WO2006008142A1 - Diagnosis and therapy of psoriasis vulgaris - Google Patents

Diagnosis and therapy of psoriasis vulgaris Download PDF

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Publication number
WO2006008142A1
WO2006008142A1 PCT/EP2005/007870 EP2005007870W WO2006008142A1 WO 2006008142 A1 WO2006008142 A1 WO 2006008142A1 EP 2005007870 W EP2005007870 W EP 2005007870W WO 2006008142 A1 WO2006008142 A1 WO 2006008142A1
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Prior art keywords
mgst2
nucleic acid
agent
est
polypeptide
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PCT/EP2005/007870
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French (fr)
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WO2006008142A8 (en
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Andreas Tzschach
Vera Kalscheuer
Hans-Hilger Ropers
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MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V.
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Publication of WO2006008142A1 publication Critical patent/WO2006008142A1/en
Publication of WO2006008142A8 publication Critical patent/WO2006008142A8/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/45—Transferases (2)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1137—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1085—Transferases (2.) transferring alkyl or aryl groups other than methyl groups (2.5)
    • C12N9/1088—Glutathione transferase (2.5.1.18)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y205/00—Transferases transferring alkyl or aryl groups, other than methyl groups (2.5)
    • C12Y205/01—Transferases transferring alkyl or aryl groups, other than methyl groups (2.5) transferring alkyl or aryl groups, other than methyl groups (2.5.1)
    • C12Y205/01018—Glutathione transferase (2.5.1.18)
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2217/00—Genetically modified animals
    • A01K2217/05—Animals comprising random inserted nucleic acids (transgenic)
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2217/00—Genetically modified animals
    • A01K2217/07—Animals genetically altered by homologous recombination
    • A01K2217/075—Animals genetically altered by homologous recombination inducing loss of function, i.e. knock out
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2330/00—Production
    • C12N2330/10—Production naturally occurring
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/20—Dermatological disorders
    • G01N2800/205—Scaling palpular diseases, e.g. psoriasis, pytiriasis

Definitions

  • the present invention relates to the use of MGST2 nucleic acids, MGST2 polypeptides, MGST2 effectors, EST CR742434 nucleic acids and EST CR742434 effectors for the diagnosis, prevention and/or therapy of psoriasis vulgaris and related diseases.
  • Psoriasis vulgaris is a chronically inflammatory and hyperproliferative skin disease from which approximately 2-5% of the Caucasian population is affected (Nevitt and Hutchinson, 1996). It represents a considerable impairment for the persons affected and mostly requires a lifelong ambulant and partially also stationary therapy due to the chronic character. Thus, psoriasis vulgaris involves considerable costs in medical care, which is estimated to about $ 3 billion per year in the U.S. (Bowcock and Cockson, 2004).
  • psoriasis vulgaris represents a multifactorial disease which is caused by the interaction of genetic predisposition factors and environmental influences (Elder et al., 2001 ).
  • the participation of several different genes explains a part of the clinical variability of this disease.
  • the exact knowledge of these genes could enable individual therapeutic strategies which are adapted to the genetic status of an individual patient.
  • MGST2 plays an important role in the metabolism of the leucotrienes which are strong inflammatory mediators. MGST2 is located in a chromosomal region for which the relation to psoriasis was found (Sagoo et al. 2004). Thus, the identification of this gene defect has important consequences for the diagnosis and therapy of psoriasis vulgaris and related diseases.
  • a first aspect of the present invention relates to the use of a MGST2 nucleic acid, a MGST2 polypeptide or a MGST2 effector for the manufacture of an agent for the diagnosis, prevention and/or therapy of psoriasis vulgaris or related diseases.
  • a further aspect of the present invention relates to a screening method for identifying agents for the diagnosis, prevention and/or therapy of psoriasis vulgaris and related disorders comprising determining the interaction of a test compound with a MGST2. nucleic acid or polypeptide.
  • Still a further aspect of the present invention relates to a method for diagnosing psoriasis vulgaris and related disorders or a predisposition thereof in a subject comprising determining the amount and/or activity of a MGST2 nucleic acid or polypeptide in said subject or in a sample therefrom.
  • Still a further aspect of the present invention relates to a method for preventing or treating psoriasis vulgaris and related disorders in a subject comprising administering a pharmaceutically effective amount of MGST2 nucleic acid, a MGST2 polypeptide or a MGST2 effector to said subject.
  • the present invention has diagnostic and therapeutic applications in human and veterinary medicine, particularly in human medicine.
  • the diagnosis particularly relates to the determination of the presence of altered MGST2 nucleic acids and/or polypeptides or altered amounts of MGST2 nucleic acids and/or polypeptides which are most usually associated with an alteration of at least one genomic copy of the MGST2 gene.
  • Such changes may also be part of a specific haplotype of DNA markers around the causative mutation, which may enable its indirect diagnosis.
  • the subjects to be treated according to the present invention are particularly subjects with a genomic alteration as described above, more particularly with a haplotypic gene defect.
  • the MGST2 nucleic acids and proteins are obtainable from vertebrate species. Preferred are nucleic acids encoding the human MGST2 polypeptide and variants thereof (GenBank Accession Number AA H25416 for the polypeptide and BC 025416 for the nucleic acid).
  • MST2 nucleic acid particularly relates to
  • MST2 polypeptide preferably relates to
  • (c) a partial sequence of the amino acid sequences of (a) to (b) having a length of at least 6, preferably 8 and more preferably of at least 12 amino acids.
  • the term "MGST2 effector” relates to any molecule which is capable of detecting MGST2 nucleic acids and/or polypeptides in a biological sample, e.g. in a tissue or body fluid sample of a human patient.
  • the term “MGST2 effector” preferably relates to any molecule capable of modulating the amount, location and/or activity of MGST2 nucleic acids and/or polypeptides or nucleic acids and/or polypeptides interacting with MGST2 nucleic acids and/or polypeptides.
  • the term “effector” encompasses both direct modulators of MGST2 and indirect modulators of MGST2 which modulate molecules interacting with MGST2.
  • the diagnosis relates to the determination of the presence of altered EST CR742434 nucleic acids or altered amounts of EST CR742434 nucleic acids.
  • EST CR742434 exhibits an RNA gene, most of which have a regulatory function on the expression of other genes. Since EST CR742434 and the MGST2 gene overlap in their 5 1 regions in antisense-direction/ a regulatory function of EST CR742434 on the expression of MGST2 is very likely in the present case.
