EP2791681A2 - Procédé d'identification de séquences de marqueurs pour le cancer gynécologique - Google Patents

Procédé d'identification de séquences de marqueurs pour le cancer gynécologique

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Publication number
EP2791681A2
EP2791681A2 EP12816261.7A EP12816261A EP2791681A2 EP 2791681 A2 EP2791681 A2 EP 2791681A2 EP 12816261 A EP12816261 A EP 12816261A EP 2791681 A2 EP2791681 A2 EP 2791681A2
Authority
EP
European Patent Office
Prior art keywords
homo sapiens
protein
sequences
malignancy
gynecological
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP12816261.7A
Other languages
German (de)
English (en)
Inventor
Angelika LÜKING
Axel Kowald
Annabel HÖPNER
Peter Schulz-Knappe
Christian Scheer
Heidelinde Fiegl
Günter Daxenbichler
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Protagen GmbH
Original Assignee
Protagen GmbH
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 Protagen GmbH filed Critical Protagen GmbH
Priority to EP12816261.7A priority Critical patent/EP2791681A2/fr
Publication of EP2791681A2 publication Critical patent/EP2791681A2/fr
Withdrawn legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G01N33/57411—Specifically defined cancers of cervix

Definitions

  • the present invention relates to a method for
  • diagnostic devices containing marker sequences for gynecological malignancy, in particular an assembly and a protein array and their use. Furthermore, the
  • Protein arrays are gaining increasing industrial importance in analytics and diagnostics as well as in pharmaceutical development. Protein arrays have become established as screening tools.
  • Protein arrays make it necessary to have the required proteins available.
  • protein expression libraries have been established for this purpose. High throughput z cloning of defined open reading frames is one
  • Affinity epitopes or proteins on the one hand for the specific detection of the recombinant fusion proteins by means of a directed against the affinity epitope
  • Antibody on the other hand becomes the specific one
  • antibody-presenting arrays are also described (Lal et al (2002) Antibody arrays: An embryonic but growing technology, DDT, 7, 143-149, Kusnezow et al., (2003) Antibody microarrays: An evaluation of production parameters, proteomics, 3, 254-264).
  • Cervical or cervical cancer is a malignant tumor of the cervix (cervix uteri). It is the second most common malignant tumor in women worldwide. Histologically, the majority of cases are squamous cell carcinoma. The most common cause of cervical cancer is infection with certain types of human papillomavirus (HPV). The cervical carcinoma initially causes no pain, only occasionally occur slight spotting. Only when the tumor grows larger and decays with ulceration, it comes to meaty, sweet-smelling discharge. In the early stages is the
  • Cervical cancer is most commonly diagnosed between the ages of 45 and 55 years, but precursors can occur in 20 to 30-year-old women.
  • the mean age of first diagnosis of cervical carcinoma has dropped by 14 years in the last 25 years and is currently around 52 years. In the
  • the age distribution was changed because the diagnosis was much more common in women between 25 and 35 years of age than in women over 65 years of age.
  • the disease can also occur during pregnancy.
  • the incidence is 1.2 per 10,000 pregnancies. It is believed that a large part of the
  • Cervical cancer is caused by the human papillomavirus (HPV).
  • HPV human papillomavirus
  • An early detection test is the Papa test. However, only 2 to 8 percent of HIV-infected women develop cell changes that are a precursor to cancer or even a carcinoma.
  • tissue pieces These are either by a targeted sampling from a noticeable in colposcopy area on the cervix, a conization after repeated conspicuous Papa test or obtained a scraping on suspicion of a change in the cervical canal.
  • US 2005/221342 Al discloses SEQ ID. No. 538 of the present invention and generally also the use of this sequence, but not the specific application in gynecologic malignancy.
  • gynecological malignancies especially cervical carcinoma is an early diagnosis for the rest Disease course and prognosis crucial.
  • Gynecological malignancy especially for cervical carcinoma. It is the object of the present invention to provide improved means for early detection and therapy control in gynecological malignancy.
  • the invention relates to a method for the identification of marker sequences for gynecological malignancy, characterized in that a. Marker sequence candidates for gynecological malignancy
  • Identifying proteins that interact with the serum (marker sequence candidates), and b. the interaction of one or more marker sequence candidates from a. is determined with the serum of patients with gynecologic malignancy compared to the interaction of / the marker sequence candidate from a. with the serum of patients with benign
  • Marker sequences are identified as having a different interaction with the serum of gynecological malignant patients than with serum from Patients with benign changes and serum from healthy controls.
  • the method of the invention identifies marker sequences that are more specific, e.g. because they are also suitable for the discrimination of gynecologic malignancy of benign changes of the tissue (benign change). Furthermore, those with the aid of the method according to the invention
  • the invention also relates to the
  • the invention provides marker sequences for gynecological malignancy obtainable by a method according to the invention and selected from
  • Sequences comprising SEQ ID o. 1-1467 and partial sequences of SEQ ID No. 1-1467 with at least 90%, preferably 95% of the length of the sequences SEQ ID NO. 1 - 1467 and homologs of SEQ ID No. 1- 1467 and their partial sequences with an identity of at least 95%, preferably 98% or more to the
  • the invention also provides an arrangement comprising one or more marker sequences according to the invention.
