecting an animal model of disease for having desired genotypic properties includes testing the animal for the presence of a parentally imprinted quantitative trait locus (QTL) that at least partially controls blood pressure and/or at least partially is indicative of a potential for developing a hypertensive disease.
  • the method further includes testing a nucleic acid sample from the animal for the presence of QTL.
  • the animal can be a rat and the QTL is located at chromosome 17.
  • the QTL is mapping at around position IpI 1.
  • the animal is a mouse and the QTL is located at chromosome- 13.
  • the QTL is related to the potential to develop hypertension.
  • the QTL comprises at least a part of Adamtsl ⁇ gene.
  • the QTL of the rat can comprise a marker marked with an asterisk (*), as identified in Table 1.
  • an isolated and/or recombinant nucleic acid sequence comprising a quantitative trait locus (QTL) or functional fragment derived thereof, where the QTL at least partially controls blood pressure and/or at least partially is indicative of a potential for developing a hypertensive disease.
  • QTL quantitative trait locus
  • an isolated and/or recombinant nucleic acid sequence comprising a synthetic quantitative trait locus (QTL) derived from at least one chromosome or functional fragment derived thereof where the QTL at least partially controls blood pressure and/or at least partially is indicative of a potential for developing a hypertensive disease.
  • the isolated and/or recombinant nucleic acid sequence can be at least partly derived from a rat chromosome 1, a mouse chromosome, 13, or a human chromosome 5.
  • the QTL comprises at least a part of an Adamtsl ⁇ gene.
  • a method for selecting an animal for having desired genotypic properties includes: testing the animal for the presence of a quantitative trait locus (QTL) with an isolated and/or recombinant nucleic acid sequence comprising a QTL or functional fragment derived therefrom, where the QTL at least partially controls blood pressure and/or at least partially is indicative of a potential for developing a hypertensive disease.
  • QTL quantitative trait locus
  • the method can include selecting a non-human animal having desired genotypic or potential phenotypic properties.
  • an animal selected for by the methods described herein.
  • the animal is homozygous for an allele at a QTL
  • the QTL can be at least partially controls blood pressure and/or at least partially is indicative of a potential for developing a hypertensive disease, and/or wherein the QTL comprises at least a part of an Adamtsl ⁇ allele.
  • an animal which is transgenic, where the animal comprising the nucleic acid of claim 11.
  • the animal can be a male or is a female.
  • a sperm or an embryo from the animal.
  • an isolated and/or recombinant nucleic acid sequence where the isolated and/or recombinant nucleic acid sequence is at least partly derived from a rat chromosome 1 from a region mapping at around position IpI 1.
  • a method for determining the predisposition of an individual to hypertension includes analyzing at least part of the DNA sequence of at least one allelic variant, fragment or derivative of the Adamtsl ⁇ gene of the individual for the presence of at least one single nucleotide polymorphism (SNP) in the Adamtsl ⁇ gene, wherein the SNP is selected from the group consisting of one or more of SNPs as described herein.
  • the predisposition is a predisposition to essential hypertension.
  • the genomic sequence of the Adamtsl ⁇ gene of the individual is analyzed.
  • the genomic sequence of a part of the Adamtsl ⁇ gene of the individual is analyzed.
  • the determination of at least a part of the Adamtsl ⁇ gene can be performed by hybridization of a nucleic acid to the Adamtsl 6 gene of the individual. Also, the hybridization can be performed with an allele-specif ⁇ c oligonucleotide probe. The analysis can also be carried out by sequence analysis. The determination of the Adamtsl ⁇ gene can be carried out by substitution mapping analysis.
  • nucleic acid probe which specifically hybridizes to one or more SNPs in the Adamtsl ⁇ gene wherein the SNP is selected from the group consisting of one or more of the SNPs described herein.
  • a method for determining whether an individual has, or is predisposed to developing, hypertension associated with a hypertensive haplotype includes analyzing at least part of the DNA sequence of the Adamtsl ⁇ gene of the individual for the presence of an allelic pattern, where each allele comprises at least one SNP selected from one or more of the SNPs described herein; and where the presence of the allelic pattern indicates that the individual is predisposed to the development of, or has, hypertension.
  • the predisposition is a predisposition to essential hypertension.
  • the genomic sequence of at least one allele of the Adamtsl ⁇ gene of the individual is analyzed.
  • the analysis can be performed by hybridization of at least one nucleic acid to the Adamtsl ⁇ gene of the individual. Also, the hybridization can be performed with an allele-specific oligonucleotide probe. Further, the analysis can be carried out by sequence analysis.
  • the determination of the Adamtsl ⁇ gene is carried out by substitution mapping analysis.
  • a part of the genomic sequence of at least one allele of the Adamtsl ⁇ gene of the individual is analyzed.
  • the analysis can be carried out by hybridization of a nucleic acid probe to at least one allele of the Adamtsl ⁇ gene.
  • the analysis can be determined hybridization is with an allele-specific oligonucleotide probe, and analysis can be carried out by substitution mapping analysis.
  • a part of the genomic sequence of the Adamtsl ⁇ gene of the human is analyzed.
  • a method of determining the predisposition of an individual to hypertension which includes analyzing at least part of the DNA sequence of the Adamtsl ⁇ gene of the individual for the presence of at least one haplotype for the Adamtsl ⁇ gene.
  • the predisposition is a predisposition to essential hypertension.
  • the genomic sequence of at least one allele of the haplotypes for the Adamtsl ⁇ gene of the individual is analyzed.
  • a part of the genomic sequence at least two alleles of the haplotypes for the Adamtsl ⁇ gene of the individual are analyzed.
  • the analysis can be performed by hybridization of at least one nucleic acid to the Adamtsl ⁇ gene of the individual.
  • hybridization can be performed with at least one allele-specific oligonucleotide probe.
  • the analysis can be carried out by sequence analysis or by substitution mapping analysis.
  • a part of the genomic sequence of the haplotypes for the Adamtsl ⁇ gene of the individual is analyzed. Also, the analysis can be carried out by hybridization of a nucleic acid probe to at least one allele of the haplotypes for the Adamtsl ⁇ gene.
  • a method of diagnosis of hypertension in a human includes determining the genotype of at least one micro satellite region of the Adamtsl ⁇ gene in the human, identifying whether the genotype is a risk genotype and, if a risk genotype is so identified, then diagnosing the human as hypertensive.
  • a method of identifying a human predisposed or susceptible to hypertension includes determining the genotype of at least one microsatellite region of the Adamtsl ⁇ gene in the human, identifying whether the genotype is a risk genotype and, if a risk genotype is so identified, then diagnosing the human as susceptible or predisposed to hypertension.
  • the method can include one or more of: determining whether the human is homozygous or heterozygous for polymorphisms of the microsatellite region of the Adamtsl ⁇ gene; and where the determination of the genotype of the region is accomplished by screening the region to identify a risk polymorphism in the region, the risk polymorphism being indicative of risk of hypertension or of predisposition or susceptibility to hypertension.
  • the determining step can be accomplished by amplification of a nucleic acid sequence located within the microsatellite region; and the amplification can be accomplished by the polymerase chain reaction using one or more primers adapted to amplify a nucleic acid sequence located within the microsatellite.