  • Still a further aspect of the present invention relates to the use of an EST CR742434 nucleic acid or an EST CR742434 effector for the manufacture of an agent for the diagnosis, prevention and/or therapy of psoriasis vulgaris or related diseases.
  • Still a further aspect of the present invention relates to a screening method for identifying agents for the diagnosis, prevention and/or therapy of psoriasis vulgaris and related disorders comprising determining the interaction of a test compound with an EST CR742434 nucleic acid.
  • Still a further aspect of the present invention relates to a method for diagnosing psoriasis vulgaris and related disorders or a predisposition thereof in a subject comprising determining the amount and/or activity of an EST CR742434 nucleic acid in said subject or in a sample therefrom.
  • Still a further aspect of the present invention relates to a method for preventing or treating psoriasis vulgaris and related disorders in a subject comprising administering a pharmaceutically effective amount of EST CR742434 nucleic acid or an EST CR742434 effector to said subject.
  • EST CR742434 nucleic acid particularly relates to
  • EST CR742434 effector relates to any molecule which is capable of detecting EST CR742434 nucleic acids in a biological sample, e.g. in a tissue or body fluid sample of a human patient.
  • EST CR742434 effector relates to any molecule capable of modulating the amount, location and/or activity of EST CR742434 nucleic acids or nucleic acids interacting with EST CR742434 nucleic acids.
  • psoriasis vulgaris or related diseases relates to diseases selected from psoriasis vulgaris, and other T-cell mediated autoimmune diseases, particularly rheumatic diseases such as rheumatoid arthritis or Morbus Crohn.
  • the diagnostic aspect of the present invention may comprise the detection of MGST2 nucleic acid, e.g. a genomic MGST2 sequence or a MGST2 transcript.
  • the diagnosis may comprise a sequence determination of a MGST2 nucleic acid, e.g. the presence of alterations, e.g. disruptions, polymorphisms etc. of a MGST2 nucleic acid sequence in a patient.
  • the sequence determination may comprise the determination of a single nucleotide, e.g. in a single nucleotide polymorphism or the determination of longer sequence portion.
  • the diagnosis may comprise the determination of a MGST2 nucleic acid expression level, e.g. a qualitative and/or quantitative determination.
  • MGST2 nucleic acid detection molecules such as primers and probes or corresponding nucleic acid analogues such as LNA or PNA molecules for primer extension, sequencing and/or amplification reactions may be employed.
  • the diagnostic aspect of the present invention also may comprise the detection of EST CR742434 nucleic acid in the embodiments described above for MGST2.
  • the diagnostic aspect of the present invention may comprise determination of a MGST2 polypeptide, e.g. the amount, location and/or activity of a MGST2 polypeptide.
  • This aspect of the present invention preferably comprises an immunoassay wherein the presence of an MGST2 polypeptide is detected by using suitable antibodies.
  • the antibodies may be specific for MGST2 wild-type molecules or MBST2 variants according to the respective test format. Methods of determining polypeptides in biological samples are known in the art.
  • the therapeutic agent may be a therapeutic nucleic acid or nucleic acid analogue, e.g. a MGST2 nucleic acid molecule, EST CR742434 nucleic acid, an aptamer, an antisense molecule, an RNA interference molecule or a ribozyme which is capable of directly or indirectly modulating the MGST2 activity in a subject to be treated.
  • the nucleic acid may be a nucleic sequence encoding a MGST2 polypeptide, e.g.
  • a genomic or a cDNA sequence which is transferred into target cells, particularly cells which are involved in psoriatic inflammatory reactions, such as blood cells, keratinocytes and/or fibroblasts, in order to substitute a lack of MGST2 activity in the target cells.
  • target cells particularly cells which are involved in psoriatic inflammatory reactions, such as blood cells, keratinocytes and/or fibroblasts, in order to substitute a lack of MGST2 activity in the target cells.
  • a gene therapeutic protocol which is used for the treatment of Xeroderma pigmentosum, a monogenic, autosomal recessive inherited skin disorder (Magnaldo 2004).
  • the therapeutic agent may be a nucleic acid or nucleic acid analogue which is a MGST2 effector.
  • Suitable acid or nucleic acid analogue effector molecules are preferably selected from EST CR742434 nucleic acid, aptamers, antisense molecules, RNA interference molecules or ribosomes.
  • Aptamers e.g. nucleic acid or nucleic acid analogue molecules which are capable of binding to MGST2 or a protein interacting therewith and modulating its activity, may be generated by a screening and selection procedure involving the use of combinatorial nucleic acid libraries.
  • Antisense molecules, RNAi molecules or ribozymes directed against the MGST2 mRNA sequence or against the sequence of mRNA molecules encoding polypeptides which interact with MGST2 may be used to modulate transcription of MGST2 mRNA or mRNA molecules encoding polypeptides interacting with MGST2.
  • antisense molecules may be used in the case of an increased MGST2 activity, in order to lower expression of MGST2.
  • the therapeutic agent also may be a nucleic acid or nucleic acid analogue which is an EST CR742434 effector.
  • nucleic acids may be carried out according to known methods, e.g. by introduction of naked DNA or by the use of transfer vehicles such as viruses, e.g. adenoviruses, adeno-associated viruses, retroviruses etc. as known in the art.
  • transfer vehicles such as viruses, e.g. adenoviruses, adeno-associated viruses, retroviruses etc. as known in the art.
  • the therapeutic agent may be a polypeptide or a fragment thereof.
  • the therapeutic agent may comprise a polypeptide, e.g. MGST2 polypeptide or an biologically active fragment which may be introduced into target cells in order to increase the amount of MGST2 activity.
  • the polypeptide may be a transcription factor capable of stimulating MGST2 transcription, particularly transcription from an unaltered, i.e. active MGST2 genomic sequence.
  • the therapeutic polypeptide may also be an antibody directed against a molecule interacting with MGST2.
  • the antibody is preferably a chimeric, humanized, or human antibody or an antigen-binding fragment of such an antibody. The production of such antibodies is known in the art.
  • the agent may be a low molecular weight organic compound with the capability of altering, e.g. stimulating or inhibiting the function of MGST2 polypeptide or polypeptides interacting therewith.
  • the low molecular weight organic compound has a molecular weight of about 100- 2500 Da.
  • the therapeutic agent is a stimulator of MGST2 activity.
  • the stimulation is a direct stimulation wherein an increased MGST2 amount and/or activity in the target cells is provided, e.g. by introduction of a MGST2 polypeptide or nucleic acid into the target cell.
  • the therapeutic agent may be an indirect stimulator of MGST2 activity, i.e. a molecule which interacts with physiologic compounds which may modulate MGST2 activity.
