EP2673379A2

EP2673379A2 - Marker sequences for diagnosing prostate cancer, and use thereof

Info

Publication number: EP2673379A2
Application number: EP12708501.7A
Authority: EP
Inventors: Angelika Lueking; Axel Kowald; Georg Bartsch; Helmut Klocker
Original assignee: Protagen GmbH
Current assignee: Protagen GmbH
Priority date: 2011-02-13
Filing date: 2012-02-13
Publication date: 2013-12-18
Also published as: WO2012107596A2; WO2012107596A3; EP2487251A1; US20140179557A1

Abstract

Use of a marker sequences for the diagnosis of prostate cancer, excluding inflammatory prostate diseases, diabetes, or polymorbidity, is claimed, where: at least one marker sequence of a cDNA comprising SEQ ID NOs 1-246 and/or SEQ ID NOs 247-452 or their coded proteins or their subsequence or fragments, is determined from a patient to be examined; and the sequences are not defined. Independent claims are included for: (1) a method for diagnosing prostate cancer, excluding inflammatory prostate diseases, diabetes, or polymorbidity comprising applying the marker sequence on a solid support and contacting with a body fluid or tissue extract of a patient and determining the interaction of the body fluid or tissue extract containing the marker sequences; (2) assay, protein biochip consisting of an assembly comprising the marker sequence, where the marker sequences are applied on a solid support; (3) use of an assay for identifying and characterizing a substance for prostate cancer comprising means for detecting a binding success, where the assembly or assay is contacted with the substance, and a binding performance is detected; and (4) target for the treatment and therapy of prostate cancer, comprising the marker sequences. - ACTIVITY : Cytostatic. - MECHANISM OF ACTION : None given.

Description

Marker sequences for the diagnosis of prostate cancer and their use

description

The present invention relates to new marker sequences for prostate cancer (syn: prostate cancer) excluding prostate inflammatory diseases or diabetes or

Polymorbidity and its diagnostic use, including a method of screening for potential drugs for such prostate diseases by means of these marker sequences. Furthermore, the invention relates to a diagnostic device

containing such marker sequences for prostate cancer, in particular a protein biochip and its use. Protein biochips are gaining an increasing industrial

Meaning in analytics and diagnostics as well as in the

Pharmaceutical development. Protein biochips have proven to be

Established screening tools.

Here, the fast and highly parallel detection of a variety of specific binding analysis molecules in one

single experiment allows. For production of

Protein biochips require the necessary proteins to be available. In particular

Protein expression libraries established. High throughput z cloning of defined open reading frames is one

Possibility (Heyman, JA, Cornthwaite, J., Foncerrada, L., Gilmore, JR, Gontang, E., Hartman, KJ, Hernandez, CL, Hood, R., Hull, HM, Lee, WY, Marcil, R. , Marsh, EJ, Mudd, KM, Patino, MJ, Purcell, TJ, Rowland, JJ,

Sindici, M.L. and Hoeffler, J.P. (1999) Genome-scale cloning and expression of individual open reading frames using

topoisomerase I-mediated ligation. Genome Res, 9, 383-392; Kersten, B., Feilner, T., Kramer, A., Wehrmeyer, S., Possling, A., Witt, I., Zanor, M.I., Stracke, R., Lueking, A.,

Kreut zberger, J., Lehrach, H. and Cahill, DJ (2003) Generation of Arabidopsis protein chip for antibody and serum screening. Plant Molecular Biology, 52, 999-1010; Reboul, J., Vaglio, P., Rual, JF, Lamesch, P., Martinez, M., Armstrong,

C.M., Li, S., Jacotot, L. , Bertin, N. , Janky, R., Moore, T., Hudson, J.R., Jr. Hartley, J.L., Brasch, M.A., Vandenhaute,

J., Boulton, S., Endress, G.A., Jenna, S., Chevet, E.,

Papasotiropoulos, V., Tolias, P.P., Ptacek, J., Snyder, M., Huang, R., Chance, M.R., Lee, H., Doucette strain, L., Hill,

D.E. and Vidal, M. (2003) C. elegans ORFeome ersion 1.1:

experimental verification of the genome annotation and

resource for proteome-scale protein expression. Nat Genet, 34, 35-41; Walhout, AJ, Temple, GF, Brasch, MA, Hartley, JL, Lorson, MA, van den Heuvel, S. and Vidal, M. (2000) GATEWAY recombinational cloning: application to the cloning of large numbers of open reading frames ORFeomes. Methods

Enzymol, 328, 575-592). However, such an approach is strongly related to the progress of genome sequencing projects and the annotation of these gene sequences. In addition, the determination of the expressed sequence is due

differential splicing is not always clear. This problem can be overcome by the use of cDNA

Büssow, K., Cahill, D., Nietfeld, W., Bancroft, D., Scherzinger, E., Lehrach, H. and Walter, G. (1998) A method for global protein expression and antibody screening Nucleic Acids Research, 26, 5007-5008, Büssow, K., Nordhoff, E., Lübbert, C., Lehrach, H. and Walter, G.

(2000) A human cDNA library for high-throughput protein expression screening. Genomics, 65, 1-8; Holz, C., Lueking, A., Bovekamp, L., Gutjahr, C., Bolotina, N., Lehrach, H. and Cahill, DJ (2001). A human cDNA expression library in yeast enriched for open reading frames. Genome Res, 11, 1730-1735; Lueking, A., Holz, C, Gotthold, C, Lehrach, H. and Cahill, D. (2000) A system for dual protein expression in Pichia pastoris and Escherichia coli, protein Expr. Purif. , 20, 372-378). Here, the cDNA of a particular tissue in a bacterial or a eukaryotic expression vector, such as yeast, cloned. The vectors used for the expression are generally characterized by the fact that they carry inducible promoters, with which the timing of protein expression can be controlled. In addition, expression vectors have sequences for so-called

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

Purification via affinity chromatography (IMAC) allows.

For example, the gene products of a cDNA expression library of human fetal brain tissue in the bacterial expression system Escherichia coli in high density format were placed on a membrane and successfully screened with different antibodies. It could be shown that the proportion of full-length proteins is at least 66%. The recombinant proteins out

In addition, expression libraries have been published in

High throughput can be expressed and purified (Braun P., Hu, Y., Shen, B., Halleck, A., Koundinya, M., Harlow, E. and LaBaer, J. (2002) Proteome-scale purification of human

proteins from bacteria. Proc Natl Acad. See U S A, 99, 2654-2659; Büssow (2000) supra; Lueking, A., Horn, M., Eickhoff, H., Büssow, K., Lehrach, H. and Walter, G. (1999) Protein microarrays for gene expression and antibody screening.