  • the primers have a nucleotide sequence selected from the group of nucleotide sequences described herein.
  • a method of diagnosis and treatment of a human susceptible or predisposed to hypertension includes determining the genotype of one or more of the genomic regions of the Adamtsl ⁇ gene that contain single nucleotide polymorphisms as genotypes and identifying whether the genotype is a risk genotype and, if a risk genotype is so identified, then diagnosing the human as susceptible or predisposed to hypertension, and administering treatment to reduce or delay or prevent hypertension.
  • the treatment is selected from the group consisting of administration of an effective amount of antihypertensive pharmaceutical, administration of an effective anti-hypertension therapy or administration of both an effective anti-hypertension therapy and an effective amount of antihypertensive pharmaceutical.
  • the risk genotype can be one or more of the alleles described herein having a length 1 or >1 bp.
  • a method of diagnosing hypertension or susceptibility to hypertension includes determining the genotype of Adamtsl ⁇ , identifying whether the genotype is a risk genotype and, if a risk genotype is so identified, then diagnosing the human as hypertensive or susceptible to hypertension, wherein an allele of 1 or >1 bp indicates a risk of hypertension or of susceptibility to hypertension and wherein an allele of 1 or >1 bp indicates the absence of risk of hypertension or of susceptibility to hypertension.
  • the risk genotype of the microsatellite region includes an allele of 1 or >1 bp.
  • the non-risk genotype of the D5S207microsatellite region includes an allele of 1 or >lbp.
  • a method for prevention or treatment of a hypertensive disease includes administering to a mammalian subject in need of such treatment an effective amount of a compound in a pharmaceutically acceptable carrier enhancing or reducing biological activity of one or several polypeptides encoded by Adamtsl ⁇ or a variant, fragment or derivative thereof; and/or enhancing or reducing activity of one or several biological networks and/or metabolic pathways related to the polypeptides.
  • the treatment can be gene therapy or gene transfer. Also, the treatment can includes treating regulator ⁇ ' regions and/or gene containing region of one or more the Adamtsl ⁇ gene or variants, fragments or derivatives thereof in somatic cells of the subject.
  • the treatment includes treating regulatory regions and/or gene containing region of one or more the Adamtsl ⁇ gene or variants, fragments or derivatives thereof in stem cells.
  • the compound can be a recombinant polypeptide encoded by the Adamtsl ⁇ gene or variant, fragment or derivative thereof.
  • the treatment can be based on siRNA hybridizing to mRNA and/or to hnRNA of the Adamtsl ⁇ gene.
  • the method of treating is a dietary treatment or a vaccination.
  • the method includes a therapy restoring, at least partially, the observed alterations in biological activity of one or several polypeptides encoded by the Adamtsl ⁇ gene in the subject, when compared with HT free healthy subjects.
  • a method of identifying subjects at risk of developing a hypertensive disease includes: a) providing nucleic acid from a subject, wherein the nucleic acid comprises an Adamtsl ⁇ gene; b) detecting the presence or absence of one or more variations in the Adamtsl ⁇ gene, wherein the variation is selected using at least one marker from the group shown in Table 1 ; and, c) determining if the subject is at risk of developing a hypertensive disease based on the presence or absence of the one or more variations.
  • the detecting in step b) is accomplished by substitution mapping analysis or by hybridization analysis.
  • the detecting in step b) comprises comparing the sequence of the nucleic acid to the sequence of a wild-type Adamtsl ⁇ nucleic acid.
  • a method of identifying a variant, fragment or derivative of Adamtsl ⁇ gene includes: a) providing nucleic acid from a subject, wherein the nucleic acid comprises a Adamtsl ⁇ gene; and b) detecting the presence or absence of one or more variations in the Adamtsl ⁇ gene, wherein the variation is selected using at least one marker shown in Table 1.
  • the method further includes: step c) determining if the subject is at risk of carrying a hypertensive disease allele based on the presence or absence of the one or more variations.
  • the test substance can be administered to the animal, and the blood pressure level of the animal is measured and estimated.
  • the disease caused by the overexpression of Adamtsl ⁇ is a dysfunction of the kidney.
  • the disease can be caused by the overexpression of regucalcin is essential hypertension.
  • the test substance can be administered to the animal, and the blood pressure level of the animal is measured and estimated, hi a particular embodiment, the disease is a hypertensive disease.
  • an animal model where the animal model is a non-human animal that over- or under- expresses the Adamtsl ⁇ gene or a variant, fragment or derivative thereof, and shows a hypertensive pathology.
  • the animal model can be selected by a morphological measurement estimation of blood pressure levels.
  • the animal has characteristics of hypertensive pathology that are stable through many generations.
  • the non-human animal can be a female or a male.
  • the non-human animal is a rat.
  • a screening method of preventive and therapeutic agents for hypertensive diseases where a test substance is administered to an animal model, and a morphological measurement estimation of blood pressure and/or a biochemical measurement estimation of over- or under- expression of Adamtsl ⁇ or a variant, fragment or derivative thereof are performed.
  • the ' screening method of preventive and therapeutic agents is useful where the hypertensive disease is essential hypertension.
  • a method for identification of an individual who has an altered risk of or susceptibility for developing a hypertensive disease includes: providing a biological sample taken from the individual; collecting personal and clinical information of the individual; determining the nucleotides present in one or several of the polymorphic sites described herein in the individual's nucleic acid; and, combining the SNP marker data with personal and clinical information to assess the risk of an individual to develop the hypertensive disease.
  • the altered risk is an increased risk of the hypertensive disease.
  • the altered risk is a decreased risk of the hypertensive disease.
  • the polymorphic sites are those present in the haplotypes described herein.
  • the polymorphic sites are associated with the SNP markers set forth herein.
  • the polymorphic sites are in complete linkage disequilibrium with the SNP markers set forth herein.
  • a method for identification of an individual who has an altered risk of or susceptibility for developing a hypertensive disease includes: providing a biological sample taken from a subject; determining the nucleotides present in one or several of the polymorphic sites as set forth herein in the individual's nucleic acid: and, combining the SNP marker data to assess the risk of an individual to develop the hypertensive disease.
  • the altered risk is an increased risk of the hypertensive disease.
  • the altered risk is a decreased risk of the hypertensive disease.
  • the polymorphic sites are those present in one or more of the haplotypes presented herein.
  • the polymorphic sites are associated with one or more of the SNP markers set forth herein.
  • the polymorphic sites are in complete linkage disequilibrium with one or more of the SNP markers set forth herein.
  • one or several polymorphic sites reside within a hypertensive risk gene or genes as set forth herein.
  • one or several of the SNP markers are selected from the group shown in Table 2.
  • a method for assessing susceptibility or predisposition to a hypertensive disease in an individual includes determining alteration of expression levels of one or several of the markers of Table 1 in the individual, wherein a difference in expression is indicative of susceptibility to a hypertensive disease.