  • the agent may be an inhibitor of LTA4 synthesis, e.g. a 5-lipoxygenase inhibitor or a 5- lipoxygenase activating protein (FLAP) inhibitor.
  • the agent may be an inhibitor of leukotriene A4 (LTA4) hydrolase.
  • the agent may be a stimulator of leukotriene C4 (LTC4) synthesis.
  • the agent may be an inhibitor of an leukotriene B4 (LTB4) receptor, e.g. an inhibitor of the LTB4 receptors BLT1 or BLT2.
  • Inhibition of MGST2 activity may be achieved by administering suitable pharmacological substances in order to decrease enzymatic activity of MGST2.
  • the therapeutic aspect of the present invention preferably comprises the administration of a pharmaceutical composition which comprises as an active ingredient a MGST2 nucleic acid, a MGST2 polypeptide, a MGST2 effector, an EST CR742434 nucleic acid or an EST CR742434 effector together with pharmaceutically acceptable carriers, diluents and/or adjuvants.
  • the pharmaceutical composition may be administered alone or in combination with at least one other agent.
  • the pharmaceutical composition may be administered by any known route of administration, e.g. oral, topical, parenteral, transdermal, subcutaneous means. Topical or transdermal administration to the skin areas involved in psoriasis is preferred.
  • the pharmaceutical composition comprises the active ingredient in a therapeutically effective amount.
  • an effective dose for a specific active ingredient is known in the art.
  • the therapeutic effective dose can be estimated in cell culture assays or in experimental animal models.
  • the exact dosage, route and frequency of administration will be determined inter alia by the type of active ingredient, the severity of disease and the condition of the subject to be treated.
  • the present invention further provides a screening method for identifying novel agents for the diagnosis, prevention and/or therapy of psoriasis vulgaris and related disorders.
  • the screening method may comprise the use of a non-cellular screening system, a cellular screening system or a transgenic non-human animal.
  • the interaction of a test compound or a plurality of test compounds, e.g. from a test compound library with a MGST2 nucleic acid or polypeptide is determined wherein the interaction may comprise a direct modulation of MGST2 nucleic acid or polypeptide or an indirect modulation of an MGST2 nucleic acid or polypeptide as described above, e.g. inhibition of LTA4 synthesis, inhibition of LTA4 hydrolase, stimulation of LTC4 synthesis and/or inhibition of LTB4 receptors.
  • the screening method may comprise determination of an interaction of a test compound with an EST CR742434 nucleic acid.
  • the interaction may comprise direct or indirect modulation of EST CR742434 nucleic acid.
  • the present invention also relates to an isolated cell, preferably a mammalian cell and more preferably a human cell, capable of over- or underexpressing the MGST2 gene or the EST CR742434 gene.
  • Such cell may provide a suitable in vitro model system for further characterizing the molecular mechanisms of psoriasis or related diseases.
  • these molecular mechanisms may be elucidated in a non-human transgenic animal, preferably a transgenic non-human mammal, which is capable of over- or underexpressing the MGST2 gene or the EST CR742434 gene.
  • the non-human animal has at least one deficient MGST2 gene copy.
  • the starting point of the tests was a 43-year old patient suffering from psoriasis vulgaris and a balanced chromosomal translocation 46,XY,t(2;4) (p25;q31.1) (Fig. 1).
  • the already deceased and cytogenetically not tested father of the patient also suffered from psoriasis, very likely, he also carried the translocation, as evidenced by repeated miscarriages of two female partners.
  • BAC-clone RP11-79601 (GenBank Accession Number AQ514709) was identified as the break point covering clone. Since no genes are known in this region, no further analysis was carried out therein.
  • Leucotrienes are inflammatory mediators.
  • the leucotriene with the strongest pro-inflammatory effect in psoriatic cutaneous areas is LTB4, which arises from LTA4 by hydrolysis (Iversen et al., 1997) (Fig. 3).
  • MGST2 The gene product of MGST2, the microsomal glutathione S-transferase 2, catalyses the formation of LTC4 by the conjugation of glutathione with LTA4 (Jakobsson et al., 1996). This reaction can also be catalysed by two further enzymes, i.e. the leucotriene-C4-synthease (LTC4S) and MGST3.
  • LTC4S leucotriene-C4-synthease
  • MGST2 In the only cell type which has been tested with regard to that so far, i.e. endothelial cells of umbilical veins, MGST2 is the main and probably even the only active enzyme for the synthesis of LTC4 (Sj ⁇ str ⁇ m et al., 2001 )-.
  • LTB4 and/or iv) a stimulation of the synthesis of LTC4.
  • a further therapeutic approach would be the blocking of LTB4 receptors.
  • the haplo-insufficiency of MGST2 could also be treated by the substitution of the gene or the gene product.
  • the skin as a surfacial organ allows an access for gene therapeutical methods, which is less problematic compared to other tissues.
  • Such strategies are already pursued with Xeroderma pigmentosum, a monogenic, autosomal recessively inherited skin disease (Magnaldo 2004).
  • An increased expression of the MGST2 gene due to a decreased expression of EST CR742434, however, may be treated by inhibiting the activity of MGST2.
  • Therapeutic strategies could comprise i) an inhibition of the expression of MGST2, ii) an inhibition of the enzymatic activity of MGST2.
  • MGST2 is the first known risk gene for psoriasis. This assumption is supported by its genomic position in the coupling interval PSORSP9, its function in the leucotriene metabolism and the detection of an inactivating mutation in combination with a mutation in its regulatory gene EST
  • CR742434 in a patient suffering from familial psoriasis vulgaris. Alterations in MGST2 and/or EST CR742434 can be assumed in a considerable part of the patients suffering from psoriasis.
  • Jakobsson PJ Mancini JA 1 Ford-Hutchinson AW. Identification and characterization of a novel human microsomal glutathione S-transferase with leukotriene C4 synthase activity and significant sequence identity to 5- lipoxygenase-activating protein and leukotriene C4 synthase. J Biol Chem. 1996 Sep 6;271(36):22203-10.
  • Sagoo GS Tazi-Ahnini R, Barker JW, Elder JT, Nair RP, Samuelsson L, Traupe H, Trembath RC, Robinson DA, lies MM. Meta-Analysis of Genome- Wide Studies of Psoriasis Susceptibility Reveals Linkage to Chromosomes 6p21 and 4q28-q31 in Caucasian and Chinese Hans Population. J Invest Dermatol. 2004 Jun;122(6):1401-5. Sjostrom M, Jakobsson PJ, Heimburger M, Palmblad J, Haeggstrom JZ.