Analytical Biochemistry, 270, 103-111). Such protein biochips are based on cDNA expression libraries

in particular the subject matter of WO 99/57311 and WO 99/57312. Further, in addition to antigen-presenting protein biochips, antibody-presenting arrangements 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). However, there is a great need to provide indication-specific diagnostic devices, such as a protein biochip.

Laboratory parameters for the diagnosis of prostate cancer include acid phosphatase (SP) and prostate-specific antigen (PSA). Especially the PSA currently has a high

Importance in diagnostics. It is specific to the

Prostate, but not for a tumor, but may also be increased in inflammation, benign prostatic hyperplasia, a urinary retention or for no apparent reason. A value above 4 ng / ml is already in need of clarification.

W02010 / 000874 of the applicant already describes the diagnosis of prostate cancer and prostatectivitis by means of a protein biochip and provides certain diagnostic

Marker sequences available.

A disadvantage, however, is that these markers are also suitable for the diagnosis of prostatectomy, so that there is no discrimination within the patient group of these marker sequences known in the prior art

Prostate cancer patients is possible. Furthermore, it is disadvantageous in the prior art that due to other indications, such as prostatectivitis or diabetes, false-positive

Marker sequences can be identified.

WO 2009/080017 discloses marker sequences for neurodegenerative disorders and their use. The sequences disclosed in WO 2009/080017 were correspondingly obtained by a method without using samples of prostate cancer patients

receive . US 2004/259086 relates to compositions, kits and methods for the detection, characterization, prevention and treatment of prostate cancer in humans. A number of marker sequences whose expression levels correlate with prostate cancer were used. However, US 2004/259086 does not disclose Marker sequences, for the diagnosis of prostate cancer excluding prostate inflammatory diseases, diabetes, polymorbidity. The marker sequences in US 2004/259086 were obtained by subtracting cDNA libraries prepared from the mRNA of patients with benign prostatic hyperplasia and the cDNA prepared from the mRNA of patients with prostate cancer ([0339] et seq. In US 2004/259086). Another

Validation did not take place. These marker sequences are thus not suitable for identifying patient subgroups within the group of prostate cancer patients.

The DNA sample arrays HG-U95, HG-U133 and HG-U133 (Affymetrix) already disclose those in the present application

used sequences SEQ ID No. 1 and 247. However, the DNA sample arrays HG-U95, HG-U133, and HG-U133 were not validated for specific diagnostic uses.

The object of the present invention is to provide improved marker sequences and their diagnostic use for the treatment of prostate cancer, wherein

in particular discrimination or exclusion of one

Prostatex inflammation and / or diabetes disease occurs.

The provision of specific marker sequences allows a safe diagnosis and stratification of patients with

Prostate carcinoma, in particular by means of a protein biochip.

Therefore, the invention relates to the use of

Marker sequences for the diagnosis of prostate carcinoma, wherein at least one marker sequence of a cDNA selected from the group SEQ 1 - 246 or in each case a protein encoded thereby or in each case a partial sequence or fragment thereof

(hereinafter: marker sequences according to the invention) to or from a patient to be examined. The marker sequences of the invention could by means of differential screening of samples and healthy

Subjects with patient samples identified with prostate cancer on a protein biochip. For demarcation and

Discrimination against prostate inflammatory disease (s) and / or diabetes, the marker sequences of the invention are tested against patient samples of prostate inflammatory diseases (e.g., all forms of prostatic hyperplasia, prostatitis) or diabetes on a protein biochip. One or more different prostate inflammatory diseases or

Forms of diabetes are excluded. Only such marker sequences according to the invention are considered as marker sequences for prostate carcinoma, which are not at the same time also as marker sequences for

Prostate inflammatory diseases or diabetes were identified.

This approach also advantageously allows for the elimination of comorbidity, e.g. in the presence of prostate inflammatory diseases or diabetes. On the other hand, in the course of this procedure false-positive

Marker sequences excluded.

This was the first time using protein biochips (see

Examples) of these marker sequences of the invention are sensitively identified. Thus, the present invention for the first time

Marker sequences available with a special

Patient group within the group of prostate cancer

Patients can be determined (hereinafter "subgroup") .These marker sequences can selectively be the subgroup - i.e. the patients within the group of patients with

Prostate cancer in which no diabetes and / or

Prostate inflammatory disease is present - identified

(diagnosed). In the context of individualized medicine, patients can thus be found subgroups within the group of prostate cancer patients and the appropriate Treatment / Therapy for this subgroup can be selected while patients with prostate cancer who are not

Subgroup belong to be excluded from inappropriate therapies. The invention is therefore a use of

Marker sequences to identify a subgroup of

Patients with prostate cancer, the subgroup none

Prostate inflammatory disease (s) and / or diabestes, and wherein at least one marker sequence of a cDNA selected from the group SEQ 1 - 246 and / or SEQ 247-452 or each protein encoded thereby or in each case a partial sequence or fragment thereof to or from one too

determined by the examining patient

The invention therefore also relates to the use of the marker sequences for the diagnosis of prostate cancer

Exclusion of prostate inflammatory diseases or diabetes or polymorbidity, wherein at least one marker sequence of a cDNA selected from the group SEQ 1 - 246 and / or SEQ 247-452 or each one protein encoded thereby or in each case a partial sequence or fragment thereof to or determined by a patient to be examined is and where the

Marker sequence (s) has been identified with a method comprising the a) identification of marker sequence candidates by differential screening with protein biochips, for example two protein biochips, each from a cDNA expression library of a patient with

Prostate cancer and a subject without

Prostate cancer, b) validation of marker sequence candidates using

Samples from patients with

Prostatectomy for inflammation (s) and / or samples from patients with diabetes. The samples used for validation come from

Patients who have one or more

Prostate inflammatory diseases and / or have diabetes but no prostate cancer. One embodiment of the invention relates to the use of the marker sequences according to the invention for the diagnosis of

Prostate cancer excluding

Prostate inflammatory diseases or diabetes or

Polymorbidity wherein 2 or 3, preferably 4 or 5,

more preferably 6, 7 or 8 or more different

Marker sequences, for example 10 to 20 or 30 or more different marker sequences on or from one to

be determined by examining patients.

