US20100129797A1 - Use of genetic modifications in human gene chk1 which codes for checkpoint kinase 1 - Google Patents

Use of genetic modifications in human gene chk1 which codes for checkpoint kinase 1 Download PDF

Info

Publication number
US20100129797A1
US20100129797A1 US12/312,474 US31247407A US2010129797A1 US 20100129797 A1 US20100129797 A1 US 20100129797A1 US 31247407 A US31247407 A US 31247407A US 2010129797 A1 US2010129797 A1 US 2010129797A1
Authority
US
United States
Prior art keywords
chk1
patients
genotype
gene
polymorphisms
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.)
Abandoned
Application number
US12/312,474
Other languages
English (en)
Inventor
Kathrin Riemann
Winfried Siffert
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.)
Universitaet Duisburg Essen
Original Assignee
Universitaet Duisburg Essen
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 Universitaet Duisburg Essen filed Critical Universitaet Duisburg Essen
Assigned to UNIVERSITAT DUISBURG-ESSEN reassignment UNIVERSITAT DUISBURG-ESSEN ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: RIEMANN, KATHRIN, SIFFERT, WINFRIED
Publication of US20100129797A1 publication Critical patent/US20100129797A1/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/112—Disease subtyping, staging or classification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/118—Prognosis of disease development
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/172—Haplotypes

