[go: up one dir, main page]

WO2011078495A2 - Procédé de prédiction chimiosensible pour un polymorphisme mononucléotidique (snp) - Google Patents

Procédé de prédiction chimiosensible pour un polymorphisme mononucléotidique (snp) Download PDF

Info

Publication number
WO2011078495A2
WO2011078495A2 PCT/KR2010/008640 KR2010008640W WO2011078495A2 WO 2011078495 A2 WO2011078495 A2 WO 2011078495A2 KR 2010008640 W KR2010008640 W KR 2010008640W WO 2011078495 A2 WO2011078495 A2 WO 2011078495A2
Authority
WO
WIPO (PCT)
Prior art keywords
snp
seq
nucleotide
base
anticancer
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.)
Ceased
Application number
PCT/KR2010/008640
Other languages
English (en)
Korean (ko)
Other versions
WO2011078495A3 (fr
Inventor
김진천
조동형
김용성
김선영
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.)
Asan Foundation
Original Assignee
Asan Foundation
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 Asan Foundation filed Critical Asan Foundation
Publication of WO2011078495A2 publication Critical patent/WO2011078495A2/fr
Publication of WO2011078495A3 publication Critical patent/WO2011078495A3/fr
Anticipated expiration legal-status Critical
Ceased 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
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
    • 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
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
    • C12Q1/6837—Enzymatic or biochemical coupling of nucleic acids to a solid phase using probe arrays or probe chips
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/15—Medicinal preparations ; Physical properties thereof, e.g. dissolubility
    • 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
    • C12Q2600/156—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 present invention relates to a method for screening SNPs for anticancer susceptibility prediction, an SNP for predicting susceptibility to anticancer drugs, and a method for predicting susceptibility to specific anticancer drugs using the same.
  • SNPs monobasic polymorphisms
  • the present inventors first confirmed tumor drug responsiveness through in vitro tumor reactivity analysis, and selected and analyzed the results from lymphocyte DNA of the same subject. Through the analysis of the most useful anticancer drug susceptibility prediction SNP genotype was discovered and the present invention was completed.
  • the present invention provides a method for screening anti-cancer drug susceptibility prediction SNP.
  • Another object of the present invention to provide an anticancer drug susceptibility prediction SNP marker.
  • Still another object of the present invention is to provide a kit for predicting anticancer drug sensitivity.
  • Still another object of the present invention is to provide a method for predicting susceptibility to anticancer drugs using the SNP marker.
  • One aspect of the invention relates to a method for screening SNPs for anticancer drug susceptibility prediction.
  • the nominal P-value of the selected genotype is 0.1% or less, the frequency of allele in the Asian genotype, genotype located in the linkage disequilibrium block, htSNP (haplotype tagging SNP), functional SNP and normal Selecting the SNP genotype in consideration of the Hardy-Wineberg equilibrium P value in the population;
  • It relates to a method for screening anticancer drug susceptibility prediction SNP comprising a.
  • step (1) of the above method can be carried out by the method described in the application number 10-2008-0115296.
  • In vitro tumor reactivity test
  • the linkage disequilibrium block (LD block) region is a region that is short enough such that a section on a particular genome is passed over generation and cross-over rarely occurs, or is a result of dense genes. Thus, the genetic information in this section is nearly identical and preserved over generations.
  • htSNP haplotype tagging SNP refers to a minimum set of SNPs that can distinguish a specific haplotype among all haplotypes, and represents a representative SNP. Therefore, LD block SNP and htSNP can reduce the time and expense by eliminating the effort of repeatedly verifying similar SNPs.
  • Hardy-Weinberg equilibrium refers to a genomic equilibrium state when randomly combined in a particular population and is generally expressed as a P value. When this value is less than 0.01, it is out of equilibrium. False positive SNPs resulting from these results can be excluded because they indicate errors in analysis or population selection.