Abstract

The present invention relates to the use of MGST2 nucleic acids, MGST2 polypeptides, MGST2 effectors, EST CR742434 nucleic acid and EST CR742434 effectors for the diagnosis, prevention and /or therapy of psoriasis vulgaris and related diseases.

Description

Diagnosis and Therapy of Psoriasis vulgaris
Description
The present invention relates to the use of MGST2 nucleic acids, MGST2 polypeptides, MGST2 effectors, EST CR742434 nucleic acids and EST CR742434 effectors for the diagnosis, prevention and/or therapy of psoriasis vulgaris and related diseases.
Psoriasis vulgaris is a chronically inflammatory and hyperproliferative skin disease from which approximately 2-5% of the Caucasian population is affected (Nevitt and Hutchinson, 1996). It represents a considerable impairment for the persons affected and mostly requires a lifelong ambulant and partially also stationary therapy due to the chronic character. Thus, psoriasis vulgaris involves considerable costs in medical care, which is estimated to about $ 3 billion per year in the U.S. (Bowcock and Cockson, 2004).
The causes for psoriasis vulgaris are still not known despite intensive research. The infection of mechanically claimed sites of predilection (e.g. elbow and knee), which is typical for psoriasis vulgaris, indicates to a defect in the regulation of local inflammatory reactions of the skin.
Besides the role of environmental factors (e.g. climate, physical and mental stress) genetic factors play an important role for the development of psoriasis. Psoriasis often occurs familial. In identical twins, concordance rates of 65-70% were determined compared to 15-20% in fraternal twins (Wuepper et al., 1990). Genome-wide linkage analyses have identified a total of 19 chromosome regions until now, wherein psoriasis-related genes are possibly localized. In a meta-analysis of these tests, only for two regions, i.e. the MHC region in 6p21.3 (PSORS1 , OMIM 177900) as well as region 4q31 (PSORS9, OMIM 607857), a significant linkage with psoriasis could be confirmed (Sagoo et al., 2004). However, for none of the genes a significant association with psoriasis vulgaris could be shown, neither in these two regions nor elsewhere in the genome.
There is no causal therapy for psoriasis vulgaris until now. The therapeutic possibilities, which are currently available, only act symptomatically and often have unsatisfactory success or accompanied by severe side effects (e.g. therapy with glucocorticoids, methotrexate or cyclosporine) (Ortonne 2004).
In summary, psoriasis vulgaris represents a multifactorial disease which is caused by the interaction of genetic predisposition factors and environmental influences (Elder et al., 2001 ). The participation of several different genes explains a part of the clinical variability of this disease. The exact knowledge of these genes could enable individual therapeutic strategies which are adapted to the genetic status of an individual patient.
According to the present invention, it was found that a patient with severe psoriasis vulgaris had a mutation in EST CR742434 which is localized on the complementary DNA-strand of MGST2 and constitutes a putative regulator of MGST2. MGST2 plays an important role in the metabolism of the leucotrienes which are strong inflammatory mediators. MGST2 is located in a chromosomal region for which the relation to psoriasis was found (Sagoo et al. 2004). Thus, the identification of this gene defect has important consequences for the diagnosis and therapy of psoriasis vulgaris and related diseases.
A first aspect of the present invention relates to the use of a MGST2 nucleic acid, a MGST2 polypeptide or a MGST2 effector for the manufacture of an agent for the diagnosis, prevention and/or therapy of psoriasis vulgaris or related diseases.
A further aspect of the present invention relates to a screening method for identifying agents for the diagnosis, prevention and/or therapy of psoriasis vulgaris and related disorders comprising determining the interaction of a test compound with a MGST2. nucleic acid or polypeptide.
Still a further aspect of the present invention relates to a method for diagnosing psoriasis vulgaris and related disorders or a predisposition thereof in a subject comprising determining the amount and/or activity of a MGST2 nucleic acid or polypeptide in said subject or in a sample therefrom.
Still a further aspect of the present invention relates to a method for preventing or treating psoriasis vulgaris and related disorders in a subject comprising administering a pharmaceutically effective amount of MGST2 nucleic acid, a MGST2 polypeptide or a MGST2 effector to said subject.
The present invention has diagnostic and therapeutic applications in human and veterinary medicine, particularly in human medicine. The diagnosis particularly relates to the determination of the presence of altered MGST2 nucleic acids and/or polypeptides or altered amounts of MGST2 nucleic acids and/or polypeptides which are most usually associated with an alteration of at least one genomic copy of the MGST2 gene. Such changes may also be part of a specific haplotype of DNA markers around the causative mutation, which may enable its indirect diagnosis. The subjects to be treated according to the present invention are particularly subjects with a genomic alteration as described above, more particularly with a haplotypic gene defect.
The MGST2 nucleic acids and proteins are obtainable from vertebrate species. Preferred are nucleic acids encoding the human MGST2 polypeptide and variants thereof (GenBank Accession Number AA H25416 for the polypeptide and BC 025416 for the nucleic acid).
The term "MGST2 nucleic acid" according to the present invention particularly relates to
(a) the nucleoctide sequence of human MGST2 and/or a sequence complementary thereto, e.g. the genomic or cDNA sequence of human MGST2,
(b) a nucleotide sequence which hybridizes at 500C, preferably at 6O0C and more preferably at 65°C in a solution containing 1 x SSC and 0.1% SDS to a sequence of (a),
(c) the sequence corresponding to the sequences of (a) and/or (b) within the degeneration of the genetic code, or
(d) a partial sequence of any of the nucleotide sequences of (a) to (c) having a length of at least 15, preferably of at least 20 bases.
The term "MGST2 polypeptide" according to the present invention preferably relates to
(a) the amino acid sequence of human MGST2,
(b) an amino acid sequence which is at least 85%, preferably at least 90%, more preferably at least 95% and still more preferably 98% identical to the amino acid sequence of (a), or
(c) a partial sequence of the amino acid sequences of (a) to (b) having a length of at least 6, preferably 8 and more preferably of at least 12 amino acids.
For diagnostic applications, the term "MGST2 effector" relates to any molecule which is capable of detecting MGST2 nucleic acids and/or polypeptides in a biological sample, e.g. in a tissue or body fluid sample of a human patient. For therapeutic applications, the term "MGST2 effector" preferably relates to any molecule capable of modulating the amount, location and/or activity of MGST2 nucleic acids and/or polypeptides or nucleic acids and/or polypeptides interacting with MGST2 nucleic acids and/or polypeptides. Thus, the term "effector" encompasses both direct modulators of MGST2 and indirect modulators of MGST2 which modulate molecules interacting with MGST2.