Definitions

  • the invention relates to an in vitro method for predicting disease risks, progression of diseases, drug risks and for finding drug targets.
  • Cancer cells are characterized by loss of contact inhibition and uncontrolled cell growth. Such modifications are triggered spontaneously or by noxae, so-called cancero-genes, which damage the genetic make-up. Such noxae include many chemicals, tobacco smoke, but also UV light. Besides that, genetic factors play a prominent role in the formation of cancer. Characteristic for cancer cells, beside their uninhibited growth, is also the tendency to form metastases in other organs.
  • prognosis factors for the progression of cancers, which provide information about the response to certain forms of treatment or are generally predictive for the occurrence of metastases, tumor progression and survival.
  • prognosis factors generally known to the person skilled in the art are used in medicine. These include, for example, the size of the tumor, its penetration depth into the surrounding tissue, cross-organ growth, the penetration into blood or lymphatic vessels or into lymph nodes, as well as the degree of differentiation of the tumor cells. In addition, some relatively unspecific serological markers exist.
  • the cell cycle of eukaryotic cells is generally subdivided into four phases: the G1-phase, in which the preparation for replication takes place, the S-phase, in which the DNA is synthesized and the actual cell functions take place, the G2-phase, in which the preparation for mitosis takes place, and the M-phase, the mitosis ( FIG. 1 ).
  • G1-phase in which the preparation for replication takes place
  • S-phase in which the DNA is synthesized and the actual cell functions take place
  • the G2-phase in which the preparation for mitosis takes place
  • M-phase the mitosis
  • FIG. 1 So far, several cell cycle checkpoints were characterized. The best investigated checkpoints in mammals are shown in FIG. 1 .
  • the DNA damage checkpoint which can be activated by a damage of the DNA in different cell cycle phases. This damage can be caused by exogenous causes, like radiation, as well as by endogenous processes, e.g. spontaneous mutations.
  • the replication checkpoint is activated by an incomplete or defective replication of the DNA.
  • the spindle checkpoint monitors the formation of the bipolar spindle, the attachment of the kinetochores and the new formation of the centromere structures.
  • the most important task of a cell is to maintain genomic identity.
  • Checkpoint kinase 1 is involved in essential control mechanisms in the cell cycle, which ensure that the transfer of defects to the daughter cell is minimized.
  • the significant CHK1 reaction cascade at the G2/M checkpoint is shown in FIG. 2 .
  • the activation of CHK1 takes place due to DNA damages, which are mainly detected by the chromatin-bound Rad17 complex.
  • the Rad17 complex recruits the Rad9-Hus1-Rad1 complex, which together with the ATR-Atrip complex activates CHK1, which is partially present in a chromatin-associated form, by phosphorylation.
  • ATR Ataxia-telangiectasia- and Rad3-related
  • CHK1 phosphorylation by the protein Claspin
  • the activated CHK1 protein migrates from the cell nucleus into the cytoplasm, where in its turn it activates CDC25C (cell division cycle 25C) by phosphorylation.
  • This process enables the 14-3-3 protein (tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein) to bind to CHK1, so that it can return into the nucleus and remains there.
  • CDC2 cell division controller 2
  • cyclin B complex are inhibited, which inhibits the entrance into mitosis.
  • the DNA repair system can be initiated to eliminate the DNA damage (Jiang et al., Regulation of Chk1 includes chromatin association and 14-3-3 binding following phosphorylation on Ser345. 2003, J. Biol. Chem. 278:25207-17; Jeong et al., Phosphorylated claspin interacts with a phosphate-binding site in the kinase domain of Chk1 during ATR-mediated activation. 2003, J. Biol. Chem. 278:46782-8).
  • CHK1 involvement in a checkpoint in the S-phase could also be verified.
  • ATM Ataxia-telangiectasia mutated
  • Claspin additional activation by Claspin is required.
  • the completely activated CHK1 now activates DNA protein kinases, together with which they phosphorylate p53 and thus can increase its activity.
  • CHK1 is likewise able to phosphorylate TLK1 (tousled like kinase 1). This protein plays a decisive role in chromatin condensation, which, however, is inhibited by CHK1 to prevent progression in the cell cycle.
  • CHK1 phosphorylates CDC25A (cell division cycle 25A) and thus initiates its degradation.
  • the CDC protein is no longer able to activate the Cyclin complexes, due to which neither the S-phase can be advanced nor the M-phase started.
  • the invention is therefore based on the object to provide a means, which enables a better prognosis of the natural progression of a cancer and the response to any form of treatment.
  • this means is to be able to detect those patients, in which increased DNA repair mechanisms aggravate a cancer treatment.
  • the invention is further based on the object to provide a means to generally predict disease risks and progression of diseases, since DNA repair mechanisms are also very important for other diseases.
  • Such polymorphisms are, for example, (-1143)G>T, ( ⁇ 1400)C>T, (-1453)T>C, and (-1154)insC.
  • the human gene CHK1 is localized on chromosome 11q23 (Accession No. NM — 033899 of the Gene Bank of the National Center for Biotechnology Information (NCBI)) and codes for a 54 kD protein, which is expressed in the nucleus.
  • the gene has the designation “CHK1” as well as the designation “CHEK1”.
  • CHK1 shall be used here.
  • FIG. 3 A schematic representation of the gene structure is shown in FIG. 3 .
  • the active promoter region of CHK1 has already been characterized.
  • the promoter sequence contains numerous binding sites for the transcription factor E2F1, the binding of which enhances the transcriptional activity.
  • CHK1 A positive regulation of CHK1 is likewise observed by an isoform of the p53-dependent p73 (Carrassa et al., Characterization of the 5′-flanking region of the human Chk1 gene. 2003, Cell Cycle 2:604-9).
  • FIG. 1 shows a schematic representation of the cell cycle with the most important checkpoints.
  • FIG. 2 shows a graphical representation of the reaction cascade at the DNA damage checkpoint.
  • FIG. 3 shows the intron/exon structure of the human gene CHK1 (not to scale).
  • FIG. 4 shows the structural relationship of some CHK1 inhibitors with staurosporine.
  • FIG. 5 shows a schematic representation of the polymorphisms in the CHK1 gene (not to scale).
  • FIG. 7 shows putative binding sites for transcription factors in the promoter of the CHK1 gene; the numbers on the left-hand side represent the relation to the ATG.
  • FIG. 8 shows the result of the Electrophoretic Mobility Shift Assay (EMSA) with constructs containing the various alleles of the -1143G>T polymorphism of CHK1. Following the addition of cell nucleus extract, increased binding of core protein to the “G-construct” can be detected. The binding is specifically inhibited by the presence of a displacing oligonucleotide.
  • ESA Electrophoretic Mobility Shift Assay
  • FIG. 9 shows the result of the Electrophoretic Mobility Shift Assay (EMSA) with constructs containing the various alleles of the -1400C>T polymorphism of CHK1. Following the addition of cell nucleus extract, increased binding of core protein to the “T-construct” can be detected. The binding is specifically inhibited by the presence of a displacing oligonucleotide.
