WO2009130465A2 - Marqueurs - Google Patents

Marqueurs Download PDF

Info

Publication number
WO2009130465A2
WO2009130465A2 PCT/GB2009/001036 GB2009001036W WO2009130465A2 WO 2009130465 A2 WO2009130465 A2 WO 2009130465A2 GB 2009001036 W GB2009001036 W GB 2009001036W WO 2009130465 A2 WO2009130465 A2 WO 2009130465A2
Authority
WO
WIPO (PCT)
Prior art keywords
tacrl
adhd
gene
subject
bipolar
Prior art date
Application number
PCT/GB2009/001036
Other languages
English (en)
Other versions
WO2009130465A3 (fr
Inventor
Hugh Malcolm Douglas Gurling
Andrew Mcquillan
Susan Clare Stanford
Stephen Hunt
Original Assignee
Ucl Business Plc
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 Ucl Business Plc filed Critical Ucl Business Plc
Publication of WO2009130465A2 publication Critical patent/WO2009130465A2/fr
Publication of WO2009130465A3 publication Critical patent/WO2009130465A3/fr

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
    • 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
    • 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

  • the invention relates to genetic markers that may be used to indicate an increased likelihood of a subject to suffer from certain disorders, particularly alcoholism, depression, anxiety, aggression and attention deficit hyperactivity disorder (ADHD). Further the invention relates to a method of using the markers to identify a subset of subjects to be screened for DNA mutations that cause susceptibility to such disorders.
  • certain disorders particularly alcoholism, depression, anxiety, aggression and attention deficit hyperactivity disorder (ADHD).
  • ADHD attention deficit hyperactivity disorder
  • Diagnosis of disorders such as conduct disorders, alcoholism, excessive drinking, ADHD, depression and related panic or general anxiety disorders has, until now, been carried out by using clinical criteria only. It would be particularly useful to be able to genetically identify those people more likely to suffer from such disorders for diagnosis and prognostic purposes. For example, a positive genetic test might help to resolve the diagnosis in a difficult or unclear case.
  • the inventors have identified a genetic association between the TACRl gene (also known as the substance P or NKl receptor) and the incidence of ADHD, bipolar disease and co-morbid disorders such as depression, anxiety, conduct disorder, personality disorder, aggression and alcoholism.
  • mice lacking functional TACRl receptors display the characteristics of humans having ADHD, specifically they are hyperactive and this hyperactivity is prevented by psychostimulants d-amphetamine and methylphenidate.
  • the inventors then screened the equivalent gene, TACRl, in humans with ADHD and discovered a strong allelic and haplotypic association between polymorphisms in the TACRl gene and ADHD.
  • a marker for increased susceptibility to one or more of ADHD, alcohol addiction, bipolar and unipolar affective disorders, conduct disorder, self harm and suicidality wherein the marker is selected from rs3771829, rs3771833 and rsl7011370, as shown in Table 1 or rs59099335, rsl477157, rs34374747, rsl477156, rsl3387833, rs2111375, rsl3384011, rs2193405, rsl0210648, rs6715729, rs2024512, rs881, rsl7010664, rsl2713828, INDELl 5 INDEL2, and INDEL3 as shown in table 5 or the presence of the short isoform of the TACRl protein:
  • a method for identifiying one or more genetic mutations as candidate mutations for increasing susceptibility to ADHD, alcohol addiction, bipolar and unipolar affective disorders, conduct disorder, self harm or suicidality comprising the step of analysing a sample for genetic mutations, the sample being obtained from a subject having one or more of the markers shown in table 1 or table 5, the genetic mutations identified being candidate mutations.
  • the subject has at least two, more preferably at least three, even more preferably at least four of the markers.
  • the subject has at least two, more preferably at least three, even more preferably all four of the markers in table 1.
  • the subject preferably has at least one of markers rs3771829 and rs3771833.
  • the subject preferably has at least one marker from each of table 1 and table 5.
  • the subject preferably has displayed or is displaying clinical signs or symptoms of one or more of ADHD, alcohol addiction, bipolar and unipolar affective disorders, conduct disorder, self harm or suicidality.