Alternatively, the diagnosis relates to the determination of the presence of altered EST CR742434 nucleic acids or altered amounts of EST CR742434 nucleic acids. EST CR742434 exhibits an RNA gene, most of which have a regulatory function on the expression of other genes. Since EST CR742434 and the MGST2 gene overlap in their 51 regions in antisense-direction/ a regulatory function of EST CR742434 on the expression of MGST2 is very likely in the present case.
Still a further aspect of the present invention relates to the use of an EST CR742434 nucleic acid or an EST CR742434 effector for the manufacture of an agent for the diagnosis, prevention and/or therapy of psoriasis vulgaris or related diseases.
Still a further aspect of the present invention relates to a screening method for identifying agents for the diagnosis, prevention and/or therapy of psoriasis vulgaris and related disorders comprising determining the interaction of a test compound with an EST CR742434 nucleic acid.
Still a further aspect of the present invention relates to a method for diagnosing psoriasis vulgaris and related disorders or a predisposition thereof in a subject comprising determining the amount and/or activity of an EST CR742434 nucleic acid in said subject or in a sample therefrom.
Still a further aspect of the present invention relates to a method for preventing or treating psoriasis vulgaris and related disorders in a subject comprising administering a pharmaceutically effective amount of EST CR742434 nucleic acid or an EST CR742434 effector to said subject.
The term "EST CR742434 nucleic acid" according to the present invention particularly relates to
(a) the nucleoctide sequence of human EST CR742434 and/or a sequence complementary thereto, e.g. the genomic or cDNA sequence of human
EST CR742434,
(b) a nucleotide sequence which hybridizes at 500C, preferably at 600C and more preferably at 650C in a solution containing 1 x SSC and 0.1% SDS to a sequence of (a),
(c) the sequence corresponding to the sequences of (a) and/or (b) within the degeneration of the genetic code, or
(d) a partial sequence of any of the nucleotide sequences of (a) to (c) having a length of at least 15, preferably of at least 20 bases.
For diagnostic applications, the term "EST CR742434 effector" relates to any molecule which is capable of detecting EST CR742434 nucleic acids in a biological sample, e.g. in a tissue or body fluid sample of a human patient. For therapeutic applications, the term "EST CR742434 effector" relates to any molecule capable of modulating the amount, location and/or activity of EST CR742434 nucleic acids or nucleic acids interacting with EST CR742434 nucleic acids.
Further, according to the present invention, the term "psoriasis vulgaris or related diseases" relates to diseases selected from psoriasis vulgaris, and other T-cell mediated autoimmune diseases, particularly rheumatic diseases such as rheumatoid arthritis or Morbus Crohn.
The diagnostic aspect of the present invention may comprise the detection of MGST2 nucleic acid, e.g. a genomic MGST2 sequence or a MGST2 transcript. The diagnosis may comprise a sequence determination of a MGST2 nucleic acid, e.g. the presence of alterations, e.g. disruptions, polymorphisms etc. of a MGST2 nucleic acid sequence in a patient. The sequence determination may comprise the determination of a single nucleotide, e.g. in a single nucleotide polymorphism or the determination of longer sequence portion. Further, the diagnosis may comprise the determination of a MGST2 nucleic acid expression level, e.g. a qualitative and/or quantitative determination. The detection of a MGST2 nucleic acid may be carried out according to known methods, comprising nucleic acid amplification, nucleic acid sequencing, genotyping, gene expression analysis etc. and combinations thereof. For this purpose, MGST2 nucleic acid detection molecules such as primers and probes or corresponding nucleic acid analogues such as LNA or PNA molecules for primer extension, sequencing and/or amplification reactions may be employed.
With EST CR742434 being a regulatory gene of MGST2, however, the diagnostic aspect of the present invention also may comprise the detection of EST CR742434 nucleic acid in the embodiments described above for MGST2.
Alternatively, the diagnostic aspect of the present invention may comprise determination of a MGST2 polypeptide, e.g. the amount, location and/or activity of a MGST2 polypeptide. This aspect of the present invention preferably comprises an immunoassay wherein the presence of an MGST2 polypeptide is detected by using suitable antibodies. The antibodies may be specific for MGST2 wild-type molecules or MBST2 variants according to the respective test format. Methods of determining polypeptides in biological samples are known in the art.
A further aspect of the present invention relates to the prevention and/or therapy of psoriasis vulgaris and related diseases. In one embodiment, the therapeutic agent may be a therapeutic nucleic acid or nucleic acid analogue, e.g. a MGST2 nucleic acid molecule, EST CR742434 nucleic acid, an aptamer, an antisense molecule, an RNA interference molecule or a ribozyme which is capable of directly or indirectly modulating the MGST2 activity in a subject to be treated. For example, the nucleic acid may be a nucleic sequence encoding a MGST2 polypeptide, e.g. a genomic or a cDNA sequence which is transferred into target cells, particularly cells which are involved in psoriatic inflammatory reactions, such as blood cells, keratinocytes and/or fibroblasts, in order to substitute a lack of MGST2 activity in the target cells. In this context, it is referred to a gene therapeutic protocol which is used for the treatment of Xeroderma pigmentosum, a monogenic, autosomal recessive inherited skin disorder (Magnaldo 2004).
Alternatively, the therapeutic agent may be a nucleic acid or nucleic acid analogue which is a MGST2 effector. Suitable acid or nucleic acid analogue effector molecules are preferably selected from EST CR742434 nucleic acid, aptamers, antisense molecules, RNA interference molecules or ribosomes. Aptamers, e.g. nucleic acid or nucleic acid analogue molecules which are capable of binding to MGST2 or a protein interacting therewith and modulating its activity, may be generated by a screening and selection procedure involving the use of combinatorial nucleic acid libraries. Antisense molecules, RNAi molecules or ribozymes directed against the MGST2 mRNA sequence or against the sequence of mRNA molecules encoding polypeptides which interact with MGST2 may be used to modulate transcription of MGST2 mRNA or mRNA molecules encoding polypeptides interacting with MGST2. Preferably, antisense molecules may be used in the case of an increased MGST2 activity, in order to lower expression of MGST2. The therapeutic agent, however, also may be a nucleic acid or nucleic acid analogue which is an EST CR742434 effector.
The administration of nucleic acids may be carried out according to known methods, e.g. by introduction of naked DNA or by the use of transfer vehicles such as viruses, e.g. adenoviruses, adeno-associated viruses, retroviruses etc. as known in the art.