  • ESA Electrophoretic Mobility Shift Assay
  • FIG. 10 shows constructs for measuring the genotype-de-pendent regulatory activity of the promoter polymorphisms after 24 h following the transfection of HELA cells using secreted alkaline phosphatase (SEAP).
  • SEAP secreted alkaline phosphatase
  • FIG. 11 shows an expression of CHK1 mRNA in urinary bladder carcinoma tissue depending on the alleles of the promoter polymorphisms. Represented is the quotient CHK1/ ⁇ -actin mRNA.
  • FIG. 12 shows an expression of CHK1 mRNA in colorectal carcinoma tissue depending on the alleles of the promoter polymorphisms. Represented is the quotient CHK1/ ⁇ -actin mRNA.
  • FIG. 13 shows the Kaplan-Meier analysis on the survival of patients with urinary bladder carcinoma depending on the genotype of the -1143G>T polymorphism.
  • A all patients, B: only patients older than 54 years; *: p ⁇ 0.05, **: p ⁇ 0.01
  • FIG. 14 shows the Kaplan-Meier analysis on the survival of patients with colorectal carcinoma depending on the genotype of the promoter polymorphisms.
  • A Dependency on the -1143G>T SNP
  • B only patients with colon carcinoma depending on the -1400C>T SNP
  • C only patients with colon carcinoma depending on the -1153T>C SNP and -1154insC SNP;
  • FIG. 15 shows the Kaplan-Meier analysis on the survival of patients with chronic lymphatic leukemia depending on the genotype of the -1143G>T polymorphism.
  • FIG. 16 shows the Kaplan-Meier analysis of patients with melanoma depending on the genotype of the -1400C>T promoter polymorphism; A: Time from initial treatment to start of further treatment; B: Survival; *: p ⁇ 0.05.
  • FIG. 17 shows the Kaplan-Meier analysis on the survival of patients with cholangiocellular carcinoma depending on the genotype of the -1400C>T polymorphism; *: p ⁇ 0.05.
  • FIG. 18 shows a Kaplan-Meier analysis on the survival of patients with cholangiocellular carcinoma depending on the genotype of the -1400C>T polymorphism. *: p ⁇ 0.05.
  • FIG. 19 shows a Kaplan-Meier analysis on the survival of patients with mammary carcinoma depending on the genotype of the -1143G>T polymorphism. *: p ⁇ 0.05
  • CHK1 is considered a potential tumor suppressor gene, since a defect in the regulation of the cell cycle results in cumulation of defective DNA and an increased cellular proliferation rate, both of which are characteristics of tumor cells. So far, somatic mutations in this gene could be verified in some patients with sporadic tumors, e.g. stomach and mammary carcinoma as well as microsatellite-instable colorectal tumors showed alterations. Unlike single nucleotide polymorphisms (SNPs), these mutations are, for example, not found in peripheral blood cells in the respective patients. Disease-specific associations for SNPs have not been described yet.
  • checkpoints are involved in many regulatory cascades, they are a suitable target for cancer therapeutic agents. Certain characteristics of the checkpoint proteins contribute to that: (1) the complex signal transduction system of checkpoints offers a multitude of targets, (2) in healthy cells, some checkpoints seem to be relatively insignificant, which highly reduces the toxicity of the inhibitors, (3) the restoration of defective checkpoints could result in a slow-down of the cell growth, (4) as a signal transduction system, checkpoints are subject to adaption, which could be interrupted, and (5) the restoration of affected checkpoints could increase the apoptosis rate of cancer cells and thus increase their sensitivity towards certain substances (Hartwell et al., Cell cycle control and cancer. 1994, Science 266:1821-8).
  • CHK1 inhibitors are already known or have already been developed. Based on staurosporine, originally identified as protein kinase C inhibitor, which likewise is a potent CHK1 inhibitor, various substances were derived. These include, for example, the CHK1 inhibitors UCN-01 (7-hydroxy-staurosporine), Gö6976, SB-218078, ICP-1, XL844, and CEP-3891, which block the G2/M checkpoint ( FIG. 4 ). Furthermore, debromohymenialdisine and the synthetic peptide TAT-S216A can inhibit CHK1 as well as a further checkpoint protein, CHK2. It has to be assumed, that in the near future even far more substances will be available, which inhibit CHK1.
  • UCN-01 (7-hydroxy-staurosporine) is the clinically best characterized substance.
  • UCN-01 has already passed through several clinical phase I studies and is currently tested in phase II.
  • the inhibition or down regulation of CHK1 had a positive effect on the response of cytostatics, like topoisomerase inhibitors, antimitotics and antimetabolites, since due to the lack of CHK1, the toxicity of these chemotherapeutic agents was potentiated especially in aggressive tumors, which even following preceding conventional treatment showed progressive growth.
  • UCN-01 increases the sensitivity towards radiation, whereby CHK1 inhibitors also have potential as radio-sensitizers.
  • CHK1 Due to the fundamental significance of CHK1 for the maintenance of genomic integrity, such polymorphisms are suited to generally predict disease risks or progression of diseases in tumor diseases or to predict treatment responses/treatment failure or undesired side-effects for all pharmaceuticals or non-pharmacological treatments.
  • Coupling imbalance means the occurrence of allele combinations (haplotypes), which statistically clearly occur more frequently or less frequently together, than this was to be expected in relation to their frequency.
  • polymorphisms -1153T>C and -1154insC link up completely.
  • Polymorphisms -1143G>T and -1400C>T do not link up, and they only restrictedly couple with the two other variants ( FIGS. 6A and B).
  • the quality of the coupling is marked with the values D′ and r 2 .
  • CHK1 or neighboring genes which for example are in coupling imbalance with genotypes in the CHK1 gene.
  • genes which are likewise located on chromosome 11, but far away from the CHK1 gene. For that, the procedure is as follows:
  • all promoter SNPs mentioned above are located in a consensus sequence for binding sites of different transcription factors (e.g. E74A, CF2-II or bZIP910), the binding of which can be affected by the polymorphisms. The occurrence of certain genotypes results in an omission of these binding sites by the modification of their consensus sequences.
  • EMSA electrospray mobility shift assay
  • FIG. 8 shows the result of this test with specific constructs, which either contain the G- or the T-allele of the -1143G>T polymorphism.
  • the presence of the G-construct band proves binding of a transcription factor to this region.
  • the T-construct has no band, which shows that no transcription factor binds to this allele.
  • the weakening of the band intensity by a specific oligonucleotide shows, that the binding transcription factor is a specific binding.
  • FIG. 9 shows the result of this test with specific constructs, which either contain the C- or the T-allele of the -1400C>T polymorphism.
  • the constructs are cloned in front of a gene, which codes for secreted alkaline phosphatase (SEAP). If the construct has a gene-regulating activity, the transcription of the SEAP gene is increased and the increased secretion of alkaline phosphatase into the cell culture medium is measurable. As shown in FIG. 10
  • the reporter activity of the individual alleles of this SNP is likewise different.
  • the T-allele shows a higher activity (3.82 ⁇ 0.6) than the G-allele (2.36 ⁇ 0.3).
  • CHK1 Since only the -1143G>T polymorphism of the CHK1 gene shows a reporter activity, next it was investigated, how the regulation in vivo takes place, because reporter assays are an artificial cell system. For that, the expression of CHK1 at the mRNA level was investigated using real-time PCR in human tissue.
