EP1080195A1 - Endometriose-assoziiertes gen - Google Patents

Endometriose-assoziiertes gen

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Publication number
EP1080195A1
EP1080195A1 EP99927792A EP99927792A EP1080195A1 EP 1080195 A1 EP1080195 A1 EP 1080195A1 EP 99927792 A EP99927792 A EP 99927792A EP 99927792 A EP99927792 A EP 99927792A EP 1080195 A1 EP1080195 A1 EP 1080195A1
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EP
European Patent Office
Prior art keywords
polypeptide
nucleic acid
seq
endometriosis
sequence
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EP99927792A
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German (de)
English (en)
French (fr)
Inventor
Anna Starzinski-Powitz
Silvia Kotzian
Heike Handrow-Metzmacher
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Individual
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4748—Tumour specific antigens; Tumour rejection antigen precursors [TRAP], e.g. MAGE
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • 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

Definitions

  • the present invention relates to an invasive process, e.g. Endometriosis-associated gene, a polypeptide encoded thereof, an antibody directed against the polypeptide and the pharmaceutical application of the nucleic acid, the polypeptide and the antibody.
  • an invasive process e.g. Endometriosis-associated gene, a polypeptide encoded thereof, an antibody directed against the polypeptide and the pharmaceutical application of the nucleic acid, the polypeptide and the antibody.
  • Endometriosis is the second most common female disease and is defined as the presence of endometrial cells outside the uterus. Endometriosis affects about one in five women of reproductive age, and even one in two in women with fertility problems.
  • uterine lining is found exclusively in the uterus.
  • tissue outside of the uterus that histologically looks like uterine mucosa can be found, for example on the outside of the uterus, on the intestine, or even in the pancreas or lungs.
  • these foci of endometriosis are located outside the uterus, they also bleed during menstruation, so they are influenced by the hormones of the female cycle. Since the focus of the endometriosis, like the endometrium, changes in volume in the cycle, depending on the location, these changes can cause pain.
  • the body responds with an inflammatory reaction to the endometriosis cells, which in turn triggers pain.
  • the inflammation also leads to adhesions in the area of the ovaries and fallopian tubes and is therefore responsible for the so-called mechanical sterility of the women concerned.
  • messenger substances e.g. cytokines, prostaglandins
  • cytokines e.g. cytokines, prostaglandins
  • endometriosis cells could be classified between normal cells and tumor cells: on the one hand, they do not show any neoplastic behavior, on the other hand, however, like metastatic tumor cells, they have the ability to move across organ boundaries and to grow into other organs, i.e. they show invasive behavior. For this reason, endometriosis cells are defined in the literature as "benign tumor cells", although no tumor-specific mutations in proto-oncogenes have been found in such cells.
  • the invention was based on the task of identifying new genes which play a role in invasive processes and which may be associated with the pathophysiological phenotype of endometriosis.
  • This object is achieved according to the invention by identifying, cloning and characterizing a gene which is referred to as an endometriosis-associated gene and which codes for a polypeptide.
  • This gene sequence was discovered using differential display RT-PCR (Liang and Pardee, Science 257 (1992), 967-971). For this purpose, invasive and non-invasive variants of an endometriosis cell line were compared with each other. This led to a cDNA sequence that is specific for the invasive variant of the endometriosis cells. An associated RNA of 4 kb in length was found. A corresponding cDNA isolated from a cDNA phage bank has an open reading frame (ORF) of 302 amino acids.
  • An object of the present invention is a nucleic acid which
  • (c) comprises a nucleotide sequence hybridizing with the sequences from (a) and / or (b) under stringent conditions.
  • nucleic acids preferably encode a polypeptide associated with invasive processes, or a portion thereof.
  • nucleotide sequences with the following accession numbers are stored in the EMBL EST database: Z98886, Ac003017, AL023586, Aa452993, Aa452856. These sequences do not represent nucleic acids according to the invention.