On the other hand, the therapeutic agent may be a polypeptide or a fragment thereof. For example, the therapeutic agent may comprise a polypeptide, e.g. MGST2 polypeptide or an biologically active fragment which may be introduced into target cells in order to increase the amount of MGST2 activity. Alternatively, the polypeptide may be a transcription factor capable of stimulating MGST2 transcription, particularly transcription from an unaltered, i.e. active MGST2 genomic sequence. The therapeutic polypeptide may also be an antibody directed against a molecule interacting with MGST2. For therapeutic purposes, the antibody is preferably a chimeric, humanized, or human antibody or an antigen-binding fragment of such an antibody. The production of such antibodies is known in the art. Suitable methods for administering polypeptides are known in the art. On the other hand, the agent may be a low molecular weight organic compound with the capability of altering, e.g. stimulating or inhibiting the function of MGST2 polypeptide or polypeptides interacting therewith. Preferably, the low molecular weight organic compound has a molecular weight of about 100- 2500 Da.
Preferably, the therapeutic agent is a stimulator of MGST2 activity. In one embodiment, the stimulation is a direct stimulation wherein an increased MGST2 amount and/or activity in the target cells is provided, e.g. by introduction of a MGST2 polypeptide or nucleic acid into the target cell. On the other hand, the therapeutic agent may be an indirect stimulator of MGST2 activity, i.e. a molecule which interacts with physiologic compounds which may modulate MGST2 activity. For example, the agent may be an inhibitor of LTA4 synthesis, e.g. a 5-lipoxygenase inhibitor or a 5- lipoxygenase activating protein (FLAP) inhibitor. Furthermore, the agent may be an inhibitor of leukotriene A4 (LTA4) hydrolase. Furthermore, the agent may be a stimulator of leukotriene C4 (LTC4) synthesis. Furthermore, the agent may be an inhibitor of an leukotriene B4 (LTB4) receptor, e.g. an inhibitor of the LTB4 receptors BLT1 or BLT2.
Inhibition of MGST2 activity, on the other hand, may be achieved by administering suitable pharmacological substances in order to decrease enzymatic activity of MGST2.
The therapeutic aspect of the present invention preferably comprises the administration of a pharmaceutical composition which comprises as an active ingredient a MGST2 nucleic acid, a MGST2 polypeptide, a MGST2 effector, an EST CR742434 nucleic acid or an EST CR742434 effector together with pharmaceutically acceptable carriers, diluents and/or adjuvants. The pharmaceutical composition may be administered alone or in combination with at least one other agent. The pharmaceutical composition may be administered by any known route of administration, e.g. oral, topical, parenteral, transdermal, subcutaneous means. Topical or transdermal administration to the skin areas involved in psoriasis is preferred.
The pharmaceutical composition comprises the active ingredient in a therapeutically effective amount. The determination of an effective dose for a specific active ingredient is known in the art. For example, the therapeutic effective dose can be estimated in cell culture assays or in experimental animal models. The exact dosage, route and frequency of administration will be determined inter alia by the type of active ingredient, the severity of disease and the condition of the subject to be treated.
The present invention further provides a screening method for identifying novel agents for the diagnosis, prevention and/or therapy of psoriasis vulgaris and related disorders. The screening method may comprise the use of a non-cellular screening system, a cellular screening system or a transgenic non-human animal. In such model, the interaction of a test compound or a plurality of test compounds, e.g. from a test compound library with a MGST2 nucleic acid or polypeptide is determined wherein the interaction may comprise a direct modulation of MGST2 nucleic acid or polypeptide or an indirect modulation of an MGST2 nucleic acid or polypeptide as described above, e.g. inhibition of LTA4 synthesis, inhibition of LTA4 hydrolase, stimulation of LTC4 synthesis and/or inhibition of LTB4 receptors.
Alternatively, the screening method may comprise determination of an interaction of a test compound with an EST CR742434 nucleic acid. The interaction may comprise direct or indirect modulation of EST CR742434 nucleic acid.
The present invention also relates to an isolated cell, preferably a mammalian cell and more preferably a human cell, capable of over- or underexpressing the MGST2 gene or the EST CR742434 gene. Such cell may provide a suitable in vitro model system for further characterizing the molecular mechanisms of psoriasis or related diseases. Alternatively, these molecular mechanisms may be elucidated in a non-human transgenic animal, preferably a transgenic non-human mammal, which is capable of over- or underexpressing the MGST2 gene or the EST CR742434 gene. Preferably, the non-human animal has at least one deficient MGST2 gene copy.
Further, the present invention is explained in more detail by the following examples.
Examples
1. Patient and Methods of the Tests
The starting point of the tests was a 43-year old patient suffering from psoriasis vulgaris and a balanced chromosomal translocation 46,XY,t(2;4) (p25;q31.1) (Fig. 1). The already deceased and cytogenetically not tested father of the patient also suffered from psoriasis, very likely, he also carried the translocation, as evidenced by repeated miscarriages of two female partners.
Since the breaking point in the chromosomal band 4q31.1 is within the region of the gene locus for psoriasis susceptibility PSORS9 (OMIM 607857; Zhang et al.; 2002; Sagoo et al., 2004), a molecular cytogenetic break point analysis was carried out in the patient. First, two break point covering BAC (bacterial artificial chromosome) clones were identified [BAC RP11-1079H4 (GenBank Accession Number AQ681984) and BAC RP11-90M18 (GenBank Accession Number AC019049)] according to the method described by Wirth et al. (1999). Then, a genomic DNA of the patient was digested with restriction enzymes. Fragments of deviating size compared to control persons were detected by means of Southern-Blot-hybridization. Thereby, the break point on chromosome 4 was localized about 6 kb upstream the 5' end of exon 1 of the MGST2 gene, and within intron 3 of EST CR742434 on the complementary DNA-strand (Fig. 2). The disruption of the regulatory gene EST CR742434 may lead to an altered expression of MGST2 in this patient.
On chromosome 2, the BAC-clone RP11-79601 (GenBank Accession Number AQ514709) was identified as the break point covering clone. Since no genes are known in this region, no further analysis was carried out therein.
2. The Function of MGST2 in the Leucotriene Metabolism
Leucotrienes are inflammatory mediators. The leucotriene with the strongest pro-inflammatory effect in psoriatic cutaneous areas is LTB4, which arises from LTA4 by hydrolysis (Iversen et al., 1997) (Fig. 3).