  • mRNA was obtained from human surgery tissue from urinary bladder and colon surgeries and transcribed into cDNA using reverse transcriptase.
  • the person skilled in the art is familiar with this method.
  • the expression level was determined using real-time PCR (Taqman method) and matched with the expression level of the housekeeping gene ⁇ -actin.
  • FIGS. 11 and 12 It is shown in section 11A, that the GG-genotype of the -1143G>T SNP has a significantly higher mRNA expression than the TT-genotype.
  • the values of the heterozygote genotype are located in-between, which indicates a gene dosage effect.
  • FIGS. 11B and 12B show an increased mRNA expression for the GG-genotype.
  • the two other polymorphisms also show an allele-dependent difference in the gene expression.
  • FIGS. 11B and 12B illustrate, C-allele carriers of the -1400C>T polymorphism have clearly more CHK1 mRNA than carriers of the TT-genotype.
  • the real-time PCR results for the SNPs -1153T>C and -1154insC are shown in FIGS. 11C and 12C .
  • Carriers of the T-allele which do not have an additional insertion, show a clearly lower mRNA expression.
  • this genotype-dependent gene expression of CHK1 can also affect the response to treatment with CHK1 inhibitors. It has to be expected that a low gene expression predisposed by a certain genotype, e.g. the TT-genotype of the -1143G>T polymorphism, responds stronger to CHK1 inhibitors than other genotypes.
  • genetic modifications in the CHK1 gene can be used to predict the response to a cancer treatment to discriminate, for example, responder versus non-responder. These genetic modifications can also be used for dosage finding or for predicting the occurrence of undesired drug effects, respectively.
  • Such cancer treatments can take place as drug treatments in the broadest sense, i.e. by supplying substances into the body, or these cancer therapeutic agents can have a physical effect (radiation, warmth, cold).
  • checkpoint kinase 1 Due to the key function of checkpoint kinase 1 for the regulation of the cell cycle, it is an essential component of the invention that using genetic modifications in CHK1, disease risks and progression of diseases can be generally predicted.
  • the multistep development of cancer reflects the accumulation of genetic modifications, which result in the transformation of normal cells into cancer cells and of normal tissue to benign and possibly malignant, invasive tumors.
  • the accumulation of alterations in tumor suppressor genes and proto-oncogenes accelerates tumorgenesis and can influence radio- as well as chemotherapy.
  • disturbed DNA repair mechanisms as well as checkpoints are the reason for the increased genomic instability of tumors (Hoeijmaker J. H., Genome maintenance mechanisms for preventing cancer. 2001 Nature, 411:366-74; Khanna et al., DNA double-strand breaks: signaling, repair and the cancer connection. 2001, Nat. Gent. 27:247-54).
  • checkpoints play a central role in the maintenance of genomic integrity, it has to be directly expected that the progression of varied and completely different tumor diseases with a genetically determined, reduced activatability is influenced by checkpoints. That means that with changes in the expression of proteins, which are expressed in all human body cells and protect the cell from DNA damage, cell functions are regulated, which decisively influence or at least modulate all physiological and pathophysiological processes. Besides that, responses to pharmaceuticals are also influenced in a particular manner. This affects desired, but also undesired drug effects.
  • checkpoint proteins have a sustained influence on varied diseases or on the progression of varied diseases, respectively, since these are phylogenetically highly conserved pathways.
  • Such genetic modifications can be structure-modifying mutations in the checkpoint proteins, which, for example, reduce the activation of the proteins by phosphorylation or the substrate selectivity.
  • the expression level can be modified, whereby the initiation of the subsequent reaction cascades, which e.g. induce apoptosis, is reduced, or splicing variants with a changed function can occur. All these modifications are considered a genetic predisposition for cancer.
  • CHK1 neoplasias of any tissue of origin and malignant neoplasias of any tissue of origin.
  • neoplasias comprise, for example, tumor diseases like tumors of the urogenital tract: urinary bladder carcinoma, kidney cell carcinoma, prostate carcinoma and seminoma;
  • tumors of the female genitals mammary carcinoma, corpus carcinoma, ovarian carcinoma, cervix carcinoma
  • tumors of the gastrointestinal tract oral cavity carcinoma, esophagus carcinoma, stomach carcinoma, liver carcinoma, bile duct carcinoma, pancreas carcinoma, colon carcinoma, rectum carcinoma
  • tumors of the respiratory tract larynx carcinoma, bronchial carcinoma
  • tumors of the skin malignant melanoma, basalioma, T-cell lymphoma
  • tumor diseases of the hematopoietic system Hodgkin and non-Hodgkin lymphomas, acute and chronic leukemias, plasmocytoma
  • tumor diseases of the brain or the nerve tissue respectively: glioblastoma, neuroblastoma, medulloblastoma, meningeal sarcoma, astrocytoma
  • soft tissue tumors for example sarcomas and head-neck tumors.
  • CHK1 Since the essential functions of CHK1 are known, genetic modifications in the CHK1 gene can increase the risk for tumor diseases or influence the progression of diseases. It is generally impossible to investigate all human tumor diseases and their progression. However, we have demonstrated this here by way of example for five different carcinomas: urinary bladder carcinoma, colorectal carcinoma, chronic lymphatic leukemia, malignant melanoma and cholangiocellular carcinoma. These data clearly prove the usability of genetic modifications in the CHK1 gene for the purpose described here. These diseases are a priori not associated at all.
  • CHK1 inhibition has only little influence on the cell cycle and the survival of the cell, as long as no DNA-damaging substances are present.
  • the G2 checkpoint is securely inhibited and the apoptosis increased. Since the discovery and development of the CHK1 inhibitors, it could be verified, that by using them, the effect of chemo- and radio-therapeutic measures could be increased.
  • CHK1 inhibitors If genetic modifications occur in CHK1, which influence the gene expression, then this has an impact on the effectiveness of these CHK1 inhibitors. It has to be expected, that patients with a genotype-dependent lower CHK1 expression respond better to the inhibitors than patients with a higher CHK1 expression. Additionally, it means that the combined treatment of CHK1 inhibitors with chemo- and immunotherapeutic agents and/or radiation can be influenced. From this results the possibility of individual diagnostics of the general responsiveness to these cancer therapeutic agents and therapy measures as well as an individual prediction of the risk of undesired effects by these therapies.
  • Chemotherapy uses such substances, which exert their damaging effect on certain cancer cells as targeted as possible and kill them or inhibit them in their growth.
  • a certain cytostatics dosage can always only kill a certain portion of the target cells, which remains the same with proceeding treatment. Therefore, chemotherapy must not be reduced within the course of the treatment, even if the tumor is not even detectable anymore. It rather has to be assumed, that with a weak treatment, especially the resistant tumor cell clones are selected.