  • an isolated nucleic acid comprising a mutation identified by the method of the invention. Additionally, the invention provides the use of a mutation identified by the method of the invention as a marker for increased susceptibility to one or more of ADHD, alcohol addiction, bipolar and unipolar affective disorders, conduct disorder, self harm and suicidality.
  • Table 1 Single nucleotide polymorphisms needed for selecting a genus of individuals who have DNA mutations increasing susceptibility to alcoholism, depression, anxiety, conduct disorder, aggression and attention deficit hyperactivity disorder (ADHD)
  • Two or more markers may be used in combination.
  • rs3771829 and rs3771853 may be used in combination.
  • ADHD attention deficit hyperactivity disorder. Included in the definition of ADHD used herein is ADD, attention deficit disorder. ADHD is a common development and behavioural disorder, seen in children, but which can remain into adulthood. It is characterized by poor concentration, distractibility, hyperactivity, and impulsiveness that are inappropriate for the child's age. Children and adults with ADHD are easily distracted by sights and sounds in their environment, cannot concentrate for long periods of time, are restless and impulsive, or have a tendency to daydream and be slow to complete tasks.
  • TACRl tachykinin 1 receptor
  • Alcoholism or alcohol addiction means a dependence on alcohol, characterized by repeated excessive use of alcoholic beverages, tolerance to the effects of alcohol, the development of withdrawal symptoms on reducing or ceasing alcohol intake and salience of drink seeking behaviour.
  • Conduct disorder is a type of disruptive behaviour disorder generally seen in childhood and adolescence involving a persistent pattern of behaviour in which the rights of others of the norms or rules of society are violated, with misconduct including aggression to people or animals, destruction of property, deceitfulness or theft, and serious violation of rules.
  • Unipolar and bipolar affective disorders are mood disorders characterised by extremes of mood. Unipolar affective disorder involves depression, whereas bipolar affective disorder involves both depression and mania. Loss of appetite, diurnal shift of mood, weight loss and initial insomnia as well as early morning wakefulness characterise major depression. Cyclothymia or mood lability may also be found in association with bipolar disorder and depression and in ADHD. In these cases mood can change rapidly from low to high and vide versa within a few hours or days. Self harm or injury is when someone deliberately hurts or injures him or herself. This can take a number of forms including, but not limited to, cutting, taking overdoses of tablets or medicines, punching, scratching, picking or tearing at one's skin causing sores and scarring, burning and inhaling or sniffing harmful substances.
  • Suicidality means suicidal thinking or behaviour and includes considering or attempting self- destructive actions.
  • the markers mentioned above are single nucleotide polymorphisms.
  • the polymorphisms are known in the art, but their association with ADHD and the other conditions mentioned herein has not previously been identified.
  • rs3771829 is in intron 1 of the TACRl gene at base pair position 75217653 on chromosome 2 in the March 2006 release of the UCSC Genome Browser.
  • rs3771833 is in intron 1 of the TACRl gene at base pair position 75220445 on chromosome 2 in the March 2006 release of the UCSC Genome Browser.
  • rs3771856 is in intron 1 of the TACRl gene at base pair position 75267522 on chromosome 2 in the March 2006 release of the UCSC Genome Browser.
  • rsl7011370 is in the 3 prime region of the TACRl gene at base pair position 75546131 on chromsome 2 in the March 2006 release of the UCSC Genome Browser.
  • a single nucleotide polymorphism is a DNA sequence variation that involves a change in one nucleotide at a particular site.
  • Predisposition is used to mean an increased likelihood of a subject displaying a certain pattern of behaviour compared to a subject without the polymorphism. Preferably it means that the subject is at least 20%, more preferably at least 30%, more preferably at least 40%, more preferably at least 50% more likely to display the pattern of behaviour.