  • the invention also provides a protein array comprising one or more marker sequences according to the invention.
  • the invention also provides a diagnostic agent comprising one or more marker sequences according to the invention and
  • the invention also provides a test kit comprising one or more marker sequences according to the invention and
  • the invention is also an inventive
  • the invention also provides an inventive
  • the invention is also an inventive
  • the invention also provides a test kit according to the invention, characterized in that 2 or 3, preferably 4 or 5, particularly preferably 7 or 8 or more different marker sequences for gynecological malignancy are used simultaneously.
  • the invention also provides the use of one or more marker sequences according to the invention, one
  • Protein arrays Protein arrays, a diagnostic agent according to the invention or a test kit according to the invention for early detection, diagnosis, prognosis, therapy control and / or aftercare
  • the invention also provides the use of one or more marker sequences according to the invention, one
  • Protein arrays a diagnostic agent according to the invention or a test kit according to the invention for distinguishing gynecological malignancy from benign changes.
  • the invention also provides the use of one or more marker sequences according to the invention, one
  • Protein arrays Protein arrays, a diagnostic agent according to the invention or a test kit according to the invention for individualized
  • the invention also provides the use of one or more marker sequences according to the invention, one
  • Protein arrays a diagnostic agent according to the invention or a test kit according to the invention for detecting and / or determining the amount of one or more autoantibodies associated with gynecological malignancy,
  • the invention also provides the use of one or more marker sequences according to the invention, one
  • Protein arrays Protein arrays, a diagnostic agent of the invention or a test kit according to the invention for the analysis of
  • Autoantibodies and / or for monitoring changes in autoantibody profiles for example in body fluids such as serum, tissue or tissue extracts of the patient.
  • the invention also provides the use of one or more marker sequences according to the invention, one
  • Protein arrays Protein arrays, a diagnostic agent of the invention or a test kit according to the invention for the screening of substances (drugs) for gynecological malignancy.
  • the invention also relates to a target for the treatment and / or therapy of gynecological malignancy, wherein the target is obtained from the marker sequences according to the invention SEQ ID No. 1-1467 and partial sequences of SEQ ID No. 1-1467 with at least 90%, preferably 95% of the length of the sequences SEQ ID NO. 1 - 1467 and homologs of SEQ ID No. 1-1467 and theirs
  • Nucleic acid and / or protein sequences is selected.
  • the invention also provides a method for
  • one or more marker sequence (s) selected from the group comprising the sequences SEQ ID NO. 1 - 1467 and
  • the invention relates to the use of one or more marker sequence (s) for gynecological malignoma for
  • the invention relates to the use of one or more gynecological marker sequence (s) according to the invention
  • the invention relates to the use of one or more gynecological marker sequence (s) according to the invention
  • Malignant are associated, for example, in body fluid or tissue of a patient.
  • the invention also relates to the marker sequence for gynecological malignoma selected from the sequences
  • SEQ ID No. 1 - 489 comprising SEQ ID No. 1 - 489 and partial sequences of SEQ ID No. 1-489 with at least 90%, preferably 95% of the length of the sequences SEQ ID NO. 1 - 489 and homologs of SEQ ID No. 1-489 and their partial sequences with an identity of at least 95%, preferably 98% or more, to the corresponding ones
  • the invention relates to an arrangement of one or more marker sequence (s) for gynecological malignancy on a support for the early detection, diagnosis, prognosis and / or therapy control in gynecological malignancy wherein the
  • Marker sequence (s) for gynecological malignancy is / are selected from the group of proteins SEQ ID No. 979-1467 and the proteins encoded by sequences SEQ ID NO. 1-978 and encoded by partial sequences of SEQ ID NO. 1-978 with at least 90%, preferably 95% or more of the length of the sequences SEQ ID No. 1-978 and encoded by homologues of SEQ ID NO. 1-978 and their partial sequences with an identity of at least 95%, preferably 98% or more to the
  • the invention relates to an arrangement according to the invention, wherein the marker sequence (s) for gynecological malignancy is / are applied to a solid carrier, in particular a filter, a membrane, a magnetic or fluorophore-labeled bead, a silicon wafer, glass, metal, Plastic, a chip, a mass spectrometric target or a matrix.
  • a solid carrier in particular a filter, a membrane, a magnetic or fluorophore-labeled bead, a silicon wafer, glass, metal, Plastic, a chip, a mass spectrometric target or a matrix.
  • the invention relates to an arrangement according to the invention or an inventive use of one or more marker sequence (s) for gynecological malignancy, wherein the marker sequence (s) for gynecological malignancy is / are present as clone (s).