The gene product of MGST2, the microsomal glutathione S-transferase 2, catalyses the formation of LTC4 by the conjugation of glutathione with LTA4 (Jakobsson et al., 1996). This reaction can also be catalysed by two further enzymes, i.e. the leucotriene-C4-synthease (LTC4S) and MGST3. In the only cell type which has been tested with regard to that so far, i.e. endothelial cells of umbilical veins, MGST2 is the main and probably even the only active enzyme for the synthesis of LTC4 (Sjδstrδm et al., 2001 )-. It is assumed that the altered activity of MGST2 results in a changed synthesis of LTC4 in some tissues. This could lead to a surplus of LTA4, which is thus available as a substrate in surplus for the increased formation of LTB4, which would stimulate the expression of psoriatic reactions.
A deficiency of the regulator CR742434, however, may result in an increased expression of the MGST2 gene. The leucotriene metabolism also may be affected in this case, again resulting in an increased inflammation tendency typical for psoriasis. 3. Diagnostic Consequences
Data obtained from the previous linkage and association studies allows the conclusion that psoriasis vulgaris is etiologically heterogeneous and two important risk genes are localized in the chromosome sections 6p21.3 and 4q31.1 (Sagoo et al., 2004). It is highly probable that defects of these genes also result in a different reaction to these therapeutic measures. Due to this reason, every therapy should preferably be preceded by a molecular genetic diagnostic in future in order to choose the therapy appropriate for the genetic status of the individual patient. MGST2 and its regulatory gene EST CR742434 are the first and only genes so far which are available for such a diagnosis.
4. Therapeutic Consequences
The results allow the assumption that the inactivation of a copy of MGST2 results in an increased formation of LTB4. Thus, both alternative metabolic pathways of LTA4 would be suitable for starting points for a therapeutic intervention. These therapeutic strategies could comprise i) an increase of the amount and/or activity of MGST2, ii) an inhibition of the LTA4-synthesis iii) an inhibition of the LTA4-hydrolase, which catalyses the formation of
LTB4, and/or iv) a stimulation of the synthesis of LTC4.
A further therapeutic approach would be the blocking of LTB4 receptors. The haplo-insufficiency of MGST2 could also be treated by the substitution of the gene or the gene product. The skin as a surfacial organ allows an access for gene therapeutical methods, which is less problematic compared to other tissues. Such strategies are already pursued with Xeroderma pigmentosum, a monogenic, autosomal recessively inherited skin disease (Magnaldo 2004). An increased expression of the MGST2 gene due to a decreased expression of EST CR742434, however, may be treated by inhibiting the activity of MGST2. Therapeutic strategies could comprise i) an inhibition of the expression of MGST2, ii) an inhibition of the enzymatic activity of MGST2.
5. Outlook to Further Studies
MGST2 is the first known risk gene for psoriasis. This assumption is supported by its genomic position in the coupling interval PSORSP9, its function in the leucotriene metabolism and the detection of an inactivating mutation in combination with a mutation in its regulatory gene EST
CR742434 in a patient suffering from familial psoriasis vulgaris. Alterations in MGST2 and/or EST CR742434 can be assumed in a considerable part of the patients suffering from psoriasis.
References
Bowcock AM, Cookson WO. The genetics of psoriasis, psoriatic arthritis and atopic dermatitis. Hum MoI Genet. 2004 Apr 1 ;13 Spec No 1 :R43-55.
Elder JT, Nair RP, Henseler T, Jenisch S, Stuart P, Chia N, Christophers E1 Voorhees JJ. The genetics of psoriasis 2001 : the odyssey continues. Arch Dermatol. 2001 Nov;137(11):1447-54.
Iversen L, Kragballe K, Ziboh VA. Significance of Ieukotriene-A4 hydrolase in the pathogenesis of psoriasis. Skin Pharmacol. 1997;10(4):169-77.
Jakobsson PJ, Mancini JA1 Ford-Hutchinson AW. Identification and characterization of a novel human microsomal glutathione S-transferase with leukotriene C4 synthase activity and significant sequence identity to 5- lipoxygenase-activating protein and leukotriene C4 synthase. J Biol Chem. 1996 Sep 6;271(36):22203-10.
Magnaldo T. Xeroderma pigmentosum: from genetics to hopes and realities of cutaneous gene therapy. Expert Opin Biol Ther. 2004 Feb;4(2): 169-79.
Nevitt GJ, Hutchinson PE., Psoriasis in the community: prevalence, severity and patients' beliefs and attitudes towards the disease. Br J Dermatol. 1996 Oct;135(4):533-7.
Ortonne JP. Redefining clinical response in psoriasis: targeting the pathological basis of disease. J Drugs Dermatol. 2004 Jan-Feb;3(1 ):13-20.
Sagoo GS, Tazi-Ahnini R, Barker JW, Elder JT, Nair RP, Samuelsson L, Traupe H, Trembath RC, Robinson DA, lies MM. Meta-Analysis of Genome- Wide Studies of Psoriasis Susceptibility Reveals Linkage to Chromosomes 6p21 and 4q28-q31 in Caucasian and Chinese Hans Population. J Invest Dermatol. 2004 Jun;122(6):1401-5. Sjostrom M, Jakobsson PJ, Heimburger M, Palmblad J, Haeggstrom JZ. Human umbilical vein endothelial cells generate leukotriene C4 via microsomal glutathione S-transferase type 2 and express the Cysl_T(1) receptor. Eur. J. Biochem. 2001 May; 268(9): 2578-86.
Wirth J, Nothwang HG, van der Maarel S, Menzel C, Borck G, Lopez-Pajares I, Brondum-Nielsen K, Tommerup N1 Bugge M, Ropers HH, Haaf T. Systematic characterisation of disease associated balanced chromosome rearrangements by FISH: cytogenetically and genetically anchored YACs identify microdeletions and candidate regions for mental retardation genes. J Med Genet. 1999 Apr;36(4):271-8.
Wuepper KD, Coulter SN, Haberman A. Psoriasis vulgaris: a genetic approach. J Invest Dermatol. 1990 Nov;95(5):2S-4S.
Zhang XJ, He PP, Wang ZX, Zhang J, Li YB, Wang HY, Wei SC1 Chen SY, Xu SJ1 Jin L, Yang S, Huang W. Evidence for a major psoriasis susceptibility locus at 6p21(PSORS1 ) and a novel candidate region at 4q31 by genome- wide scan in Chinese hans. J Invest Dermatol. 2002 Dec;119(6):1361-6.

Claims

Claims
1. Use of a MGST2 nucleic acid, a MGST2 polypeptide or a MGST2 effector for the manufacture of an agent for the diagnosis, prevention and/or therapy of psoriasis vulgaris or related diseases.