  • Identification of the SNP may be carried out by methods known in the art, as discussed in the description. In particular polymerase chain reaction (PCR) based methods, comprising amplifying the region of DNA comprising the polymorphism followed by direct sequencing of the amplicon may be used.
  • PCR polymerase chain reaction
  • the polymorphisms may be identified using the "Amplifluor” method which uses the polymerase chain reaction to amplify the SNP from the SNA of a case or control combined with a third oligonucleotide labelled fluorimetrically so that it can discriminate between the SNP alleles.
  • it may be genotyped using the "TaqMan assays on demand” method which uses a quencher and a reporter fluorimetric signal on the same oligonucletide to discriminate between alleles.
  • the subject may be any subject, but is preferably a human.
  • the subject may be displaying characteristics of a particular disease or condition, such as ADHD.
  • the method may be combined with observing the behaviour of the subject, or of the subject's relatives for signs of the condition of interest.
  • Candidate mutation is a genetic mutation identified as being likely to increase susceptibility to one or more of ADHD, alcohol addiction, bipolar and unipolar affective disorders, conduct disorder, self harm or suicidality. The effect of such a mutation may be tested further, to confirm its effect on a subject's susceptibility.
  • the marker and/or method may also be used for predicting a subject's likelihood to respond to a particular course of treatment or to predict the likely results of other tests on that subject, such as brain imaging.
  • the TACRl protein is well known in the art. It has short and long isoforms. The inventors have found that the short isoform of the TACRl receptor protein is involved in the genetic aetiology of ADHD and TACRl related disorders such as alcoholism and affective disorders.
  • a variety of methods can be employed for the diagnostic and prognostic evaluation of TACRl gene disorders and neuropsychiatric disorders, such as conduct disorder, attention deficit hyperactivity disorder and alcoholism, and for the identification of subjects having a predisposition to such disorders.
  • the methods described herein may be performed, for example, by utilizing pre-packaged diagnostic kits comprising at least one specific TACRl gene nucleic acid or anti-TACRl gene antibody reagent described herein, which may be conveniently used, e.g., in clinical settings, to diagnose patients exhibiting abnormalities of a TACRl gene disorder or a neuropsychiatric disorder, such as conduct disorder, attention deficit hyperactivity disorder and alcoholism.
  • any nucleated cell can be used as a starting source for genomic nucleic acid.
  • any cell type or tissue in which the TACRl gene is expressed may be utilized.
  • a variety of methods can be employed to screen for the presence of the markers and to detect and/or assay levels of TACRl gene nucleic acid sequences.
  • Markers within the TACRl gene can be detected by utilizing a number of techniques. Nucleic acid from any nucleated cell can be used as the starting point for such assay techniques, and may be isolated according to standard nucleic acid preparation procedures that are well known to those of skill in the art.
  • TACRl gene nucleic acid sequences may be used in hybridization or amplification assays of biological samples to detect abnormalities involving TACRl gene structure, including point mutations, insertions, deletions, inversions, translocations and chromosomal rearrangements.
  • assays may include, but are not limited to, Southern analyses, single-stranded conformational polymorphism analyses (SSCP), and PCR analyses.
  • Diagnostic methods for the detection of TACRl gene-specific mutations can involve for example, contacting and incubating nucleic acids including recombinant DNA molecules, cloned genes or degenerate variants thereof, obtained from a sample, e.g., derived from a patient sample or other appropriate cellular source, with one or more labelled nucleic acid reagents including recombinant DNA molecules, cloned genes or degenerate variants thereof, under conditions favourable for the specific annealing of these reagents to their complementary sequences within the TACRl gene.
  • the lengths of these nucleic acid reagents are at least 15 to 30 nucleotides.