  • Chemotherapy is applied in fast succession, and almost always two or more cytostatics are combined to increase effectiveness.
  • the therapy thus also causes side-effects, which are classified according to the common toxicity criteria. These criteria include: number of leukocytes and thrombocytes, sickness, vomiting, diarrhea and stomatitis.
  • Radiotherapy means the use of ionizing high-energy radiation to heal malignant tumor diseases. Such malignant tumors are often also treated in combination with chemo- and radiotherapy. A multitude of tumor diseases can thus also be healed in advanced stages. In order to keep the side-effects low, the radiation is divided into many daily single doses and administered over several weeks. Still, side-effects like redness, sickness, diarrhea, or hair loss occur, depending on the dosage, penetration depth and number of single doses.
  • the invention is now based on the fact that a method has been developed, which is generally suited for diagnostics of the activatability of checkpoint kinase 1 and, associated with it, the G2 checkpoint. For that, one or more polymorphisms in the CHK1 gene are investigated. With high expression, there predictably is an increased activatability of the G2 checkpoint and thus sufficient time to perform repair mechanisms in the DNA after damaging of the same. With low CHK1 expression, the G2 checkpoint is less activatable and DNA damage is not or not sufficiently repaired.
  • lymphocytes of healthy subjects were cultivated and stimulated for cleavage. After 72 hours, these cells were radiated with a dose of 1 Gy and subsequently arrested in the M-phase by the mitosis inhibitor colchicine. From these cells, chromosomes were prepared using methods the person skilled in the art is familiar with and, depending on the -1143G>T polymorphism, evaluated for damage by radiation in 50 metaphases each. In this manner, only those cells were included into the evaluation, which at the time of radiation were in the G2-phase and until the chromosome preparation reached the M-phase.
  • the activity of the G2/M checkpoint can be assessed.
  • the G2/M checkpoint is the most active and the respective DNA repair mechanisms could work best.
  • the weak checkpoint for the TT-genotype allows only few repair mechanisms to work, and more damage can accumulate.
  • a determination of the presence of polymorphisms in CHK1 allows for diagnostics of the effectiveness and undesired effects of drugs, in particular cytostatics, as well as other forms of treatment, which damage the genetic make-up of the tumor cells, e.g. radiation.
  • polymorphisms in CHK1 can be used to diagnose the effects of pharmaceuticals used in combination with a CHK1 inhibitor.
  • the diagnostics of the allele or haplotype status in CHK1 can be used to determine the individually optimal and tolerated dosage of drugs.
  • For diagnostics of an increased or reduced activatability of checkpoint kinase and the G2 checkpoint serves in particular the verification of the CHK1 promoter polymorphisms described here, either alone or in any perceivable combinations.
  • any further genetic modifications in CHK1 can be used for diagnostics, which are in a coupling imbalance to these polymorphisms and/or additionally promote or inhibit the alternative splicing process or the expression.
  • the method stated is particularly suited for diagnostics of the effect of substances, which damage the DNA of the tumor cells.
  • substances include oxaliplatin, 5-fluorouracil, folinic acid, irinotecan, capecitabine and cisplatin, whereat the list could be randomly extended.
  • immunotherapeutic agents e.g. interferons or interleukins
  • inhibitors of signal transduction in tumor cells, respectively can be predicted.
  • radiotherapeutic measures like gamma radiation, X-ray radiation, electrons, neutrons, protons and carbon ions, whereat the list could be randomly extended.
  • radiation therapy also implies the medical application of microwaves and heat rays, light and UV therapy as well as the treatment with ultrasound radiation.
  • a substantial subject of the invention is the provision of diagnostically relevant genetic modifications in the CHK1 gene as prognosis factor for all human tumor diseases. Naturally, not all tumor diseases can be described in that. The principle will therefore be further explained in selected examples, which demonstrate general usability without restricting the scope of patent to the exemplary embodiments.
  • Bladder cancer is a malignant tumor of the mucous membrane of the urinary bladder and most frequently occurs between the age of 60 and 70. Men are affected by it three times as often as women. In men, bladder cancer is the third most frequent type of cancer after lung and prostate cancer. Bladder cancer can be caused by external influences. The risk factors include smoking, permanent strain on the organism by chemicals, like for example colorants or analgesic misuse. In many patients, the examinations show that it is a superficial tumor. This can be removed by surgery using a cystoscope. More than 70% of the patients treated because of a superficial bladder carcinoma show tumor rescrudescence in progression. In that, in more than half the patients, rescrudescence tumors with non-muscle-invasive disease occur.
  • FIG. 14A shows the survival depending on the -1143G>T SNP.
  • the median time until death is only 48 months for carriers of the TT-genotype, while for G-allele carriers, no median time can be stated, since up to the end of the study, less than half of these patients died.
  • the median time of survival is 87 months (GG-genotype) or 50 months (GT-genotype), respectively, in patients with the G-allele; for the homozygous T-allele carriers, on the other hand, only 33 months.
  • the colorectal carcinoma is the most frequent type of tumor in the gastrointestinal tract and one of the main causes for tumor-related death worldwide (12-15% of the total cancer mortality). In Germany, the incidence is about 51,000 new cases per year. The average 5-year survival rate after tumor resection is only approx. 50%. Eating habits, cancer-promoting metabolites, exogenous carcinogens and certain predisposing diseases are included in the risk factors for the formation of a colorectal carcinoma.
  • the standard method for predicting the progression of the disease is the TNM or UICC stage system, respectively. Patients with UICC stages III or IV generally have a worse prognosis than patients with UICC stages I or II.
  • An adjuvant chemotherapy is performed for metastasized colorectal carcinomas (UICC stages III and IV) and can enhance the local effect of radiation therapy.
  • a majority of these patients develops recrudescences or metastases, which makes intensive aftercare necessary.
  • a further component of the invention consists in using genetic modifications in CHK1 to predict the further progression of the colorectal carcinoma.
  • the median survival for the TT-genotype is 70 months, it is only 34 months for the CT-genotype and only 26 months for the CC-genotype.
  • a significant difference in regards to survival can like-wise be detected ( FIG. 15C ).
  • Patients, who have the TT-genotype at position -1153 and have no insertion at position -1154, survive longer than patients, who have the -1154C allele and the insertion (p 0.007).
  • the median survival is 70 months for the TT-genotype without insertion compared to 21 and 15 months for the other genotypes.