2. The use of claim 1 for human medicine.
3. The use of claim 1 or 2 for the manufacture of an agent for diagnosis.
4. The use of any one of claims 1-3 wherein the diagnosis comprises the detection of MGST2 nucleic acid.
5. The use of claim 4 wherein the MGST2 nucleic acid is a genomic sequence.
6. The use of claim 4 wherein the MGST2 nucleic acid is a transcript.
7. The use of any one of claims 4-6 wherein the diagnosis comprises a determination of MGST2 nucleic acid sequence.
8. The use of any one of claims 4-7 wherein the diagnosis comprises a determination of MGST2 nucleic acid expression level.
9. The use of any one of claims 1-3 wherein the diagnosis comprises the determination of a MGST2 polypeptide.
10. The use of claim 9 wherein the diagnosis comprises determination, amount and/or activity of a MGST2 polypeptide.
11. The use of claim 1 or 2 for the manufacture of an agent for prevention and/or therapy.
12. The use of claim 11 wherein the agent is a nucleic acid or nucleic acid analogue.
13. The use of claim 12 wherein the agent is a nucleic acid encoding a MGST2 polypeptide.
14. The use of claim 12 wherein the agent is a nucleic acid analogue which is a MGST2 effector.
15. The use of claim 11 wherein the agent is a polypeptide.
16. The use of claim 11 wherein the agent is a low-molecular weight organic compound.
17. The use of claim 14 wherein the agent is a modulator of MGST2 activity.
18. The use of any one of claim 11-17 wherein the agent is an inhibitor of LTA4 synthesis.
19. The use of claims 11-17 wherein the agent is an inhibitor of LTA4 hydrolase.
20. The use of claims 11-17 wherein the agent is a stimulator of LTC4 synthesis.
21. The use of claims 11-17 wherein the agent is an inhibitor of a LTB4 receptor.
22. Use of an EST CR742434 nucleic acid or an EST CR742434 effector for the manufacture of an agent for the diagnosis, prevention and/or therapy of psoriasis vulgaris or related diseases.
23. A screening method for identifying agents for the diagnosis, prevention and/or therapy of psoriasis vulgaris and related disorders comprising determining the interaction of a test compound with a MGST2 nucleic acid or polypeptide.
24. The method of claim 23 wherein the interaction comprises a direct modulation of a MGST2 nucleic acid or polypeptide.
25. The method of claim 23 wherein the interaction comprises an indirect modulation of a MGST2 nucleic acid or polypeptide.
26. The method of claim 25 wherein the indirect modulation comprises (i) inhibition of LTA4 synthesis,
(ii) inhibition of LTA4 hydralase, (iii) stimulation of LTC4 synthesis and/or
(iv) inhibition of a LTB4 receptor.
27. A screening method for identifying agents for the diagnosis, prevention and/or therapy of psoriasis vulgaris and related disorders comprising determining the interaction of a test compound with an EST CR742434 nucleic acid.
28. A method for diagnosing psoriasis vulgaris and related disorders or a predisposition thereof in a subject comprising determining the amount and/or activity of a MGST2 nucleic acid or polypeptide in said subject or in a sample therefrom.
29. A method for diagnosing psoriasis vulgaris and related disorders or a predisposition thereof in a subject comprising determining the amount and/or activity of an EST CR742434 nucleic acid in said subject or in a sample therefrom.
30. A method for preventing or treating psoriasis vulgaris and related disorders in a subject comprising administering a pharmaceutically effective amount of MGST2 nucleic acid, a MGST2 polypeptide or a MGST2 effector to said subject.
31. A method for preventing or treating psoriasis vulgaris and related disorders in a subject comprising administering a pharmaceutically effective amount of EST CR742434 nucleic acid or an EST CR742434 effector to said subject.
32. The method of any one of claims 28-31 wherein said subject is a human.
33. A cell capable of over- or underexpressing the MGST2 gene.
34. A cell capable of over- or underexpressing the EST CR742434 gene.
35. The cell of claim 33 or 34 which is a human cell.
36. A non-human transgenic animal capable of over- or underexpressing the MGST2 gene.
37. A non-human transgenic animal capable of over- or underexpressing the EST CR742434 gene.
38. The cell of claim 33 or 35 or the non-human animal of claim 36 which has at least one deficient MGST2 gene copy.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9981926B2 (en) 2013-12-20 2018-05-29 Novartis Ag Heteroaryl butanoic acid derivatives

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003081201A2 (en) * 2002-03-21 2003-10-02 Yissum Research Development Company Of The Hebrew University Of Jerusalem Peripheral blood cell markers useful for diagnosing multiple sclerosis and methods and kits utilizing same

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003081201A2 (en) * 2002-03-21 2003-10-02 Yissum Research Development Company Of The Hebrew University Of Jerusalem Peripheral blood cell markers useful for diagnosing multiple sclerosis and methods and kits utilizing same

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
DATABASE GENBANK [online] 21 December 2004 (2004-12-21), XP002344422, retrieved from STN Database accession no. CR742434 *
GOSWAMI SUMANTA ET AL: "Spectrum and range of oxidative stress responses of human lens epithelial cells to H2O2 insult.", IOVS, vol. 44, no. 5, May 2003 (2003-05-01), pages 2084 - 2093, XP009053378 *
JAKOBSSON P-J ET AL: "IDENTIFICATION AND CHARACTERIZATION OF A NOVEL HUMAN MICROSOMAL GLUTATHIONE S-TRANSFERASE WITH LEUKOTRIENE C4 SYNTHASE ACTIVITY ANDSIGNIFICANT SEQUENCE IDENTITY TO 5-LIPOXYGENASE-ACTIVATING PROTEIN AND LEUKOTRIENE C4 SYNTHASE", JOURNAL OF BIOLOGICAL CHEMISTRY, AMERICAN SOCIETY OF BIOLOGICAL CHEMISTS, BALTIMORE, MD, US, vol. 271, no. 36, 6 September 1996 (1996-09-06), pages 22203 - 22210, XP002044920, ISSN: 0021-9258 *
SJOSTROM, MATTIAS ET AL: "Human umbilical vein endothelial cells generate leukotriene C4 via microsomal glutathione S-transferase type 2 and express the CysLT1 receptor", EUROPEAN JOURNAL OF BIOCHEMISTRY , 268(9), 2578-2586 CODEN: EJBCAI; ISSN: 0014-2956, 2001, XP009053368 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9981926B2 (en) 2013-12-20 2018-05-29 Novartis Ag Heteroaryl butanoic acid derivatives
US11453651B2 (en) 2013-12-20 2022-09-27 Novartis Ag Heteroaryl butanoic acid derivatives

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