  • nucleic acid:TACRl gene molecule hybrid After incubation, all non-annealed nucleic acids are removed from the nucleic acid:TACRl gene molecule hybrid. The presence of nucleic acids that have hybridized, if any such molecules exist, is then detected. Using such a detection scheme, the nucleic acid from the cell type or tissue of interest can be immobilized, for example, to a solid support such as a membrane, or a plastic surface such as that on a microtitre plate or polystyrene beads. In this case, after incubation, non-annealed, labelled nucleic acid reagents are easily removed. Detection of the remaining, annealed, labelled TACRl gene nucleic acid reagents is accomplished using standard techniques well-known to those in the art. The TACRl gene sequences to which the nucleic acid reagents have annealed can be compared to the annealing pattern expected from a normal TACRl gene sequence in order to determine
  • Alternative diagnostic methods for the detection of TACRl markers, in patient samples or other appropriate cell sources may involve their amplification, e.g., by PCR (the experimental embodiment set forth in MuIUs, 1987, U.S. Pat. No. 4,683,202), followed by the detection of the amplified molecules using techniques well known to those of skill in the art.
  • the resulting amplified sequences can be compared to those that would be expected if the nucleic acid being amplified contained only normal copies of the TACRl gene in order to determine whether a TACRl marker exists.
  • genotyping techniques can be performed to identify individuals carrying TACRl markers. Such techniques include, for example, the use of restriction fragment length polymorphisms (RFLPs), which involve sequence variations in one of the recognition sites for the specific restriction enzyme used.
  • RFLPs restriction fragment length polymorphisms
  • Caskey et al. U.S. Pat. No. 5,364,759 describe a DNA profiling assay for detecting short tri and tetra nucleotide repeat sequences. The process includes extracting the DNA of interest, such as the TACRl gene, amplifying the extracted DNA, and labelling the repeat sequences to form a genotypic map of the individual's DNA.
  • TACRl gene expression assays “in situ", i.e., directly upon tissue sections (fixed and/or frozen) of patient tissue obtained from biopsies or resections, such that no nucleic acid purification is necessary.
  • Nucleic acid reagents may be used as probes and/or primers for such in situ procedures (see, for example, Nuovo, G. J., 1992, “PCR In Situ Hybridization: Protocols And Applications", Raven Press, NY).
  • Antibodies directed against gene products of TACRl genes containing the markers, or conserved variants or peptide fragments thereof, may also be used as diagnostics and prognostics for a TACRl gene disorder or a neuropsychiatric disorder, such as conduct disorder, attention deficit hyperactivity disorder and alcoholism, as described herein. Such methods may be used to detect abnormalities in the level of TACRl gene product synthesis or expression, or abnormalities in the structure, temporal expression, and/or physical location of TACRl gene product.
  • the antibodies and immunoassay methods described below have, for example, important in vitro applications in assessing the efficacy of treatments for TACRl gene disorders or neuropsychiatric disorders, such as conduct disorder, attention deficit hyperactivity disorder and alcoholism .
  • the tissue or cell type to be analyzed will generally include those that are known, or suspected, to express the TACRl gene.
  • the protein isolation methods employed herein may, for example, be such as those described in Harlow and Lane (1988, "Antibodies: A Laboratory Manual", Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York).
  • the isolated cells can be derived from cell culture or from a patient.
  • the analysis of cells taken from culture may be a necessary step in the assessment of cells to be used as part of a cell-based gene therapy technique or, alternatively, to test the effect of compounds on the expression of the TACRl gene.
  • Preferred diagnostic methods for the detection of TACRl gene products or conserved variants or peptide fragments thereof may involve, for example, immunoassays wherein the TACRl gene products or conserved variants or peptide fragments are detected by their interaction with an anti-TACRl gene product-specific antibody.