  • the subject of the invention is also an assay or
  • Protein biochip comprising an arrangement according to the invention or one or more marker sequence (s) for gynecological malignancy according to the invention.
  • the invention also provides a diagnostic (test kit) for the early detection and / or diagnosis of gynecological
  • An assay or protein array according to the invention or one or more gynecological malignoma marker sequence (s) selected from the group comprising SEQ ID Nos. 1-1467 and partial sequences of SEQ ID Nos. 1-1467 with at least 90%, preferably 95% of the length of the sequences SEQ ID No. 1 - 1467 and homologs of SEQ ID No. 1-1467 and theirs
  • the invention also provides a method for
  • gynecologic malignancy by comparative analysis of signals resulting from contact of the marker sequence candidates with body fluid or tissue extract of a patient with gynecologic malignancy and Body fluid or tissue extract of a patient without gynecological malignancy,
  • Gynecological malignancy using a protein array d. Selection of marker sequences for gynecologic malignancy, characterized in that they include
  • the invention relates to an arrangement of marker sequences for gynecological malignancy on a support for early detection, diagnosis, prognosis, therapy control with one or more different marker sequences for gynecological malignoma selected from the group of the proteins SEQ ID No. 979 to 1467 and / or partial sequences of these proteins and / or encoded by sequences SEQ ID no. 1 - 489 (clone sequences, cDNA) and / or encoded by sequences SEQ ID. No.
  • RNA sequences and / or encoded by partial sequences of SEQ ID. No. 1-978.
  • proteins SEQ ID No. 979 to 1467 and / or partial sequences of these proteins and / or proteins encoded by sequences SEQ ID NO. 1 - 489 and / or proteins encoded by sequences SEQ ID. No. 490-978 are proteins SEQ ID No. 979 to 1467 and / or partial sequences of these proteins and / or proteins encoded by sequences SEQ ID NO. 1 - 489 and / or proteins encoded by sequences SEQ ID. No. 490-978
  • the invention thus provides a panel of marker sequences for gynecologic malignancies that can be used as part of an individualized diagnosis and therapy to treat different patients, patient groups, cohorts, population groups, variants of gynecological Malignoma, etc. specifically and individually adjusted
  • the invention also provides the use of one or more marker sequences for gynecological malignoma selected from the group comprising SEQ ID o. 1-1467 and partial sequences of SEQ ID No. 1-1467 with at least 90%, preferably 95% of the length of the sequences SEQ ID No. 1 - 1467 and homologs of SEQ ID No. 1- 1467 and their partial sequences with an identity of at least 95%, preferably 98% or more to the corresponding nucleic acid and / or
  • Protein sequences in particular of the proteins SEQ ID No. 979 to 1467 and / or partial sequences of these proteins and / or encoded by sequences SEQ ID no. 1 - 489 and / or encoded by sequences SEQ ID. No. 490-978 and / or coded by
  • the arrangement / use according to the invention comprises 2 or 3, preferably 4 or 5, more preferably 7 or 8 or more different marker sequences for gynecological malignancy.
  • the arrangement / use can be 9 or 10 or more
  • the invention thus also relates to an arrangement / use where at least 2 to 5 or 10, preferably 30 to 50, marker sequences for gynecological malignancy or 50 to 100 or more marker sequences for
  • a preferred embodiment of the invention relates to an arrangement / use, characterized in that the
  • Marker sequences for gynecological malignancy are applied to a solid support, in particular a filter, a membrane, a bead, for example a magnetic or fluorophore-labeled bead, a silicon wafer, glass, metal, plastic, a chip, a mass spectrometric target or a matrix ,
  • a filter or a bead is preferred according to the invention.
  • the filter is PVDF, nitrocellulose or nylon (e.g., Immobilon P Millipore, Protran Whatman, Hybond N + Amersham).
  • the invention relates to the use of
  • Marker sequences for gynecological malignancy for the diagnosis of gynecological malignancy, wherein at least one marker sequence for gynecological malignoma of a DNA, in particular cDNA selected from the group SEQ ID No. 1-489 or RNA selected from the group 490-978 or a partial sequence or a homologous sequence thereof on or to a patient to be examined.
  • marker sequences for gynecological malignancy also called marker sequences according to the invention
  • the marker sequences according to the invention for gynecological malignancy could by means of differential screening of samples and healthy subjects with patient samples with
  • Protein array (see examples) can be identified.
  • gynecologic malignancy includes a group of diseases that may be precursors to gynecologic malignancy and their establishment as gynecological
  • Malignancy Included are malignant diseases of the genital tract in women, especially malignant diseases of the
  • Cervix such as cervical carcinoma, in particular invasive cervical carcinoma (definition eg according to Pschyrembel, de Gruyter, 263th edition (2012), Berlin). Variants of gynecologic malignancy and stages of gynecologic malignancy are also defined in the Pschyrembel. In a further embodiment of the invention (eg
  • Marker sequences for gynecological malignancy may also be combined, augmented or augmented with known biomarkers for this indication. However, at least 50%, preferably 60%, more preferably 70% or more
  • the arrangement according to the invention, the assay and protein array according to the invention and the use according to the invention represent at least 75%, preferably 80% or 85%, particularly preferably 90% or 95% of marker sequences according to the invention.