  • the first two of these sequences are DNAs which have been isolated from the human brain and which, with the sections from nucleotide 970 to approx. 2000 and from 760 to approx. 1450, show a 90% base identity with SEQ ID NO. 1, or with the sections from nucleotide 1054 to 2084 and from 844 to approx. 1534 based on SEQ ID NO. 3, which has 84 additional bases at the 5 'end.
  • ALO23586 is also a human sequence, very similar to Z98885 and also with SEQ ID NO. 1 in the range from 970 to approx. 2000 homology.
  • sequences Aa452993 and Aa452856 originate from mouse embryos and show base identity with the nucleotides (nt) from approx. 1060 to approx. 1450 and from approx. 24 to 440 of SEQ ID NO. 1 or from about 1 144 to about 1534 or from about 108 to about 524 according to the nucleotide positions in SEQ ID NO. 3. None of these 4 sequences has been assigned a reading frame or a function.
  • the in SEQ ID NO. 1 nucleotide sequence contains an open reading frame, which corresponds to a polypeptide with a length of 302 amino acids. This polypeptide is in SEQ ID NO. 2 amino acid sequence shown.
  • SEQ ID NO. shows a nucleotide sequence as in SEQ ID NO.
  • SEQ ID NO. 3 encoded polypeptides thus has 28 additional amino acids at the N-terminus and, with a total of 330 amino acids, is in SEQ ID NO. 4 shown.
  • SEQ ID NO. 2 and 4 represent a portion of the C-terminal end of the native polypeptide.
  • FIG. 1 shows a schematic representation of the cDNA of the endometriosis-associated gene according to the invention.
  • 5 exons E1 to E5 and the position of fragment 1 (394 nt) used as a probe in the DDRT-PCR can be seen.
  • the positions of the PCR primers (see Example 4, Table 1) which were used for the RT-PCR are also shown.
  • exon 4a whose nucleotide sequence is shown in SEQ ID NO. 5 is shown. This exon 4a can be present. If it exists, it is located between exon 4 and exon 5. This corresponds to the position between nt1054 and nt 1055 in SEQ ID NO.3.
  • a combination of the sequences SEQ ID NO. 1/3 with SEQ ID NO. 5 is thus e.g. a sequence which contains the sequence of exon 4a at the position mentioned.
  • nucleotide sequences that hybridize with one of the aforementioned sequences.
  • hybridization according to the present invention is used in Sambrook et al. (Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press (1 989), 1 .101-1 .104).
  • One under such washing conditions with one or more of those in SEQ ID NO. 1, 3 and 5 nucleotide sequences shown or a nucleotide sequence hybridizing to these sequences in the context of the degeneration of the genetic code hybridizing nucleotide sequence is a nucleotide sequence according to the invention.
  • the nucleotide sequence according to the invention is preferably a DNA. However, it can also comprise an RNA or a nucleic acid analog, such as a peptidic nucleic acid.
  • the nucleic acid according to the invention particularly preferably comprises a protein-coding section of the sequence shown in SEQ ID NO. 1, 3 or / and 5 nucleotide sequences shown or a sequence which has a homology of more than 80%, preferably more than 90% and particularly preferably more than 95% to that in SEQ ID NO. 1, 3 or 5 nucleotide sequences shown or a preferably at least 20 nucleotides (nt) and particularly preferably at least 50 nt long section thereof.
  • nucleic acids which, as described above, have the sequence of SEQ ID NO. 5 in addition to those of SEQ ID NO. 1 or 3.
  • the homology is given in percent of identical positions when comparing two nucleic acids (or peptide chains), whereby 100% homology means the complete identity of the compared chain molecules (Herder: Lexicon of Biochemistry and Molecular Biology, Spektrum Akademischer Verlag 1 995).
  • Nucleic acids according to the invention are preferably obtainable from mammals and in particular from humans. They can be made using known techniques using short sections of the SEQ ID NO. 1, 3 or / and 5 nucleotide sequences shown are isolated as hybridization probes and / or as amplification primers.