  • Chronic lymphatic leukemia is a chronic form of leukemia. Characteristic for the disease is a high number of abnormal lymphocytes. A total of 30 percent of all leukemic diseases are chronic lymphatic leukemias. The median disease age is 65 years. A CLL can be benign for up to 20 years, i.e. the patients show no symptoms except for enlarged lymph nodes, tiredness and lack of appetite. The treatment only starts, if the number of lymphocytes highly increases, the portion of erythrocytes and thrombocytes decreases, or other complications occur. An early treatment has no influence on the progression of the disease. The most important therapeutic measure is chemotherapy. The further the disease has progressed, the higher are the disturbances of health by the modification of the organ system.
  • the stage of a CLL is, among others, characterized by how many lymphocytes are present in blood and bone marrow, how large spleen and liver are and whether an anemia is present.
  • a CLL results in modifications in the immune system, so that humans suffering from this disease have a higher risk of developing other types of cancer.
  • patients show a completely different progression of the disease. It is an object of the invention to demonstrate that genetic modifications in the CHK1 gene are suited to predict the progression of CLL.
  • FIG. 16 shows the survival depending on the -1143G>T genotype. Patients, who are carriers of the T-allele, survive longer than patients, who are homozygous GG. For the GG-genotype, the median survival is 146 months; on the other hand, however, less than half the patients carrying the T-allele died during the observation period. Here, too, it shows that under intensive treatment, a genotype, which results in a low CHK1 expression, is most favorable for survival.
  • malignant melanoma is a malignant abnormality of the melanocytes (pigment cells of the skin), which is why it is also called “black skin cancer”. This type of tumor tends to spread metastases via the blood and lymph streams very early. The incidence of malignant melanoma is increasing, it doubles every 15 years. Particularly at risk are persons with low pigmentation. The risk factors include, above all, intensive sun exposure and a sunburn anamnesis of 5 or more episodes in the youth. In the Western world, malignant melanoma is the most frequent cancer in women between the age of 20 and 40.
  • Criteria for prognosis and therapy are provided by the stages of the TNM classification, the tumor thickness according to Breslow, the penetration depth according to Clark, the differentiation by subtypes and localization. With early diagnosis and treatment, the prognosis still is good. With late diagnosed melanomas with lymph node metastases, the 5-year survival rate lies at approx. 30%; if remote metastases are present already, it is only 0-5%. There are no molecular markers for the progression of this disease and responses to treatment. The identification of such markers would highly improve the pre- and aftercare of the patients.
  • FIG. 17A shows the genotype-dependent difference of the -1400C>T SNP from the time of initial diagnosis and initial treatment up to the time of requirement of further treatment.
  • C-allele carrier For the heterozygous or homozygous, respectively, C-allele carrier, treatment becomes necessary in median after 71 or 57 months, respectively, and for the TT-genotype after months already. Contrary to the curve progressions discussed so far, here it shows that without medical measures, those genotypes are advantaged, which have a high CHK1 expression and thus very well working checkpoints.
  • FIG. 17B represents the survival of all patients.
  • the treatment of the primary tumor is a completely surgical one. After the excision of the tumor, further treatment is considered. In 76.5% of the patients, there was no further treatment, 11.2% received an immune therapy with interferon- ⁇ , 3.4% a hyper-thermal extremities perfusion, in 5.4%, a re-resection was necessary and the remaining 3.5% were subjected to other therapeutic measures.
  • the cholangiocellular carcinoma is a malignant tumor of the bile ducts of the liver. Compared to Asia and Africa, where it represents the most frequent type of tumor with 20-30% of the malignant tumors, it is relatively seldom in Central Europe ( ⁇ 1% of all malignant diseases).
  • the risk factors include colitis ulcerosa, chronic bile duct inflammations and viral hepatitides.
  • the curative treatment of a CCC is a partial liver resection or a total hepatectomy with liver transplantation. The recrudescence rate is very high. The prognosis for CCC is therefore very unfavorable, in particular in patients with non-resectable tumors, who have a 5-year survival rate of ⁇ 10%.
  • the median survival period in patients with non-resectable CCC is 6 to 10 months. So far, no molecular markers are known for the progression of this disease, however, would substantially improve the care for patients with CCC. It is therefore a component of the invention to demonstrate that the use of genetic modifications in CHK1 is suited to predict the further progression of the disease.
  • FIG. 18 shows the survival of CCC patients.
  • the median survival of the TT-genotype is 5 months, that of the other genotypes 9 months. Since the probability of survival of the CCC patients is only very low, the time for an intensive treatment after the surgery is very short. Due to the mostly pre-damaged cirrhotic liver, the indication for radiotherapy is given only rarely; additionally, the liver is extremely radiosensitive. Thus, until achieving a tumoricidal dose, the liver might already be destroyed. So far, systemic chemotherapy with numerous cytostatics and combinations has not shown lasting effectiveness.
  • the remission rates are 20 to 30%, the median duration of remission is 4 to 6 months.
  • chemoembolizations or targeted intratumoral alcohol injections can be applied. Since for CCC neither radio- nor chemotherapy can be used to a sufficient extent, which for a low CHK1 level would show advantages, for this type of tumor, it is the genotypes showing an increased CHK1 expression, which show longer survival.
  • the mammary carcinoma is the most frequent tumor of the female population in Europe and the USA. It affects 7-10% of all women and accounts for 25% of the total female cancer mortality.
  • the etiology of the mammary carcinoma is still unknown, however, risk factors have been described, like a family disposition, radiation exposure or estrogen influence. In most patients, the examinations show that it is an invasive carcinoma. With a few exceptions, any operable mammary carcinoma even with verified remote metastatization is treated surgically.
  • the differently radical initial surgical treatment results in variations of the locoregional recrudescence rate, but not the long-term chance for survival. Furthermore, recrudescences or remote metastases quite often can become manifest after 5 or even 10 years only. It is therefore important to detect these lesions early and to closely monitor the patients in aftercare.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Engineering & Computer Science (AREA)
  • Analytical Chemistry (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Hospice & Palliative Care (AREA)
  • Oncology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
US12/312,474 2006-11-17 2007-11-19 Use of genetic modifications in human gene chk1 which codes for checkpoint kinase 1 Abandoned US20100129797A1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE102006054292.4 2006-11-17
DE102006054292A DE102006054292A1 (de) 2006-11-17 2006-11-17 Verwendung von Genveränderungen im menschlichen Gen CHK1, das für die Checkpointkinase 1 kodiert
PCT/EP2007/062519 WO2008059069A2 (de) 2006-11-17 2007-11-19 Verwendung von genveränderungen im menschlichen gen chk1, das für die checkpointkinase 1 kodiert