  • the determination of the marker sequences for gynecological malignancy outside the human body and the determination is carried out in an ex vivo / in vitro diagnosis.
  • the invention also relates to an assay or protein array comprising an arrangement / use according to the invention.
  • the invention relates to a diagnostic device and / or an assay, in particular a protein array for
  • the invention also relates to the use of a
  • the invention also provides a diagnostic (test kit) for the early detection and / or diagnosis of gynecological
  • Malignancy and / or prognosis and / or prediction of the risk of metastasis formation in gynecological malignant comprising, for example, an inventive arrangement preferably on a support or an assay according to the invention or
  • the invention is also a
  • test kit for therapy monitoring and / or follow-up in gynecological malignancy.
  • the invention relates to the use of marker sequences for
  • gynecological malignancy as a diagnostic agent, wherein at least one marker sequence of a cDNA selected from the group SEQ ID no. 1-489 (clone sequences) or SEQ ID No. 490-978 (RNA) or one each by SEQ ID No. 1-978 is a coded protein or a partial sequence or fragment thereof.
  • the invention also provides a method for
  • the invention relates to a method for the early detection and diagnosis of gynecological malignancy wherein a. ) at least one marker sequence for gynecological malignancy of a cDNA selected from the group SEQ ID No. 1-489 (clone sequences) or SEQ ID No. 490-978 (RNA) or a
  • Partial sequence of SEQ ID. No. 1-978 is applied to a carrier and b. ) is brought into contact with body fluid or tissue extract of a patient and c.) the detection of an interaction of the body fluid or tissue extract with the marker sequences from a.).
  • One or more marker sequences for gynecologic malignancy are used in a diagnostic and / or diagnostic procedure.
  • the detection of an interaction of the body fluid or the tissue extract with the marker sequences or sequences for gynecological malignancy can be done for example by a probe, in particular by an antibody.
  • the invention also provides a method for
  • a particular embodiment of the invention relates to the method, wherein the stratifying or
  • Surveillance of disease progression and therapy, etiology or classification of a disease including prognosis includes.
  • the invention relates to a method for
  • Nucleic acid for example cDNA selected from the group SEQ ID no. 1-489 (clone sequences) or SEQ ID No. 490-978 (RNA) or in each case a protein encoded thereby or in each case a partial sequence of SEQ ID No. 1 - 978 on one too
  • the invention relates to a method for stratifying, in particular for risk stratification and / or therapy control of a
  • gynecological malignancy wherein at least one marker sequence of a DNA, cDNA selected from the group SEQ ID no. 1-489 (clone sequences) or SEQ ID No. 490-978 (RNA or DNA) or a subsequence thereof is used to detect, identify, monitor, and monitor autoantibodies and / or autoantibody profiles associated with gynecologic malignancy in the patient.
  • Autoantibody profiles include the amount of one or more
  • Therapy control also includes the classification of
  • Diagnosis in the sense of this invention means the positive detection of gynecological malignancy by means of
  • Marker sequences according to the invention for gynecological malignancy and the assignment of patients to gynecological malignancy includes medical diagnostics and related investigations, in particular in vitro diagnostics and laboratory diagnostics, also proteomics and
  • diagnosis also includes the
  • Stratification also: stratification or therapy control
  • stratification means that, for example, the methods according to the invention allow decisions for the treatment and therapy of the patient, be it hospitalization of the patient, use, effect and / or dosage of one or more drugs, a therapeutic one Measure or the monitoring of a course of disease as well as course of therapy or etiology or classification of a disease, eg in a new or existing subtype or the differentiation of diseases and their patients.
  • the term "stratification" includes in particular the
  • Prognosis means the prediction of disease progression, for example the prediction of recurrence-free survival, overall survival, the risk of metastasis formation.
  • patient is understood to mean any subject - human or mammal - with the proviso that the test person is being examined for gynecological malignancy.
  • patient is understood to mean any female subject.
  • healthy or “control” or “healthy control person” is any subject,
  • marker sequence for gynecological malignancy in the sense of this invention means that the nucleic acid, for example DNA, in particular cDNA or RNA or the
  • Body fluid or tissue extract of a patient with gynecological malignancy e.g., antigen (epitope) / antibody (paratope) interaction.
  • gynecological malignancy e.g., antigen (epitope) / antibody (paratope) interaction.
  • marker sequences for gynecologic malignancy may also be distinguished by the fact that they interact with substances from the body fluid or tissue extract of patients with gynecological malignancy, because these substances no longer or at least occur in a significantly smaller amount / concentration in gynecological malignancy or
  • Gynecological malignancies may also be present in healthy volunteers, but their amount varies
  • the marker sequences for gynecological malignancy are therefore biomarkers for gynecological malignancy.