  • Alteration of expression levels can be determined by assessing transcription levels of one or several of the genes of Table 1 in the individual.
  • alteration of expression levels can be determined by assessing translation of mRNAs encoded by one or several of the genes of Table 1 in the individual.
  • test kit based on a method described herein for assessment of an altered risk of or susceptibility for the hypertensive disease in a subject.
  • the test kit can be useful for determining the nucleotides present in one or several of the SNP markers as set forth herein in the individual's nucleic acid for assessment of an altered risk of a hypertensive disease in a subject.
  • test kit for determining the nucleotides present in one or several of the SNP markers as set forth herein tables in the individual's nucleic acid for assessment of an altered risk of a hypertensive disease in a subject, containing: a) reagents and materials for assessing nucleotides present in one or several SNP markers as set forth herein; and b) software to interpret the results of the determination.
  • the test kit can further include a PCR primer set for amplifying nucleic acid fragments containing one or several SNP markers as set forth herein from the nucleic acids of the subject.
  • the test kit can include a capturing nucleic acid probe set specifically binding to one or several SNP markers present in hypertensive disease associated markers and haplotype regions as set forth herein.
  • the test kit can include a microarray or multiwell plate to assess the genotypes.
  • the test kit can include a questionnaire for obtaining patient information concerning age. gender, height, weight, waist and hip circumference, skinfold and adipose tissue thicknesses, the proportion of adipose tissue in the body, the family history of diabetes and obesity, the medical history concerning hypertension.
  • the test kit can also include a capturing nucleic acid probe set specifically binding to one or several SNP markers present in the Adamtsl ⁇ gene.
  • the test kit can include a microarray or multiwell plate to assess the genotypes.
  • an isolated nucleic acid comprising an Adamtsl6-like gene comprising a polymorphism at position 820 as defined by the position shown in Figs. 3 and 9A where the amino acid corresponding to position 274 is not a proline.
  • the nucleotide at position 820 is T.
  • the amino acid at position 274 is serine.
  • an isolated nucleic acid molecule comprising a sequence complementary to the isolated nucleic acid molecule described herein.
  • an isolated nucleic acid comprising an Adamtsl6-like gene comprising a polymorphism at position 3508 as defined by the position shown in Figs. 3 and 9B, where the amino corresponding to position 1170 is not a threonine.
  • the nucleotide at position 3508 is T.
  • the amino acid at position 1170 is a serine.
  • an isolated nucleic acid molecule comprising a sequence complementary to the isolated nucleic acid molecules described herein.
  • a method for detection of at least one single nucleotide polymorphism (SNP) in a human Adamtsl6-like gene includes determining a nucleotide at position 820 in the human Adamtsl6-like gene, and detecting absence or presence of at least one SNP.
  • the amino acid encoded by the single polymorphism at position 820 is histidine.
  • a method for detection of at least one single nucleotide polymorphism (SNP) in a human Adamtsl6-like gene includes determining a nucleotide at position 3508 in the human Adamtsl6-like gene as shown in Fig. 7, and thereby detecting absence or presence of at least one SNP.
  • the amino acid encoded by the single polymorphism at position 3508 is L.
  • an allele-specific nucleic acid primer that comprises between 100 -350 nucleotides which hybridizes to and detects an Adamtsl ⁇ - like gene polymorphism at position 820 in the Adamtsl6-like gene as defined by the positions in Figs. 3 and 9A.
  • the T allele is indicative of increased levels of blood pressure in an animal.
  • FIG. 1 Comprehensive representation of mapping BP QTL2.
  • LOD plot for BP using the F2 (S x LEW) population 41 is shown at the top followed by the congenic strains constructed to map the QTL to a 2.73Mb region shown in red.
  • Linkage map of chromosome 1 is drawn to scale except for distances marked with an asterisk. The current location of this region on the physical map of the rat genome is between the markers shown on the cytogenetic map of rat chromosome 17. The inventor believes that it should lie within IpI 1 of chromosome 1, which is also shown in the diagram.
  • substitution mapping 18 ' 42 ' 43 All the iterations of substitution mapping 18 ' 42 ' 43 are shown by congenic strains with BP lowering effect shown in green and the ones without the BP lowering effect shown in grey.
  • White boxes around the congenic strain schematics represent regions of recombination. The region highlighted in purple denotes the current fine-mapped interval of 789kb. Predicted genes within the critical interval are shown by blue arrows.
  • Figure 2 Predicted Transcript analysis.
  • the horizontal lines at the top are schematic representations of predicted transcripts (RGSC 3.4, Dec 2004) of LOC306664 at the NCBI and Ensembl databases. Primers were designed at the locations specified as A through D in the directions depicted by the arrows. Primers were used in multiple combinations to PCR amplify DNA from a mixed cDNA pool obtained from S or LEW rats.
  • Predicted candidate genes for the 2.73Mb QTL interval are shown at the top of the figure as blue colored boxes within the region flanked by the markers DlRat211 and DlRatl2, as shown in Fig. 1. This information was extracted from the NCBI database. Arrows above the boxes indicate the orientation of the gene sequence on the chromosome. The exons and introns of LOC306664, which codes for Adamtsl ⁇ 5_predicted, are shown above the corresponding transcript sequence. Locations of transcript variants detected between S and LEW are numbered on the transcript sequence.
  • transcript variations in S versus LEW are as follows: (1) T/C 820 , (2) C/T 1482 , (3) C/T 1785 , (4) A/G 1926 , (5) T/C 2226 , (6) C/T 2646 , (7) T/ A 3308 .
  • Resultant amino acid changes and their locations on the schematic diagram of the predicted polypeptide domain of Adamtsl 6 are shown at the bottom of the figure. Sequences disclosed as SEQ ID NOS 1-4, respectively, in order of appearance.
  • FIG. 1 Tissue-specific " expression of Adamtsl 6.
  • the transcript of Adamtsl 6 was amplified from cDNA obtained from various tissues of the LEW rat. RT-PCR was performed as described under materials and methods. PCR products of Adamtsl 6 and Actin as resolved on a 1.5% agarose gel and stained with ethidium bromide are shown in the top and bottom panels respectively.
  • FIG. 5 Fine-mapping the QTL.
  • (a) Congenic substrains that retain smaller regions within the 2.73Mb BP QTL are schematically represented alongside the physical map of the rat in Mbs. The 2.73MB BP QTL is shown by the red line on the physical map. Congenic strains are depicted underneath the physical map. Green colored areas indicate the introgressed homozygous LEW alleles of the congenic substrain and the white boxes flanking the green-colored boxes are the regions of cross-over, (b) Histograms represent the difference in BP of the congenic substrains compared to the BP of S rats. These measurements were obtained in separate experiments by the tail-cuff method.
  • FIG. 6 Mapping of the homologous mouse segment. An overview of Chromosome 13 for the mouse is shown, along with the DNA(contigs), markers, Ensemble Genes, ncRNA genes, and EST genes.
  • Figure 7 Mapping of the homologous human segment. An overview of Chromosome 5 for the human is shown, along with the DNA(contigs). markers. Ensemble Genes, ncRNA genes, and EST genes.