One embodiment of the invention relates to the use of the marker sequence (s) according to the invention for the diagnosis of

Prostate cancer excluding

Prostate inflammatory diseases or diabetes or

Polymorbidity, wherein at least one of the marker sequences is selected from the group SEQ ID No. 247, SEQ ID No. 250, SEQ ID NO. 290 or a by SEQ ID No. 247, SEQ ID No. 250, SEQ ID No. 290 encoded protein or a partial sequence or a fragment of SEQ ID No. 247, SEQ ID No. 250, SEQ ID No. 290. Further preferred are therefore the corresponding

Marker sequences selected from the group SEQ ID No. 1, SEQ ID no. 4 and SEQ ID No. 44. or a SEQ ID No. 1, SEQ ID No. 4, SEQ ID No. 44 encoded protein or a partial sequence or a fragment of SEQ ID No. 1, SEQ ID No. 4, SEQ ID NO. 44th

Further preferred are the marker sequences SEQ ID no. 249, SEQ ID No. 255, SEQ ID No. 271, SEQ ID No.

301, SEQ ID No. 341, wherein at least one of the marker sequences is selected from the group SEQ ID No. 249, SEQ ID No. 255, SEQ ID no. 271, SEQ ID No. 301, SEQ ID No. 341 or a by SEQ ID NO. 249, SEQ ID No. 255, SEQ ID No. 271, SEQ ID No. 301, SEQ ID No. 341 encoded protein or a partial sequence or a fragment of SEQ ID No. SEQ ID No. 249, SEQ ID No. 255, SEQ ID No. 271, SEQ ID No. 301, SEQ ID No.341. Preference is therefore further given to the corresponding marker sequences selected from the group SEQ ID No. 3, SEQ ID No. 9, SEQ ID NO. 25, SEQ ID No. 55, SEQ ID No. 95 or a by SEQ ID No. 3, SEQ ID no. 9, SEQ ID No. 25, SEQ ID No. 55, SEQ ID No. 95 encoded protein or a partial sequence or a fragment of SEQ ID No. 3, SEQ ID No. 9, SEQ ID No. 25, SEQ ID No. 55, SEQ ID NO. 95. An embodiment of the invention relates to the use of the marker sequence (s) according to the invention for the diagnosis of

Prostate cancer excluding

Prostate inflammatory diseases or diabetes or

Polymorbidity, wherein at least one marker sequence of a protein selected from the group neuro-oncological

ventral antigen 2 (NOVA2), syntaxin 18 (STX18), heat shock (Heatshock) 105kDa / 110kDa protein 1 (HSPH1) or a nucleic acid encoding it or a partial sequence or a

A fragment thereof is determined on or from a patient to be examined and wherein the marker sequence (s) with a

A method has been identified, comprising the steps of a) identification of marker sequence candidates by differential screening with protein biochips, for example two protein biochips, each from a cDNA expression library of a patient with

Prostate cancer and a subject without

Prostate cancer; b) validation of marker sequence candidates using samples from patients with

Prostatectomy for inflammation (s) and / or samples from patients with diabetes.

Another embodiment of the invention relates to

Use of the marker sequence (s) according to the invention for

Diagnosis of prostate cancer to the exclusion of Prostate inflammatory diseases or diabetes or

Polymorbidity according to one of the preceding claims, characterized in that the determination is carried out by means of in vitro diagnosis. Another embodiment of the invention relates to

Use of at least one marker sequence of a cDNA in each case selected from the group SEQ 1 - 246 and / or SEQ 247-452 or in each case a protein encoded thereby or in each case a partial sequence or fragment thereof as a diagnostic agent, wherein the marker sequence (s) was identified by a method ( n), comprising the steps of a) identification of marker sequence candidates by differential screening with protein biochips, for example two protein biochips from each cDNA expression library of a patient with

Prostate cancer and a subject without

The invention also provides a method for the diagnosis of prostate cancer excluding

Prostate inflammatory diseases or diabetes or

Polymorbidity, where

a. ) at least one marker sequence of a cDNA selected from the group SEQ 1 - 246 and / or SEQ 247-452 or in each case a protein encoded thereby or in each case a partial sequence or fragment thereof is applied to a solid support

(become) 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 excerpt with the marker sequence (s) from a.).

The invention also provides a method for

Stratify, in particular for risk stratification, or for therapy control of a patient with prostate cancer excluding prostate inflammatory diseases or diabetes or polymorbidity, wherein at least one

Marker sequence of a cDNA selected from the group SEQ 1 - 246 and / or SEQ 247 - 452 or a protein encoded thereby or in each case a partial sequence or fragment thereof is determined on or from a patient to be examined and wherein the marker sequence (s) with a method was identified (n) comprising the steps a) identification of marker sequence candidates by differential screening with protein biochips, for example two protein biochips, each from a cDNA expression library of a patient with

Prostate cancer and a subject without

Prostate cancer, b) validation of marker sequence candidates using

Samples from patients with

The invention also relates to an assay, protein biochip consisting of an arrangement containing at least one

Marker sequence of a cDNA in each case selected from the group SEQ 1 - 246 and / or SEQ 247-452 or in each case a protein encoded thereby or in each case a partial sequence or fragment thereof, characterized in that the marker sequence (s) is applied to a solid support (are ) and where the

Marker sequence (s) has been identified by a method comprising the steps a) Identification of marker sequence candidates by differential screening with protein biochips, for example two protein biochips, each from a cDNA expression library of a patient with

Prostate cancer and a subject without

The invention also provides a diagnostic for the diagnosis of prostate cancer excluding

Prostate inflammatory diseases or diabetes or

Polymorbidity containing at least one marker sequence each selected from the group SEQ 1 - 246 and / or SEQ ID 247 - 452 or in each case a protein encoded thereby or in each case a partial sequence or fragment thereof, wherein the

Marker sequence (s) has been identified by a method comprising the steps of a) identifying marker sequence candidates by differential screening with protein biochips, for example two protein biochips each from a cDNA expression library of a patient with

Prostate cancer and a subject without

Prostatectomy for inflammation (s) and / or samples from patients with diabetes. The term "prostate cancer" includes only the indication prostate cancer or prostate cancer under

Exclusion of co- or polymorbidity (definition eg according to Pschyrembel, de Gruyter, 261st edition (2007), Berlin). The If necessary, overlapping diseases of prostate inflammation or diabetes are strictly excluded according to the invention.

The term "prostate inflammatory diseases" includes

all forms of prostatitis, including chronic forms of prostatitis, including one

Prostate hyperplasia, especially benign prostate hyperplasia. Included in the invention is the presence of one or more different prostate inflammatory diseases.

In the context of this invention, diabetes is understood to mean, for example, "diabetes mellitus", in particular type II diabetes mellitus (insulin resistance), a chronic one

Metabolic disease, in which the production of insulin in the ß-cells of the islets of Langerhans is disturbed in the pancreas or insulin is indeed present on his

Destination, the cell membranes, but can not work properly. The result of this impaired insulin production or effect is increased blood sugar levels (hyperglycemia). At the

Diabetic disease is distinguished between prediabetes, in which it comes only in its final stage to a laboratory-chemically detectable "disturbed glucose tolerance", and the actual manifest diabetes mellitus.At the beginning of the prediabetic phase of the disease is the

Insulin resistance. Almost simultaneously, endothelial dysfunction, hyperlipoproteinemia and hypertensive circulatory dysregulation are already developing.