  • Table 1 regimens gene SNP ID Allele type result Reference how FL GPC5 rs553717 G A A155V Capecitabine AJAP1 rs242056 G A G263R Capecitabine ERCC4 rs4309380 C T upstream FOLFIRI TNFRSF11B rs2073618 C G N3K FOLFOX SULT1C2 rs17036104 T G S255A FOLFOX SULT1C4 rs7580171 C T upstream FOLFOX EPHA7 rs2278106 + G A R278C FOLFOX EPHA7 rs2278107 T C I138V FL, FOLFIRI, FOLFOX SSTR4 rs2567608 A G F321S SAHA OR5AC2 rs4518168 G A M200I SAHA OR5H1 rs6775533 T C upstream PXD101 DPYD rs1801265 C T R29C
  • the screening method further comprises the step of associating the SNP genotype selected in step (2) with an existing clinical course.
  • Anticancer agents can be used to validate clinical relevance in the use of anticancer agents whose treatment has been confirmed.
  • the clinical course tracking of the patient was based on the NCCN surveillance guideline (www.nccn, org), and the tumor responsiveness of the anticancer agent was determined by RECIST criteria (Therasse et al., J Natl Cancer Inst 2000; 92). : 205-16).
  • the average follow-up period was 40 months (range 4-108 months), which was judged to be sufficient for judging results. This step eliminates the difference of the in vitro tumor reactivity test with the metabolic environment in the body and allows direct entry into the clinical trial.
  • the screening method preferably further comprises a cell biological verification.
  • a cell biological verification includes gene transformation, cell survival and cytotoxicity assays, apoptosis by caspase-3 western blot and flow cytometry.
  • the cytobiological validation comprises the steps of transforming tumor cells with a wild type gene or a mutant gene; Treating the tumor cells with a specific anticancer agent; It may include the step of measuring the cell viability of the tumor cells.
  • Another aspect of the invention relates to an anticancer drug susceptibility prediction SNP.
  • the SNP of the present invention is as follows.
  • SNPs for predicting susceptibility to the anticancer agent FL are in the rs553717 (SNP id) polynucleotide (SEQ ID NOs: 1 and 13) that constitute a part of the GPC5 (glypican5) gene, 401th Polynucleotides comprising a base and consisting of 8 or more consecutive bases or complementary polynucleotides thereof. Those with alternative alleles are more responsive to anticancer FL than those with reference alleles.
  • the leucovorin is an adjuvant drug that enhances the efficacy of the anticancer drug 5-FU.
  • SNPs for predicting susceptibility to the anticancer agent capecitabine include the 401th base in the rs242056 polynucleotide (SEQ ID NO: 2, No. 1 chromosome) constituting part of the AJAP1 (adherens junctions associated protein 1) gene. Polynucleotides consisting of two or more consecutive bases or complementary polynucleotides thereof. Those with alternative alleles have a higher responsiveness to the anticancer agent capecitabine than with reference alleles.
  • SNP for predicting susceptibility to the anticancer agent capecitabine is the 201st in the rs4309380 polynucleotide (SEQ ID NOs: 3 and 16), which forms part of the ERCC4 gene (excision repair cross-complementing rodent repair deficiency, complementation group 4). At least one of polynucleotides or their complementary polynucleotides comprising a base and consisting of eight or more consecutive bases. Those with alternative alleles have a higher responsiveness to the anticancer agent capecitabine than with reference alleles.
  • Anticancer drug FOLFIRI (5-FU + leucovorin + irinotecan): A combination of anticancer drug 5-FU and irinotecan, and leukoborin is a synergistic agent for 5-FU. 301st for rs2073618 polynucleotide (SEQ ID NOs: 4, 8) that forms part of the necrosis factor receptor superfamily member 11B precursor) gene Polynucleotides comprising a base and consisting of 8 or more consecutive bases or complementary polynucleotides thereof. Those with alternative alleles are more responsive to the anticancer agent FOLFIRI than those with reference alleles.
  • Anticancer drug FOLFOX (5-FU + leucovorin + oxaliplatin): A combination of 5-FU and oxaliplatin, and leukoborin is a synergistic agent for 5-FU. 301st for the rs17036104 polynucleotide (SEQ ID NO: 5, 2) which constitutes a part of Polynucleotides comprising a base and consisting of 8 or more consecutive bases or complementary polynucleotides thereof. Those with alternative alleles are more responsive to the anticancer agent FOLFIRI than those with reference alleles.
  • Another susceptibility predictive SNP for the anticancer agent FOLFOX is a polynucleotide comprising the 201 base and consisting of 8 or more consecutive bases in the rs7580171 polynucleotide (SEQ ID NO: 6, 2 chromosome) constituting part of the SULT1C4 gene or Complementary polynucleotides thereof.