  • antibodies or fragments of antibodies useful in the present invention may be used to quantitatively or qualitatively detect the presence of TACRl gene products or conserved variants or peptide fragments thereof. This can be accomplished, for example, by immunofluorescence techniques employing a fluorescently labeled antibody coupled with light microscopic, flow cytometric, or fluorimetric detection. Such techniques are especially preferred for TACRl gene products that are expressed on the cell surface.
  • the antibodies (or fragments thereof) useful in the present invention may, additionally, be employed histologically, as in immunofluorescence or immunoelectron microscopy, for in situ detection of TACRl gene products or conserved variants or peptide fragments thereof.
  • In situ detection may be accomplished by removing a histological specimen from a patient, and applying thereto a labelled antibody of the present invention.
  • the antibody (or fragment) is preferably applied by overlaying the labeled antibody (or fragment) onto a biological sample.
  • Immunoassays for TACRl gene products or conserved variants or peptide fragments thereof will typically comprise incubating a sample, such as a biological fluid, a tissue extract, freshly harvested cells, or lysates of cells, that have been incubated in cell culture, in the presence of a detectably labeled antibody capable of identifying TACRl gene products or conserved variants or peptide fragments thereof, and detecting the bound antibody by any of a number of techniques well-known in the art.
  • the biological sample may be brought in contact with and immobilized onto a solid phase support or carrier such as nitrocellulose, or other solid support that is capable of immobilizing cells, cell particles or soluble proteins.
  • a solid phase support or carrier such as nitrocellulose, or other solid support that is capable of immobilizing cells, cell particles or soluble proteins.
  • the support may then be washed with suitable buffers followed by treatment with the detectably labeled TACRl gene specific antibody.
  • the solid phase support may then be washed with the buffer a second time to remove unbound antibody.
  • the amount of bound label on solid support may then be detected by conventional means.
  • solid phase support or carrier any support capable of binding an antigen or an antibody.
  • supports or carriers include glass, polystyrene, polypropylene, polyethylene, dextran, nylon, amylases, natural and modified celluloses, polyacrylamides, gabbros, and magnetite.
  • the nature of the carrier can be either soluble to some extent or insoluble for the purposes of the present invention.
  • the support material may have virtually any possible structural configuration so long as the coupled molecule is capable of binding to an antigen or antibody.
  • the support configuration may be spherical, as in a bead, or cylindrical, as in the inside surface of a test tube, or the external surface of a rod.
  • the surface may be flat such as a sheet, test strip, etc.
  • Preferred supports include polystyrene beads. Those skilled in the art will know many other suitable carriers for binding antibody or antigen, or will be able to ascertain the same by use of routine experimentation.
  • binding activity of a given lot of anti-TACRl gene product antibody may be determined according to well known methods. Those skilled in the art will be able to determine operative and optimal assay conditions for each determination by employing routine experimentation.
  • EIA enzyme immunoassay
  • the enzyme which is bound to the antibody will react with an appropriate substrate, preferably a chromogenic substrate, in such a manner as to produce a chemical moiety that can be detected, for example, by spectrophotometric, fluorimetric or by visual means.
  • Enzymes that can be used to detectably label the antibody include, but are not limited to, malate dehydrogenase, staphylococcal nuclease, delta-5-steroid isomerase, yeast alcohol dehydrogenase, ⁇ -glycerophosphate, dehydrogenase, triose phosphate isomerase, horseradish peroxidase, alkaline phosphatase, asparaginase, glucose oxidase, ⁇ -galactosidase, ribonuclease, urease, catalase, glucose-6-phosphate dehydrogenase, glucoamylase and acetylcholinesterase.
  • the detection can be accomplished by colorimetric methods that employ a chromogenic substrate for the enzyme. Detection may also be accomplished by visual comparison of the extent of enzymatic reaction of a substrate in comparison with similarly prepared standards.
  • Detection may also be accomplished using any of a variety of other immunoassays.
  • a radioimmunoassay RIA
  • the radioactive isotope can be detected by such means as the use of a gamma counter or a scintillation counter or by autoradiography.
  • fluorescent labeling compounds fluorescein isothiocyanate, rhodamine, phycoerythrin, phycocyanin, allophycocyanin, o-phthaldehyde and fluorescarnine.
  • the antibody can also be detectably labeled using fluorescence emitting metals such as 152 Eu, or others of the lanthanide series. These metals can be attached to the antibody using such metal chelating groups as diethylenetriaminepentacetic acid (DTPA) or ethylenediaminetetraacetic acid (EDTA).
  • DTPA diethylenetriaminepentacetic acid
  • EDTA ethylenediaminetetraacetic acid
  • the antibody also can be detectably labeled by coupling it to a chemiluminescent compound.
  • the presence of the chemiluminescent-tagged antibody is then determined by detecting the presence of luminescence that arises during the course of a chemical reaction.
  • chemiluminescent labeling compounds are luminol, isoluminol, theromatic acridinium ester, imidazole, acridinium salt and oxalate ester.
  • Bioluminescence is a type of chemiluminescence found in biological systems in which a catalytic protein increases the efficiency of the chemiluminescent reaction. The presence of a bioluminescent protein is determined by detecting the presence of luminescence. Important bioluminescent compounds for purposes of labeling are luciferin, luciferase, green fluorescent protein and aequorin.
  • the inventors carried out genetic association studies to test whether there are abnormalities in the TACRl gene of patients with a positive diagnosis of ADHD.
  • the significance for this association with ADHD remained after empirical tests of significance were applied. No evidence of inconsistent genotyping was found after 17% of samples were reduplicated and genotyped twice.
  • Samples of DNA were collected from 450 probands from NHS services in the UK., and all were of white European origin. Subjects were referred for research assessment if they were thought by experienced clinicians to have a diagnosis of the combined subtype of ADHD under DSM-IV criteria, an IQ greater than 70 and no significant Axis I comorbidity apart from oppositional defiant disorder and conduct disorder. Only individuals fulfilling the recruitment criteria after completion of research assessments were included in the study.
  • HYPESCHEME data sheets were completed with the data from the parent and school questionnaires plus review of case notes.
  • the HYPESCHEME diagnoses were checked against researcher applied DSM-IV criteria and discrepancies were reviewed by two researchers.
  • Situational pervasiveness was operationalized as the presence of 1 or more ADHD symptoms at home and school plus descriptions from the parent of impairment from ADHD symptoms in more than one setting. Where consensus could not be reached, cases were brought to case conference and final agreement was made with a senior clinical researcher.
  • Genomic DNA for the Edinburgh and Birmingham ADHD subjects was extracted from whole blood samples using standard cell lysis, proteinase K digestion and phenol/chloroform ethanol precipitation method. All SNP markers were determined using KASPar (KBioscience, Hoddesdon, UK), a modified Amplifluor SNP genotyping method. 17% of the samples from each microtitre plate were reduplicated in order to detect possible errors and to also confirm the reproducibility of genotypes.
  • HWE Hardy- Weinberg equilibrium
  • Any markers lacking HWE were repeated using an alternative Taqman method.
  • the genotype data was assessed using the SCANGROUP program in GENECOUNTING (Zhao et al 2002). This program highlights any potential genotyping errors by identifying significant differences in haplotypic frequencies between any single 96 well microtitre-plate compared to haplotypic frequencies in all other plates combined. (It is argued that genotyping errors can show up as rare haplotypes occurring on just a single plate, whereas true associations between haplotypes and disease will appear as differences in haplotype frequencies in cases and control samples across several plates.
  • the invention comprises information about the DNA sequence in and adjacent to the PACXRl gene and the consequence of these changes in causing genetic susceptibility to attention deficit hyperactivity disorder (ADHD) and other related disorders such as bipolar affective disorder, unipolar affective disorders (Major depressive disorder), cyclothymia, anxiety disorder, alcoholism and also bipolar disorders comorbid with alcoholism.
  • ADHD attention deficit hyperactivity disorder
  • Other related disorders such as bipolar affective disorder, unipolar affective disorders (Major depressive disorder), cyclothymia, anxiety disorder, alcoholism and also bipolar disorders comorbid with alcoholism.
  • SNPs single nucleotide polymorphisms
  • mutations listed in tables 1 and 5 as well as insertion/deletions found by us can down regulate the expression of the TACRl receptor and thus down-regulate the TACRl protein or reduce the function of the TACRl receptor by shortening the protein or making the protein less active due to an amino acid change.
  • rs59099335, rsl477157, rs34374747, rsl477156, rsl3387833, rs2111375, rsl3384011, rs2193405 and rsl 0210648 are mutations in the TACRl promoter with the base pair changes listed in table 1 and in the TACRl sequence TACR Seq ID. These changes can produce a reduction or loss of transcription of the TACRl mRNA and reduced translation of the TACRl protein. This happens because effects of these SNPs/mutations alter transcription factor binding sites, as well as enhancer and promoter activity of the TACRl gene.
  • the exon 1 synonymous SNP can alter mRNA stability, or harbour a cryptic splice enhancer sequence thus reducing TACRl function.
  • the intron 1 SNP (rs2024512) can alter the function of a cryptic splice enhancer site and thus reduce TACRl receptor activity.
  • the 3'UTR SNP rs881 can alter interfering micro RNA (miRNA) binding patterns that change mRNA stability and produce a hypofunctioning or loss of activity of the TACRl receptor.
  • the 3'UTR SNP rsl7010664s can also change mRNA stability and produce a hypofunctioning or loss of activity of the TACRl receptor.
  • the insertion/deletions INDELl, INDEL2 and INDEL3 at chromosome 2 base pair positions 75283217, 75282897, 75129909 in the March 2006 UCSC can alter miRNA binding patterns and change mRNA stability to cause a change in function of the TACRl receptor.
  • the missense SNP rsl 7838409 in exon 4 of TACRl codes for a Valine to Methionine amino acid change one variant of which can reduce the response of the TACRl receptor protein such that it is hypofunctional and causes ADHD and TACRl related disorders.
  • the short isoform of the TACRl receptor protein is involved in the genetic aetiology of ADHD and TACRl related disorders such as alcoholism and affective disorders.
  • the long isoform has a ten-fold higher affinity for Tachykinin (substance P or neurokinin) binding than the short form of the receptor.
  • the short form mRNA lacks the fourth intron and exon five and has an extra conserved UTR region which we predict contains an abnormal splice site which results in alternative splicing of the gene.
  • This variant uses a different splice site in the 3' coding region, compared to variant long.
  • the resulting protein (isoform short) has a shorter and distinct C-terminus when compared to the long isoform.
  • This abnormal splice site and splicing of the mRNA and subsequent shortening of the TACRl receptor explain the reduced TACRl activity in response to Tachykin and causes reduced prefrontal TACRl cortical activity which causes hyperactivity in ADHD patients as well as other TACRl related disorders such as alcoholism and bipolar disorder.
  • SNPs numbered 3, 4, 7, 8 show allelic association with ADHD.
  • haplotypes are combined into haplotypes using the programme HAPLOVIEW, which found association between two haplotypes and ADHD. Only one haplotype was positively associated with ADHD and this remained significant after an empirical test of association (9,999 iterations).
  • the associated haplotype comprised allele C from SNPs rs3771829 and allele A from rs3771833 .
  • Table 4 TACRl SNPs IN BIPOLAR-ALCOHOLICS OR ATTENTION- DEFICIT HYPERACTIVITY DISORDER PATIENTS AND NORMAL