  • the marker sequences for gynecological malignancy are therefore biomarkers for gynecological malignancy.
  • Gynecological malignancies can thus map a profile of substances from body fluid and tissue extract, such as an autoantibody profile for gynecological malignancy.
  • composition and / or the amount or concentration are specific for
  • the gynecological malignancy marker sequence is an antigen or part of an antigen or encodes an antigen or part of an antigen.
  • the marker sequence for gynecologic malignancy recognizes / binds
  • Autoantibodies present (or enhanced) to a lesser extent (or no longer) in the course of the development, establishment and therapy of gynecologic malignancy (hereafter referred to as "autoantibodies for gynecological malignancy”.) Autoantibodies are secreted by the body against endogenous antigens, for example arise in gynecological malignancy, educated. Autoantibodies are being targeted by the body
  • gynecological malignancy are formed and / or upregulated or downregulated in their expression.
  • Autoantibodies associated with gynecological malignancy can be detected using the methods of the invention and marker sequences for gynecologic malignancy and thus serve as an indication for gynecological malignancy. The detection and monitoring of the amount of autoantibodies for
  • Gynecological malignancy in the patient can be used for early detection, diagnosis and / or therapy monitoring / therapy control.
  • These autoantibody profiles associated with gynecologic malignancy can be sufficiently characterized even when using a marker sequence for gynecological malignancy. In other cases, two or more marker sequences for gynecologic malignancy will be necessary to associate with an autoantibody profile
  • the autoantibodies associated with gynecologic malignancy may be detected with gynecologic malignant marker sequences derived from another individual, for example, from a commercial cDNA library, or may be compared to a gold standard.
  • the autoantibodies for gynecologic malignancy may include
  • Autoantibodies can be formed by the patient many years before the onset of the first disease symptoms. This would be an early detection, diagnosis and also prognosis and
  • Devices and means thus allow for very early intervention compared to known methods, which significantly improves prognosis and survival rates.
  • the invention also provides for the detection and monitoring of gynecologic malignancy at any stage of development and treatment, as well as gynecologic malignancy follow-up monitoring.
  • the agents according to the invention also allow easy handling, e.g. at home, by the patient himself and the cost-effective routine care
  • Marker sequence for gynecological malignancy sufficient, while in other cases, at least two or more marker sequences for gynecological malignancy must be used together or in combination to a meaningful
  • gynecological malignant for example in the serum / plasma has the advantage over other biomarkers of a high Stability and shelf life and good traceability. Also, the presence of autoantibodies is not subject to a circadian rhythm, so sampling is independent of time of day, food intake, and the like. In addition, autoantibodies associated with gynecologic malignancy with the help of the corresponding
  • Antigens / autoantigens in known assays e.g. ELISA or Western Blot can be detected and the results are checked.
  • Marker sequence for gynecological malignancy is selected "that an interaction is detected
  • Interaction is e.g. a binding, in particular a binding substance to at least one marker sequence for
  • gynecologic malignancy or, in the case where the marker sequence for gynecologic malignancy is a nucleic acid, for example a cDNA, hybridization with a suitable
  • Hybridization conditions are: hybridization in 4 x SSC at 65 ° C (alternatively in 50% formamide and 4X SSC at 42 ° C), followed by several washes in 0.1 x SSC at 65 ° C for a total of about one hour.
  • An example of less stringent hybridization conditions is hybridization in 4 x SSC at 37 ° C, followed by several washes in 1 x SSC
  • Tissue extraction of a patient and the marker sequences for gynecological malignancy is preferably a protein-protein interaction.
  • Such substances for example antigens, autoantigens,
  • Autoantibodies associated with gynecological malignancy are part of a body fluid according to the invention.
  • blood especially blood, whole blood, blood plasma, blood serum,
  • Synovial fluid or a tissue extract for example from tumor tissue of the patient.
  • the invention relates to Synovial fluid or a tissue extract, for example from tumor tissue of the patient.
  • the marker sequences for gynecologic malignancy or the substances recognized by these marker sequences may be present at a significantly higher or lower expression rate or concentration, indicating gynecological malignancy.
  • Nucleic acid blots the relative expression rates ill / healthy of the marker sequences according to the invention for
  • the protein is preferred for a protein Detection signal an epitope and / or paratope and / or hapten and cDNA for a hybridization or
  • RNA RNA sequences for gynecological malignancy according to the invention.
  • the invention also encompasses the full-length sequences of the gynecological malignoma marker sequences according to the invention as defined above the known database entry according to Table A and in the Sequence Listing, hereinafter called SEQ. 1-1467. Furthermore, therefore, also include analog
  • the marker sequences also include such modifications of the nucleic acid sequence, in particular cDNA sequence and the corresponding amino acid sequence, such as chemical modification, such as citrullination, acetylation, phosphorylation, glycosylation or polyA strand and other modifications known to those skilled in the art.