  • Figure 8 Comparison of Rat Adamtsl ⁇ gene to Human Adamtsl ⁇ gene. Shown is a comparison of rat Adamtsl ⁇ polypeptide domain organization to human Adamtsl ⁇ polypeptide domain organization. Shown are the exons, protein domains, the furin recognition site, TS thrombospondin site, and the protease and lacunin domain (PLAC). Indicated with arrows are: the rat nonsynonymous SNP / amino acid variant location, the corresponding location of rat nonsynonymous SNP / amino acid variant location, the rat synonymous SNP / amino acid location, corresponding location of Rat synonymous SNP / amino acid location, and human nonsynonymous SNP / amino acid variant location.
  • Figures 9A and 9B Shown are amino acid sequences for human (SEQ ID NO: 5), splice (SEQ ID NO: 6), mouse (SEQ ID NO: 7), rat (S) (SEQ ID NO: 8), rat (LEW) (SEQ ID NO: 9) and cow (SEQ ID NO: 10).
  • the positions 270 and 1170 are boxed in red.
  • Table 4 shows human ADAMST 16 variants with significant (p ⁇ 0.05) associations to systolic, diastolic and mean arterial blood pressure.
  • the present invention provides a method of identifying subjects at risk of developing hypertension comprising: providing nucleic acid from a subject, where the nucleic acid comprises an Adamtsl ⁇ gene; and detecting the presence or absence of one or more variations in the Adamtsl ⁇ gene.
  • the method further comprises determining if the subject is at risk of developing hypertension based on the presence or absence of the one or more variations.
  • the variation in the method of the present invention is a single nucleotide polymorphism that causes a frameshift mutation in Adamtsl ⁇ . or the variation causes a splice mutation in Adamtsl ⁇ . or the variation causes a nonconservative amino acid substitution Adamtsl 6.
  • the variation is selected from the group consisting of the mutations shown in Table 1.
  • the detecting is accomplished by hybridization analysis.
  • the detecting step comprises comparing the sequence of the nucleic acid to the sequence of a wild-type Adamtsl ⁇ nucleic acid.
  • the present invention also provides a method of identifying subjects at risk of developing hypertension comprising: providing a blood or other biological sample from a subject, wherein the blood or other biological sample comprises an Adamtsl 6 protease; and detecting the presence or absence of one or more variants of the Adamtsl 6 protease.
  • the detecting step is accomplished by an antibody assay.
  • the invention also provides a method of identifying subjects at risk of carrying an allele for hypertension comprising: providing nucleic acid from a subject, wherein the nucleic acid comprises an Adamtsl 6 gene; and detecting the presence or absence of one or more variations in the Adamtsl ⁇ gene.
  • the method of the present invention further comprises a step of determining if the subject is at risk of carrying a hypertensive disease based on the presence or absence of the one or more variations.
  • the present invention also provides an isolated nucleic acid comprising a sequence encoding a polypeptide of Adamtsl 6.
  • the present invention also provides a method of treating a patient with a hypertensive disease, comprising administering a therapeutically effective amount of an Adamtsl 6 protease such that the symptoms of the hypertensive disease are alleviated.
  • the Adamtsl ⁇ protease is selected from the group consisting: recombinant Adamtsl ⁇ ; synthetic Adamtsl ⁇ ; mutants, variants, fragments, and fusions of recombinant Adamtsl 6; and mutants, variants, fragments, and fusions of synthetic Adamtsl 6.
  • the present invention also provides a kit for determining if a subject is at risk of developing hypertension comprising a detection assay.
  • the detection assay is capable of specifically detecting a variant Adamtsl ⁇ allele.
  • the detection assay comprises a nucleic acid probe that hybridizes under stringent conditions to a nucleic acid sequence comprising at least one mutation selected from the group consisting of the mutations shown in Table 1.
  • the invention also provides an isolated nucleic acid comprising at least one marker sequence noted with an asterisk (*) as shown in Table 1.
  • the sequence is operably linked to a heterologous promoter.
  • the invention provides a vector comprising the isolated sequence.
  • the invention provides a host cell comprising the vector.
  • the host cell is selected from the group consisting of animal and plant cells: in other embodiments, the host cell is located in an organism.
  • the invention also provides an isolated nucleic acid sequence comprising at least one sequence noted with an asterisk ⁇ *) as shown in Table 1.
  • the invention provides a computer readable medium encoding a representation of the nucleic acid sequence.
  • the invention also provides an isolated polypeptide comprising an amino acid sequence encoded by the sequence noted with an asterisk ⁇ *) as shown in Table 1.
  • the identification of the association between the Adamtsl ⁇ gene and hypertension permits the screening of individuals to determine a predisposition to hypertension. Those individuals who are identified at risk for the development of the disease may benefit from dietary sodium restriction, can have their blood pressure more closely monitored and be treated at an earlier time in the course of the disease. Such blood pressure monitoring and treatment ma ⁇ ' be performed using conventional techniques well known in the art.
  • the invention is based, in part, upon the finding that the activity the Adamtsl ⁇ gene is central to the etiology of hypertension and hypertension-related disorders in mammals. Accordingly, the invention provides methods for treating hypertension using specific inhibitors of Adamtsl ⁇ expression and/or activity, particularly the expression and/or activity of the Adamtsl ⁇ gene.
  • Metalloproteases are a class of enzymes that have been implicated in a large number of cellular processes. Despite the fact that these enzymes differ widely in their sequence and structure, they all rely on the action of a catalytically active metal atom, hi most instances this metal is a zinc atom, but examples are known which contain cobalt or nickel. Metalloproteases use the catalytic metal atom to catalyze hydrolysis of peptide bonds. Currently, metalloproteases have been divided into at least five distinct families based on structure and activity.
  • Adam A Disintegrin And Metalloprotease
  • the Adam metalloproteases consist of a prodomain that blocks protease activity; a zinc-binding metalloprotease domain: disintegrin and cysteine-rich domains with adhesion activity; an epidermal growth factor-like domain with cell fusion activity; a transmembrane domain; and a phosphorylated cytoplasmic regulatory domain.
  • a subfamily of the Adam metalloproteases can be distinguished from other the Adam metalloproteases family members based on the multiple copies of thrombospondin 1 -like repeats they carry.
  • Adamts 16 is a member of a family of metalloproteinases, the function of which is unknown 2 " . Also, none of the Adamts proteins are previously implicated in blood pressure regulation.
  • QTL2 is localized to a 2.73 Mb region, as determined on the basis of the differential segment between two congenic strains (see Fig. 1), one with the BP effect and the other without the BP effect 18 .
  • Fig. 1 provides a comparison between substitution mapping and the evidence obtained by genetic linkage analysis of the same region.
  • the location of the QTL gene is far removed from the confidence intervals of LOD peaks with suggestive linkage (Fig 1).
  • This comparison reiterates the difficulty of mapping QTL genes solely based on linkage analysis as is done in humans.
  • This comparison also exemplifies the power of substitution mapping in animal models to resolve QTLs by isolating them as 'monogenic' QTL effectors. Since substitution mapping relies on location on the genome, any bias of previous knowledge of physiological function of a gene related to the phenotype of interest is removed, allowing for identification of novel QTL genes.
  • One advantage of using this method is exemplified by the data presented herein because the fine-mapped BP QTL did not encompass any gene with previously known function related to BP.
  • Table 1 shows the physical map locations of micro satellite markers that are linked to RNOl by genetic linkage analysis of F2(S x LEW).
  • Microsatellite markers are those that are placed in order on the genetic linkage map of F2(S x LEW) (Physiol Genomics. 2001, 4:201-214). *Newly developed markers.