In the context of this invention, diabetes is also understood to mean type I diabetes.

Therefore, the invention relates to such marker sequences that co-polymorphism or polymorbidity with other indications, such as

Prostatic inflammation (s) or diabetes (in all forms), excludes.

In a further embodiment, therefore, at least 2 to 5 or 10, preferably 30 to 50 marker sequences or 50 to 100 or more marker sequences on or from one to

intended for the examining patient.

In a further embodiment of the invention, the marker sequences according to the invention can also be combined, supplemented or extended with known biomarkers for this indication. Also particularly preferred are such markers as disclosed in WO2010 / 000874.

In a preferred embodiment, the determination of the marker sequences takes place outside the human body and the determination is made in an ex vivo / in vitro diagnosis.

In a further embodiment of the invention, the invention relates to the use of marker sequences as diagnostics, wherein at least one marker sequence of a cDNA is selected from the group SEQ 1 - 246 or in each case a protein coding therefor or in each case a partial sequence or fragment thereof.

Furthermore, the invention relates to a method for the diagnosis of prostate carcinoma, wherein a.) At least one marker sequence of a cDNA selected from the group SEQ 1 - 246 or a respective coding protein or a partial sequence or fragment thereof is applied to a solid support and b. ) with body fluid or tissue extract one

C.) The detection of an interaction of the body fluid or tissue extract with the marker sequences from a.) Is done.

Therefore, the invention also relates to diagnostics for

Diagnosis of prostate cancer each selected from the

Group SEQ 1 - 246 or in each case a protein coding therefor or in each case a partial sequence or fragment thereof. In a particularly preferred embodiment, the

Marker sequences SEQ 1-18 are particularly preferred and SEQ 19-56 are preferred, again lower numerical values being preferred. The detection of such an interaction can, for example, by a probe, in particular by an antibody

respectively .

Therefore, the invention also has the object of providing a diagnostic device or assay, in particular a protein biochip, which allows for diagnosis or examination of prostate cancer.

Furthermore, the invention relates to a method for

Stratify, in particular for risk stratification and / or therapy control of a patient with a

Prostate carcinoma, wherein at least one marker sequence of a cDNA selected from the group SEQ 1 - 246 or in each case a protein coding therefor is determined on a patient to be examined.

Also included is the stratification of patients with

Prostate carcinoma in new or established subgroups of the

Prostate cancer, as well as the sensible selection of

Patient groups for the clinical development of new ones

Therapeutics. The term therapy control also includes the classification of patients in responders and non-responders with respect to a therapy or its

Therapy course.

"Diagnosis" in the sense of this invention means the positive detection of a prostate carcinoma by means of

marker sequences according to the invention and the assignment of the patients to a prostate carcinoma. The term diagnosis includes medical diagnostics and related

Studies, in particular in vitro diagnostics and

Laboratory diagnostics, also proteomics and nucleic acid blots. Further investigations can be made for protection and for

Exclusion of other diseases. Therefore, the term diagnosis also includes the differential diagnosis of prostate cancer by means of the marker sequences according to the invention and the prognosis of a prostate carcinoma. "Stratification (also: stratification) or therapy control" in the sense of this invention means that the inventive method decisions for the treatment and therapy of the

Patients are allowed, whether it is the hospitalization of the patient, use, effect and / or dosage of one or more drugs, a therapeutic measure or the

Surveillance of disease progression as well as treatment course or etiology or classification of a disease, e.g. into a new or existing subtype or the differentiation of diseases and their patients.

In a further embodiment of the invention, the term "stratification" includes in particular the

Risk stratification with the prognosis of an "outcome" of a detrimental health event For the purposes of this invention, "patient" is understood to mean any test subject - human or mammal - with the proviso that the test person is examined for prostate carcinoma.

The term "marker sequences" in the sense of this invention means that the cDNA or the respective polypeptide or protein obtainable therefrom are significant for prostate carcinoma For example, the cDNA or the respectively thereof

available polypeptide or protein interacts with a sample, e.g. Substances from the body fluid or tissue extract of a patient with

From prostate cancer to prostate cancer (e.g., antigen (epitope) / antibody (paratope))

Interaction). For the purposes of the invention, "at least one marker sequence of a cDNA selected from the group SEQ. 1-246 or in each case a protein coding therefor or in each case a partial sequence or fragment thereof to or from a patient to be examined is determined" that an interaction between the body fluid or

Tissue extract of a patient and the invention

Marker sequences is detected. Such an interaction is for example a bond, in particular a binding substance at least one marker sequence according to the invention or, in the case of a cDNA, the hybridization with a suitable substance under selected conditions, in particular stringent

Conditions (e.g., as defined in J. Sambrook, E. F. Fritsch, T. Maniatis (1989), Molecular cloning: A laboratory manual, 2nd Edition, Cold Spring Habor Laboratory Press, Cold Spring Habor, USA or Ausubel, "Current Protocols in

Molecular Biology, Green Publishing Associates and Wiley Interscience, N.Y. (1989)). An example of stringent

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

Room temperature.

Such substances are part of a component according to the invention

Body fluid, in particular blood, whole blood, blood plasma, blood serum, patient serum, urine, cerebrospinal fluid, synovial fluid or a tissue extract of the patient.

"Samples" of patients or subjects include, for example, body fluid, especially blood, whole blood, blood plasma, blood serum, patient serum, urine, cerebrospinal fluid, synovial fluid, or a tissue extract of the patient or subject.

In a further embodiment of the invention, however, the marker sequences of the invention may be present at a significantly higher or lower expression rate or concentration indicative of prostatic carcinoma. In this case, the relative. By proteomics or nucleic acid blots

Expression rates ill / healthy of the invention

Marker sequences for prostate inflammatory diseases,

Prostate carcinoma determined. In a further embodiment of the invention, the marker sequences have a recognition signal which is addressed to the substance to be bound (for example antibodies,

Nucleic acid). According to the invention, for a protein, the recognition signal is preferably an epitope and / or paratope and / or hapten and, for a cDNA, a hybridization or hybridization protein

Binding region.

The marker sequences of the invention are the subject of Table A and can by the respective cited

Database entry (also via Internet:

http://www.ncbi.nlm.nih.gov/) can be clearly identified (see Table A: Accession No. there).