  • the invention also relates to homologues of the marker sequences for gynecological malignancy and homologues of the partial sequences, For example, fragments of marker sequences for
  • Gynecological malignant marker sequences of at least 70% or 80%, preferably 90% or 95%, more preferably 96% or 97% or more, for example 98% or 99%.
  • Gynecological malignant marker sequences of at least 70% or 80%, preferably 90% or 95%, more preferably 96% or 97% or more, for example 98% or 99%.
  • At least 95%, preferably at least 97%, particularly preferably at least 99%, of the homology in the sequence region in which the antigen-antibody or antigen-autoantibody interaction takes place are for gynecological malignant antigen.
  • the invention also subsequences of
  • Partial sequences also include fragments of the marker sequences according to the invention, partial sequences are those nucleic acids or proteins / peptides which are opposite to the complete nucleic acid or the
  • shortened complete protein / peptide The deletion may be at or near the end and / or within the sequence.
  • partial sequences and / or fragments comprising 50 to 100 nucleotides, 70 to 120 nucleotides of a complete sequence, for example of SEQ ID-1467, are included. Homologs of partial sequences and fragments are also included according to the invention.
  • the marker sequences are for gynecologic malignancy
  • marker sequences for gynecological malignancy which differ from the sequences SEQ ID 1-1-1467 in that they contain one or more insertions, the insertions
  • nucleotides / amino acids for example 1 to 100 or more nucleotides / amino acids, preferably 5 to 50, particularly preferably 10 to 20
  • Nucleotides / amino acids are long and the sequences but otherwise identical or homologous to the sequences 1-1467. Preference according to the invention are partial sequences with
  • the regions may each comprise a total of gynecological malignant marker sequences, i. a sufficient number of different marker sequences for gynecological malignancy, in particular 2, 3, 4, 5, 6, 7, 8, 9 or 10 or more different and optionally further nucleic acids and / or proteins, in particular biomarkers.
  • marker sequences for gynecological malignancy and optionally further nucleic acids and / or proteins, in particular biomarkers on the carrier
  • nucleic acids and / or proteins in particular biomarkers on the carrier.
  • array synonymously means “array”, and insofar as this "array” is used to identify substances on marker sequences for gynecological malignancy, this is preferably to be understood as meaning an "assay” or a diagnostic device.
  • the arrangement is such
  • Arrangements are preferred which allow a high-density arrangement of marker sequences for gynecological malignancy, for example protein binders.
  • the marker sequences for gynecologic malignancy are spotted.
  • Such high density spotted assemblies are disclosed, for example, in WO 99/57311 and WO 99/57312, and may be advantageously used in a robotic automated high throughput method.
  • the term "assay" or diagnostic device also includes such
  • Embodiments of a device such as ELISA, bead-based assay, line assay, Western blot, immunochromatographic
  • Methods e.g., so-called lateral flow immunoassays or similar immunological single or multiplex detection methods.
  • a “protein array” (also protein biochip) in the sense of this
  • the invention is the systematic arrangement of gynaecologic malignoma marker sequences on a solid support, wherein the gynecological malignoma marker sequences are proteins or peptides or portions thereof.
  • the marker sequences for gynecological malignancy of the assembly are fixed to a solid support, but preferably spotted or immobilized even printed, ie
  • One or more marker sequences for gynecological malignancy may be present multiple times in the totality of all marker sequences for gynecological malignancy and be present in different amounts relative to one spot. Furthermore, the marker sequences for
  • the marker sequences for gynecological malignancy are clones.
  • Such clones can be obtained, for example, by means of a cDNA expression library according to the invention (Büssow et al., 1998
  • Expression vectors obtained from an expressing cDNA library consisting of the cDNA marker sequences.
  • Expression vectors preferably contain inducible
  • Promoters Induction of expression may be e.g. by means of an inductor, such as IPTG.
  • an inductor such as IPTG.
  • Expression libraries are known to the person skilled in the art, these can be prepared according to standard works, such as Sambrook et al., Molecular Cloning, Laboratory Handbook, 2nd edition (1989), CSH press, Cold Spring Harbor, New York Further preferred are such expression libraries.
  • expression libraries which can be obtained by exon trapping are also included in the invention.
  • Uniclone® library protein arrays or corresponding expression libraries which have no redundancy
  • Uniclone® library protein arrays or corresponding expression libraries which have no redundancy
  • the clones may not be such as transformed bacteria, recombinant phage or transformed cells of mammals, insects, fungi, yeasts or plants.
  • the clones are fixed on a solid support, spotted or immobilized. Therefore, the invention relates to a
  • marker sequences for gynecological malignancy in the respective form may be present in the form of a fusion protein, which may include at least one
  • the day may be one such as c-myc, his-tag, arg-tag, FLAG, alkaline
  • Phosphatase V5 tag, T7 tag or strep tag, HAT tag, NusA, S tag, SBP tag, thioredoxin, DsbA, a fusion protein,
  • green fluorescent protein maltose binding protein, Calmodulin-binding protein, glutathione S-transferase or lacZ.