  • nucleic acids according to the invention can also be produced by chemical synthesis, it being possible, instead of the usual nucleotide building blocks, to use modified nucleotide building blocks, for example 2′-O-alkylated nucleotide building blocks.
  • nucleic acids or sections thereof according to the invention can thus be used for the production of primers and probes which are preferably provided with markers or marking groups.
  • Intron-spanning oligonucleotide primers are also preferred, which are particularly suitable for identifying different mRNA species.
  • polypeptides encoded by the nucleic acids as defined above are the polypeptides encoded by the nucleic acids as defined above. These polypeptides preferably contain (a) those in SEQ ID NO. 2 or 4 amino acid sequence shown or
  • the invention also relates to muteins, variants and fragments thereof. These are to be understood as sequences which differ from those in SEQ ID NO. By substitution, deletion and / or insertion of individual amino acids or short amino acid segments. Distinguish 2 or 4 amino acid sequences shown.
  • variant includes both naturally occurring allelic variations or splice variations of the endometriosis protein, as well as recombinant DNA technology (in particular in w ' tr ⁇ mutagenesis with the help of chemically synthesized oligonucleotides) produced proteins which, in terms of their biological and / or immunological activity, correspond to those described in SEQ IN NO. 2 or 4 shown proteins essentially correspond.
  • This term also includes chemically modified polypeptides. These include polypeptides that are modified on the termini and / or on reactive amino acid side groups by acylation, for example acetylation or amidation.
  • the amino acid sequences according to the invention also include polypeptide fragments (peptides) which have a section of at least 10 amino acids long in the sequence shown in SEQ ID NO. Represent 2 or 4 amino acid sequence shown.
  • Another object of the present invention is a vector which contains at least one copy of a nucleic acid according to the invention.
  • This vector can be any prokaryotic or eukaryotic vector on which the DNA sequence according to the invention is preferably located in connection with expression signals such as a promoter, operator, enhancer etc.
  • prokaryotic vectors are chromosomal vectors, such as bacteriophages, and extrachromosomal vectors, such as plasmids, with circular plasmid vectors being particularly preferred.
  • Suitable prokaryotic vectors are e.g. in Sambrook et al., supra, Chapters 1-4.
  • the vector according to the invention is particularly preferably a eukaryotic vector, e.g.
  • yeast vector or a higher cell vector, e.g. a plasmid vector, viral vector or plant vector.
  • a yeast vector or a higher cell vector, e.g. a plasmid vector, viral vector or plant vector.
  • Such vectors are familiar to the person skilled in the field of molecular biology, so that there is no need to go into them here. In this context, reference is made in particular to Sambrook et al., Supra, Chapter 1 6.
  • the invention also relates to a vector which has a section of at least 21 nucleotides long in SEQ ID NO. 1, 3 or / and 5 sequences shown or a combination thereof.
  • This preferably has Section of a nucleotide sequence which originates from the protein-coding region of the said sequences or from a region essential for the expression of the protein or polypeptide.
  • These nucleic acids are particularly suitable for the production of therapeutically usable antisense nucleic acids, which are preferably up to 50 nucleotides long.
  • Another object of the present invention is a cell which is transformed with a nucleic acid according to the invention or a vector according to the invention.
  • the cell can be both a eukaryotic and a prokaryotic cell. Methods for transforming cells with nucleic acids are state of the art and therefore need not be explained in more detail. Examples of preferred cells are eukaryotic cells, in particular animal and particularly preferably mammalian cells.
  • Another object of the present invention is an antibody or a fragment of such an antibody against the polypeptide (s) encoded by the endometriosis gene or variants thereof.
  • Such antibodies are particularly preferably directed against the entire polypeptides encoded therein or against a peptide sequence which corresponds to the amino acids 1 -330 of the amino acids shown in SEQ ID NO. 4 corresponds to the amino acid sequence shown.
  • the identification, isolation and expression of a gene according to the invention which is specifically associated with invasive processes and in particular with endometriosis provides the prerequisites for the diagnosis, therapy and prevention of diseases which are based on the above-mentioned disorders.