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2007/062519 A-371-Of-International WO2008059069A2 (de) 2006-11-17 2007-11-19 Verwendung von genveränderungen im menschlichen gen chk1, das für die checkpointkinase 1 kodiert

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US13/666,606 Continuation US9074259B2 (en) 2006-11-17 2012-11-01 Use of genetic modifications in human gene CHK1 which codes for checkpoint kinase 1

Publications (1)

Publication Number Publication Date
US20100129797A1 true US20100129797A1 (en) 2010-05-27

Family

ID=39402042

Family Applications (3)

Application Number Title Priority Date Filing Date
US12/312,474 Abandoned US20100129797A1 (en) 2006-11-17 2007-11-19 Use of genetic modifications in human gene chk1 which codes for checkpoint kinase 1
US13/666,606 Active 2028-08-28 US9074259B2 (en) 2006-11-17 2012-11-01 Use of genetic modifications in human gene CHK1 which codes for checkpoint kinase 1
US14/718,755 Abandoned US20150337389A1 (en) 2006-11-17 2015-05-21 Use Of Genetic Modifications In Human Gene CHK1 Which Codes For Checkpoint Kinase 1

Family Applications After (2)

Application Number Title Priority Date Filing Date
US13/666,606 Active 2028-08-28 US9074259B2 (en) 2006-11-17 2012-11-01 Use of genetic modifications in human gene CHK1 which codes for checkpoint kinase 1
US14/718,755 Abandoned US20150337389A1 (en) 2006-11-17 2015-05-21 Use Of Genetic Modifications In Human Gene CHK1 Which Codes For Checkpoint Kinase 1

Country Status (5)

Country Link
US (3) US20100129797A1 (de)
EP (1) EP2094864B1 (de)
AT (1) ATE550442T1 (de)
DE (1) DE102006054292A1 (de)
WO (1) WO2008059069A2 (de)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20120171671A1 (en) * 2009-03-06 2012-07-05 Universitaet Duisburg-Essen Chk2 polymorphism as a cancer marker
CN103374606A (zh) * 2012-04-12 2013-10-30 广州益善生物技术有限公司 Chek1基因突变检测特异性引物和液相芯片

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030087847A1 (en) * 2000-02-03 2003-05-08 Onyx Pharmaceuticals, Inc. Method and reagent for the inhibition of checkpoint kinase-1 (Chk1) enzyme
US20060147936A1 (en) * 2003-02-19 2006-07-06 Ulrich Frey Use of a gene mutation in the human gnas gene for predicting risks of diseases, courses of the disease and for predicting the response to disease therapies
US20080020385A1 (en) * 2004-05-26 2008-01-24 Ulrich Frey Use Of A Genetic Modification In The Human Gnaq Gene For Predicting Risk Of Disease, The Course Of Disease, And Reaction To Treatments