Therefore, the invention also relates to the full-length sequences of the markers according to the invention and indeed as defined in Table 1 on the known database entry according to Table A, hereinafter called SEQ ID NOS 247-452, see also that

associated sequence protocol.

Furthermore, therefore, also include analog

Embodiments of SEQ 247-452 to the marker sequences SEQ 1-246, e.g. set forth in the claims, since the

SEQ ID Nos. 4,646, in turn, represent partial sequences, at least with high homology. The specific ones

However, marker sequences SEQ 1-246 are in accordance with the invention

prefers . Further preferred are SEQ 247-260 and SEQ 261-294.

In a further embodiment of the invention

Marker sequences preferred that have P values less than or equal to 0.2, preferably less than or equal to 0.15, more preferably less than or equal to 0.1. In another

Embodiment of the invention are the marker sequences SEQ ID No. 247, 250, 290, partial sequences, fragments or homologs and the peptides / proteins encoded thereby.

These marker sequences have particularly suitable P values: SEQ ID no. 247 (P value: 0.1053), SEQ ID No. 250 (P value:

0.0310) and SEQ ID No. 290 (P value: 0.0254). The P value indicates the probability that a match was found in the database. For the definition of the P-value, see, for example, http://www.ncbi.nlm.nih.gov/books/NBK62051/.

According to the invention, the marker sequences also include such

Modifications of the cDNA sequence and the corresponding

Amino acid sequence, such as chemical modification, such as

Citrullination, acetylation, phosphorylation,

Glycation or polyA strand and other modifications known to those skilled in the art.

In a further embodiment of the invention are also

Partial sequences or fragments of the invention

Includes marker sequences. In particular, those partial sequences having an identity of 99% or more, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, especially 85%, 80% or 70% with have the marker sequences according to the invention and for the inventive use - the detection

Prostate cancer excluding

Prostate inflammatory diseases or diabetes or

Polymorbidity - are suitable. (so-called "homologues", homologous marker sequences) .Hiomologists may use protein or

Nucleic acid sequences.

Partial sequences are also those sequences which have 50 to 100 nucleotides, 70-120 nucleotides of a sequence of SEQ 1-452, or peptides obtainable therefrom.

In a further embodiment, the respective

Marker sequence can be represented in different amounts in one or more areas on a solid support. This allows a variation of the sensitivity. The regions can each have an entirety of marker sequences, ie a sufficient number of different marker sequences, in particular 2 to 5 or 10 or more and, if appropriate, further nucleic acids and / or proteins, in particular biomarkers. However, at least 96 to 25,000 (numerically) or more from different or identical marker sequences and further nucleic acids and / or proteins, in particular, are preferred

Biomarkers. Further preferred are more than 2,500, more preferably 10,000 or more different or the same

Marker sequences and optionally other nucleic acids and / or proteins, in particular biomarkers.

Another object of the invention relates to an array of marker sequences containing at least one marker sequence of a cDNA selected from the group SEQ 1-452 or in each case a protein coding therefor. Preferably, the

Arrangement of at least 2 to 5 or 10, preferably 30 to 50 marker sequences or 50 to 100 or more marker sequences.

In the context of this invention, "arrangement" synonymously means "array" and insofar as this "array" is used to identify substances on marker sequences, this is to be understood as meaning an "assay" or a diagnostic device. In a preferred embodiment, the arrangement is designed such that those represented on the assembly

Marker sequences are in the form of a grid on a solid support. Further, such arrangements are preferred which allow a high density array of protein binders and spotting the marker sequences. 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.

In the context of this invention, however, 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 (eg so-called lateral flow immunoassays) or similar immunological single or multiplex detection methods. A protein biochip in the sense of this Invention is the systematic arrangement of proteins on a solid support.

The marker sequences of the assembly are fixed to a solid support, but preferably spotted or immobilized even printed, i. applied reproducibly. One or more marker sequences can be duplicated in the totality of all

Marker sequences are present and available in different quantities related to a spot. Furthermore, the

Marker sequences on the solid support may be standardized (e.g., by serial dilution series of e.g.

Human globulins as internal calibrators for

Data normalization and quantitative evaluation).

Therefore, the invention relates to an assay or protein biochip consisting of an array containing marker sequences according to the invention.

In a further embodiment, the marker sequences are present as clones. Such clones can be obtained, for example, by means of a cDNA expression library according to the invention (Büssow et al., 1998 (supra)). In a preferred embodiment, such expression libraries become

containing clones by means of expression vectors from a

expressing cDNA library consisting of the cDNA

Obtained marker sequences. These expression vectors preferably contain inducible promoters. The induction of

Expression can e.g. by means of an inductor, such as IPTG. Suitable expression vectors are described in Terpe et al. (Terpe T Appl Microbiol Biotechnol 2003 Jan; 60 (5): 523-33). 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

are tissue specific (eg human tissue, in particular human organs). Furthermore, such expression libraries are also included according to the invention, which can be obtained by exon trapping. Instead of expression library can be spoken synonymously from an expression bank. Also preferred are protein biochips or corresponding expression libraries which have no redundancy (so-called: Uniclone® library) and can be prepared, for example, according to the teachings of WO 99/57311 and WO 99/57312. These preferred Uniclone libraries have a high content of non-defective, fully expressed proteins of a cDNA expression library.

Also within the scope of this invention, 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 an arrangement, wherein the

Marker sequences are present as clones. In addition, the marker sequences may be in the form of a fusion protein in the particular form

For example, at least one affinity epipope or "tag" contains. The tag 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, preferably a cellulose-binding one

Domain, green fluorescent protein, maltose binding

Protein, calmodulin-binding protein, glutathione S-transferase or lacZ. In all embodiments, the term "solid support" includes embodiments such as a filter, a membrane, a magnetic or fluorophore-labeled bead

Silicon wafer, glass, metal, plastic, a chip, a Mass spectrometric target or a matrix. However, a filter is preferred according to the invention.

Also preferred as the filter is PVDF, nitrocellulose or nylon (e.g., Immobilon P Millipore, Protran Whatman, Hybond N + Amersham).

In a further preferred embodiment of the

According to the invention, this corresponds to a grid having the order of a microtiter plate (8-12 wells strips, 96 wells, 384 wells or more), a silicon wafer, a chip, a mass spectrometric target or a matrix.

In a further embodiment, the invention relates to an assay or protein biochip for identifying and

Characterizing a substance for prostate cancer, characterized in that an inventive arrangement or

Assay with a.) Is brought into contact at least one substance to be examined and b.) A binding success is detected.

The invention further relates to a method for identifying and characterizing a substance for prostate carcinoma, 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 tested may be any native or non-native biomolecule, a synthetic chemical

Be a molecule, a mixture or a substance library.