  • Antibodies can be produced in a customary manner by immunizing experimental animals with the complete polypeptide or fragments thereof and subsequently obtaining them of the resulting polyclonal antisera. According to the Köhler and Milstein method and their further developments, monoclonal antibodies can be obtained from the antibody-producing cells of the experimental animals in a known manner by cell fusion. Human monoclonal antibodies can also be produced by known methods. Such antibodies could then be used both for diagnostic tests, in particular of endometriosis cell tissue, or for therapy.
  • samples such as body fluids, especially human body fluids can be examined for the presence of a polypeptide encoded by the endometriosis gene and for the presence of autoantibodies against such a polypeptide .
  • Polypeptides or fragments thereof encoded by the endometriosis gene can then be detected in such samples with the help of a specific antibody, e.g. of an antibody according to the invention can be detected.
  • recombinant fusion proteins can preferably be used which contain part or a whole or all of the polypeptide which is encoded by the endometriosis gene and which are fused to a protein domain which enables detection, for example the maltose-binding Protein (MBP).
  • MBP maltose-binding Protein
  • Diagnostic tests can also be carried out using specific nucleic acid probes for detection at the nucleic acid level, e.g. at the gene or transcript level.
  • the provision of the nucleotide and amino acid sequences and antibodies according to the invention enables a targeted search for effectors of the polypeptides / proteins.
  • Effectors are substances which have an inhibitory or activating effect on the polypeptide according to the invention and which are able to control the cell functions controlled by the polypeptides selectively influence. These can then be used in the therapy of corresponding clinical pictures, such as those based on invasive processes.
  • the invention thus also relates to a method for identifying effectors of the endometriosis proteins, in which cells which express the protein are brought into contact with various potential effector substances, for example low-molecular substances, and the cells are checked for changes, for example cell-activating, cell-inhibiting. cell proliferative and / or cell genetic changes, analyzed. In this way, binding targets of the endometriosis proteins can also be identified.
  • the discovery of the gene according to the invention provides additional possibilities for the diagnosis, prevention and therapy of cancerous diseases.
  • the present invention thus also relates to a pharmaceutical composition which comprises, as active components, nucleic acids, vectors, cells, polypeptides, peptides and / or antibodies, as stated above.
  • the pharmaceutical composition according to the invention can furthermore contain pharmaceutically customary excipients, auxiliaries and / or additives, and optionally further active components.
  • the pharmaceutical composition can be used in particular for the diagnosis, therapy or prevention of diseases involving invasive Processes are associated.
  • the composition according to the invention can also be used for the diagnosis of a predisposition to such diseases, in particular for the diagnosis of a risk for endometriosis.
  • FIG. 1 shows a schematic representation of the cDNA of the endometriosis-associated gene, only exons E1 to E5 being shown.
  • SEQ ID NO. 1 represents a nucleotide sequence which contains genetic information coding for the endometriosis-associated gene, an open reading frame ranging from nucleotide 3 to 91
  • SEQ ID NO. 2 represents the amino acid sequence of the open reading frame of the one in SEQ ID NO. 1 shown nucleotide sequence, wherein the amino acid sequence of the open reading frame ranges from amino acid 1 to 302.
  • SEQ ID NO. 3 represents a nucleotide sequence like that of SEQ ID NO. 1, but it contains an additional 84 nucleotides at the 5 'end, the open reading frame extends from nucleotide 3 to 995.
  • SEQ ID NO. 4 represents the amino acid sequence of the open reading frame of the one in SEQ ID NO. 3 shows the nucleotide sequence shown, this amino acid sequence having 320 amino acids, of which the C-terminal 302 are identical to those in SEQ ID NO. 2.
  • SEQ ID NO. 5 shows the nucleotide sequence of the additional exon 4a which may be present, consisting of the 21 8 nt shown, with exon 4a between nucleotide 1 054 and 1055 (based on SEQ ID NO. 3), if it exists.
  • invasive and non-invasive cells from the epithelial endometriosis cell line EEC1 45T + were used.