  • Adamtsl ⁇ was highly expressed in the kidney, which is a major organ involved in BP homeostasis. While not wishing to be held to theory, the inventor believes that, based on the roles of the Adamts family members and based on the presence of a metalloproteinase domain within Adamtsl ⁇ , it is likely that Adamtsl ⁇ functions as a metalloproteinase in vivo.
  • the single nucleotide polymorphisms described herein do not reside within the metalloproteinase domain of Adamtsl ⁇ . They are located within a potential furin- cleavage site and a thrombospondin-motif, respectively, which may or may not influence the metalloproteinase activity of Adamtsl ⁇ . Furin-cleavage is known to relieve enzyme latency of other members of the Adamts family of proteins 31 ⁇ 36 . Alternately, thrombospondin-motifs have protein-protein interactions with integrins, which in turn are implicated in vascular remodeling 37 . Identifying substrates and/or binding partners for Adamtsl ⁇ is an important step in linking the physiological role of Adamtsl ⁇ to blood pressure regulation.
  • MHS Milan hypertensive strain
  • SHR spontaneously hypertensive strain
  • R Dahl salt-resistant strain
  • WKY Wistar Kyoto rat
  • LN Lyon normotensive rat
  • LL Lyon low-tensive or hypotensive rat
  • MNS Milan normotensive strain
  • BN Brown Norway rat
  • SBN Sabra hypertension resistant
  • SBH Sabra hypertension prone
  • MWF Munich Wistar Fromter rat
  • SD Sprague Dawley rat. Further details on the strains are available at the rat genome database. Wild rat samples were from Prof.
  • Cowley indicates that the gold standard for validation of a potential candidate gene for hypertension is that the rescue of the phenotype must be done in the same species and with the same genetic background that was used to identify the QTL.
  • the present invention provides a 'gold standard' because (1) The QTL is fine-mapped to a small region solely based on natural recombination events, thus closely mimicking the performance of a natural allele in the rat as opposed to artificially genetically manipulated systems such as transgenics and knock-ins; and (2) it is now possible to map the QTL to a gene-sparse region as confirmed by comparative mapping in rats, mice and humans 18 , thus allowing for testing of the two expressed candidate transcripts, only one of which had coding sequence variants.
  • Congenic substrains S.LEW(D1MCO4X1X3BX1) and S.LEW(D1MCO4X1X3BX2) were derived from S.LEW(D1MCO4X1X3B) as the progenitor with LEW alleles at the QTL region.
  • the congenic substrains were constructed by crossing the progenitor congenic strain to the S rat to obtain a heterozygous Fl population. An Fl X Fl intercross then yielded an F2 population.
  • the recombinant F2 animals were selected for varying sizes of the LEW RNOl introgressed region and backcrossed to the S rats.
  • the resulting heterozygous progeny were genotyped to identify LEW alleles and then intercrossed to duplicate the recombinant chromosome. Finally the animals were selectively bred to generate a homozygous congenic rat strain containing the desired LEW chromosomal segment on the S genomic background.
  • Microsatellite repeats in the region of interest were identified on the rat genomic sequence (version 3.1) at the NCBI website.
  • primers were designed using the PrimerS software available and synthesized by Integrated DNA technologies.
  • DNA was isolated with the QIAamp Tissue Kit (Qiagen Inc.) from S, LEW and congenic substrains.
  • PCR reactions were set up using the primers designed to amplify microsatellite repeats from S and LEW rats.
  • Polymorphic repeats were used as markers to genotype DNA from tail biopsies of the relevant rat populations.
  • Predicted gene sequences within the QTL region were downloaded from the rat genome sequence at the NCBI website. Primers to amplify exons were designed using the ExonPrimer software. Primers tagged with M- 13 sequences were synthesized by Integrated DNA technologies (IDT). All primer sequences mentioned in our work are available. Genomic DNA was amplified using these primers and sequenced by MWG Biotech Inc.
  • Predicted transcript sequences were extracted from the NCBI database and/or Ensembl. Primers were designed encompassing the start and stop codons of each predicted transcript using Primer3 or Oligonucleotide properties calculator.
  • Messenger RNA from S and LEW kidney, liver, heart, lung, thyroid, pancreas, and spleen was extracted using TRIzol Reagent (Life Technologies).
  • cDNA of the predicted transcripts within the QTL region was obtained by reverse transcription using the Superscript III First-Strand Synthesis System for RT-PCR (Invitrogen). Standard polymerase chain reaction, using Platinum Taq DNA Polymerase High Fidelity (Invitrogen) and gene- specific primers, was used to amplify the cDNA.
  • BP was also collected using a telemetry system (Data Sciences International, St. Paul, MN) as explained in detail previously .
  • BP QTL2 was fine-mapped to a 2.73Mb region flanked by the microsatellite markers DlRat211 and Dl Rat 12.
  • Table 1 lists the microsatellite markers in and around this 2.73Mb region that are linked to the map of chromosome 1.
  • Fig. 1 is a comprehensive representation of markers genotyped for linkage analysis and for the construction and genetic analysis of congenic strains, which lead to the fine-mapping of BP QTL2 to 2.73Mb.
  • the 2.73Mb BP QTL2 region is represented as a red highlighted band on rat chromosome 17 instead of on rat chromosome 1.
  • the 2.73 Mb QTL region had 19 annotated genes 18 . Because homologous regions of this QTL in mice and humans encompass BP QTLs 19 ' 29 , we chose to focus on sequencing Rat genes that have orthologues in both mouse and human. Two out of the four orthologous gene sequences annotated on the rat genome database at NCBI (LOC306664 and LOC290935) were predicted as associated with Adamts 16, an orphan disintegrin-like metalloproteinase with thrombospondin motif 16 18 . We sequenced all the exons predicted by the NCBI database. Seven nucleotide substitutions within predicted exon sequences of Adamts 16 were detected. Since the exon predictions of Adamts 16 were different between the NCBI and Ensembl databases, the issue of whether these polymorphic variants resulted in amino acid substitutions was not immediately discernable.
  • the PCR amplified samples from both S and LEW rats were cloned and sequence confirmed as the transcript of Adamtsl, which confirmed the NCBI predicted transcript. Only two out of the seven polymorphisms detected by genomic DNA sequence analysis of LOC306664 were nonsynonymous variants that resulted in amino acid substitutions of the predicted mature polypeptide chain of Adamts 16 from Pro 274 and Thr 1170 in LEW rats to Ser 274 and Ser 1170 in S rats (Fig. 3).
  • the variant at position 274 maps to no particular domain of Adamtsl ⁇ , but lies in between the propeptide and metalloproteinase domains (Fig. 3).
  • the variant at 1 170 maps to the C-terminal thrombospondin domain of Adamts 16 (Fig.
  • a gene Underlying a QTL may either have coding-sequence differences that change the function of the protein it encodes or may have differences in regulatory- sequences that cause a difference in gene expression between the parental strains or both.
  • Reverse-transcription PCR of cDNA samples from various tissues of the LEW rat were probed with primers designed to amplify the full length Adamtsl 6 transcript. (Fig. 4).
  • Expression of Adamtsl 6 was prominent in the rat kidney, but detected in other tissues including the heart, pituitary, brain and lung (Fig. 4).
  • Real-time PCR of kidney and heart samples revealed that Ad ⁇ mtsl ⁇ is not differentially expressed in these organs (data not shown).
  • Negative values indicate a decrease in the congenic effect compared to the S rat.
  • Number of rats in each group ranged from 20 to 30.
  • a first aspect of the present invention is directed to methods for identifying individuals predisposed to essential hypertension.
  • the methods can be conducted using a sample of cells that express the Adamts 16 gene or a promoter thereof, isolated from the individual.