After the substance to be examined contacts a marker sequence, the evaluation of the binding success, for example, using commercially available image analysis software (GenePix Pro (Axon Laboratories), Aida

(Raytest), ScanArray (Packard Bioscience). The visualization of protein-protein interactions according to the invention (eg protein on marker sequence, such as

Antigen / antibody) or corresponding "means for detecting the binding success" can, for example, by means of

Fluorescent labeling, biotinization, radio-isotopic labeling or colloidal gold or latex particle marking in a conventional manner. Detection of bound antibodies takes place with the help of secondary antibodies

Antibodies with commercial reporter molecules

(e.g., Cy, Alexa, Dyomics, FITC or similar fluorescent dyes, colloidal gold or latex particles) or with reporter enzymes such as alkaline

Phosphatase, horseradish peroxidase, etc. and the

corresponding colorimetric, fluorescents or

chemoluminescent substrates. A readout is e.g. by means of a microarray laser scanner, a CCD camera or visually.

In a further embodiment, the invention relates to a drug / drug or prodrug designed for prostate cancer and obtainable through the use of the prostate cancer

Assays according to the invention or protein biochip.

Therefore, the invention also relates to the use of an inventive arrangement or an assay for screening prostate cancer drugs. Therefore, in another embodiment, the invention also relates to a target for the treatment and therapy of

Prostate carcinoma, each selected from the group SEQ 1 - 246 or each protein coding therefor.

In a further embodiment, the invention also relates to the use of the invention

Marker sequences, preferably in the form of an arrangement, as an affinity material for performing an apheresis or iwS. a blood wash, wherein substances from bodily fluids of a patient with prostate cancer, such as blood or plasma, to bind the marker sequences according to the invention and consequently the body fluid can be selectively withdrawn.

Corresponding devices are known in the art, e.g. chromatographic devices containing beads, spheres or chromatographic material, e.g. in a column having the marker sequences of the invention and therefore e.g. (Auto) can selectively withdraw antibodies.

Examples and figures:

Ten or more patient samples were individually screened against a cDNA expression library. The prostate carcinoma - specific expression clones were identified by comparison with ten or more healthy samples. Furthermore, the identified markers were used against samples of

Checked prostate inflammatory or diabetic patients.

False-positive markers were removed. The subsequent

Identity of the remaining marker sequences was determined by DNA sequencing.

FIG. 1 shows the differential screening between two protein biochips from in each case one cDNA expression bank of a patient and one healthy subject. The

Differential clones are detected by fluorescence labeling and evaluated bioinformatorisch.

In the context of biomarker identification various bioinformatic analyzes are carried out. For each serum, microarray reactivities against about 2000

measured different antigens. These data are used for a ranking of the spotted antigens regarding their

Differentiation ability between healthy and diseased sera used. This evaluation is by means of not

parametric Mann-Whitney tests on normalized

Intensity data performed. For normalization, 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. When In a very conservative approach, a Bonferroni correction is performed and, in addition, the less restrictive False Discovery Rate (FDR) according to Benjamini & Hochberg is calculated.

Furthermore, the data are used to classify the sera. Here, different multivariate methods are used. These are methods from the statistical

Learning methods such as support vector machines (SVM), neural networks or classification trees, as well as a

Threshold method, which is suitable for both classification and visual representation of the data.

To avoid overfitting, a 10-fold cross-validation of the data is performed.

Table A:

SEQ gi name DNA

ID Accession Accession

247 gil5902723 Neuro-oncological ventral antigen 2 (NOVA2) NM_002516

248 gill 13411825 Similar to Cyclin L2 (Paneth cell-enhanced expression protein) NM_030937

transcript variant 1 (LOC727877)

249 cast 1543652 Signal recognition particle 14kDa (homologous Alu RNA binding NM_003134

protein) (SRP14)

250 _g il42544158 Heat shock 105kDa / l lOkDa protein 1 (HSPH1) NM_006644

251 gill 13428756 Zinc finger protein 154 (pHZ-92) (ZNF154) NM_0010853

84

252 gil32490571 Erythrocyte membrane protein band 4.1-like 3 (EPB41L3) NM_012307

253 gil77628146 Endoplasmic reticulum protein 29 (ERP29) transcript variant 1 NM_006817

254 gil22749232 Zinc finger protein 579 (ZNF579) NM_152600

255 gill 94097404 Peptide chain release factor 3 NM_018094

256 gil217330633 serine / threonine kinase 36 NM_015690

257 gill 6507207 Capicua homolog (Drosophila) (CIC) NM_015125

258 gil32261293 Protein kinase interferon-inducible double stranded RNA dependent NM_003690

activator (PRKRA)

259 gil49574506 Neugrin neurite outgrowth associated (NGRN) transcript variant 1 NM_0010330

88

260 gil89031690 Chromosomes 10 genomic contig, reference assembly NM_005876

261 gill 13427652 Alveolar soft part sarcoma chromosomes region candidate 1 (ASPSCRl) NM_024083

262 gil40353201 OTU domain containing 5 (OTUD5) NM_017602

263 gil66932901 SREBP cleavage-activating protein (SCAP) NM_012235

264 gil33624820 Septin 6 (SEPT6) transcript variant V NM_145800

265 gil40807365 Dihydrouridine synthase 1-like (S. Cerevisiae) (DUS 1L) NM_022156 gil70609878 Ribosomal protein S2 (RPS2) NM_002952 gil83267865 Dynein, light chain, LC8-type 1 (DYNLL1), transcript variant 1 NM_0010374

94

gill 13422143 Similar to 60S ribosomal protein L21, transcript variant 2 (LOC731567) XM_0011335

19

High Mobility Group nucleosomal binding domain 4 (HMGN4) NM_006353 gil39725675 CDK2-associated protein 2 (CDK2AP2) NM_005851 gil46389548 endosulfine alpha (ENSA ), transcript variant 3 NM_004436 gil46389553 endosulfine alpha (ENSA), transcript variant 4 NM_207044 gil46389549 endosulfine alpha (ENSA), transcript variant 1 NM_207042 gil47078237 G protein pathway suppressor 1 (GPS 1), transcript variant 1 NM_212492 gil47078280 Family with sequence similarity 53, member B (FAM53B) NM_014661 gil4757715 Sperm associated antigen 7 (SPAG7) NM_004890 gil50557645 Zinc finger, FYVE domain containing 27 (ZFYVE27), transcript variant NM_0010022

1 61

gil5174742 Ubiquinol-cytochrome c reductase, Rieske iron-sulfur polypeptides 1 NM_006003