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)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Pathology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne des marqueurs permettant d’identifier une susceptibilité accrue à l’un ou à plusieurs des problèmes suivants : troubles déficitaires de l’attention avec hyperactivité (ADHD), alcoolisme, troubles affectifs bipolaires et unipolaires, trouble de la conduite, autodestruction et suicidabilité chez des individus.
PCT/GB2009/001036 2008-04-23 2009-04-23 Marqueurs WO2009130465A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB0807421A GB0807421D0 (en) 2008-04-23 2008-04-23 Markers
GB0807421.3 2008-04-23

Publications (2)

Publication Number Publication Date
WO2009130465A2 true WO2009130465A2 (fr) 2009-10-29
WO2009130465A3 WO2009130465A3 (fr) 2010-01-28

Family

ID=39494122

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2009/001036 WO2009130465A2 (fr) 2008-04-23 2009-04-23 Marqueurs

Country Status (2)

Country Link
GB (1) GB0807421D0 (fr)
WO (1) WO2009130465A2 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001071029A1 (fr) * 2000-03-21 2001-09-27 Hormos Medical Corporation Diagnostic du risque de developper un alcoolisme chez une personne
WO2002090541A1 (fr) * 2001-05-03 2002-11-14 Murdoch Childrens Research Institute Determination d'une predisposition genetique a des troubles comportementaux
WO2007052166A2 (fr) * 2005-10-14 2007-05-10 Institut Pasteur Variations genetiques associees a des troubles psychiatriques

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001071029A1 (fr) * 2000-03-21 2001-09-27 Hormos Medical Corporation Diagnostic du risque de developper un alcoolisme chez une personne
WO2002090541A1 (fr) * 2001-05-03 2002-11-14 Murdoch Childrens Research Institute Determination d'une predisposition genetique a des troubles comportementaux
WO2007052166A2 (fr) * 2005-10-14 2007-05-10 Institut Pasteur Variations genetiques associees a des troubles psychiatriques

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
DATABASE SNP [Online] 8 October 2004 (2004-10-08), XP002544231 retrieved from NCBI Database accession no. rs3771829 *

Also Published As

Publication number Publication date
GB0807421D0 (en) 2008-05-28
WO2009130465A3 (fr) 2010-01-28

Similar Documents

Publication Publication Date Title
US20210222245A1 (en) Autism associated genetic markers
Flint et al. The use of telomere probes to investigate submicroscopic rearrangements associated with mental retardation
WO2004020968A2 (fr) Mutations de nod2 associees a la stenose fibreuse chez des patients souffrant de la maladie de crohn
EP2585824A2 (fr) Analyse du risque génétique dans le syndrome du déficit de récompense
Bidari et al. Common MEFV mutation analysis in 36 Iranian patients with familial Mediterranean fever: clinical and demographic significance
Monaghan et al. Technical standards and guidelines for reproductive screening in the Ashkenazi Jewish population
Elce et al. Three novel CFTR polymorphic repeats improve segregation analysis for cystic fibrosis
JP5706612B2 (ja) 加齢黄斑変性症易罹患性の判定マーカー並びに判定方法及び判定キット
Chiu et al. Circulating Nucleic Acids in Plasma/Serum III and Serum Proteomics Recent Developments in Fetal DNA in Maternal Plasma
EP2174139A1 (fr) Procédé permettant de tester un sujet supposé souffrir d'asthme ou être prédisposé à l'asthme
JP6128654B2 (ja) 関節リウマチの新規遺伝因子としてのミエリン塩基性蛋白の利用
WO2009130465A2 (fr) Marqueurs
TWI351436B (en) Method for detecting a risk of the development of
US20100255468A1 (en) Method of assessing gene examination data, program therefor and apparatus of the same
US20070037194A1 (en) Allelic variation in the serotonin transporter (SERT) as an indicator of autism
WO2010007063A2 (fr) Procédé de diagnostic de la dyslexie
CA2349127A1 (fr) Diagnostic moleculaire de galactosemie
WO2009070764A1 (fr) Une variation dans le gène chi3l1 affecte les teneurs sériques en ykl-40, le risque d'asthme et la fonction pulmonaire
Strom et al. Tay-Sachs carrier screening in the genomics age: gene sequencing versus enzyme analysis in non-Jewish individuals
WO2002090585A2 (fr) Marqueur genetique
US8216787B2 (en) Biomarker for successful aging without cognitive decline
Shahbazian et al. Identification of MeCP2 Mutations in a Series of Females with Autistic Disorder
JP2008141961A (ja) 第１０番染色体長腕領域における２型糖尿病感受性遺伝子

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: 09734084

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: 09734084

Country of ref document: EP

Kind code of ref document: A2