  • the cells were cultivated in Dulbecco medium (DMEM) with 10% fetal calf serum and diluted 1: 5 twice a week (passage).
  • DMEM Dulbecco medium
  • invasive cells from passage 1 7 and non-invasive cells from passage 33 were used.
  • the cells were transformed with SV40 and analyzed by differential display reverse transcription polymerase chain reaction (DDRT-PCR).
  • This method developed by Liang and Pardee is a method for distinguishing the expression pattern of different cell types or the changing expression pattern of a cell type under different living conditions or during changing developmental stages (Liang and Pardee (1 992), Science 257, 967-971).
  • the basis of the DDRT-PCR technique is based on the consideration that approximately 1,500 genes are expressed in each cell and that in principle each individual mRNA molecule can be represented by means of reverse transcription and amplification with random primers.
  • the cellular PolyA + RNA was first transcribed into cDNA with the aid of several different dT ⁇ VX primers (downstream primers, anchor primers).
  • the resulting cDNA populations were then PCR-amplified with 4 downstream and 20 upstream primers from the RNA-Map TM kit from Genhunter, Nashville (1 994) with the addition of a radioactively labeled nucleotide.
  • the reaction mixtures were concentrated in vacuo and the cDNA fragments obtained were separated in a six percent native PAA gel (polyacrylamide). DNA detection was carried out by autoradiography. PCR approaches which showed clear differences in the band pattern for the two cell variants to be examined were repeated twice in order to check the reproducibility. If the differences found previously were confirmed, the bands were eluted from the gel, reamplified, cloned and sequenced according to known methods.
  • Northern blot analyzes were carried out to check the expression pattern for the DDRT-PCR fragment 1.
  • 20 ⁇ g total RNA or 4 ⁇ g polyA + RNA were separated in 1% denaturing agarose gels and transferred to a nylon membrane overnight.
  • the RNA was fixed on the membrane by irradiation with UV light.
  • the hybridization with 32 P-labeled probes (labeling using the RPL kit from Amersham) was carried out overnight in a hybridization solution containing formamide at 42 ° C.
  • the membrane was then washed with increasing stringency until a selective intensity of the radioactive radiation was measurable.
  • the hybridization pattern was displayed by placing an X-ray film (NEF-NEN: DuPont) and exposure for several days.
  • the Northern blot analyzes were carried out using two human Multiple Tissue Northern (MTN) blots from Clontech according to the manufacturer's protocol. The expression was checked in the following tissues: large intestine, small intestine, heart, brain, testes, liver, lungs, spleen, kidney, ovaries, pancreas, peripheral blood leukocytes, placenta, prostate, skeletal muscle, thymus.
  • MTN Multiple Tissue Northern
  • mRNA was carried out in situ hybridizations on 10 ⁇ m paraffin sections from different tissues.
  • the "DDRT-PCR fragment1" was used as a digoxigenin-labeled RNA probe.
  • the detection reaction was carried out using a digoxigenin-specific antibody coupled to alkaline phosphatase (A).
  • A alkaline phosphatase
  • BM Purple served as the substrate for the AP, from which a blue precipitate is formed after dephosphorylation. The results are shown in the table below and show predominant expression in invasive / migrating cells.
  • RT-PCR Reverse Transcription PCR
  • the PCR primers P1 to P7 used are shown in Table 1 (see Figure 1).
  • EJ28 invasive bladder carcinoma cell line
  • RT1 1 2 non-invasive bladder carcinoma cell line
  • RT-PCR results confirmed the expression specific for fragment 1 in the early passages (passage 17, passage 20) of the endometriosis cell line EEC145T "1" . In contrast to the Northern blot analyzes, weak expression in the endometrium was also shown.
  • the cDNA phage bank EEC14 was developed according to the method of Short, J.M. et al. (1 988) Nucleic Acids Res. 1 6: 7583-7600.
  • the reverse transcription of polyA + RNA of invasive cells (passage 1 7) of the epithelial endometriosis cell line EEC145T "1" was carried out .