  • the cells are assayed to determine the extent of post-translational modification, where a change relative to cells isolated from a normotensive individual is indicative of predisposition to essential hypertension.
  • a nucleic acid sample can be isolated from the individual in order to analyze the Adamts 16 gene or fragment thereof to detect Adamts 16 associated with essential hypertension.
  • Also provided is a method for identifying a human subject as having an increased risk of developing essential hypertension comprising correlating the presence of a nucleic acid sequence at nucleotides -820 through -3508 in a human Adamts 16 gene with an increased risk of developing essential hypertension; and determining the nucleic acid sequence of nucleotides -820 through -3508 of the subject's Adamtsl ⁇ gene, whereby a subject having a nucleic acid sequence at nucleotides -820 through -3508 in the Adamtsl ⁇ gene correlated with an increased risk of developing essential hypertension is identified as having an increased risk of developing essential hypertension.
  • a method of identifying an allele in the promoter region of the human Adamtsl ⁇ gene correlated with an increased risk of developing essential hypertension comprising determining the nucleic acid sequence of the nucleotides at positions -820 through -3508 in the human Adamtsl ⁇ gene from a subject; and correlating the presence of the nucleic acid sequence of the subject with the diagnosis of essential hypertension in the subject, whereby the nucleic acid sequence of the nucleotides at positions -820 through -3508 in the promoter region of the human Adamtsl ⁇ gene identifies an allele correlated with an increased risk of developing essential hypertension.
  • a method of identifying an allele in the promoter region of a human Adamtsl ⁇ gene correlated with an increased risk of developing essential hypertension comprising determining the nucleic acid sequence of the nucleotides at positions -820 through -3508 in the promoter region of the human Adamtsl ⁇ gene; cloning the nucleic acid sequence encoding the promoter region of the human Adamtsl ⁇ gene into an expression vector having a reporter gene; expressing the reporter gene to produce a gene product under the direction of the promoter region of the human Adamtsl ⁇ gene; and determining the amount of reporter gene product produced, an amount of reporter gene product less than the amount of reporter gene product produced under the direction of a Adamtsl ⁇ gene promoter containing an allele of a nucleic acid sequence of the nucleotides at position -820 through -3508 of the human Adamtsl ⁇ gene where an allele correlated with an increased risk of developing essential hypertension.
  • a method for identifying a human subject as having an increased risk of developing essential hypertension comprising cloning the nucleic acid sequence encoding the promoter region of the human Adamtsl ⁇ gene of the subject into an expression vector having a reporter gene; expressing the reporter gene to produce a gene product under the direction of the promoter region of the human Adamts 16 gene; and determining the amount of reporter gene product produced, an amount of reporter gene product less than the amount of reporter gene product produced under the direction of a Adamts 16 gene promoter containing the nucleotide sequence of one or more alleles that identify a subject as having an increased risk of developing essential hypertension.
  • a method for identifying an allele in the promoter region of the human Adamts 16 gene is provided, comprising determining the nucleic acid sequence of the nucleotides at positions -820 through -3508.
  • the present invention further provides an isolated nucleic acid consisting of the promoter region of the human Adamts 16 gene, the complementary sequence of the isolated nucleic acid, vectors containing the nucleic acid and cells expressing the vector.
  • the present invention also provides a method of identifying a subj ect at reduced risk of having essential hypertension, comprising determining the presence in the subject of an allele in the promoter region of the subject's Adamtsl6 gene correlated with a reduced risk of developing essential hypertension, the presence of the allele identifying the subject as having a reduced risk of developing essential hypertension.
  • a method for identifying a human subject as having a reduced risk of developing essential hypertension comprising correlating the presence of a nucleic acid sequence at nucleotides -820 through -3508 in a human Adamts 16 gene with a reduced risk of developing essential hypertension; and determining the nucleic acid sequence of nucleotides -820 through -3508 of the subject's Adamts 16 gene, whereby a subject having a nucleic acid sequence at nucleotides -820 through -3508 in the Adamtsl6 gene correlated with a reduced risk of developing essential hypertension is identified as having a reduced risk of developing essential hypertension.
  • a method for identifying an allele in the promoter region of the human Adamts 16 gene correlated with a reduced risk of developing essential hypertension comprising determining the nucleic acid sequence of the nucleotides at positions -820 through -3508 in the human Adamtsl6 gene from a subject; and correlating the presence of the nucleic acid sequence of the subject with the absence of essential hypertension in the subject, whereby the nucleic acid sequence of the nucleotides at positions -820 through -3508 in the promoter region of the human Adamts 16 gene identifies an allele correlated with a reduced risk of developing essential hypertension.
  • the present invention further provides a method of identifying an allele in the promoter region of a human Adamts 16 gene correlated with a reduced risk of developing essential hypertension, comprising: determining the nucleic acid sequence of the nucleotides at positions -820 through -3508 in the promoter region of the human Adamts 16 gene; cloning the nucleic acid sequence encoding the promoter region of the human Adamts 16 gene into an expression vector having a reporter gene; expressing the reporter gene to produce a gene product under the direction of the promoter region of the human Adamts 16 gene; determining i) the amount of reporter gene product produced, ii) an amount of reporter gene product greater than the amount of reporter gene product produced under the direction ' of a Adamts 16 gene promoter containing the nucleotide sequence of one or more alleles thereof; and identifying a nucleic acid sequence of the nucleotides at position -820 through - 3508 of the human Adamts 16 gene as an allele
  • Also provided is a method for identifying a human subject as having a reduced risk of developing essential hypertension comprising cloning the nucleic acid sequence encoding the promoter region of the human Adamts 16 gene of the subject into an expression vector having a reporter gene; expressing the reporter gene to produce a gene product under the direction of the promoter region of the human Adamts 16 gene; and determining the amount of reporter gene product produced, an amount of reporter gene product greater than the amount of reporter gene product produced under the direction of a Adamts 16 gene promoter, and identifying a subject as having a reduced risk of developing essential hypertension.
  • the present invention provides a method of identifying a human subject as having an increased likelihood of regulating blood pressure with dietary sodium intake, comprising determining the presence in the subject of a Adamts 16 promoter allele genotype correlated with an increased likelihood of regulating blood pressure with dietary sodium intake, whereby the presence of the genotype identifies the subject as having an increased likelihood of regulating blood pressure by dietary sodium intake.
  • a method of identifying a human subject as having an increased likelihood of regulating blood pressure with dietary sodium intake comprising a) correlating the presence of a Adamts 16 promoter allele genotype with an increased likelihood of regulating blood pressure with dietary sodium intake; and b) determining the Adamts 16 promoter allele genotype of the subject, whereby a subject having a Adamts 16 promoter allele genotype correlated with an increased likelihood of regulating blood pressure with dietary sodium intake is identified as having an increased likelihood of regulating blood pressure with dietary sodium intake.