  • SEQ ID NO: 6, 2 chromosome a polynucleotide comprising the 201 base and consisting of 8 or more consecutive bases in the rs7580171 polynucleotide (SEQ ID NO: 6, 2 chromosome) constituting part of the SULT1C4 gene or Complementary polynucleotides thereof.
  • Those with alternative alleles are more responsive to the anticancer agent FOLFIRI than those with reference alleles.
  • Another susceptibility predictive SNP for the anticancer agent FOLFOX is the 301th in the rs2278107 (SEQ ID NO: 7, Chromosome No. 7) constituting part of the EPHA7 (ephrin type-A receptor 7 precursor) gene.
  • Polynucleotides comprising a base and consisting of 8 or more consecutive bases or complementary polynucleotides thereof. Those with alternative alleles are more responsive to the anticancer agent FOLFIRI than those with reference alleles.
  • the SNP for predicting susceptibility to at least one of the anticancer agent FL, FOLFIRI or FOLFOX is the 401st of the rs2567608 polynucleotide (SEQ ID NOs: 8 and 20) that constitutes a part of the somatostatin receptor type 4 (SSTR4) gene.
  • Polynucleotides comprising a base and consisting of 8 or more consecutive bases or complementary polynucleotides thereof. Those with alternative alleles are more responsive to at least one of the anticancer agents FL, FOLFIRI or FOLFOX compared to those with the reference allele.
  • SNPs for predicting susceptibility to the anticancer drug SAHA include the 401th base and the 8th base of the rs4518168 polynucleotide (SEQ ID NOs: 9 and 3) that constitute a part of the olfactory receptor 5AC2 (OR5AC2) gene.
  • SNPs for predicting susceptibility to the anticancer drug SAHA include the 251 st base for the rs6775533 polynucleotide (SEQ ID NO: 10, chromosome 3) that forms part of the OR5H1 (olfactory receptor 5H1) gene. At least one of a polynucleotide consisting of two or more consecutive bases or a complementary polynucleotide thereof. Those with alternative alleles are more responsive to anticancer drug SAHA than those with reference alleles.
  • the SNP for predicting susceptibility to the anticancer agent PXD101 is the 401th of rs1801265 polynucleotide (SEQ ID NO: 11, No. 1) which forms part of the DPYD (Dihydropyrimidine dehydrogenase [NADP +] precursor) gene.
  • DPYD Dihydropyrimidine dehydrogenase [NADP +] precursor
  • Polynucleotides comprising a base and consisting of 8 or more consecutive bases or complementary polynucleotides thereof. Those with alternative alleles are more responsive to anticancer agent PXD101 than those with reference alleles.
  • SNP stands for single nucleotide polymorphism, where rs is an abbreviation for the reference sequence, and the number after it is an accession number that distinguishes each single nucleotide polymorphism provided by the database dbSNP.
  • the present invention is the 401 nucleotide of SEQ ID NO: 1,2,8,9,11, the 201 nucleotide of SEQ ID NO: 3,6, the 301 nucleotide of SEQ ID NO: 4,5,7 and the 251 nucleotide of SEQ ID NO: 10
  • one or more anticancer drug sensitivity predictive polynucleotides selected from polynucleotides consisting of 8 or more consecutive bases comprising a complementary polynucleotide thereof.
  • the eight or more consecutive bases are preferably 8 to 100 consecutive bases.
  • susceptibility prediction for an anticancer agent means the prediction of the therapeutic effect when the patient is treated with the anticancer agent. If the sensitivity to the anticancer agent is high, excellent therapeutic effect can be expected, while if the sensitivity to the anticancer agent is low, the therapeutic effect is expected to be low.
  • SNP Single Nucleotide Polymorphism
  • the polynucleotides of SEQ ID NOs. 1-11 described above are polymorphic sequences.
  • the polymorphic sequence refers to a sequence including a polymorphic site representing SNP in the polynucleotide sequence.
  • the polynucleotide can be DNA or RNA.
  • the polynucleotides of SEQ ID NOs. 1-11 described above are allelic-specific.
  • the allele specific polynucleotide means to hybridize specifically to each allele. That is, it means hybridizing so that the base of the polymorphic site in each polymorphic sequence of SEQ ID NO: 1-11 can be distinguished specifically.
  • hybridization is usually carried out under stringent conditions, for example, salt concentrations below 1 M and temperatures above 25 ° C.
  • stringent conditions for example, salt concentrations below 1 M and temperatures above 25 ° C.
  • 5 ⁇ SSPE 750 mM NaCl, 50 mM Na Phosphate, 5 mM EDTA, pH7.4
  • 25-30 ° C. may be suitable for allele specific probe hybridization.
  • the allele-specific polynucleotide may be a primer.
  • primers refer to single-stranded oligonucleotides that can act as a starting point for template-directed DNA synthesis under appropriate conditions (eg, dNTP or dUTP, DNA, RNA polymerase or reverse transcriptase) in appropriate buffers and at appropriate temperatures.
  • the appropriate length of the primer may vary depending on the purpose of use, but is usually 15 to 30 nucleotides. In general, short primer molecules require lower temperatures to form stable hybrids with the template.
  • the primer has its 3 'end aligned with the polymorphic site of SEQ ID NOs: 1-11.
  • the primer hybridizes to the target DNA comprising the polymorphic site and initiates amplification of an allelic form in which the primer shows complete homology.
  • This primer is used in pairs with a second primer that hybridizes to the other side. By amplification the product is amplified from two primers, which means that certain allelic forms are present.
  • the allele specific polynucleotide may be a probe.
  • the probe refers to a hybridization probe, and refers to an oligonucleotide capable of sequence-specific binding to the complementary strand of a nucleic acid.
  • the probe of the present invention is an allele-specific probe, in which polymorphic sites exist in nucleic acid fragments derived from two members of the same species, and hybridize to DNA fragments derived from one member, but not to fragments derived from other members. .
  • Hybridization conditions in this case show significant differences in hybridization strength between alleles and should be sufficiently stringent to hybridize to only one of the alleles.
  • the center portion eg, position 7 for 15-nucleotide probes and position 8 or 9 for 16-nucleotide probes. This can lead to good formation differences between different allelic forms.
  • microarray comprising an anticancer drug susceptibility predicting SNP, a polynucleotide comprising a SNP, a polypeptide encoded by the same or a cDNA thereof according to the present invention.
  • Microarrays according to the present invention can be prepared by conventional methods known to those skilled in the art using anti-cancer drug susceptibility prediction SNP.
  • the nucleotide may be immobilized on a substrate coated with an active group selected from the group consisting of amino-silane, poly-L-lysine, and aldehyde.
  • the substrate may be selected from the group consisting of silicon wafer, glass, quartz, metal and plastic.
  • Another aspect of the present invention relates to a kit for predicting anticancer drug sensitivity comprising the microarray according to the present invention.
  • the kit according to the present invention may further comprise a primer set used to separate and amplify DNA containing the SNP from the subject in addition to the microarray of the present invention.
  • a primer set used to separate and amplify DNA containing the SNP from the subject in addition to the microarray of the present invention.
  • Another aspect of the invention relates to a method for predicting anticancer drug susceptibility using SNPs for anticancer drug susceptibility prediction.
  • DNA is isolated from tissues, body fluids, or cells of a subject, and then amplified by PCR, followed by SNP analysis.
  • SNP analysis can be performed by conventional methods known in the art.
  • the SNP analysis may be performed using a real time PCR system, or may be performed by determining the nucleotide sequence of the nucleic acid directly by the dideoxy method, or a probe including the sequence of the SNP region or a complementary probe thereof.
  • step (2) the base of the SNP position corresponding to the 401th nucleotide of SEQ ID NO: 1 is identified as A (identified by G / A or A / A heterologous type)
  • the anticancer drug FL (5-FU + leucovorin) can be expected to be highly susceptible.
  • the clinical correlation analysis of the FL formulation using group shows a homozygous replacement allele (homozygous substitution allele) in the SNP id rs553717 of the gene GPC 5 having a reference allele (reference allele)
  • the relapse was about 2 times higher, and the average disease-free survival was significantly reduced (Example 3-1). This suggests that the addition of another medicament is recommended for those with alternative allele types.
  • the anticancer drug capcitabine has high sensitivity.
  • the base of the SNP position corresponding to 301 nucleotide of SEQ ID NO: 5 is G
  • the base of the SNP position corresponding to 201 nucleotide of SEQ ID NO: 6 is T
  • the base of the SNP position corresponding to 301 nucleotide of SEQ ID NO: 7 is C
  • the results of clinical association analysis of the FOLFIRI or FOLFOX® drug use group showed a significantly lower anti-cancer drug responsiveness when the reference allele group of EPHA7 rs2272107 and SSTR4 rs2567608 was used (Example 3-2).
  • the base of the SNP position corresponding to the 401 nucleotide of SEQ ID NO: 9 is identified as A or the base of the SNP position corresponding to the 251 nucleotide of SEQ ID NO: 10 can be predicted to have high susceptibility to the anticancer drug SAHA.
  • the base of the SNP position corresponding to the 401 nucleotide of SEQ ID NO: 11 is identified as T, it can be predicted that the susceptibility to the anticancer agent PXD101 is high.
  • the present invention allows the use of small blood samples from patients to predict in advance useful drugs for tumors that are resistant to drugs, enable the selection of the most appropriate anticancer drugs throughout the course of the patient, and develop appropriate genetic diagnostic tools. Was done.
  • the present invention can extend the scope of application to the technology that can newly discover the SNP markers that can predict the reactivity of the anticancer agent and the general treatment using the drug, including the new drug in the future.
  • 1 shows 12 SNP candidates of 11 selected genes showing susceptibility to 6 anticancer agents.
  • Figure 2 shows the overall survival (OS) and disease-free survival (DFS) in the FL-assisted use group according to the GPC5 rs553717 type, showing a significant DFS reduction in the homozygous allele.
  • a total of 766 SNPs out of a total of 344,048 SNPs were obtained by correlating high-intensity high-speed SNP analysis (using Affymetrix SNP Array 5.0) and in vitro tumor reactivity analysis in 104 patients with colorectal cancer.
  • the nominal P-value is 0.001 (0.1%) or less, SNP genotype showing a frequency of 5% or more in the existing Japanese and Chinese analysis data (), and linkage disequilibrium. Candidates were selected and 12 candidate SNPs were further identified through non-synonymous, haplotype-tagging and functional SNP screening (FIG. 1).
  • the cDNA clone of the gene (GPC5 was obtained from Korea Research Institute of Bioscience and Biotechnology; SSTR4 and EPHA7 were obtained from OriGene Technologies, Rockville, MD, USA).
  • SNP-type mutant plasmids were prepared using a site-directed mutagenesis kit and wild-type and mutated plasmids were transferred to RKO cells, a colorectal cancer cell line, by sequencing method, followed by 10 days G418 selection. Cell lines were constructed in which each gene was overexpressed. Primers used for the mutagenesis are listed in Table 4.
  • FIG. 1 The apoptosis of the corresponding agents against the cell lines was indicated by a trypan blue exclusion assay, and cell viability was determined using a cytotoxicity assay kit (CCK8).
  • Figure 3 shows the results of treatment with the corresponding anticancer agent at 24 hours, three repeats showed the same low sensitivity as the clinical association analysis in the reference allele genotype (P ⁇ 0.001).
  • flow cytometry was performed on the corresponding anticancer agents of the imported cell lines of the SSTR4 and EPHA7 genotypes to observe cell death along the cell cycle. Measurement was performed using a FACscaliber flow cytometer (Becton Dickinson, SanJose, CA, USA) using the Annexin V-FITC Apoptosis Detection kit.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Immunology (AREA)
  • Analytical Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Pathology (AREA)
  • Hospice & Palliative Care (AREA)
  • Oncology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention porte sur un procédé de criblage chimiosensible pour prédire un polymorphisme mononucléotidique (SNP) à l'aide de marqueurs à polymorphisme mononucléotidique, et prédire une sensibilité à un cancer à l'aide de tels marqueurs à SNP. Le procédé fournit également un moyen pour sélectionner le meilleurs cours de traitement et la médication anticancer la plus appropriée même dans des cas de résistance aux médicaments après que les patients ont été identifiés de façon sélective conformément à la présente invention.
PCT/KR2010/008640 2009-12-24 2010-12-03 Procédé de prédiction chimiosensible pour un polymorphisme mononucléotidique (snp) Ceased WO2011078495A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR10-2009-0130540 2009-12-24
KR1020090130540A KR20110073799A (ko) 2009-12-24 2009-12-24 항암제 감수성 예측용 ｓｎｐ

Publications (2)

Publication Number Publication Date
WO2011078495A2 true WO2011078495A2 (fr) 2011-06-30
WO2011078495A3 WO2011078495A3 (fr) 2011-11-10

Family

ID=44196248

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2010/008640 Ceased WO2011078495A2 (fr) 2009-12-24 2010-12-03 Procédé de prédiction chimiosensible pour un polymorphisme mononucléotidique (snp)

Country Status (2)

Country Link
KR (1) KR20110073799A (fr)
WO (1) WO2011078495A2 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112195247A (zh) * 2020-11-06 2021-01-08 南京普恩瑞生物科技有限公司 一种folfox药物方案有效性检测方法及试剂盒
CN112680517A (zh) * 2020-12-28 2021-04-20 广东南芯医疗科技有限公司 奥沙利铂个体化用药基因的指导方法及试剂盒

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
CHA, PC ET AL.: 'Single nucleotide polymorphism in ABCG2 is associated with irinotecan- induced severe myelosuppression.' JOURNAL OF HUMAN GENETICS. vol. 54, no. 10, October 2009, pages 572 - 580 *
DATABASE NCBI 04 December 2007 Database accession no. rs553717 *
GAMELIN, E ET AL.: 'Pharmacogenetics of anti-cancer drugs' ANNALES PHARMACEUTIQUES FRANCḉAISES vol. 65, no. 6, November 2007, pages 390 - 401 *
KIM, JC ET AL.: 'Genome-wide identification of chemosensitive single nucleotide polymorphism markers in colorectal cancer' CANCER SCIENCE vol. 101, no. 4, 08 December 2009, pages 1007 - 1013 *
LIU, C ET AL.: 'Genome-wide association scan identifies candidate polymorphisms associated with differential response to anti-TNF treatment in rheumatoid arthritis' MOLECULAR MEDICINE vol. 14, no. 9-10, September 2008, pages 575 - 581 *
NORDGARD, SH ET AL.: 'Pathway based analysis of SNPs with relevance to 5-FU therapy: relation to intratumoral mRNA expression and survival' INTERNATIONAL JOURNAL OF CANCER vol. 123, no. 3, 01 August 2008, pages 577 - 585 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112195247A (zh) * 2020-11-06 2021-01-08 南京普恩瑞生物科技有限公司 一种folfox药物方案有效性检测方法及试剂盒
CN112195247B (zh) * 2020-11-06 2024-02-23 南京普恩瑞生物科技有限公司 一种folfox药物方案有效性检测方法及试剂盒
CN112680517A (zh) * 2020-12-28 2021-04-20 广东南芯医疗科技有限公司 奥沙利铂个体化用药基因的指导方法及试剂盒

Also Published As

Publication number Publication date
KR20110073799A (ko) 2011-06-30
WO2011078495A3 (fr) 2011-11-10

Similar Documents

Publication Publication Date Title
WO2016167408A1 (fr) Procédé permettant de prédire le rejet d'une greffe d'organe au moyen d'un séquençage de nouvelle génération
WO2015156519A1 (fr) Procédé de prédiction d'une réaction au traitement par le sorafénib à l'aide du polymorphisme génique
WO2021029473A1 (fr) Kit de diagnostic pharmacogénomique à grande vitesse pour pharmacothérapie personnalisée et prédiction des effets secondaires de multiples médicaments de prescription associés au cancer et à des maladies chroniques
WO2019151751A1 (fr) Panel de gènes pour médicament personnalisé, son procédé de formation et procédé de traitement personnalisé l'utilisant
JP2011217753A (ja) 腎細胞癌に対するインターフェロン治療反応性識別マーカー
KR101359782B1 (ko) 간세포암종의 재발 진단용 단일 염기 다형성
WO2011159004A1 (fr) Snp destiné à prédire la sensibilité à une formulation thérapeutique anticancéreuse ciblée
WO2011078495A2 (fr) Procédé de prédiction chimiosensible pour un polymorphisme mononucléotidique (snp)
WO2005123951A2 (fr) Procedes de typage de l'antigene leucocytaire humain en mettant a proximite des haplotypes individuels de polymorphisme nucleotidique
Grzemski et al. FTO and IRX3 genes are not promising markers for obesity in Labrador retriever dogs
WO2017007275A1 (fr) Composition pour la détermination d'un phénotype nasal
WO2019132581A1 (fr) Composition de diagnostic du cancer, tel que du cancer du sein et du cancer de l'ovaire, et son utilisation
KR20110034297A (ko) 내인성 아토피 피부염 진단용 마커 및 그의 용도
Mifsud et al. ABO genotyping—identification of O, O, and O 2 alleles using the polymerase chain reaction–sequence specific oligonucleotide (PCR-SSO) technique
WO2018026039A1 (fr) Kit et procédé de détection de polymorphisme nucléotidique simple
KR100754398B1 (ko) 심혈관 질환 진단용 다중 ｓｎｐ, 그를 포함하는마이크로어레이 및 키트 및 그를 이용한 심혈관 질환 진단방법
KR101174823B1 (ko) 개인식별용 단일염기다형성 마커 및 그의 용도
Okumura et al. Sequences and mapping of genes encoding porcine tyrosinase (TYR) and tyrosinase-related proteins (TYRP1 and TYRP2).
WO2020096247A1 (fr) Procédé de préparation d'une sonde permettant de détecter une mutation dérivée de cellules dans des tissus d'un cancer du sein et procédé de détection
WO2019045431A1 (fr) Marqueur réactif antibiotique macrolide d'helicobacter pylori
WO2011118967A2 (fr) Polymorphisme mononucléotidique pour pronostiquer un carcinome hépatocellulaire
KR100908125B1 (ko) 심근 경색에 관련된 유전자 다형성 및 그의 용도
WO2011118968A2 (fr) Polymorphisme mononucléotidique pour diagnostiquer la récurrence d'un carcinome hépatocellulaire
WO2017222247A2 (fr) Marqueur de prédiction de l'efficacité d'un agent thérapeutique contre l'hémophilie et son utilisation
KR101381227B1 (ko) 항암 표적치료제제 감수성 예측용 ｓｎｐ

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 10839692

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 10839692

Country of ref document: EP

Kind code of ref document: A2