  • this invention corresponds to a grid that the order of a microtiter plate (8-12 wells strips, 96 wells, 384 wells or more), one
  • the arrangement is on a bead or a small plate.
  • the invention relates to an assay or protein array for identification and
  • Malignancy characterized in that an arrangement or assay according to the invention is brought into contact with a.) At least one substance to be investigated and b.) A binding success is detected.
  • the substance to be investigated may be any native or non-native biomolecule, a (synthetic) chemical molecule, a natural product, a mixture or a
  • Substance library After the substance under investigation has contacted a marker sequence for gynecological malignancy, the
  • Binding results interactions such as protein-protein interactions (e.g., protein to marker sequence for
  • gynecological malignancy such as antigen / antibody
  • corresponding "means for detecting binding success” can be performed, for example, by fluorescence labeling, biotinization, radioisotope labeling or colloidal gold or latex particle labeling in a conventional manner using secondary antibodies labeled with commercial reporter molecules (eg Cy, Alexa, Dyomics, FITC or similar fluorescent dyes, colloidal gold or latex particles), or reporter enzymes such as alkaline
  • a readout is e.g. by means of a microarray laser scanner, a CCD camera or visually.
  • the invention relates to a drug / drug or prodrug for gynecological malignancy developed and obtainable by the use of a marker sequence according to the invention, an inventive arrangement, a use according to the invention, a
  • the invention is also the use of a
  • Marker sequence for gynecological malignoma selected from the group of proteins SEQ ID No. 979-1467, partial sequences of these proteins and those coding for these proteins Sequences, the sequences SEQ ID No. 1 - 489 and SEQ ID. No. 490-978 and partial sequences of SEQ ID No 1 - 978 as well as of SEQ ID No. 1 - 978 coded proteins as
  • the invention therefore relates to the use of the marker sequences according to the invention, preferably in the form of an arrangement, as an affinity material for carrying out a blood wash in a broader sense, substances from body fluids of a patient with gynecological malignancy, such as blood or plasma, binding to the marker sequences according to the invention and consequently to the body fluid selectively can be withdrawn.
  • the examples are performed using the UNIarray technology platform based on quantitative analysis of autoantibody profiles in serum of gynecological malignant patients.
  • the aim is to systematically identify antigens associated with gynecological malignancy and autoantigens associated with gynecological malignancies (so-called biomarkers), which provide early detection of
  • Marker sequences for gynecological malignancy by means of bioinformatic analysis.
  • the candidates for gynecological malignancy marker sequences are evaluated as discriminating between different subjects (e.g., healthy / unhealthy) / patient groups (e.g., low / high risk of metastasis) / cohorts (e.g., particular history).
  • the marker sequence candidates will be on a
  • Biochip applied and validated.
  • the data analysis takes place via statistical analyzes, for example
  • This biochip includes one or more
  • Cohort II Clinical findings: gynecological malignancy ⁇ negative group (control group), age-matched. Patients are selected after inclusion and
  • Anti-estrogen therapy adjuvant chemotherapy or adjuvant aromatase inhibitor therapy.
  • biochips are used for diagnosis, predicting the course of therapy and predicting metastasis
  • Example 4 For the development of a protein biochip for the diagnosis of gynecological malignancy, the results of the
  • Example 5 In the development of a protein biochip for predicting the
  • gynecologic malignancies which interact with autoantibodies for gynecologic malignancy, which are suitable as an indicator of metastasis.
  • bioinformatics analyzes are carried out. For each serum, microarray reactivities of about 2000 different antigens are measured. These data are used for a ranking of the spotted antigens regarding their
  • Intensity data are performed.
  • an internal standard is used, which is spotted on each chip. Since a p-value is calculated for each antigen, methods for correcting the multiple testing are used. As a very conservative approach, a Bonferroni correction is performed and, in addition, the less restrictive False Discovery Rate (FDR) is calculated according to Benjamini & Hochberg.
  • FDR False Discovery Rate
  • SVM Support Vector Machines
  • Threshold method which is suitable for both classification and visual representation of the data. To avoid overfitting, for example, a 10-fold cross-validation of the data is performed.
  • the erfindungsgermä call sequences are listed in the attached sequence listing.
  • the clone sequences (cDNA) SEQ ID no. 1 - 489, the RNA sequences SEQ ID. No. 490-978 and the protein sequences SEQ ID No. 979-1467).