  • a method for identifying a Adamts 16 promoter allele genotype correlated with an increased likelihood of regulating blood pressure with dietary sodium intake comprising a) determining the Adamts 16 promoter allele genotype of a subject; and b) correlating the presence of the genotype of step (a) with the ability of the subject to regulate blood pressure by dietary sodium intake, thereby identifying a genotype correlated with an increased likelihood of regulating blood pressure by dietary sodium intake.
  • the present invention provides a method for identifying a human subject as having a reduced likelihood of regulating blood pressure with dietary sodium intake, comprising determining the presence in the subject of a Adamts 16 promoter allele genotype correlated with a reduced likelihood of regulating blood pressure with dietary sodium intake, whereby the presence of the genotype identifies the subject as having a reduced likelihood of regulating blood pressure with dietary sodium intake.
  • a method of identifying a human subject as having a reduced likelihood of regulating blood pressure with dietary sodium intake comprising a) correlating a Adamts 16 promoter allele genotype with a reduced likelihood of regulating blood pressure with dietary sodium intake; and b) determining the Adamts 16 promoter allele genotype of the subject, whereby a subject having a Adamts 16 promoter allele genotype correlated with a reduced likelihood of regulating blood pressure with dietary sodium intake is identified as having a reduced likelihood of regulating blood pressure with dietary sodium intake.
  • a method is provided of identifying a Adamts 16 promoter allele genotype correlated with a reduced likelihood of regulating blood pressure with dietary sodium intake, comprising a) determining the Adamts 16 promoter allele genotype of a subject: and b) correlating the presence of the genotype of step (a) with the inability of the subject to regulate blood pressure by dietary sodium intake, thereby identifying the genotype correlated with a reduced likelihood of regulating blood pressure by dietary sodium intake.
  • the present invention provides a method of identifying a human subject as having an increased likelihood of regulating blood pressure with a diuretic, comprising determining the presence in the subject of a.
  • Adamtsl 6 promoter allele genotype correlated with an increased likelihood of regulating blood pressure with a diuretic whereby the presence of the genotype identifies the subject as having an increased likelihood of regulating blood pressure with a diuretic.
  • a method of identifying a human subject as having an increased likelihood of regulating blood pressure with a diuretic comprising a) correlating the presence of &Adamtsl6 promoter allele genotype with an increased likelihood of regulating blood pressure with a diuretic; and b) determining the Adamtsl 6 promoter allele genotype of the subject, whereby a subject having a Adamtsl 6 promoter allele genotype correlated with an increased likelihood of regulating blood pressure with a diuretic is identified as having an increased likelihood of regulating blood pressure with a diuretic.
  • Also provided herein is a method of identifying a Adamtsl 6 promoter allele genotype correlated with an increased likelihood of regulating blood pressure with a diuretic, comprising a) determining the Adamtsl 6 promoter allele genotype of a subject; and b) correlating the presence of the genotype of step (a) with the ability of the subject to regulate blood pressure with a diuretic, thereby identifying a genotype correlated with an increased likelihood of regulating blood pressure with a diuretic.
  • a method of identifying a human subject as having a reduced likelihood of regulating blood pressure with a diuretic comprising determining the presence in the subject of a Adamtsl 6 promoter allele genotype correlated with a reduced likelihood of regulating blood pressure with a diuretic, whereby the presence of the genotype identifies the subject as having a reduced likelihood of regulating blood pressure with a diuretic.
  • the present invention provides a method of identifying a human subject as having a reduced likelihood of regulating blood pressure with a diuretic, comprising a) correlating a. Adamtsl 6 promoter allele genotype with a reduced likelihood of regulating blood pressure with a diuretic; and b) determining the Adamtsl 6 promoter allele genotype of the subject, whereby a subject having a Adamtsl 6 promoter allele genotype correlated with a reduced likelihood of regulating blood pressure with a diuretic is identified as having a reduced likelihood of regulating blood pressure with a diuretic.
  • a method of identifying a Adamtsl 6 promoter allele genotype correlated with a reduced likelihood of regulating blood pressure with a diuretic comprising a) determining the Adamtsl 6 promoter allele genotype of a subject; and b) correlating the presence of the genotype of step (a) with the inability of the subject to regulate blood pressure with a diuretic, thereby identifying the genotype as correlated with a reduced likelihood of regulating blood pressure with a diuretic.
  • the present invention relates to identification of specific polymorphisms of the Adamtsl 6 gene associated with human hypertension.
  • the analysis of the Adamtsl 6 gene for these polymorphisms will identify subjects with a genetic predisposition to develop essential hypertension or pregnancy-induced hypertension. The management of hypertension in these subjects could then be more specifically managed, e.g., by dietary sodium restriction, by carefully monitoring blood pressure and treating with conventional drugs, by the administration of certain inhibitors or by the administration of drugs to inhibit the synthesis of Adamtsl 6.
  • the analysis of the Adamtsl 6 gene is performed by comparing the DNA sequence of an individual's Adamtsl 6 gene with the DNA sequence of the native, non- variant Adamtsl 6 gene.
  • the invention provides several new polymorphisms as described herein that can be can be used to determine the predisposition to hypertension.
  • the invention provides a method of that which can be used in place of, or in addition to, an analysis based upon the previously known molecular variants.
  • DNA sequencing of the entire Adamtsl 6gene in a series of study subjects can lead to the identification of a number single nucleotide polymorphisms (SNPs) in the Adamtsl 6 gene.
  • SNPs single nucleotide polymorphisms
  • Typing of a number of these SNPs in larger series of subjects can provide the definition of the haplotype structure of the Adamtsl ⁇ gene, that is, the observed distribution of these genetic variants on human chromosomes.
  • These data can provide the most common haplotypes that are sufficient to describe the majority of the variation observed in the Adamtsl 6 gene in either population.
  • the invention provides a reduced set of SNPs that can be used to characterize such haplotypes by conventional DNA typing methods. Further evaluation of this variation aids in assessing predisposition for hypertension.
  • the analysis of the Adamtsl 6 gene for molecular variants will identify subjects with a genetic predisposition to develop essential hypertension or pregnancy-induced hypertension.
  • the present invention also relates to the identification of haplotypes of the Adamtsl ⁇ gene which can also be used to determine predisposition to hypertension.
  • the haplotype of an individual is analyzed for the alleles described herein and the presence of a particular haplotype is then associated with a predisposition to hypertension.
  • Shimkets, ' R.A. et al. Liddle's syndrome Heritable human hypertension caused by - mutations in the beta epithelial sodium channel. Cell 79, 407-414 (1994).
  • Tortorella, M.D. et al. ADAMTS-4 (aggrecanase-1): N-terminal activation mechanisms. Arch Biochem Biophys 444, 34-44 (2005).