(UQCRFS 1)

gil53759121 Adenomatosis polyposis coli (APC) NM_000038 gil57242754 calsyntenin 1 (CLSTN1), transcript variant 2 NM_014944 gil7705480 ubiquitin-fold modifier conjugating enzyme 1 (UFC1) NM_016406 gil82659090 Staufen, RNA binding protein, homologue 1 (Drosophila) (STAU1), NM_0010373

transcript variant T5 28

gil83641890 glyceraldehyde-3-phosphate dehydrogenase (GAPDH) NM_002046 gil57222567 myeloid / lymphoid or mixed-lineage leukemia (trithorax homologue, NM_005937

Drosophila); translocated to, 6 (MLLT6)

gill4249519 Hypothetical protein FLJ 14668 (FLJ 14668) NM_032822 gil38201669 Eukaryotic translation elongation factor 2 (EEF2) NM_001961 gil20127557 Chromatin-modifying protein 5 (CHMP5) NM_016410 gil20302149 palmitoyltransferase 1B (muscle) (CPT1B), NM_152247

nuclear gene encoding mitochondrial protein, transcript variant 4

gil33286421 Pyruvate kinase, muscle (PKM2), transcript variant 3 NM_182471 gil34147626 Zinc finger protein 447 (ZNF447) NM_023926 gil38176162 Ring finger protein 130 (RNF130) NM_018434 gil47933378 N-ethylmaleimide-sensitive factor attachment protein, alpha (NAPA) NM_003827 gil49472815 Fascin homolog 1 , actin-bundling protein (Strongylocentrotus NM_003088

purpuratus) (FSCN1)

gil67189547 Ribosomal protein L6 (RPL6), transcript variant 2 NM_000970 gil6912539 Nucleotide binding protein 2 (MinD homologue, E. coli) (NUBP2) NM_012225 gil73622128 Caspase 6, apoptosis-related cysteine peptidase (CASP6), transcript NM_001226

variant alpha

gil83641894 Heterogeneous nuclear ribonucleoprotein AI (HNRPA1), transcript NM_031157

variant 2 gil94721349 islet cell autoantigen 1, 69kDa (ICA1), transcript variant 2 NM_004968 gil42794610 6-phosphogluconolactonase (PGLS), NM_012088 gil 113420239 Similar to block of proliferation 1 (LOC727967) XM_0011262

55

gi | 14149778 Chromosomes 1 open reading frame 160 (Clorfl60) NM_032125 gill6306547 seryl-tRNA synthetase (SARS) NM_006513 gil20336240 proprugase convertase subtilisin / kexin type 1 inhibitor (PCSK1N) NM_013271 gil22027621 TNF receptor-associated factor 4 (TRAF4), transcript variant 1 NM_004295 1543190 Chromosomes 10 open reading frame 58 (C10orf58) NM_032333 gil32129198 Cytokines induced protein 29 kDa (CIP29) NM_033082 gil32455235 Helicase (DNA) B (HELB) NM_033647 gil34452680 Ring finger protein 10 (RNF10) NM_014868 gil34996518 Galectin-3 internal gene (GALIG) NM_194327 gil40804743 Sema domain, immunoglobulin domain (Ig), transmembrane domain NM_017789

(TM)

and short cytoplasmic domain, (semaphorin) 4C (SEMA4C)

gil41393582 EGF-like domain, multiple 7 (EGFL7), transcript variant 2 NM_201446 _g il46430498 V-rel reticuloendotheliosis viral oncogene homolog A, NM_021975

nuclear factor of kappa light polypeptide gene enhancer in B-cells 3,

p65 (avian) (RELA)

gil47519746 mi togen-activated protein kinase 11 (MAPK11) NM_002751 gil50233802 NDRG family member 4 (NDRG4) NM_020465 gil56090145 Hypothetical LOC339123 (LOC339123), NM_0010059

20

gil71164877 Ribosomal protein S 12 (RPS 12) NM_001016 gil72534683 Phospholipase D family, member 3 (PLD3), transcript variant 1 NM_0010316

96

gil7305502 stomatin (EPB72) -like 2 (STOML2) NM_013442 gil90193629 septin 5 (SEPT5) NM_002688 gil50592995 tubulin, beta 3 (TUBB3) NM_006086 gill6554608 mitochondrial ribosomal protein S 11 (MRPS 11), NM_022839

nuclear gene encoding mitochondrial protein, transcript variant 1

giss0581139 Proteasome (prosome, macropain) activator subunit 1 (PA28 alpha) NM_006263

(PSME1), transcript variant 1

gil31317308 phosphatidylinositol 4-phosphate 5-kinase, type I, gamma (PIP5K1C) NM_012398 gil46048184 Sterile alpha motif domain containing 10 (S AMD 10) NM_080621 gil56549124 Dynamin 2 (DNM2), transcript variant 4 NM_0010053

62

gil70166553 GRINL1A combined protein (Gcoml), transcript variant 9 NM_0010180

97

gil40795668 Solute carrier family 38, member 3 (SLC38A3) NM_006841 giss0410780 Hypothetical protein FLJ 12949 (FLJ 12949), transcript variant 2 NM_178159 giss 1361946 Stathmin-like 4 (STMN4) NM_030795 gill 13413590 Similar to deoxythymidylate kinase (thymidylate kinase), transcript XM_0011262

variant 4 (LOC727761) 11

gil 113430465 Similar to ataxin 7-like 3 (LOC392485) X _018762 gil22219473 Fas (TNFRSF6) -associated via death domain (FADD) NM_003824 gil71361681 Nuclear mitotic apparatus protein 1 (NUMA1) NM_006185 gil20336312 B-cell CLL / lymphoma 11A (zinc finger protein) (BCL11A), transcript NM_138559

variant 3

gil34147391 Coiled-coil domain containing 130 (CCDC130) NM_030818 gil 12056467. Function plakoglobin (JUP), transcript variant 2 NM_021991 gil21464122 Methyl CpG binding domain protein 3 (MBD3) NM_003926 _g i | 42544170 Excision repair cross-complementing rodent repair deficiency, NM_001983

complementation group 1

(includes overlapping antisense sequence) (ERCCl), transcript variant 2 gil83523747 Hypothetical protein FLJ 13305 (FLJ 13305) NM_032180 gil46358416 Glutamate receptor, metabotropic 3 (GRM3) NM_000840 gil39812062 Mitochondrial ribosomal protein L28 (MRPL28), nuclear gene encoding NM_006428

mitochondrial protein

gil24308369 Nudix (nucleoside diphosphate linked moiety X) -type motif 16 NM_152395

(NUDT16)

Claims

claims

Use of marker sequences for the diagnosis of

Prostate cancer excluding

Prostate inflammatory diseases or diabetes or polymorbidity, wherein at least one marker sequence of a cDNA selected from the group SEQ 1 - 246 and / or SEQ 247-452 or a protein encoded thereby or in each case a partial sequence or fragment thereof is determined to or from a patient to be examined, and wherein the marker sequence (s) has been identified by a method comprising the c) identification of marker sequence candidates by differential screening with protein biochips from each of a cDNA expression library of a patient with prostate cancer and a subject without

Prostate cancer, d) validation of marker sequence candidates by means of

Samples from patients with

Use of marker sequences for the diagnosis of

Prostate cancer excluding

Prostate inflammatory diseases or diabetes or polymorbidity according to claim 1, characterized in that 2 or 3, preferably 4 or 5, especially

preferably 6, 7 or 8 or more different

Marker sequences, for example 10 to 20 or 30 or more different marker sequences are determined on or from a patient to be examined.