  • gi 1282847377 NM_080592.3 apoptosis-related protein 3 isoform b [Homo sapiens]
  • isoform 1 [Homo sapiens] gi 1116014343 NM_001624.2 absentee in melanoma 1 protein
  • domain family B member 1 isoform a [Homo sapiens]
  • taxl-binding protein 3 isoform 1 [Homo sapiens] gi
  • IIF subunit 1 [Homo sapiens]
  • sirtuin-7 [Homo sapiens] gi 116554603 NM_016070.2 28S ribosomal protein S23, mitochondrial [Homo sapiens] gi 1210147465 NM_022164.2 tubulointerstitial nephritis antigen-like isoform 1 precursor [Homo sapiens] gi 1282400941 NM_022737.2 lipid phosphate phosphatase-related protein type 2 isoform 1 [Homo sapiens] gi 1154426254 NM_032361.2 THO complex subunit 3 [Homo sapiens]
  • regulator 3 isoform 2 [Homo sapiens]
  • transcription factor 1 isoform 3 [Homo sapiens]
  • transcription factor HIB 50 kDa subunit [Homo sapiens]
  • mitochondrial precursor [Homo sapiens]
  • nGAP isoform 1 [Homo sapiens]
  • RNA polymerase I I -associated protein 1 [Homo sapiens] 649 gi 187829711 NM_014417.3 bcl-2-binding component 3 isoform 4 [Homo sapiens]
  • RNA-binding protein 4B [Homo sapiens]
  • neuropathy target esterase isoform b [Homo sapiens]
  • B receptor subunit 1 isoform b precursor [Homo sapiens]
  • inhibitor 1 isoform a [Homo sapiens]
  • DNA ligase 3 isoform beta precursor [Homo sapiens] gi
  • a isoform b [Homo sapiens] gi 1219879812 NM_002696.2 DNA-directed RNA polymerase
  • mitochondrial isoform 2 precursor [Homo sapiens]

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Hospice & Palliative Care (AREA)
  • Biotechnology (AREA)
  • Food Science & Technology (AREA)
  • Oncology (AREA)
  • Microbiology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
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  • Peptides Or Proteins (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

La présente invention concerne un procédé d'identification de séquences de marqueurs pour le cancer gynécologique, les séquences de marqueurs identifiées par ce procédé ainsi que leur utilisation diagnostique, les dispositifs diagnostics contenant les séquences de marqueurs pour le cancer gynécologique, notamment un agencement et une puce à protéines ainsi que leur utilisation. L'invention concerne également un procédé d'identification de principes actifs potentiels pour le traitement et la prévention du cancer gynécologique à l'aide de ces séquences de marqueurs.
EP12816261.7A 2011-12-16 2012-12-17 Procédé d'identification de séquences de marqueurs pour le cancer gynécologique Withdrawn EP2791681A2 (fr)

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EP12816261.7A EP2791681A2 (fr) 2011-12-16 2012-12-17 Procédé d'identification de séquences de marqueurs pour le cancer gynécologique

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EP11194099.5A EP2605017A1 (fr) 2011-12-16 2011-12-16 Séquences de marqueurs pour le malignome gynécologique et leur utilisation
PCT/EP2012/075778 WO2013087929A2 (fr) 2011-12-16 2012-12-17 Procédé d'identification de séquences de marqueurs pour le cancer gynécologique
EP12816261.7A EP2791681A2 (fr) 2011-12-16 2012-12-17 Procédé d'identification de séquences de marqueurs pour le cancer gynécologique

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WO2010094499A1 (fr) 2009-02-20 2010-08-26 Ganymed Pharmaceuticals Ag Methodes et compositions de diagnostic et de traitement du cancer
CN102741289B (zh) 2009-11-11 2016-08-03 加尼梅德药物公司 密蛋白6(cldn6)特异性的抗体
EP2404936A1 (fr) 2010-07-06 2012-01-11 Ganymed Pharmaceuticals AG Thérapie du cancer utilisant des anticorps in vivo dirigés sur la cible CLDN6
PL3026064T3 (pl) 2011-05-13 2019-05-31 Ganymed Pharmaceuticals Gmbh Przeciwciała do leczenia nowotworu z ekspresją klaudyny 6
WO2015014376A1 (fr) 2013-07-31 2015-02-05 Biontech Ag Diagnostic et thérapie du cancer impliquant des cellules souches cancéreuses
ES2693465T3 (es) * 2014-06-06 2018-12-11 Uroimmun Medizinische Labordiagnostika Ag Diagnóstico de una enfermedad neurológica
GB201511196D0 (en) * 2015-06-25 2015-08-12 Cytosystems Ltd Monoclonal antibodies
WO2021030293A1 (fr) * 2019-08-09 2021-02-18 Arizona Board Of Regents On Behalf Of The University Of Arizona Méthodes de surveillance ou de prédiction de réponse à des immunothérapies pour un cancer gynécologique

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DE69921982T2 (de) 1998-04-30 2005-12-29 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Neuartiges verfahren zur identifizierung von klonen mit einer gewünschten biologischen eigenschaft, ausgehend von einer expressionsgenbank
EP1073771B1 (fr) 1998-04-30 2004-12-01 Max-Planck-Gesellschaft Zur Förderung Der Wissenschaften E.V. Nouveau procede permettant la selection de clones dans une banque d'expression et comprenant un rearrangement
US20050221342A1 (en) * 2000-04-18 2005-10-06 Nuvelo, Inc. Nucleic acids and polypeptides

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2013087929A2 *

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EP2605017A1 (fr) 2013-06-19
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