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Pathology (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Measuring Pulse, Heart Rate, Blood Pressure Or Blood Flow (AREA)

Abstract

La présente invention concerne un système destiné à évaluer un locus à caractère quantitatif (QTL) ou un fragment fonctionnel dérivé de celui-ci, le QTL contrôlant au moins partiellement la pression sanguine et/ou étant au moins partiellement indicatif d'une possibilité de développer de l'hypertension.
PCT/US2008/002938 2007-03-06 2008-03-05 Système de régulation de la pression sanguine Ceased WO2008109110A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US90518607P 2007-03-06 2007-03-06
US60/905,186 2007-03-06

Publications (2)

Publication Number Publication Date
WO2008109110A2 true WO2008109110A2 (fr) 2008-09-12
WO2008109110A3 WO2008109110A3 (fr) 2008-12-11

Family

ID=39738985

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2008/002938 Ceased WO2008109110A2 (fr) 2007-03-06 2008-03-05 Système de régulation de la pression sanguine

Country Status (1)

Country Link
WO (1) WO2008109110A2 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013159085A1 (fr) * 2012-04-20 2013-10-24 University Of Virginia Patent Foundation Compositions et procédés d'identification et de diagnostic de la sensibilité au sel de la pression sanguine
CN114636774A (zh) * 2022-04-22 2022-06-17 苏州市疾病预防控制中心 一种预测青少年高血压患病风险的生物代谢标志物组合物

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
CAL S. ET AL.: 'Cloning, Expression Analysis, and Structural Characterization of Seven Novel Human ADAMTSs, a Family of Metalloproteinases with Disintegrin and Thrombospondin-1 Domains' GENE vol. 283, 2002, pages 49 - 62, XP002207625 *
GARRETT M. ET AL.: 'Identification of Blood Pressure Quantitative Trait Loci that Differentiate Tow Hypersensitive Strains' JOURNAL OF HYPERTENSION vol. 20, 2002, pages 2399 - 2406 *
GOERTSCHES R. ET AL.: 'Genetic Association Between Polymorphisms in the ADAMTS14 Gene and Multiple Sclerosis' JOURNAL OF NEUROIMMUNOLOGY vol. 164, 2005, pages 140 - 147, XP004941875 *
HILLS C. ET AL.: 'Cri du Chat Syndrome and Congential Heart Disease: A Review of Previously Reported Cases and Presentation of an Additional 21 Cases From the Pediatric Cardiac Care Consortium' PEDIATRICS vol. 117, May 2006, pages E924 - E927 *
JONES G.C. ET AL.: 'ADAMTS Proteinases: a Multi-Domain, Multi-Functional Family with Roles in Extracellular Matrix Turnover and Arthritis' ARTHRITIS RESEARCH AND THERAPY vol. 7, 2005, pages 160 - 169, XP009058555 *
SANADA H. ET AL.: 'Single-Nucleotide Polymorphisms for Diagnosis of Salt-Sensitive Hypertension' CLINICAL CHEMISTRY vol. 52, 2006, pages 1 - 9 *
ST. LEZIN E. ET AL.: 'Effect of Renin Gene Transfer on Blood Pressure in the Spontaneously Hypersensitive Rat' HYPERTENSION vol. 31, no. PART 2, 1998, pages 373 - 377 *
SUGIYAMA F. ET AL.: 'Concordance of Murine Quantitative Trait Loci for Salt-Induced Hypertension with Rat and Human Loci' GENOMICS vol. 71, 2001, page 7077 *
WILSON T.W. ET AL.: 'Effects of Lipid-Lowering Agents in the Dahl Salt-Sensitive Rat' HYPERTENSION no. PART 3, 1998, pages 225 - 231 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013159085A1 (fr) * 2012-04-20 2013-10-24 University Of Virginia Patent Foundation Compositions et procédés d'identification et de diagnostic de la sensibilité au sel de la pression sanguine
US9708664B2 (en) 2012-04-20 2017-07-18 University Of Virginia Patent Foundation Compositions and methods for identifying and diagnosing salt sensitivity of blood pressure
CN114636774A (zh) * 2022-04-22 2022-06-17 苏州市疾病预防控制中心 一种预测青少年高血压患病风险的生物代谢标志物组合物
CN114636774B (zh) * 2022-04-22 2024-05-17 苏州市疾病预防控制中心 一种预测青少年高血压患病风险的生物代谢标志物组合物

Also Published As

Publication number Publication date
WO2008109110A3 (fr) 2008-12-11

Similar Documents

Publication Publication Date Title
Azzedine et al. Mutations in MTMR13, a new pseudophosphatase homologue of MTMR2 and Sbf1, in two families with an autosomal recessive demyelinating form of Charcot-Marie-Tooth disease associated with early-onset glaucoma
US6525185B1 (en) Polymorphisms associated with hypertension
Joe et al. Positional identification of variants of Adamts16 linked to inherited hypertension
Watzka et al. Thirteen novel VKORC1 mutations associated with oral anticoagulant resistance: insights into improved patient diagnosis and treatment
US20030092013A1 (en) Diagnosis and treatment of vascular disease
CN102317470A (zh) 促进前列腺癌风险的遗传性变型
Samollow et al. First-generation linkage map of the gray, short-tailed opossum, Monodelphis domestica, reveals genome-wide reduction in female recombination rates
CN106957907B (zh) 用于狗中的肝铜积累的遗传检测
WO2003020118A2 (fr) Diagnostic et traitement des maladies vasculaires
EP1852505B1 (fr) Mutations dans des canaux d'ion
Rolland et al. Leptin receptor gene in a large cohort of massively obese subjects: no indication of the fa/fa rat mutation. Detection of an intronic variant with no association with obesity
WO2008109110A2 (fr) Système de régulation de la pression sanguine
Yang et al. Molecular analysis in newborns from Texas affected with galactosemia
Zee et al. Frequencies of variants of candidate genes in different age groups of hypertensives
WO2003020120A2 (fr) Diagnostic et traitement de maladies vasculaires
JP2005529585A (ja) Ｉｌ−１遺伝子群ならびに関連する炎症性多型およびハプロタイプ
Yoon et al. Analysis of VNTRs in the solute carrier family 6, member 18 (SLC6A18) and lack of association with hypertension
WO2000000647A1 (fr) Procede pour determiner une predisposition a l'hypertension
KR101447098B1 (ko) 플라스미노겐 활성억제효소-1 유전자 다형성 인자를 이용하여 원인불명 반복자연유산 발병을 예측 또는 진단하는 방법
WO2000071751A1 (fr) Gene du diabete
EP2129801B1 (fr) Tbc1d1 comme marqueur diagnostique de l'obesite ou du diabete
EP1259649B1 (fr) Procedes et compositions pour le diagnostic et le traitement de la pseudoxanthoma elasticum et des pathologies associees
WO2003007801A2 (fr) Diagnostic et traitement d'une maladie vasculaire
EP1283889A1 (fr) 2 alpha kinase 3 facteur mute d'initiation de traduction eucaryote (eif2ak3), chez des patients a diabete neonatal insulino-dependant et a dysplasie epihysaire (syndrome de wolcott-rallison)
KR20060096437A (ko) 비만증에 대한 유전적 표식형질

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 08742025

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase in:

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 08742025

Country of ref document: EP

Kind code of ref document: A2