3. Use of marker sequences for the diagnosis of

Prostate cancer excluding Prostate inflammatory diseases or diabetes or polymorbidity according to one of the preceding claims wherein at least one of the marker sequences is selected from the group SEQ ID No. 247, SEQ ID No. 250, SEQ ID NO. 290 or a by SEQ ID No. 247, SEQ ID No. 250, SEQ ID No. 290 coded protein or one

Partial sequence or a fragment of SEQ ID No. 247, SEQ ID no. 250, SEQ ID No. 290, and / or

wherein at least one of the marker sequences is selected from the group SEQ ID No. 249, SEQ ID No. 255, SEQ ID no. 271, SEQ ID No. 301, SEQ ID No.341 or SEQ ID NO. 249, SEQ ID No. 255, SEQ ID No. 271, SEQ ID NO. 301, SEQ ID No.341 encoded protein or a partial sequence or a fragment of SEQ ID No. 249, SEQ ID No. 255, SEQ ID No. 271, SEQ ID No. 301, SEQ ID No.341.

Use of marker sequences for the diagnosis of

Prostate cancer excluding

Prostate inflammatory diseases or diabetes or polymorbidity, wherein at least one marker sequence of a protein selected from the group neuroncological ventral antigen 2 (NOVA2), Syntaxin 18 (STX18), heat shock (Hockock) 105kDa / 11 OkDa protein 1 (HSPH1) or a coding for it Nucleic acid or a partial sequence or a fragment thereof to or from a patient to be examined, and wherein the marker sequence (s) has been identified by a method comprising the steps of a) identifying marker sequence candidates by differential screening with protein biochips from each a cDNA expression library of a patient with prostate cancer and a subject without

Prostate cancer; b) validation of marker sequence candidates using samples from patients with Prostatectomy for inflammation (s) and / or samples from patients with diabetes.

Use of the marker sequence (s) for the diagnosis of

Prostate cancer excluding

Prostate inflammatory diseases or diabetes or polymorbidity according to one of the preceding claims, characterized in that the determination is carried out by means of in vitro diagnosis.

Use of a marker sequence of a cDNA in each case selected from the group SEQ 1 - 246 and / or SEQ ID 247-452 or in each case a protein encoded thereby or in each case a partial sequence or fragment thereof as

A diagnostic agent, wherein the marker sequence (s) has been identified by a method comprising the

Steps a) Identification of marker sequence candidates by differential screening with protein biochips from each of a cDNA expression library of a patient with prostate cancer and a subject without

Prostate inflammatory diseases and / or samples from patients with diabetes.

Use of the marker sequence (s) for the diagnosis of

Prostate cancer excluding

Prostate inflammatory diseases or diabetes or polymorbidity according to any one of the preceding claims, characterized in that the marker sequence (s) is (are) applied to a solid support,

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.

Procedure for the diagnosis of prostate cancer under

Exclusion of prostate inflammatory diseases or diabetes or polymorbidity, wherein

a. ) at least one marker sequence of a cDNA selected from the group SEQ 1 - 246 and / or SEQ ID 247-452 or in each case a protein encoded thereby or in each case a partial sequence or fragment thereof is (are) applied to a solid carrier and

b. ) with body fluid or tissue extract one

Patients are (and will) be contacted

c. ) the proof of an interaction of the

Body fluid or tissue extract with the (the) marker sequence (s) from a.) Is done.

Process for stratifying, in particular for

Risk stratification, or therapy control of a patient with prostate cancer excluding prostate inflammatory diseases or diabetes or polymorbidity, wherein at least one marker sequence of a cDNA selected from the group SEQ 1 - 246 and / or SEQ 247 - 452 or a protein encoded thereby or each a partial sequence or Fragment thereof to or from a patient to be examined.

The method of claim 9, wherein the stratifying or therapy control decisions for

Treatment and therapy of the patient, in particular hospitalization of the patient, use, effect and / or dosage of one or more drugs, a therapeutic measure or the monitoring of a disease course and therapy course, etiology or classification of a disease including prognosis. Assay, protein biochip consisting of an assembly containing at least one marker sequence of a cDNA selected from the group SEQ 1 - 246 and / or SEQ ID 247-452 or each protein encoded thereby or each of a partial sequence or fragment thereof, characterized in that

Marker sequence (s) are (are) mounted on a solid support and wherein the marker sequence (s) has been identified by a method (s) comprising the

Use of an assay according to claim 11 for

Identifying and characterizing a substance for prostate carcinoma comprising means for detecting a binding success, characterized in that an arrangement or assay is brought into contact with a.) At least one substance to be investigated and b.) A binding success is detected.

Use of an assay according to any one of claims 11 or 12 for the screening of drugs for

Prostate cancer inom.

Diagnostics for the diagnosis of prostate cancer excluding prostate inflammatory diseases or diabetes or polymorbidity containing at least one marker sequence each selected from the group SEQ 1 - 246 and / or SEQ ID No. 247-452 or a protein encoded thereby or in each case a partial sequence or fragment thereof, wherein the marker sequence (s) has been identified by a method comprising the

Target for treatment and therapy of

Prostate carcinoma, each selected from the group SEQ 1 - 246 and / or SEQ ID 247 - 452 or in each case a protein coding therefor or in each case a partial sequence or fragment thereof.

Use of a marker sequence of a cDNA selected from the group SEQ 1-246 and / or SEQ 247-452 or in each case a protein encoded thereby or in each case a partial sequence or fragment thereof as

Affinity material for performing apheresis or blood lavage on patients with prostate cancer.

Use of at least one marker sequence of a cDNA selected from the group SEQ 1 - 246 and / or SEQ

247-452 or a protein encoded thereby or each a partial sequence or fragment thereof for identifying a subgroup of patients within the group of patients with prostate cancer, wherein the patients of the subgroup none Prostatectomy and / or have no diabetes.