  • the primer used for this is composed of an X / 7 ⁇ / interface and a 1 8 nucleotide long poly (dT) sequence.
  • An adapter containing a £ co /? / Site was ligated to the resulting cDNA fragments.
  • the two restriction sites allow a directed insertion of the cDNA fragments into the ZAP Express TM vector. Inserts can be cut out of the phage in the form of a kanamycin-resistant pBK CMV phagemid.
  • the DDRT-PCR fragment 1 (394 bp) was used as a probe to screen 10 6 pfu (plaque forming units) of the cDNA phage bank EEC14 according to the manufacturer's protocol (company Stratagene).
  • the probe was labeled with digoxigenin (Boehringer Mannheim) using PCR.
  • the plaques formed after infection of the XL Iblue MRF 'bacterial strain were transferred to a nylon membrane and hybridized thereon with the above-mentioned probe.
  • the hybridized, digoxigenin-labeled probe was detected according to the Boehringer Mannheim chemiluminescence protocol.
  • plaques were selected and rescreened.
  • the plaques positive in the rescreening were used for the excision.
  • kanamycin-resistant pBK CMV phagemids were created, which after propagation in the bacterial strain XL0LR TM isolated and sequenced could become.
  • the isolated phagemid clone Q2A with a size of 2.3 kb contained the longest insert, the sequence of which was determined and SEQ ID NO. 1 is shown.
  • the sequence of the DDRT-PCR fragment 1 is found in nucleotides 1 235 to 1 628 based on SEQ ID NO. 1 .
  • genomic DNA from female and male subjects was cut with different restriction endonucleases. The fragments were separated in an agarose gel and transferred to a nylon membrane. Hybridization with the digoxigenin-labeled DDRT-PCR fragment1 took place on this membrane.
  • Example 7 The genomic clones obtained in Example 7 were localized to chromosome 1 (1 p36) by means of fluorescence in situ hybridization (Lichter et al. (1 990), Science 247: 64-69).
  • the nucleotides 584 to 909 of the cDNA sequence mentioned above were cloned into the expression vector pMAL cRI via suitable restriction sites.
  • the construct was transformed into E.coli DH5 ⁇ cells.
  • the translated protein fragment was cut out of the SDS polyacrylamide gel and used to immunize rabbits.
  • the cDNA first strand synthesis was carried out with a gene-specific primer, which Exon hybridizes, and then a homopolymeric nucleotide tail is attached using the enzyme terminal transferase. This attached sequence allowed an amplification of the sequence region, which is between the gene-specific primer and the homopolymeric nucleotide tail.
  • the following additional sequence was obtained, which is 5 'of the Q2A sequence and belongs to the first exon: cc egg ccg cec ega gtg gag egg atc cac ggg cag atg cag atg cet 47
  • the underlined sequence represents the first nucleotides of the Q2A sequence, the sequence before that corresponds to the new one obtained by 5'RACE Sequence.
  • the open reading frame fits into the one already derived for fragment and contains two putative start codons (underlined).
  • nucleotide acid sequence which the previously obtained and in SEQ ID NO. 1 sequence shown and the additional 84 nt at the 5 'end is in SEQ ID NO. 3 shown.

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EP99927792A 1998-05-29 1999-05-28 Endometriose-assoziiertes gen Withdrawn EP1080195A1 (de)

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DE19824230A DE19824230A1 (de) 1998-05-29 1998-05-29 Neues Endometriose-assoziiertes Gen
DE19824230 1998-05-29
PCT/EP1999/003716 WO1999063079A1 (de) 1998-05-29 1999-05-28 Endometriose-assoziiertes gen

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US20030109018A1 (en) 2003-06-12
CA2329527A1 (en) 1999-12-09
CN1307636A (zh) 2001-08-08
AU765515B2 (en) 2003-09-18
WO1999063079A1 (de) 1999-12-09
JP2002517193A (ja) 2002-06-18
AU4502499A (en) 1999-12-20
NZ508439A (en) 2004-02-27
DE19824230A1 (de) 1999-12-02

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