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB9718952D0 (en) * 1997-09-05 1997-11-12 Medical Res Council Mammalian chk1 effector cell cycle checkpoint protein kinase materials and methods

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030087847A1 (en) * 2000-02-03 2003-05-08 Onyx Pharmaceuticals, Inc. Method and reagent for the inhibition of checkpoint kinase-1 (Chk1) enzyme
US20060147936A1 (en) * 2003-02-19 2006-07-06 Ulrich Frey Use of a gene mutation in the human gnas gene for predicting risks of diseases, courses of the disease and for predicting the response to disease therapies
US20080020385A1 (en) * 2004-05-26 2008-01-24 Ulrich Frey Use Of A Genetic Modification In The Human Gnaq Gene For Predicting Risk Of Disease, The Course Of Disease, And Reaction To Treatments

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
Carrassa et al. (Cell Cycle 2:6, 604-609; November/December 2003) *
Hegele. Arterioscler Thromb Vasc Biol 2002;22;1058-1061 *
Riemann et al. (The FASEB Journal, 2007:21:567.5; published April 2007) *
Zill et al.Molecular Psychiatry (2004) 9, 1030-1036 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20120171671A1 (en) * 2009-03-06 2012-07-05 Universitaet Duisburg-Essen Chk2 polymorphism as a cancer marker
US9255297B2 (en) * 2009-03-06 2016-02-09 Universitaet Duisburg-Essen CHK2 polymorphism as a cancer marker
CN103374606A (zh) * 2012-04-12 2013-10-30 广州益善生物技术有限公司 Chek1基因突变检测特异性引物和液相芯片

Also Published As

Publication number Publication date
EP2094864B1 (de) 2012-03-21
US20130189676A1 (en) 2013-07-25
DE102006054292A1 (de) 2009-01-08
ATE550442T1 (de) 2012-04-15
US20150337389A1 (en) 2015-11-26
US9074259B2 (en) 2015-07-07
WO2008059069A3 (de) 2008-09-12
WO2008059069A2 (de) 2008-05-22
EP2094864A2 (de) 2009-09-02

Similar Documents

Publication Publication Date Title
Kim et al. Frequent amplification of CENPF, GMNN and CDK13 genes in hepatocellular carcinomas
Ratner et al. A KRAS-variant in ovarian cancer acts as a genetic marker of cancer risk
Nord et al. Focal amplifications are associated with high grade and recurrences in stage Ta bladder carcinoma
Schiewer et al. DNA damage response in prostate cancer
Mosrati et al. Association between TERT promoter polymorphisms and acute myeloid leukemia risk and prognosis
Cipollini et al. MicroRNA binding site polymorphisms as biomarkers in cancer management and research
Lei et al. Identification of MSRA gene on chromosome 8p as a candidate metastasis suppressor for human hepatitis B virus-positive hepatocellular carcinoma
Wan et al. Telomere length in circulating serum DNA as a novel non‐invasive biomarker for cirrhosis: a nested case–control analysis
Tang et al. ARHGEF10L contributes to liver tumorigenesis through RhoA-ROCK1 signaling and the epithelial-mesenchymal transition
Alhadheq et al. The effect of poly (ADP-Ribose) polymerase-1 Gene 3′ Untranslated region polymorphism in colorectal cancer risk among Saudi cohort
Heidari et al. The effect of TP53 and P21 gene polymorphisms on papillary thyroid carcinoma susceptibility and clinical/pathological features
Sugawara et al. Association of germline or somatic TP53 missense mutation with oncogene amplification in tumors developed in patients with Li‐Fraumeni or Li‐Fraumeni‐like syndrome
US20150337389A1 (en) Use Of Genetic Modifications In Human Gene CHK1 Which Codes For Checkpoint Kinase 1
Huszno et al. A comparison between CHEK2* 1100delC/I157T mutation carrier and noncarrier breast cancer patients: A clinicopathological analysis
Shinoura et al. Identification and partial sequence of a cDNA that is differentially expressed in human brain tumors
Wang et al. A functional polymorphism C‐1310G in the promoter region of Ku70/XRCC6 is associated with risk of renal cell carcinoma
Yeh et al. An intron 4 VNTR polymorphism of the endothelial nitric oxide synthase gene is associated with early‐onset colorectal cancer
Hur et al. Gene expression profiling of human gastrointestinal stromal tumors according to its malignant potential
Li et al. Association of the hypoxia inducible factor-1α gene polymorphisms with gastric cancer in Tibetans
Schick et al. Comprehensive genomic analysis identifies MDM2 and AURKA as novel amplified genes in juvenile angiofibromas
Cumova et al. Genetic variations in 3′ UTRs of SMUG1 and NEIL2 genes modulate breast cancer risk, survival and therapy response
Lyng et al. Response of malignant B lymphocytes to ionizing radiation: gene expression and genotype
Mehra et al. Comprehensive analysis of ETS family members in melanoma by fluorescence in situ hybridization reveals recurrent ETV1 amplification
Chae et al. PPP1R13L variant associated with prognosis for patients with rectal cancer
JP5688379B2 (ja) 癌マーカーとしてのｃｈｋ２多型

Legal Events

Date Code Title Description
AS Assignment

Owner name: UNIVERSITAT DUISBURG-ESSEN, GERMANY

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:RIEMANN, KATHRIN;SIFFERT, WINFRIED;SIGNING DATES FROM 20100116 TO 20100118;REEL/FRAME:023863/0639

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION