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WO2010020619A2 - Susceptibilité au dasatinib - Google Patents

Susceptibilité au dasatinib Download PDF

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Publication number
WO2010020619A2
WO2010020619A2 PCT/EP2009/060641 EP2009060641W WO2010020619A2 WO 2010020619 A2 WO2010020619 A2 WO 2010020619A2 EP 2009060641 W EP2009060641 W EP 2009060641W WO 2010020619 A2 WO2010020619 A2 WO 2010020619A2
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cell
src
epha3
frk
treatment
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WO2010020619A3 (fr
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Roman Thomas
Martin Sos
Jonathan Weiss
Thomas Zander
Peter Frommolt
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Universitaet zu Koeln
Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
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Universitaet zu Koeln
Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
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    • 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
    • C12Q1/6841—In situ hybridisation
    • 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
    • C12Q2537/00—Reactions characterised by the reaction format or use of a specific feature
    • C12Q2537/10—Reactions characterised by the reaction format or use of a specific feature the purpose or use of
    • C12Q2537/157—A reaction step characterised by the number of molecules incorporated or released
    • 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
    • C12Q2600/136—Screening for pharmacological compounds

Definitions

  • the present invention relates to a method of selecting (a) cell(s). (a) tissue(s) or (a) cell culture(s) with susceptibility to dasatinib. Also a method for determining the responsiveness of a mammalian tumor cell or cancer cell to treatment with dasatinib is described herein.
  • an in vitro method for the identification of a responder for or a patient sensitive to an dasatinib uses of an oligo- or polynucleotide capable of detecting (an) the amplification status of at least one gene selected from the group consisting of SRC, EPHA3, FRX. EPHA5. EPHA8. YES, ABL2, LCK, and BLK gene are provided.
  • the present invention also relates to a method of diagnosing non-small cell lung cancer and a method of monitoring the efficacy of a treatment of said cancer.
  • a method of predicting the efficacy of a cancer treatment is described, in particular a non-small cell lung cancer.
  • a (transgenic) non-human animal or a (transgenic) cell having at least one amplified marker gene as defined herein for screening and/or validation of a medicament for the treatment of said cancer is described and a kit useful for carrying out the methods described herein is provided.
  • the ERBB2/Her2-targeted antibody trastuzumab shrinks tumors in women with £ ⁇ J3J?2-amplified breast cancer (Slamon et al., 2001);
  • the ABL/KIT/ PDGFR inhibitor imatinib induces responses in patients with chronic myeloid leukemia carrying the BCRJABL translocation (Druker et a!.. 2001 a; Druker et al., 2001b) as well as in gastrointestinal stromal tumors and melanomas bearing (see Hodi et al..
  • Non-small cell lung cancer is one of the two main types of lung carcinoma, non- small cell (80.4%) and small-cell (16.8%) lung carcinoma, the classification being based on histological criteria.
  • the non-small cell lung carcinomas have a similar prognosis and similar management and comprise three sub-types: squamous cell lung carcinoma, adenocarcinoma and large cell lung carcinoma.
  • Squamous ceil lung carcinoma (31.1% of lung cancers) often starts near a central bronchus and commonly shows cavitation and necrosis within the center of the cancer.
  • Adenocarcinoma (29,4% of lung cancers) mostly originates in peripheral lung tissue and is usually associated with smoking.
  • Large cell lung carcinoma (10.7% of lung cancers) is a fast- growing form that develops near the surface of the lung.
  • Common treatments of NSCLC include surgery, chemotherapy, and radiation therapy, fn particular, NSCLC is treated with adjuvant chemotherapy (i.e. chemotherapy after surgery).
  • Targeted therapies for NSCLC have also been developed.
  • gefitinib which targets the tyrosine kinase domain of EGFR (epidermal growth factor) is used in the treatment of NSCLC.
  • Erlotinib another tyrosine kinase inhibitor, has been shown to increase survival in lung cancer patients.
  • the angiogenesis inhibitor bevacizumab (in combination with paclitaxel and carboplatin) is known to improve the survival of patients with advanced non-small cell lung carcinoma.
  • Further drugs under evaluation in the treatment of NSCLC are cyclo-oxygenase-2 inhibitors, proteasome inhibitors and bexarotene.
  • dasatinib a BCR/ABL and Src family tyrosine kinases inhibitor
  • BMS-354825 a drug produced by Bristol-Myers Squibb and sold under the trade name Sprycel.
  • Dasatinib is used in the treatment of patients with chronic myelogenous leukemia (CML) after imatinib treatment and Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph 4 - ALL). It is also being assessed for use in metastatic melanoma.
  • CML chronic myelogenous leukemia
  • Ph 4 - ALL Philadelphia chromosome-positive acute lymphoblastic leukemia
  • NSCLC tumor-specific tumor entity
  • a specific tumor entity e.g "NSCLC " '
  • NSCLC tumor-specific tumor entity
  • a specific tumor entity e.g "NSCLC " '
  • NSCLC tumor-specific tumor entity
  • NSCLC tumor-specific tumor entity
  • many patients with tumors belonging to the same tumor entity would profit from an identification of markers/predictors for susceptibility of the specific tumor type to (a) certain drug(s)/compound(s).
  • the identification prior to initiation/completion of clinical trials is desirable.
  • Cancer cell lines may be used in corresponding in vitro experiments for identification of drugs to which NSCLC tumors/tumor cell are susceptible; yet. the validity and clinical interpretability of these widely used models have been questioned. In addition, cell lines are frequently thought to be genomically disarrayed and unstable and therefore likely poorly representative of primary tumors. Furthermore, the genetic diversity of histopathologically defined classes of tumors is often substantial: e.g., the clinical tumor entity non-small cell lung cancer (NSCLC) comprises EGFR- and KRAS-irmtant lung adenocarcinomas as well as KRA S-mutant squamous-cell lung cancers. Thus, any representative pre-clinical model would need to capture the nature of lesions of primary tumors as well as their distribution in the histopathologically defined cohort.
  • NSCLC clinical tumor entity non-small cell lung cancer
  • somatic genetic alterations (lesions) in cancer have been causally linked with response to targeted therapeutics as they frequently expose a specific dependence on activated oncogenic signaling pathways.
  • an identification of compounds/drugs to which tumors are susceptible is often time-consuming and cost-intensive since these compounds/drugs may only be identified after completion of clinical trials.
  • the technical problem underlying the present invention is the provision of means and methods for the evaluation of cells, in particular tumor cells, for their susceptibility or responsiveness to anti-cancer treatment,
  • the present invention relates to a method of selecting (a) cell(s). (a) tissue(s) or (a) cell cult ⁇ jre(s) with susceptibility to dasatinib, comprising the steps'
  • the method may additionally comprise the steps (i) contacting said cell(s). tissue(s) or cell culture(s) with dasatinib; and (ii) evaluating viability of said cell(s), tissue(s) or cell culture(s) contacted with dasatinib. It is of note that steps (I) and (ii) may be performed prior to step (a) but also after step (a) or, optionally after step (b). Said steps (i) and (ii) may in particular serve as further experimental proof that the selected cell, tissue or cell culture that comprises (a) gene amplification is susceptible in its viability to dasatinib.
  • cell, tissue and cell culture is not only limited to isolated cells, tissues and ceil cultures but also comprises the use of samples, i.e. biological, medical or pathological samples that consist of fluids that comprise such cells, tissues or cell cultures.
  • a fluid may be a body fluid or also excrements and may also be a culture sample, like the culture medium from cultured cells or cultured tissues.
  • the body fluids may comprise, but are not limited to blood, serum, plasma, urine, saliva, synovial fluid, spinal fluid, cerebrospinal fluid, tears, stool and the like.
  • the gist of the present invention lies in the fact that a method is provided that allows for the determination of the susceptibility of a given cell, tissue or cells in a tissue, (or a cell culture or individual cells in such a cell culture, or as will be explained below, (a) cell(s) in a biological/medical/palhological sample) for the anti-cancer or antiproliferative treatment with dasatinib.
  • a method is provided that allows for the determination of the susceptibility of a given cell, tissue or cells in a tissue, (or a cell culture or individual cells in such a cell culture, or as will be explained below, (a) cell(s) in a biological/medical/palhological sample) for the anti-cancer or antiproliferative treatment with dasatinib.
  • a biological/medical/palhological sample for the anti-cancer or antiproliferative treatment with dasatinib.
  • the present invention does not only provide for a method for selecting cells/tissues/cell cultures which are susceptible to dasatinib. but also for an in vitro method for assessing an individual, i.e. a human or animal patient, for its potential responsiveness to an anti-cancer or anti-proliferate treatment with dasatinib.
  • the present invention provides not only for the possibility to select cells, tissues and cell cultures that are susceptible for dasatinib treatment (i.e. the selection of e.g.
  • dasatinib drugs with a structural similarity to dasatinib may be tested or which are useful in screening methods for compounds that are suspected to function like dasatinib) but also for a method to evaluate whether a given patient, preferably a human patient, in need of treatment but also prevention of a proliferative disease, is a responder for dasatinib treatment. Most preferably, the responsiveness of a given patient to dasatinib is tested. Dasatinib is described herein below in more detail.
  • the selection method of a dasatinib responding cell or a responding patient comprises a step, wherein (a) cell(s), (a) tissue(s) or (a) cell culture(s) with (a) gene amplification(s) above normal of at least one of SRC, EPH ⁇ 3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK is selected.
  • Said cell/tissue may also be derived from a human sample or from a body fluid that comprises such a cell, for example a cancer cell.
  • EPHA8, YES, ABL2, LCK and'or BLK is indicative for susceptibility to dasatinib.
  • the present invention relates in particular to a method for determining the responsiveness of a mammalian tumor cell or cancer cell to treatment with dasatinib, said method comprising determining the amplification status of at least one gene selected from SRC, EPHA3, FRK, EPHA5. EPHA8. YES, ABL2, LCK and/or BLK in said tumor cell, wherein said amplification status is indicative of whether the cell is likely to respond or is responsive to the treatment.
  • a determination may take place on an individual, isolated tumor cell.
  • Such an evaluation may also be carried out on biological/medical/pathological samples, like body fluids, isolated body tissue samples and the like, wherein said samples preferably comprise cells or cell debris to be analyzed.
  • markers which can predict the outcome of an anti-cancer therapy with dasatinib prior to and during treatment.
  • Subject of the present invention is a method for diagnosing an individual who is to be subjected to or is being subjected to an anti -cancer treatment or an anti-pro liferative treatment to asses the responsiveness to dasatinib prior, during and/or after dasatinib treatment which comprises the steps of (a) detection of the gene amplification status of at least one gene selected from the group consisting of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK, and BLK in a biological/medical/pathological sample wherein the gene amplification above normal of at least one of said genes is indicative for the responsiveness to dasatinib treatment prior, during and after treatment with dasatinib; and (b) sorting the individual into responder or Non-responder based on detection of said gene amplification of at least one of said genes.
  • the invention provides for markers which can predict the outcome of an anti- cancer/anti-pro liferative treatment with dasatinib prior to treatment in addition to during and/or after treatment.
  • the present invention solves the above identified technical problem since, as documented herein below and in the appended examples, it was surprisingly found that the presence of (a) gene amplification above normal of at least one of SRC, EPHA3, FRK. EPHA5, EPHA8, YES. AB L2, LCK, and/or BLK in (a) cell(s), (a) tissue(s) or (a) cell culture (or in a biological sample comprising cells or cell debris) is highly predictive for susceptibility of said cell(s), tissue(s) or cell culture(s) (or the individual who provided said biological sample) to dasatinib.
  • the presence of (a) gene amplif ⁇ cation(s) above normal of at least one of SRC is highly predictive for susceptibility of said cell(s), tissue(s) or cell culture(s) (or the individual who provided said biological sample) to dasatinib.
  • EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK was surprisingly identified as a marker /predictor for responsiveness to treatment with dasatinib or for susceptibility to dasatinib.
  • the terms "marker for responsiveness to treatment with dasatinib”7'"marker for susceptibility to dasatinib " and "predictor for responsiveness to treatment with an dasatinib'V'predictor for susceptibility to dasatinib” can be used interchangeably and refer to (a) gene amplification(s) of said genes, whereby the amplification status is indicative for susceptibility to dasatinib.
  • a gene amplification is defined herein as amplification of the gene above normal.
  • “'Normal amplification * 7"Normal amplification status” used herein refers to the presence of two copies of a gene in the genome. "Amplification above normal' * refers accordingly to the presence of at least three copies of a gene in a genome. The presence of (a) gene amplification(s) of the genes defined herein above correlates significantly (p ⁇ 0.05) with a responsiveness to treatment with dasatinib or susceptibility to dasatinib.
  • LCK and/or BLK as markers for susceptibility of tumor cell(s) to an dasatinib allows for the first time a reliable identification of subjects/patients which can be specifically and efficiently treated with dasatinib.
  • the likelihood for susceptibility of patients having no gene amplification above normal of at least one of said genes to treatment with dasatinib is below 10 %.
  • the probability for susceptibility of patients having (a) gene amplification(s) above normal of one of said genes to treatment with dasatinib rises 5-fold to about 50 %.
  • the probability for susceptibility of patients having (a) gene amplification(s) above normal of two of said genes to treatment with dasatinib rises 8-fold to about 80 %.
  • the gene-expression signatures described by Huang was in no way predictive of responsiveness to dasatinib.
  • the chromosomal gene copy number signature i.e. the gene amplification above normal of at least one of the following genes SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK ) described herein represents an important and unexpected contribution to the art and a significant improvement.
  • the present invention is illustrated by the experiments described in the appended Example.
  • the gene amplification status above normal of at least one of the following genes SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK was surprisingly identified as predictor/marker for susceptibility to dasatinib.
  • the gene amplification status of said gene(s) was identified as predictor/marker for susceptibility to dasatinib using the representative NSCLC (non small cell lung cancer) cell line collection as demonstrated in the appended example.
  • One advantage of the present method is the fact that it allows for an in vitro selection of (a) cell(s), (a) tissue(s) or (a) cell culture with susceptibility to dasatinib/responsive to treatment with dasatinib.
  • genomically annotated NSCLC cell lines are used that are representative of the genetic diversity, the transcriptional profile and the phenotypic properties of primary NSCLC (non-small cell lung cancer) tumors. It is shown herein by integrated genomic profiling on a global scale that the genomes of non-small cell lung cancer (NSCLC) ceil lines are highly representative of several primary NSCLC tumors isolated by surgery from patients in gene copy number, oncogene mutation and gene expression space.
  • using said NSCLC cell line collection in context of the present invention may avoid animal tests or voluntary tests with, cancer patients; at the same time use of said cell line collection allows, in contrast to methods known in the art.
  • EPHA5, EPHA8. YES, ABL2, LCK and/orBLK as markers for susceptibility to dasatinib.
  • NSCLC non-small cell lung cancer
  • EGFR mutations were confirmed to predict sensitivity to EGFR inhibitors (erlotinib. PD168393, vandetanib) (Arao et al., 2004; Lynch et al, 2004; Paez et al., 2004; Pao et ah, 2004; Sos et al., 2008) which is in accordance with prior ail observations.
  • EGFR inhibitors in EGFR-mutant NSCLC cell lines has been described in the prior art (McDermott et al. 2007; Paez et ah, 2004; Tracy et al.. 2004).
  • EGFff-routations were identified as markers/predictors of susceptibility to EGFR mhibition (p ⁇ 0.0001) using the systematic cell-based compound screening followed by computational prediction of sensitivity based on lesion profiles, EGFR mutations were also identified herein as predictors/markers for susceptibility to the SRC/ ABL inhibitor dasatinib. These findings suggest that mutant EGFR might be a target of dasatinib (Song et al, 2006). However, not only EGFR-mutations were identified and confirmed as predictors/markers for susceptibility to EGFR inhibitors but also and unexpectedly the amplification status of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2. LCK and/or BLK as predictors/markers for susceptibility to dasatinib.
  • SRC is the relevant target in cells with SRC amplification by showing growth inhibition and cell death induced by sliRNA- mediated knockdown of SRC.
  • the finding that SRC is indeed the relevant target of dasatinib has been further confirmed in a chemical genetics approach that can be considered as the most stringent assay for target validation (Du et al., 2009).
  • This assay exploits the fact that dasatinib is an ATP-competitive hinge region-binding compound that is highly sensitive to modifications of amino acids regulating the entrance to the ATP binding pocket.
  • a change of the threonin to a stericall '"bulky" methionin at the amino acid position 341 lead to a physical barrier that acts as a "gatekeeper” and prevents the binding of dasatinib to the hinge region.
  • the gatekeeper region is a unique residue in the entrance of the ATP-binding pocket that is occupied by competitive inhibitors and thus a common position for the occurance of resistance mutations that abrogate the activity of such compounds.
  • SRC has been ectopically expressed with and without the above described gatekeeper mutation at the amino acid position 341 (i.e., threonin to methionin [T341M]) and it was found herein that the cells stably expressing the gatekeeper mutation were fully resistant to dasatinib treatment, This demonstrates that SRC is indeed the relevant target of dasatinib in SRC-amplified cells.
  • susceptibility to dasatinib(s)/ responsiveness to treatment with dasatinib are well known in the art.
  • susceptibility to dasatinib/responsiveness to treatment with dasatinib may be determined by contacting (a) cell(s) 5 (a) tissue(s), or (a) cell culture(s) with dasatinib and determining the viability of said (a) cell(s), (a) tissue(s), or (a) cell culture(s) after contacting.
  • These above-mentioned methods for determining the susceptibility to dasatinib(s)/ responsiveness to treatment with dasatinib may. for example, comprise an evaluation/determination step, which may.
  • tissue(s) or cell culture(s) contacted with/exposed to an dasatinib or fa) mammalian cellfs) or cancer cell treated with an dasatinib For example, (a) cell(s), (a) tissue(s) or (a) cell culture(s) described herein above may show decreased viability upon contacting/exposing/treating with a dasatinib.
  • the ce ⁇ l(s). tissue(s) or cell culture(s) may show an at least 10 %, 20 %, 30 % 5 40 %.
  • control cell(s), tissue(s) or cell culture(s) not contacted/exposed/treated with an dasatinib Preferably, the control cell(s).
  • tissue(s) or cell culture(s) will be identical to the cell(s).
  • ce ⁇ l(s), (a) tissue(s) or (a) cell culture(s) contacled/exposed/treated with dasatinib and showing, for example, a decreased viability as described herein above, can be considered as being susceptible to dasatinib.
  • tissue(s) or cell culture(s) contacled/exposed/treated with dasatinib and showing, for example, a decreased viability as described herein above can be considered as being susceptible to dasatinib.
  • mammalian tumor cellfs) or (a) cancer cell(s) treated with dasatinib showing such a decreased viability can be considered as responsive to treatment with dasatinib.
  • a reduction in viability may. for example, be reflected in a decreased proliferation, such as 10 %, 20 %, 30 %, 40 %, 50 %, 60 %, 70 %. 80 %, and most preferably, 90 % reduction in proliferation compared to control cell(s), tissue(s) or cell culture(s) not contacted/exposed/treated with dasatinib.
  • the decreased proliferation may be quantitated, for example, by measuring the total cell volume, tissue volume or cell culture volume using standard techniques.
  • the difference in proliferation between contacted/exposed/treated cell(s), tissue(s) or cell culture(s) and corresponding controls as defined herein may, for example, be evaluated/determined by measuring the volume of the cell(s), tissue(s) or cell culture(s) taking advantage of standard techniques. Said evaluation/determination may be performed in various points in time, for example, 15 minutes, 30 minutes. 60 minutes. 2 hours, 5 hours, 18 hours. 24 hours. 2 days, 3 days, 4 days, 5 days, six days and/or seven days after contacting/treating said cell(s), tissue(s) or cell culture(s) with an dasatinib or exposing said cell(s), tissue(s) or cell culture(s) to an dasatinib.
  • said evaluation/determination may be performed repeatedly, for example, at 15 minutes, 30 minutes and 60 minutes after said contacting/exposing/treating.
  • said cell(s). tissue(s) or cell culture(s) may be contacted /treated not oniy once with said dasatinib or exposed to said dasatinib but several times (e.g. 2 times, 3 times. 5 times, 10 times or 20 times) under various conditions (e.g. same concentration of inhibitor, different concentration of inhibitor, inhibitor comprised in a composition with different stabilizers, diluents, and/or carriers and the like).
  • said optionally repeated evaluation/determination may be performed after the final contacting/treating with or exposing to said dasatinib or in between said above-mentioned various contacting/exposing/treating steps.
  • These tests may include but are not limited to, measurements of Annexin-V exposure on the outer membrane, cell cycle analyses, propidium iodide staining, TUNEL assay, DNA fragmentation assays, nuclear condensation assays, KI-67 staining, resazurm staining, protein cleavage assays (e.g., PARP or Caspase-3) and others.
  • dasatinib is a known tyrosine kinase inhibitor.
  • the respective formula is given herein below:
  • HTS high throughput screening
  • tissue(s) and/or cell culture(s) for responsiveness/sensitivity to dasatinib.
  • Suitable (HTS) approaches are known in the art.
  • An exemplary protocol for such a screening method is also provided in the appended examples; a person skilled in the art is readily in the position to adapt this protocol or known HTS approaches to the performance of the methods of the present invention.
  • Screening-assays are usually performed in liquid phase, wherein for each cell/tissue/cell culture to be tested at least one reaction batch is made.
  • Typical containers to be used are micro titer plates having for example, 384, 1536. or 3456 wells (i.e. multiples of the "original" 96 reaction vessels).
  • Robotics, data processing and control software and sensitive detectors are further commonly used components of a HTS device.
  • robot system which transport micro titer plates from station to station for addition and mixing of sample(s) and reagent(s), incubating the reagents, and final readout (detection).
  • ITTS can be used in the simultaneous preparation, incubation, and analysis of many plates.
  • the assay can be performed in a singly reaction (which is usually preferred), may, however, also comprise washing and/or transfer steps. Detection can be performed taking advantage of radioactivity, luminescence or fluorescence, like fiuorescence-resonance-energytransfer (FRET) and fluorescence polarisation (FP) and the like.
  • FRET fiuorescence-resonance-energytransfer
  • FP fluorescence polarisation
  • the biological samples described herein can also be used in such a context.
  • cellular assays and in vivo assays can be employed in HTS.
  • Cellular assays may also comprise cellular extracts, i.e. extracts from cells, tissues and the like.
  • cell(s) or tissue(s) as biological sample (in particular a sample obtained from a patient/subject suffering or being prone to suffer from cancer), whereas in vivo assays (wherein suitable animal models are employed, e.g. the herein described mouse models) are particularly useful in the validation/monitoring of the treatment with dasatinib.
  • in vivo assays wherein suitable animal models are employed, e.g. the herein described mouse models
  • follow up assays can be performed by re-running the experiment to collect further data on a narrowed set (e.g. samples found "positive" in the first assay), confirming and refining observations.
  • a suitable readout in animal (in vivo) models is tumor growth (or respectively the complete or partial inhibition of tumor growth and/or its remission).
  • the herein described HTS methods for the detection of copy number changes include but are not limited to techniques such as single nucleotide polymorphism (SNP)-array and Comparative Genomic Hybridization (CGH)-arrays.
  • SNP-array technology allows parallel interrogation of up to two million genomic locations in high-throughput. After DNA labelling and hybridization, fluorescence intensities are measured for each allele of each SNP and genomic copy number changes can be inferred.
  • the CGH technique allows the detection of "tumor” and "normal” tissue extracts, differently fluorescence labeled, on the same glass slide and the relative copy number changes (amplifications and deletions) can thus be inferred.
  • cell(s) refers to a single cell or a plurality of cells.
  • plural of cells means in the context of the present invention a group of cells comprising more than a single cell. Thereby, the cells out of said group of cells may have a similar function. Said cells may be connected cells and/or separate cells.
  • tissue in the context of the present invention particularly means a group of cells that perform a similar function.
  • culture(s) means in context of the present invention cells as defined herein above which are grown/cultured under controlled conditions.
  • Cell culture(s) comprise in particular cells (derived/obtained) from multicellular eukaryotes, preferably animals as defined elsewhere herein. It is to be understood that the term "cell culture(s)” as used herein refers also "tissue culture(s)” and/or “organ culture(s)", an “organ” being a group of tissues which perform the same function.
  • the cell(s), tissue(s) or cell culture(s) to be selected comprise/are derived from or are (a) tumor cell(s).
  • the tumor cells may, for example, be obtained from a biopsy, in particular a biopsy/biopsies from a patient/subject suffering from or being prone to suffering from non-small cell lung cancer. It is preferred herein that said subject is a human.
  • the term "mammalian tumor cell(s)" used herein refers to (a) tumor cell(s) which is derived from or is a tumor cell from a mammal, the term mammal being derived herein below.
  • tumor cells may be obtained from a biopsy, in particular a biopsy/biopsies from a patient/ subject suffering from non-small cell lung cancer or a patient/subject being prone to suffer from said disorders.
  • tissue cell also relates to "cancer cells”
  • said tumor cell or cancer cell may be obtained from any biological source/organism, particularly any biological source/organism, suffering from or being prone to suffer from the above-mentioned non-small cell lung cancer.
  • the (tumor) cell(s) or (cancer) cell to be contacted is (are) obtained/derived from an animal. More preferably, said (turnor)/cancer cell(s) is (are) derived from a mammal
  • animal or “mammal”
  • Non-limiting examples for mammals are even-toed ungulates such as sheep, cattle and pig, odd-toed angulates such as horses as well as carnivores such as cats and dogs.
  • DNA samples are derived from organisms that are economically, agronomically or scientifically important.
  • Scientifically or experimentally important organisms include, but are not limited to. mice, rats, rabbits, guinea pigs and pigs.
  • the tumor cell(s) may also be obtained from primates which comprise lemurs, monkeys and apes.
  • the meaning of the terms “primate”, “lemur”, “monkey * ' and “ape” is known and may, for example, be deduced by an artisan from Wehner und Gehring (1995, Thieme Veriag).
  • the tumor or cancer cell(s) is (are) most preferably derived from a human being suffering from the above-mentioned non-small cell lung cancer, pancreatic cancer, colorectal cancer, breast cancer, leukemias.
  • particular useful cells, in particular- tumor or cancer cells are, accordingly, human cells. These cells can be obtained from e.g. biopsies or from biological samples but the term "cell” also relates to in vitro cultured cells.
  • the present invention relates to an in vitro method for the identification of a responder for or a patient sensitive to dasatinib, said method comprising the following steps: (a) obtaining a sample from a patient suspected to suffer from or being prone to suffer from non- small cell lung cancer; and
  • Said sample may, for example, be obtained by (a) biopsy (biopsies).
  • said sample is obtained from a patient suspected to suffer from or being prone to suffer from non-small cell lung cancer. It is preferred herein that said sample is obtained from (a) tumor(s) and, accordingly, is (a) tumor cell(s) or (a) tumor tissue(s).
  • tumor, sample(s) may be obtained from subjects/patients suffering from non-small cell lung cancer.
  • SRC single as marker in context of the present invention
  • EPHA3 or FRK alone as marker.
  • the amplification status of at least two genes selected from the group of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and BLK is assessed or evaluated.
  • the amplification status of SRC in combination with at least one further of the above 8 remaining genes is assessed or evaluated.
  • Exemplary combinations which are preferred in this context are: SRC and EPHA3; SRC and FRK; SRC. EPHA3 and FRK.
  • SRQ EPHA3 and EPHA5 SRC, EPHA3 and EPHA8: SRC, EPHA3 and YES; SRC 5 EPHA3 and ABL2; SRC, EPHA3 and LCK; SRC. EPHA3 and BLK; SRC. FRK and EPHA5: SRC. FRK and EPHA8; SRC, FRK and YES; SRC 5 FRK and ABL2;
  • SRC EPHA5 and EPHA8; SRC, EPHA5 and YES; SRC. EPHA5 and ABL2; SRC, EPHA5 and LCK; SRC, EPHA5 and BLK;
  • SRC EPHA8 and YES; SRC. EPHA8 and ABL2; SRC, EPHA8 and LCK; SRC, EPHA8 and BLK;
  • SRC SRC, EPHA3, and two further markers
  • SRC SRC.
  • SRC EPHA3. EPHA5 and EPHA8; SRC, EPHA3, EPHA5 and YES; SRC, EPHA3, EPHA5 and ABL2; SRC, EPHA3, EPHA5 and LCK; SRC, EPHA3, EPHA5 and BLK; SRC, EPHA3, EPHA8 and YES; SRC, EPHA3, EPHA8 and ABL2; SRC, EPHA3, EPHA8 and LCK; SRC, EPHA3, EPHA8 and BLK;
  • SRC 5 EPHA3, YES and ABL2; SRC, EPHA3, YES and LCK; SRC, EPHA3, YES and BLK: SRC, EPHA3, ABL2 and LCK; SRC, EPHA3, ABL2 and BLK; or SRC, EPHA3, LCK and BLK; wherein combinations which comprise SRC, EPHA3 and FRK are particularly preferred.
  • Preferred combinations of 4 genes are SRC, FRK, EPHA5 and EPHA8; SRC, FRK, EPHA5 and YES; SRC, FRIC, EPHA5 and
  • ABL2 ABL2; SRC, FRK, EPHA5 and LCK; SRC, FRK, EPHA5 and BLK;
  • SRC SRC, FRK, EPHA8 and YES; SRC. FRK, EPHA8 and ABL2; SRC, FRK, EPHA8 and LCK;
  • SRC SRC with any of the remaining markers EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK, and BLK are easily deduclble by a skilled person, wherein SRC can be combined with at least one, two, three, four, five, six, seven, or eightfurther markers is envisaged. Again, combinations comprising SRC and EPHA3; SRC and FRK; or SRC, EPFIA3 and FRK are preferred.
  • EPHA3 or FRK alone can be used as marker in context of the present invention.
  • the amplification status of EPHA3 and/or FRK in combination with at least one further of the above remaining genes is assessed or evaluated.
  • the amplification status of EPFIA3 can also be assessed/evaluated in combination with FRK, EPHA5, EPHA8, YES, ABL2, LCK, and/or BLK, wherein combinations of EPHA3 and FRK with at least one further of the remaining markers are preferred.
  • the amplification status of FRK can also be assessed/evaluated in combination with EPHA3, EPHA5, EPHA8, YES, ABL2, LCK, and/or BLK; wherein combinations of FRK and EPFIA3 with at least one further of the remaining markers are preferred.
  • EPHA 3 The following combinations of EPHA 3 are preferred: EPHA3 and SRC; EPHA3 and FRK; EPHA3 and EPHA5; EPHA3 and EPHA8; EPHA3 and YES; EPHA3 and ABL2; EPHA3 and LCK; EPHA3 and BLK. Also preferred herein are combinations of FRK and SRC; FRK and EPHA3; FRK and EPHA5; FRK and EPFIA8; FRK and YES; FRK and ABL2; FRK and LCK: FRK and BLK.
  • said at least two amplified genes are selected from the group consisting of SRC and EPHA3.
  • EPHA3 and FRK, and EPHA3 and ABL2 and, as mentioned above, it is preferred herein that said at least one gene/said at least two genes is/are present in at least 3 copies.
  • the gene amplification status may, for example, be detected, assessed or evaluated by an in situ hybridization method, comparative genomic hybridisation and smgie-nucleotide polymorphism arrays.
  • exemplary in situ hybridisations are, inter alia, fluorescent in situ hybridisation (FISH), chromogenic in situ hybridisation (CISH) and silver in situ hybridisation (SISH).
  • the present invention relates to the use of an oligo- or polynucleotide capable of detecting the amplification status of at least one gene selected from the group consisting of SRC. EPHA3. FRK. EPHA5, EPHA8. YES, ABL2, LCK 5 and BLK for diagnosing sensitivity to dasatinib.
  • said oligonucleotide is about 15 to 50 nucleotides in length.
  • a person skilled in the art is, based on his general knowledge and the teaching provided herein, easily in the position to identify and/or prepare (a) an oligo- or polynucleotide capable of detecting the amplification status of at least one gene selected from the group consisting of SRC, EPHA3, FRK. EPHA5, EPHA8, YES, ABL2, LCK and BLK for diagnosing sensitivity to dasatinib.
  • these oligo- or polynucleotides may be used as probe(s) in the detection methods described herein.
  • a skilled person will know, for example, computer programs which may be useful for the identification of corresponding probes to be used herein.
  • the SRC EPHA3. FRK, EPHA5, EPHA8.
  • LCK and/or BLK nucleic acid sequences may be used in this context for identifying specific probes for detecting the amplification status.
  • Exemplary, non- limiting SRC, EPHA3, FRK, EPHA5, EPHA8, YES 5 ABL2, LCK and/or BLK nucleic acid sequences are also available on corresponding databases, such as the NCBI database (www.ncbi.nlm. nih.gov/sites/entrez).
  • the present invention relates to a method of diagnosing (non-small cell lung cancer) in a subject/patient suspected of suffering from non-small cell lung cancer or suspected of being prone to suffering from non-small ceil lung cancer comprising the steps of a) determining in a cell or tissue sample obtained from said subject/patient the activity of at least one marker gene selected from the group consisting of SRC, EPHA3, FRK, EPHA5, EPHA8, YES 5 ABL2, LCK and BLK ; and b) comparing the activity of said at least one marker gene determined in a) with a reference activity of said at least one marker gene determined in (a sample from) a control subject/patient (healthy subject), wherein said non-small cell lung cancer is diagnosed when said activity determined in a) differs from said reference activity.
  • the present invention also relates to a method of monitoring the efficacy of a treatment of a non-small cell lung cancer in a subject/patient suffering from said disorder or being prone to suffering from said disorder comprising the steps of a) determining in a cell or tissue sample obtained from said subject/patient the activity of at least one marker gene selected from the group consisting of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and BLK ; and b) comparing the activity of said at least one marker gene determined in a) with a reference activity of said at least one marker gene, optionally determined in (a sample from) a control subject/patient (responder and/or non-responder), wherein the extent of the difference between said activity determined in a) and said reference activity is indicative for said efficacy of a treatment of a non-small cell lung cancer.
  • the term "activity" as used herein refers to the activity of a protein as described elsewhere herein.
  • the method of monitoring the efficacy of a treatment of a cancer may comprise a step of determining in a cell or tissue sample obtained from a subject/patient suffering from Non- small cell lung cancer (e.g. a biopsy) the gene amplification status of at least one gene selected from the group consisting of SRC, EPHA3, FRK. EPHA5, EPHA8. YES, ABL2, LCK and BLK .
  • a gene amplification above normal of at least one gene selected from the group consisting of SRC, EPHA3. FRK, EPHA5. EPHA8, YES, ABL2. LCK and BLK may be present in a sample before start of the treatment of a cancer. During or after treatment of the cancer, the tumor cells having said gene amplification above normal are erased or otherwise depleted. Thus, the absence of a detectable gene amplification above normal of at least one of said genes in a sample (cell samples/biopsy samples and the like) obtained from a subject/patient during or after treatment of a cancer is indicative of the efficacy of the treatment.
  • the present invention relates to a method of predicting the efficacy of a treatment of a non-small cell lung cancer for a subject/patient suffering from said disorder or being prone to suffering from said disorder comprising the steps of a) determining in a cell or tissue sample obtained from said subject/patient the activity of at least one marker gene selected from the group consisting of SRC, EPHA3, FRK. EPHA5, EPHA8. YES. ABL2.
  • the treatment of non-small cell lung cancer may comprise the administration of dasatinib.
  • the non-small cell lung cancer may, inter alia, be a squamous cell lung carcinoma, an adenocarcinoma, a large cell lung carcinoma, or an anaplastic carcinoma.
  • the subject/patient suffering from said non-small cell lung cancer or being prone to suffering from said non-small cell lung cancer may also exhibit resistance (primary or secondary) against any platinum- based, taxane-based chemotherapy, vinorelbine, gemcitinibe, erlotinib, sunitinib and/or vandetanib.
  • EPHA5, EPHA8, YES, ABL2, LCK and/or BLK as disclosed herein act as markers/predictors for susceptibility to dasatinib.
  • a responder for or a patient sensitive to dasatinib may be identified in accordance with the present method.
  • the present invention provides the possibility to recognize (aberrant) changes of SRC, EPHA3. FRK, EPPIA5, EPHA8. YES, ABL2. LCK and/or BLK activity immediately once the ⁇ ' occur, for example, by determining the activity of said marker gene(s).
  • the assessment/evaluation/detection of the amplification status of any of the above marker genes is sufficient for determining whether a patient is likely to respond to or is sensitive to dasatinib. whether a (mammalian tumor or cancer) cell is likely to respond or is responsive to treatment with dasatinib.
  • the assessment/evaluation/detection of the amplification status of any of the above marker genes (and their combinations) is also sufficient for diagnosing sensitivity to dasatinib.
  • the amplification status alone of any of the above marker genes is indicative for a sensitivity/responsiveness to dasatinib and the expression level/activity of the gene products of the above marker genes need not be determined in addition to the amplification status.
  • LCK BLK may not only be determined by measuring the expression level but also, be determined, for example, by measuring substrate turnover of SRC, EPHA3. FRK. EPHA5, EPHA8, YES. ABL2, LCK. This may be particularly useful in methods described herein for diagnosing non- small cell lung cancer, monitoring non-small cell lung cancer or predicting the efficacy of a treatment of cell lung cancer. Means and methods for determining the activity of said proteins are well known in the art and may. for example, be deduced from Lottspeich (Spektrum Akademischer Verlag, 1998).
  • determining the activity may comprise determining the expression level
  • the expression status i.e. expression of gene products such as mRNA and/or proteins
  • SRC expression status
  • LCK and/or BLK can be determined by standard techniques. Amplification of SRC, EPHA3, FRK, EPH A5, EPHAS 5 YES, ABL2. LCK and/or BLK gene does not necessarily correlate with a change in the expression level of these genes. In context of the present invention it is therefore preferred that the activity of (amplified) SRC. EPHA3, FRK, EPHA5, EPHA8, YES.
  • ABL2, LCK and/or BLK is not determined by measuring the expression status of these genes but by alternative methods which reflect, for example, the enzymatic activity of the corresponding proteins. It is of note that, for example, the enzymatic activity of proteins encoded by a (amplified) SRC, EPHA3. FRK, EPHA5, EPHA8, YES, ⁇ BL2, LCK and/or BLK may differ from that of the respective proteins encoded by wild type genes (i.e. reference activity) without a change in the expression level. Preferably, (amplified) SRC. EPHA3, FRK.
  • EPHA5, EPFIA8, YES, ABL2, LCK and/or BLK show an increased activity compared to non-amplified (amplified) SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK (controls).
  • the enzymatic activity and/or the expression level described herein above are increased.
  • Exemplary ranges of changes in the activity compared to ''normal" activity i.e. activity of (non-amplified) SRC, EPHA3.
  • FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK are described herein below.
  • 'non-amplified refers to the ''normal" amplification status, i.e. two gene copies, whereas "amplified” refers to an amplification status above normal, i.e. at least three copies of the respective gene.
  • the present invention provides the particular advantages that, by determining the amplification status of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK in accordance with this invention, (aberrant) changes of (amplified) SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK activity/expression level can be recognized early, i.e. that the efficacy of a treatment NSCLC can be monitored early and that the efficacy of a treatment of said cancer can be predicted early. Hence, also a possible resistance to the treatment can be recognized early by using the means and method of this invention.
  • the present invention relates to corresponding means, methods and uses which are based on the early recognition of (aberrant) changes of (amplified) SRC, EPHA3. FRK. EPHA5, EPHA8, YES, ABL2, LCK and/or BLK activity/expression level of the respective genes.
  • the possibility of recognizing (aberrant) changes of (amplified) SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK activity/expression level early provides several advantages, like a higher lifespan/likelihood of survival of the subject/patient (for example due to the notice of possible treatment failures and a corresponding change of the treatment regimen) and the possibility of a more efficient therapy (for example due to the possibility to avoid/recognize treatment failures early and. hence, to correspondingly change the treatment regimen early in therapy, i.e. to timely switch to a more suited inhibitor, to discontinue an expensive, ineffective treatment early after diagnosis and to opt for alternative therapy) .
  • '"early particularly means prior to (the onset of) a (complete or partial) cytogenetic or haematological response or a response measured by any type of imaging technique and/or prior to the outbreak of NSCLC (or susceptibility th ereto ) .
  • "'early" monitoring the efficacy of a therapy /treatment of said cancer may be at least 1. at least 2. at least 3, at least 4, at least 5. at least 6. at least 7, at least 10, or at least 14 days prior to (the onset of) a (partial) cytogenetic or haematological response or a response measured by any type of imaging technique to said therapy/treatment and/or at least 1. at least 2. at least 3. at least 4. at least 5 5 at least 6, at least 7, at least 10, at least 12, at least 15, or at least 18 month prior a complete cytogenetic or haematological response or a response measured by any type of imaging technique to said therapy/treatment (of the patient or control patient (responder)), wherein the longer periods are preferred.
  • ''early' ' monitoring the efficacy of a therapy/treatment of said cancer may also be at most 1 , at most 2, at most 3, at most 4, at most 5, at most 6, at most 7, at most 10. or at most 14 days after (onset of) the therapy /treatment of said cancer, wherein the shorter periods are preferred.
  • EPHA5, EPHA8, YES, ABL2, LCK and/or BLK activity /expression level changes upon said therapy/treatment.
  • the reference activity/expression level may be taken at the day the therapy/treatment is initiated, from the subject/patient to be treated and/or from a corresponding control subject/patient (responder/non-responder); see below.
  • '"early 1" predicting the efficacy of a therapy/treatment of the cancer defined herein may be at least 1, at least 2. at least 3, at least 4. at least 5, at least 6, at least 7, at least 10, or at least 14 days prior to (the onset of) a (partial) cytogenetic or haematological response to said therapy/treatment and/or at least 1, at least 2. at least 3. at least 4, at least 5, at least 6. at least 7, at least 10, at least 12, at least 15, or at least 18 month prior a complete cytogenetic or haematological response or a response measured by any type of imaging technique to said therapy/treatment, wherein the longer periods are preferred.
  • "early " ' predicting the efficacy of a therapy/treatment of the cancer defined herein may also be at most 1, at most 2. at most 3, at most 4. at most 5, at most 6, at most 7, at most 10, or at most 14 days after (onset of) the therapy/treatment of the cancer defined herein, wherein the shorter periods are preferred. Most preferably, it is envisaged to already monitor the efficacy of a therapy/treatment of said cancer at the day the therapy/treatment was initiated, i.e. once the (amplified) SRC 5 EPHA3, FRK. EPHA5, EPHA8. YES, ABL2, LCK, and/or BLK activity/expression level changes upon said therapy/treatment.
  • "early" predicting the efficacy of a therapy /treatment of the cancer defined herein may also be at most L at most 2, at most 3, at most 4, at most 5, at most 6, at most 7. at most 10. or at most 14 days after diagnosis of the cancer, wherein the shorter periods are preferred. Most preferably, it is envisaged to already predict the efficacy of a therapy/treatment of said cancer at the day of diagnosis.
  • the present invention is particularly useful for monitoring the efficacy of a therapy/treatment of the cancer as defined herein.
  • Corresponding means, uses and methods are provided herein.
  • monitoring the efficacy of a certain kind of therapy/treatment is regularly applied in clinical routine.
  • the skilled person is aware of the meaning of monitoring the efficacy of a certain kind of therapy/treatment.
  • the meaning of the term “monitoring " ' encompasses the meaning of terms like "tracking", '"discovering " ' etc..
  • monitoring the efficacy of a therapy/treatment of NSCLC refers to monitoring whether a subject/patient suffering from said disorder (or being prone to suffering from said cancer) responds at all to a therapy/treatment of said disorder and/or how the course of said respond is (e.g. how fast/slow the respond is and/or to what extent the respond is).
  • the present invention is further useful for predicting the efficacy of a therapy/treatment of the cancer as defined herein.
  • predicting the efficacy of a certain kind of therapy/treatment is highly desired in clinical routine, since it allows for preventing the disorder and/or increasing the efficiency of a therapy/treatment and hence, leads to savings in cost and time and to a higher lifespan/likelihood of survival or of 'Genesung' of the affected patient.
  • the definitions given with respect to the term "efficacy of a therapy/treatment of NSCLC * provided herein apply here, mutatis mutandis.
  • the term "predicting the efficacy of a therapy/treatment of NSCLC for a subject/patient” is used in basically the same sense like determining whether, and/or to what extent, a subject/patient exhibits susceptibility to such therapy/treatment, i.e. whether said subject/patient will or would respond at all to a therapy/treatment of said disorder and/or how the course of said respond will or would be (e.g. how fast/slow the respond is and/or to what extent the respond is).
  • a subject/patient exhibits susceptibility to said cancer in accordance with this invention, when its (amplified) SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK activity/expression level is aberrant.
  • said "amplification” is already “aberrant” as defined herein.
  • the '"predicting the efficacy of a therapy/treatment of the cancer defined herein" in accordance with this invention may be performed after initiation of the therapy/treatment, i.e. during the already ongoing therapy/treatment.
  • said "predicting" may be performed during the herein described monitoring the efficacy of a therapy/treatment of said cancer, preferably early after the beginning of said monitoring.
  • the predicting may be based on results from said monitoring obtained at a certain point in time of the ongoing therapy/treatment.
  • said point in time is an early point in time, like, for example that point in time, when a first result from said monitoring has been obtained.
  • the "predicting the efficacy of a therapy/treatment of the cancer defined herein" is performed during an already ongoing therapy/treatment, it refers to the following/subsequent efficacy of said therapy/treatment.
  • the "predicting the efficacy of a therapy/treatment of the cancer defined herein" in accordance with this invention may be performed (immediately) after diagnosis but, however, prior to initiation of the therapy/treatment.
  • "predicting the efficacy of a therapy/treatment of said cancer” refers to the efficacy of a therapy/treatment which has not yet been initiated (or has been initiated substantially at the same point in time when the "predicting" w r as performed.
  • one non-limiting example of a healthy control subject/patient is one having (a) non-amplified SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK gene(s). This is in contrast to an amplification leading to an aberrant activity/expression of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK, and/or BLK.
  • the amplification of said marker genes as provided herein is considered aberrant.
  • the "reference activity" of SRC is one having (a) non-amplified SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK gene(s).
  • EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK, and/or BLK or the "reference expression level" of said marker gene(s), with respect to the means, methods and uses of monitoring the efficacy of a treatment of a cancer defined herein, is that "reference activity/reference expression level" determined in (a sample of) the corresponding healthy control subject, i.e. is the "normal" activity/expression level.
  • control subject/patient is, in one embodiment, envisaged to be a subject/patient suffering from said cancer or being prone to suffering from said cancer, i.e.
  • a subject/patient having, for example, an aberrant activity/expression level of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK and, hence, not a "normal” activity or "normal expression level" of SRC, EPHA3, FRK, EPHA5, EPPIA8, YES, ABL2, LCK, and/or BLK as described in accordance with this invention.
  • “different” means and comprises “higher” or “lower”, depending on whether the cancer defined and described herein comes along with an up- or down-regulated activity of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK.
  • “different”, “higher” or “lower” means different, higher or lower than the normal (range of) activity of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or
  • “higher” or “lower” means different, higher or lower than the normal (range of) expression level of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK.
  • different, higher or lower means at least 1,5 fold, at least 2 fold, at least 2,5 fold, at least 3 fold, at least 4 fold, at least 5 fold, at least 7 fold, at least 10 fold, at least 15 fold, at least 25 fold, at least 50 fold, at least 100 fold, at least 200 fold different, higher or lower, wherein the higher values are preferred. Whether, in which direction (i.e.
  • LCK and/or BLK differs from Its corresponding reference activity/expression level of SRC, EPHA3, FRK, EPHA5. EPHA8. YES.
  • ABL2, LCK and/or BLK can easily be deduced by the skilled person based on the teaching provided herein and the common general knowledge.
  • control subject/patient is subjected to the same treatment of the cancer subject/patient described and defined herein.
  • Said control subject/patient may be a responder (positive control) or non-responder (negative control) to this treatment.
  • a subject/patient is a "'responder * ' or "non-responder " ' with respect to a certain kind of cancer treatment/therapy can be evaluated by the skilled person on the basis of his common general knowledge and/or the teaching provided herein.
  • a ''responder may be a subject/patient whose cytological/haemato logical parameters and/or (aberrant) SRC, EPHA3, FRK, EPHA5.
  • EPHA5, EPHA8, YES. ABL2, LCK and/or BLK change towards the their "normal" activity/(expression) level(s) (in a sufficient manner) upon the cancer treatment/therapy
  • a "'responder” may be a subject/ patient not suffering from one of the herein defined resistances.
  • a "'non-responder " ' may be a subject/patient whose cytological/haematological parameters and/or (aberrant) activity/expression level of SRC. EPHA3.
  • a '"non-responder may be a subject/patient suffering from one of the herein defined resistances.
  • the patient responds to cancer treatment/therapy, if the activity/expression level of SRC. EPHA3, FRK. EPHA5, EPHA8. YES, ABL2, LCK and/or BLK is reduced upon said treatment/therapy.
  • the expression/ activity of SRC. EPHA3. FRK, EPFIA5, EPHA8. YES, ABL2, LCK and/or BLK is reduced to control expression/activity (e.g. determined in a sample obtained from a person not suffering from said cancer).
  • a reduction in expression/ activity of SRC, EPHA3, FIOC, EPHA5, EPHA8. YES, ABL2. LCK and/or BLK is indicative for a successful treatment/therapy.
  • a skilled person is readily in the position to determine whether a patient responds to cancer treatment/therapy by evaluation of the expression/ activity of SRC.
  • a person skilled in the art may also determine cytological/haematological parameters characteristic for a specific cancer in order to assess whether a patient responds to cancer treatment/therapy.
  • a patient who does not respond to cancer treatment/therapy does not show a reduced expression/ activity of SRC, EPHA3.
  • one non-limiting example of a diseased control subject/patient (responder and/or non-responder) suffering from a cancer defined herein or being prone to suffering from a susceptibility thereto is one having an amplified SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK leading to an aberrant SRC.
  • EPHA5, EPHA8. YES, ABL2. LCK and/or BLK are examples of a diseased control subject/patient (responder and/or non-responder) suffering from a cancer defined herein or being prone to suffering from a susceptibility thereto.
  • the skilled person is aware of how a typical/desired response to a known therapy/treatment of NSCLC should proceed or is intended to proceed. Moreover, the skilled person can consider how a typical/desired response to a (unknown) therapy/treatment of a NSCLC proceeds or is intended to proceed. Based on this knowledge, the means, methods and uses of this invention referring to the efficacy of a therapy/treatment of such a cancer can. for example, also be carried out without employing (a sample of) a particular control subject/patient, i.e. without comparing the activity or expression level of SRC. EPHA3, FRK, EPHA5, EPHA8.
  • the subject/patient is a " 'responder". If the response of a subject/patient is slower than the "typical/desired response", the subject/patient is a "non- responder" (when no substantial response can be seen) or "weak-responder".
  • the efficacy of a cancer treatment/therapy can be determined taking account of the change in the activity/expression level of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK during the treatment/therapy.
  • a skilled person is able to assess the efficacy of a treatment by evaluating the activity/expression level of the above marker genes at various points in time during the treatment (e.g. prior to the treatment, after start of the treatment, and subseqently in intervals during the treatment). In this particular case, it may not be necessary to compare the activity/expression level with reference values (control values) as indicated above in order to assess the efficacy of the treatment. Instead it may suffice to detect the change in the activity/expression level of the marker genes in samples obtained from a treated patient after start of the treatment.
  • a (desired) efficacy of a treatment of a cancer described herein or susceptibility thereto is indicated/predicted, when the aberrant (Le, enhanced or decreased) activity or expression level of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK is shifted back towards the "normal level” of a (healthy)control subject/patient or to "normal level” of a defined responder ("positive control") due to/in consequence of said treatment of the cancer or susceptibility thereto.
  • the efficacy of a treatment of the cancer defined herein is high, when the subject/patient (to be) treated responds as fast (or even faster) and as complete as a "responder", i.e. exhibits a 'typical/desired response".
  • the subject/patient reaches the "normal" level of the relevant cytological/haematological parameters and/or SRC, EPHA3, FRK, EPHA5, EPHA8, YES 5 ABL2, LCK and/or BLK activity (and hence of the coiTcsponding marker gene expression level(s)) of a healthy subject/patient as fast as a "responder", i.e. in the same manner as in a "typical/desired response".
  • the efficacy of a treatment of the cancer is high, if the patient treated shows a "typical/desired response".
  • the efficacy is high, when the activity/expression level of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK in said patient reach a "normal” acivity/level as rapidly as in a "typical/desired response".
  • the efficacy of a treatment of the cancer defined herein is moderate/low, when the subject/patient (to be) treated responds not as fast and/or not as complete as a "responder" ' , i.e. does not exhibit a "typical/desired response " .
  • said subject/patient does not reach the ''normal" level of the relevant cytological/haematological parameters and/or activity of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2 5 LCK and/or BLK (and hence of the corresponding marker gene expression level(s)) of a healthy subject/patient as complete and/or as fast as a "responder", i.e. not in the same manner as in a "typical/desired response".
  • the efficacy of a treatment of the cancer is low, if the patient treated does not show a ''typical/desired response".
  • the efficacy is low, when the activity/expression level of SRC, EPHA3. FRK, EPHA5, EPHA8, YES. ABL2. LCK and/or BLK in said patient reaches a "norma!' " activity/level slower than in a "typical/desired response".
  • EPHA8, YES 5 ABL2, LCK and/or BLK of a "control subject/patient'” can be replaced by a '"own" reference activity or expression level sample of SRC.
  • Such an "own' " reference sample may be obtained prior to (or at the beginning of) the treatment/therapy.
  • the '"control subject/patient would be the subject/patient to be treated itself.
  • the efficacy of the cancer treatment would then be assessed on the basis of how the activity or expression level of SRC, EPHA3, FRIC 5 EPHA5.
  • the efficacy of a treatment of the cancer is assessed in accordance with specific embodiments of this invention, on the basis that the activity/expression level of SRC.
  • FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK is different from a certain or given '"reference activity/reference expression level" of SRC, EPHA3.
  • EPHA8, YES, ABL2, LCK and/or BLK EPHA8, YES, ABL2, LCK and/or BLK.
  • the efficacy of a treatment of the cancer is assessed based on the comparison of the activity/ expression level of SRC, EPHA3, FRK 5 EPHA5, EPHA8, YES, ABL2, LCK and/or BLK in a sample obtained from a patient with a reference (control) activity/expression level.
  • '"different", ''higher” or “lower” means different, higher or lower than the normal (range of) activity/expression level of SRC.
  • LCK and/or BLK means at least 1.5 fold, at least 2 fold, at least 2.5 fold, at least 3 fold, at least 4 fold, at least 5 fold, at least 7 fold, at least 10 fold, at least 15 fold, at least 25 fold, at least 50 fold, at least 100 fold or at least 200 fold different, higher or lower, wherein the higher values are preferred.
  • a certain type of cancer can be associated with increased activity/expression level of any one of the above marker genes or with a decreased increased activity/expression level of any one of the above marker genes. Since a skilled person will be aware of reference acitivity/expression levels of the marker genes (e.g. in a healthy person), he will be readily in the position to determine whether the activity/expression level of any one of the above marker genes is increased or decreased when compared to the reference acitivity/expression level.
  • control subject/patient is a responder.
  • activity/expression level of SRC, EPHA3, FRK, EPHA5, EPHAS. YES, ABL2, LCK and/or BLK is evaluated on the basis of a 'typical/desired response " , a low difference (at a certain point in time) indicates a high efficacy.
  • a responder shows expression/ activity of SRC, EPHA3.
  • EPHA5, EPHA8, YES, ABL2, LCK and/or BLK similar to a typical/desired response, wherein a typical/desired response Is indicative for a successful cancer treatment/therapy.
  • a responder may show reduced or increased expression/ activity of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2. LCK and/or BLK 5 depending on the type of cancer.
  • the cancer is, for example, characterised by a high expression/activity of at least one of the marker genes and if expression/ activity of SRC.
  • FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK, is reduced in a responder to a similar extent as in a typical/desired response, the cancer treatment/therapy can be considered successful.
  • the cancer treatment/therapy can be considered successful if the cancer is characterised by a low expression/activity of at least one of the marker genes and if expression/ activity of SRC, EPHA3, FRK 5 EPHA5, EPHA8, YES, ABL2, LCK and/or BLK, is increased in a responder to a similar extent as in a typical/desired response, the cancer treatment/therapy can be considered successful.
  • control subject/patient is a non-responder
  • activity/expression level of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK is evaluated on the basis of a reference activity/reference expression level of SRC, EPHA3, FRK, EPFIA5, EPHA8, YES, ⁇ BL2, LCK, and/or BLK obtained form the subject to be treated prior to/at the beginning of a therapy/treatment of NSCLC, a high difference (at a certain point in time) indicates a high efficacy.
  • a control sample can be obtained from a non-responder or can be obtained prior to/at the beginning of a therapy /treatment of a cancer. Accordingly, if the difference between expression/ activity of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK in a responder (or similarly in a typical/desired response) compared to said control is high, such a high difference indicates a successful treatment/therap ⁇ * (i.e.
  • a responder shows a reduced expression/ activity of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK compared to high expression/ activity of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK in a control, when the cancer is associated with such a high expression/activity.
  • the difference between the expression/ activity of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK in a responder and the control should be high.
  • EPHA8, YES 5 ABL2, LCK and/or BLK may be taken at the day of diagnosis, once the therapy/treatment is initialed, in between and/or during therapy/treatment, either from the subject/patient to be treated itself or from a corresponding control subject/patient (healthy/re sponder/non ⁇ responder).
  • EPHA5, EPHAS 5 YES 5 ABL2, LCK and/or BLK may be determined at the same or at a different point in time than the activity/expression level of
  • SRC 5 EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK, for example with respect to the course of the therapy/treatment.
  • ABL2, LCK and/or BLK can be determined in a control sample obtained from a patient (healthy tissue) or healthy person at the same time or at a different time when the cancer sample is obtained from said patient.
  • the reference activity/reference expression level of SRC. EPHA3, FRK, EPHA5. EPHA8, YES, ABL2, LCK and/or BLK is obtained from a control subject/patient different from the subject/patient to be treated, it is preferred that the reference activity/reference expression level of SRC. EPHA3, FRIC. EPHA5. EPHA8. YES, ABL2. LCK and/or BLK is determined at the same point in time during therapy/treatment.
  • the reference activity/reference expression level of SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK is obtained from the subject/patient to be treated itself, the reference activity/reference expression level of SRC, EPHA3, FRK, EPHA5. EPHAS 5 YES. ABL2. LCK and/or BLK should be determined at a different point in time during therapy/treatment to allow comparison, for example, at the beginning of (or prior to) the therapy/treatment.
  • activity/expression level of SRC 5 EPHA3. FRK. EPHA5, EPHA8, YES. ABL2. LCK and/or BLK as described herein can be determined once or. preferably, several times. For example, activities/expression levels of SRC. EPHA3, FRK, EPHA5. EPHAS 5 YES, ABL2, LCK. and/or BLK can be determined on a daily, weekly, monthly or yearly basis during therapy/treatment. Commonly, the requirements of corresponding studies would be met, if the frequency of determining activity/expression level of SRC 5 EPHA3, FRK, EPHA5, EPHA8, YES. ABL2, LCK and/or BLK decreases during process of therapy/treatment.
  • Non- limiting examples of schemes of determining activities/expression levels of SRC, EPHA3, FRK, EPFIA5, EPHA8, YES, ABL2, LCK and/or BLK in accordance with this invention are provided herein.
  • the present invention relates to the use of a (transgenic) cell or a (transgenic) non-human animal having at least one amplified marker gene as defined herein for screening and/or validation of a medicament for the treatment of non-small cell lung cancer.
  • the term "cell" as used in this context may also comprise a plurality of cells as well as cells comprised in a tissue.
  • a cell to be used may, for example, be a primary tumor cell.
  • the tumor cell or cell to be used in the screening or validation method may be obtained from samples from a (transgenic) non-human animal suffering from non-small cell lung cancer.
  • the tumor cell or cell may also be obtained from patient samples (e.g.
  • the tumor cell or cell may be a human tumor cell or cell.
  • such a cell to be used in the present screening or validation methods may be comprised in a tissue or tissue sample, like in a sample biopsy.
  • the used non-human animal or cell may be transgenic or non transgenic.
  • Transgenic in this context particularly means that at least one of the marker genes as described or defined herein is over- or under-expressed or has a higher or lower activity. For example, if dasatmib is to be screened and/or validated, it is preferred that such marker genes as SRC. EPHA3, FRK. EPHA5, EPHA8, YES, ABL2, LCK and/or BLK are over-expressed or have a higher activity.
  • Transgenic in this context may also mean that SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK is over- or under-expressed, and/or that the SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK-activity in the transgenic non-human animal or a transgenic cell is enhanced or decreased. It is also envisaged in this context that SRC 5 EPHA3, FRK. EPHA5. EPHA8, YES, ABL2.
  • LCK and/or BLK is under-expressed, and/or that the SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK and/or BLK-activity in the transgenic non-human animal or a transgenic cell is decreased.
  • a preferred (transgenic) non-human animal or (transgenic) cell in context of the invention suffers from NSCLC for the treatment of which the medicament is to be screened and/or validated.
  • the (transgenic) non-human animal or (transgenic) cell is particularly intended to suffer from non-small lung cancer, i.e. to have, for example, an increased SRC, EPHA3, FRK. EPHA5.
  • EPHA8. YES, ABL2, LCK and/or BLK activity and/or increased expression level of, for example, SRC. EPHA3. FRK, EPHA5. EPHA8. YES 5 ABL2, LCK and/or BLK.
  • transgenic non-human animal or “transgenic cell” as used herein refers to a non- human animal or cell, not being a human, that comprises genetic material different from the genetic material of a corresponding wild-type animal/cell
  • ""Genetic material " ' in this context may be any kind of a nucleic acid molecule, or analogues thereof, for example a nucleic acid molecule, or analogues thereof as defined herein.
  • "Different' ' in this context means additional or fewer genetic material with respect Io the genome of the wild-type animal/cell and/or rearranged genetic material, i.e. genetic material present at a different locus of the genome with respect to the genome of the wild-type animal/cell.
  • the (transgenic) non-human animal or (transgenic) cell is or is derived from a mammal.
  • Non-limiting examples of the (transgenic) non-human animal or derived (transgenic) cell are selected from the group consisting of a mouse, a rat, a rabbit, a guinea pig and a Drosophila.
  • the (transgenic) cell in accordance with this invention may be an animal cell, for example, a non-human animal cell.
  • human cells are envisaged to be employed as cells in context of the present invention.
  • such cell may be an embryonic stem cell (ES cell), particularly a non-human animal ES, like, for example, a mouse or rat ES cell.
  • the (transgenic) cell as described herein may also be used for generating the (transgenic) non-human animal as described herein.
  • the ES cell technology for generating transgenic animals is well known in the art and for example is described in Pirity et.al.( Methods Cell Biol, 1998, 57:279).
  • the (transgenic) cell may be a prokaryotic or eukaryotic cell.
  • the (transgenic) cell may be a bacterial, yeast, fungus, plant or animal cell.
  • the transformation or genetically engineering of a cell with a nucleic acid construct or vector can be carried out by standard methods, as for instance described in Sambrook and Russell (2001), Molecular Cloning: A Laboratory Manual, CSH Press, Cold Spring Harbor. NY 5 USA; Methods in Yeast Genetics, A Laboratory Course Manual, Cold Spring Harbor Laboratory Press, 1990.
  • the (transgenic) non-human animal or (transgenic) cell as described or defined in context of this invention is particularly useful in methods for screening and/or validation of a medicament for the treatment of cancers as defined and described herein.
  • These screening methods may, in particular, performed in vivo using, for example, (transgenic) animals as described herein (e.g. rats, mice and the like) and/or animals comprising (a) NSCLC cell(s), (a) tissue(s) or (a) ceil culture(s).
  • Said (a) cell(s), (a) tissue(s) or (a) cell culture(s) may, for example, be obtained/derived from (a) NSCLC tumor cell(s)/tumor(s).
  • said (a) cell(s), (a) tissue(s) or (a) cell culture(s) may be obtained from a subject/patient suffering from a NSCLC.
  • These in vivo screening methods may in particular comprise measuring and determining differences in tumor volume, for example, in the (transgenic) animals described herein above.
  • the present invention also relates to a method for screening and/or validation of a medicament for the treatment of a cancer.
  • Said method may comprise the steps of a) administering to a (transgenic) non-human animal or (transgenic) cell as defined herein said medicament to be screened/validated; b) determining in (a sample from) said animal or cell the activity or expression level of SRC, EPHA3, FRK, EPHA5. EPHA8, YES.
  • LCK and/or BLK gene in accordance with this invention c) comparing the activity or expression level of said at least one marker gene determined in b) with a reference activity or reference expression level of SRC, EPHA3, FRK, EPHA5, EPHA8, YES.
  • ABL2, LCK and/or BLK said activity or said expression level optionally determined in (a sample from) a control (transgenic) non-human animal or (transgenic) cell as defined herein to which said medicament to be screened has not been administered; and d) selecting said medicament when said activity/expression level of SRC.
  • LCK and/or BLK determined in b) differs from said reference activity/expression level determined in c).
  • “'screening and/or validation of medicaments” means, on the one hand, whether a given set of compounds comprises one or more compound(s) that can function as (a) medicament(s). and/or, on the other hand, whether (a) given compound(s) can function as (a) medicament(s). It is particularly intended that the medicaments to be screened and/or ⁇ alidated in context of this invention are medicaments for the treatment, prevention and/or amelioration of a cancer as defined herein.
  • the compound(s)/medicament(s) to be screened and/or validated may be administered to the non-human (transgenic) animal or cell described herein, and. afterwards (for example after a certain period of time sufficient to allow a compound to effect on a cancer as described herein), it is analyzed whether the cancer, or a symptom thereof, of said animal/cell is ameliorated.
  • the present invention also relates to a kit useful for carrying out the method or used of this invention.
  • said kit comprises oligonucleotides or polynucleotides capable of detecting the amplification status of at least one gene selected from the group of SRC, EPHA3, FRK 3 FPHA5 EPH A8. YES. ABL2. LCK and BLK.
  • said kit may comprise (a) compound(s) required for specifically determining the amplification status of at least one gene of SRC. EPHA3, FRK. EPHA5. EPHA8, YES, ABL2. LCK and/or BLK.
  • the kit (to be prepared in context) of this invention is a diagnostic kit.
  • the kit (to be prepared in context) of this invention or the methods and uses of the invention may further comprise or be provided with (an) instruction manual(s).
  • said instruction manual(s) may guide the skilled person (how) to determine amplification status of at least one gene of SRC, EPHA3, FRK. EPHA5, EPHA8, YES. ABL2, LCK and/or BLK., i.e. (how) to diagnose the susceptibility to dasati ⁇ ib.
  • said instruction manual(s) may comprise guidance to use or apply the herein provided methods or uses.
  • the kit (to be prepared in context) of this invention may further comprise substances/chemicals and/or equipment suitable/required for carrying out the methods and uses of this invention.
  • substances/chemicals and/or equipment are solvents, diluents and/or buffers for stabilizing and/or storing (a) compound(s) required for specifically determining the amplification status of at least one gene of SRC, EPHA3.
  • FRK. EPHA5, EPHA8. YES. ABL2, LCK and/or BLK are solvents, diluents and/or buffers for stabilizing and/or storing (a) compound(s) required for specifically determining the amplification status of at least one gene of SRC, EPHA3.
  • FRK. EPHA5, EPHA8. YES.
  • ABL2, LCK and/or BLK are solvents, diluents and/or buffers for stabilizing and/or storing (a) compound(s) required for specifically determining the amplification status of at least one gene
  • the present invention also relates to a combination of cell lines selected from the group consisting of A427, A549, CaIu-I, Calu-3. Calu-6. H1299, H1355, H1395. H1437, H1563. H1568, H1648, H1650, H1666. H1734. H1755. H1770, H1781, H1792. H1819, H1838, H1915. Hl 944. H1975. Hl 993. H2009, H2030, H2052.
  • combinations of cell lines should comprise at least 3, 4, 5, 6. 7. 8 5 9, 10, 11, 12, 13. 14, 15, 20, 25. 30, 35, 40, 45. 50. 55 or 60 of the cell lines as provided herein above.
  • combination of eel! lines should comprise at least 60 cell lines as provided herein above.
  • These cell lines, and in particular their combination are particular useful as model systems for the assessment of any potential drug susceptibility. This usefulness of these specifically selected cell lines (in combination) for the assessment of drug susceptibility is demonstrated in the appended example. All of the mentioned cell lines are available to the person skilled in the ait and the public from cell depositary institutions, in particular ATCC or DMSZ as illustrated in Figure 19 where also corresponding accession numbers for these cells are provided.
  • the combination of cell lines as defined herein above for predicting susceptibility to a drug, in particular, dasatinib is disclosed herein.
  • the combination of cell lines may be useful for predicting the susceptibility to a drug, in particular to dasatinib or responsiveness of a (mammalian) tumor cell or cancer cell to treatment with a drug, in particular dasatinib. It may also be useful in predicting whether a patient is likely to respond to or is sensitive to a drug, in particular dasatinib.
  • Corresponding means and methods for predicting susceptibility/responsiveness and the like are well known in the art and also described herein above. Accordingly, a skilled person will know how to use such a combination of cell lines in this context.
  • the present invention is further described by reference to the following non-limiting figures and examples.
  • FIG. 4 Profiles of aberrations in glioma, melanoma and lung cancer
  • A Chromosomal copy number changes of KSCLC cell lines are plotted against those of primary gliomas. Two separate figures are given for deletions (left panel, NSCLC cell lines in black, gliomas in grey) and amplifications (right panel, NSCLC cell lines in light black, gliomas in dark grey).
  • B Chromosomal copy number changes of NSCLC cell lines are plotted against those of primary melanomas (NSCLC cell lines in red respectively blue as above, melanoma short term cultures in purple).
  • Genomic similarity was analyzed by computing correlations of GlSTIC q-values for each SNP between NSCLC cell lines and the indicated cancer entity primary lung cancer, ovarian cancer, glioma, melanoma cell culture samples, normal tissues and a randomly split subset of NSCLC cell lines.
  • A Hierarchical clustering of primary lung adenocarcinomas was performed using genes identified as being differentially expressed in erlotinib-sensitive versus erlotinib-resistant NSCLC cell lines. Whitebars represent EGFR-mxAsnt tumors.
  • B The EGFR mutation signature published by Choi et al. PLoS ONE 2: el 226 (2007) was used to perform hierarchical clustering of primary lung adenocarcinomas. Red bars represent £GFi?-mutant tumors.
  • Multi-Jesion predictors of sensitivity tested with the KNN method Fisher's exact test and t- test are displayed. Only significant predictors are displayed for two different GLAD thresholds.
  • the half-maximal inhibitory concentrations (y-axis; IC 50 values) for 1 1 compounds are shown for the entire collection of NSCLC cell lines (individual cell lines, x-axis). Due to the fact that rapamycin typically fails to completely abrogate cellular proliferation (O'Reilly et al, 2006) v the 25%-inhibitory concentration is shown for this compound. Bars represent IC 50 respectively IC 25 values (y-axis) throughout the cell line collection (x-axis) ranked according to sensitivity. The maximum concentration is assigned to the IC 50 resp.
  • IC 25 value (10 ⁇ M for 17- AAG, erlotinib, vandetanib, sunitinib and PDl 68393, 30 ⁇ M for SU-11274 and dasatinib. 60 ⁇ M for VX-680, 90 ⁇ M for purvaianol and UO 126) for resistant cell lines.
  • EGFR inhibitors form a distinct subcluster, where EGFR-mutaXed samples show the highest degree of sensitivity.
  • Dasatinib represented as white to dark grey ballandsticks is modeled into the ATP binding site of EGFR.
  • lower panel The T790M mutation at the gatekeeper position of the ATP pocket, associated with secondary EGFR- inhibitor resistance in patients displaces both erlotinib and dasatinib from the ATP binding pocket of the kinase domain.
  • (D) Upper panel: Ba/F3 cells expressing mutant (del Ex 19 or Exl9/T790M) EGFR were treated for 12h with the indicated concentrations of either dasatinib or erlotinib and whole-ceil Iy sales were inimunoblotted for phospho-EGFR and EGFR.
  • Lower panel Dose-dependent growth inhibition after 96h treatment with either dasatinib or erlotinib was assessed measuring cellular ATP content.
  • FIG. 12 Mutated EGFR as a target for vandeta ⁇ ib activity.
  • Vandetanib represented as ballandsticks is modeled into the ATP binding site of EGFR based on its crystallographically determined binding mode with the RET kinase
  • right panel The T790M mutation at the gatekeeper position of the ATP pocket, associated with secondary EGFR-inhibitor resistance in patients displaces the drug from the ATP- binding pocket.
  • the crystographically determined binding mode of erlotinib asballandsticks is shown in both panels as an overlay for reference.
  • B Ba/F3 cells expressing mutant (del ExI 9 or Exl9/T790M) EGFR were treated for 12h with the indicated concentrations of either vandetanib or erlotinib and whole-ceil lysates were inimunoblotted for phospho-EGFR and EGFR.
  • C Dose-dependent growth inhibition after 96h treatment with either dasatinib or erlotinib was assessed measuring cellular ATP content.
  • FIG. 13 Lesion-based prediction for activity of 17- AAG, UO126 and dasatinib.
  • A Distribution of KRAS mutations (black columns) across the 17-AAG-sensitivity profile (IC 50 values) in the NSCLC cell line collection and the NCI-60 cell line panel. Incidence of KRAS mutation and sensitivity towards 17- A AG is represented by a Fisher's exact test for both datasets.
  • B Lysates of a KRAS wildtype (wt) and a KRAS mutated (Gl 2C) cell line treated with 17AAG at different concentrations were imraunoblotted for c-RAF, KRAS, cyclinDl and Akt.
  • C Distribution of copy number gain at 1 q21.3 (black columns) across the UO126-sensitivity profile (IC JO values) in the NSCLC cell line collection and the hypothemycin- sensitivity profile in the NCI-60 cell line panel. Incidence of amplification of Iq21.3 mutation and sensitivity towards 17-AAG is represented by a Fisher's exact test for both datasets.
  • D Cell lines were sorted according to their sensitivity to dasatinib (IC50 ⁇ l ⁇ M; light grey ). Strikingly, most dasatinib-sensitive NSCLC ceil lines are found among those with highest copy number for members of the SRC and Ephrin receptor kinase families.
  • Figure 14 Validation of the target-enriched sensitivity prediction method that yielded genomic predictors of dasatin ⁇ b sensitivity.
  • AU cell lines were sorted according to their sensitivity to erlotinib (IC 5 o ⁇ l ⁇ M; greybars). Cell lines enriched for sensitivity to erlotinib are found among those with highest copy numbers for EGFR.
  • the contingency table for EGFR (dark grey bars) amplification and erlotinib sensitivity including the p-vaiue determined using fisher ' s exact test are displayed in the right panel.
  • FIG. 16 Prostate/breast cancer -gene signature associated with dasatinib sensitivity
  • the expression levels of the respective genes were analyzed by hierarchical clustering with the dasatinib sensitivity denoted using the annotation by 0 and 1.
  • the samples with a similar expression profile across these genes are found in the same subcluster.
  • Bright spots represent genes that are repressed, dark spots represent genes that are overexpressed when compared to average mRNA expression levels.
  • Figure 17 Ail genes associated with dasatinib sensitivity in prostate cancer
  • Figure 18 shows a list of cell lines in the initially tested NSCLC cell line panel, their respective KRAS mutations and the corresponding half-maximal inhibitory concentrations (IC50).
  • Figure 19 shows a set of cell lines which are to be used in accordance with the screening methods provided herein for the identification of dings, which can be used in and -cancer treatment/anti-proliferative treatment.
  • mice expressing a Gl 2D mutation of KRAS specifically in the lung were generated by intranasal application of adenoviral Cre recombinase to Lox-Stop-Lox KRASG12D mice (upper panels from left to right) as described in the literature (Jackson. E. L. et al, Genes Dev 15:3243-3248 (2001 ): Johnson L. et al. Nature. 410(6832):! 1 11-6 (2001) and Ji H. et al.. Nature. 448 (7155):807-10 (2007)). These mice develop lethal lung adenocarcinomas at high penetrance (lower panels from left to right). Note that the images were taken from the ealier publications for illustration purposes only.
  • FIG. 21 Treatment of Los-Stop-Lox KRASG12D mice with HSP90 inhibitor Transgenic mice (as described in Figure 20) with KRAS-mutant lung tumors were treated with the HSP90 inhibitor 17-DMAG for seven days. Tumor volumes were determined by magnetic resonance imaging and shown as transthoracic images (left panels), quantified and changes in tumor volume relative to the pre-therapy images are given in percent.
  • the Example illustrates the invention.
  • Cells were obtained from ATCC (www.atcc.org), DSMZ (www.dsmz.de), from own or from other cell culture collections. Details on all cell lines are listed in Figure 1. This also contains information on providers and on culture conditions. Cells were routinely controlled for infection with my ooplasm by MycoAleit (www.cambrex.com) and were treated with antibiotics according to a previously published protocol (Uphoff and Drexler, 2005) in case of infection.
  • Genomic DNA was extracted from cell lines using the PureGene kit (www.gentra.com) and hybridized to high-density oligonucleotide arrays (Affymetrix. Santa Clara, CA) interrogating 238,000 SNP loci on all chromosomes except Y, with a median intermarker distance of 5.2 kb (mean 12.2 kb; www.affymetrix.com ).
  • Array experiments were performed according to manufacturer's instructions, SNPs were genotyped by the Affymetrix Genotyping Tools Version 2.0 software.
  • SNP array data of primary samples were obtained from the Tumor Sequencing Project (http://www.genome.gov/cancereequencing/).
  • GlSTiC Genomic Identification of Significant Targets in Cancer
  • the analysis was performed computing ratios of observed vs. expected co-occurrence frequency of individual lesions.
  • Hierarchical clustering of mutation data combined to a dichotomized version of quantitative copy number changes was performed using the reciprocal co-occurrence ratio as distance measure with average linkage method.
  • the adequate threshold for occurrence of copy number lesions depends on the overall level, of copy number alteration for that specific lesion, the sum of these ratios for three distinct thresholds was used.
  • genes EGFR, BRAF 1 ERBB2, PIK3CA, KRAS, TP53, STKIl, PTEN and CDKN2A were bidirectionally sequenced following PCR-amplificatioii of all coding exons.
  • Mutation detection for choice of appropriate therapy depending on the respective mutation has been further developed to compensate for the methodological issues connected with sequencing of tumor samples with high admixture of non-tumoral cells, in our laboratory we have therefore developed the following algorithm: if the tumor content of the tumor specimens is higher or equal than 70% estimated by conventional histomorphology, we have found Sanger dideoxy-chain-termination sequencing to be optimal in terms of cost-efficiency and sensitivity. However, when the tumor content is between 70% and 20% we have found conventional pyrosequencing as, for example, implemented in the Biotage instrument, to deliver higher sensitivity and specificity at acceptable costs.
  • next-generation sequencing as for example implemented in the Roche-454 sequencing system (Thomas et al, Nature Medicine JuI; 12(7): 852-5 2006), to be the most sensitive and accurate method in this setting. Together, this algorithm provides high sensitivity in all settings combined with maximum cost- efficiency.
  • Expression arrays Expression data were obtained using Affymetrix Ul 33A arrays from 54 of the cell lines. RNA extraction, hybridization and scanning of arrays were performed using standard procedures (Bhattacharjee et al, 2001 ). CEL files from U133A arrays were preprocessed using the dChip software. We compared the cell lines to primary lung cancer, renal cell carcinomas and lymphoma specimens as well as to the respective cell lines by hierarchical clustering. For comparison with expression profiles from further entities, we used lung cancer (Lu el a!. , 2006).
  • Erlotinib, vandetanib and sunitinib were purchased from commercial suppliers, dissolved in DMSO and stored according to manufacturer's instructions. Cells were plated into sterile microtiter plates using a Multidrop instrument (www.thermo.com) and cultured overnight. Compounds were then added in serial dilutions. Cellular viability was determined after 9 ⁇ h by measuring cellular ATP content using the CellTiter-Glo assay (www.promega.com). Plates were measured on a Mithras LB940 plate reader (www.bertholdtech.com). Half-maximal inhibitory concentrations were determined from the respective preimage under the kill curve, where the latter was smoothed according to the logistic function with the parameters appropriately chosen.
  • multi-lesion predictors of sensitivity were calculated using a KNN algorithm with a leave-one-out strategy (Golub el ai , 1999). where the same choice of samples was used as above for Fisher's exact test: For all but one sample, genetic lesions strongly discriminating between sensitive and resistant cell lines were selected and the KNN algorithm was based on these. The prediction was validated by the remaining left-out sample. The collection of features where this validation had best performance was taken as the best combined predictor to the respective compound.
  • EGFR cDNA was subcloned into pBabe-hygro vectors.
  • the most prevalent NSCLC-derived mutants http://www.sanger.ac.uk/genetics/CGP/cosmic/
  • site-directed mutagenesis Quiick-Change Mutagenesis XL kit; Stratagene, La Jolla, CA, USA
  • virus was packed and produced as previously described (Greulich et al, 2005).
  • Murine Ba/F3 cells were stably transduced with the retroviruses and after IL- 3 withdrawal, independently growing cells were chosen for further experiments.
  • the piperazine moiety of the inhibitor points out of the ATP site into the solvent while the 2-amino-thiazole forms two hydrogen bonds with the hinge region of the kinase (N 3 of the thiazole ring with the amide nitrogen of Met793 , Met318 in AbI) and the 2-amino hydrogen of dasatinib with O of Met793 (Met318 in AbI).
  • An additional hydrogen bond can form between the side chain hydroxy! of the gatekeeper Thr790 (Thr315 in AbI) and the amide nitrogen of the inhibitor.
  • the chloro-melhyl-phenyl ring of dasatinib binds into a hydrophobic pocket near the gatekeeper Thr790 and helix C and would clearly clash with the Met side chain of drug resistant EGFR- T790M.
  • I vandetanib, Nl of the quinazoline scaffold forms one key hydrogen bond to the backbone of the hinge region (Met793 in EGFR, Ala807 in RET kinase).
  • the brorao-fluoro-phenylamine moiety of vandetanib adopts a conformation similar to the ethynyl-phenylamine of erlotinib being close to the side chain of Thr790 in EGFR and Val804 in RET kinase.
  • Figures of the structures were prepared using PyMoI.
  • NP40 lysis buffer 50 mmol/L Tris-HCI (pH 7.4), 150 mmol/L NaCl. 1% NP40
  • Protein concentrations were determined using the Bicinchoninic Acid Protein Assay kit (www.piercenet.com) and equivalent amounts (40-60 ⁇ g) were subjected to SDS- PAGE on 12% gels, except where indicated. Western blotting was done as described previously (Shimamura et al, 2006).
  • Anti-EGFR, anti-phospho-EGFR (Tyr 1068 ) and anti-pAkt antibodies were purchased from Cell Signaling Technology (Beverly. MA).
  • Anti c-raf and anti-cyclin Dl antibody s were purchased from Santa Cruz.
  • Anti KRAS antibody was purchased from Merck. Results
  • NSCLC cell lines were collected from various sources ( Figure 1) and formed the basis for all subsequent experiments.
  • Cell lines were derived from tumors representing all major subtypes of NSCLC tumors, including adenocarcinoma, squamous-cell carcinoma and large- cell carcinoma.
  • Genomic Identification of Significant Targets in Cancer (GlSTIC) to statistically distinguish biologically relevant lesions from background noise (Beroukhim el al., 2007). This method assigns a statistical score to each chromosomal marker reflecting both the mean amplitude and frequency of alterations at a given locus within a data set.
  • HD homozygous deletions
  • LOH loss of heterozygosity
  • EGFR mutations define phenotypic properties of lung tumors in vitro aad in vivo
  • Activated oncogenes typically cause a transcriptional signature that can be used to identify tumors carrying such oncogenes (BiId et al, 2006; Lamb el al . 2003).
  • a transcriptional signature that can be used to identify tumors carrying such oncogenes.
  • EGFR inhibitors such as erlotimb
  • mutations in the EGFR gene Lynch el al, 2004; Paez et al, 2004; Pao et al. 2004.
  • To systematically identify genetic lesions associated with sensitivity to erlotinib we determined erlotinib sensitivity in all cell lines. Then, we analyzed the distribution of genetic lesions in sensitive compared to insensitive cell lines ( Figure 7) and further compared the mean sensitivity of cell lines with and without the respective genetic lesion. In both analyses, EGFR mutations were the best single-lesion predictor of erlotinib sensitivity ( Figure 5D, p ⁇ 0.0001).
  • UO 126 is a MEK inhibitor that also showed enhanced activity in a subset of the lung cancer cell line collection.
  • hypothemycin was used as a MEK inhibitor (Solit et al, 2006).
  • Example 2 Animal data show that mice with KRAS-driven lung adenocarcinomas are susceptible to treatment with an HSP90 inhibitor
  • mice genetically engineered to develop KRAS-driven lung adenocarcinomas are susceptible to treatment with an HSP90 inhibitor.
  • mice earn,' a Lox-Stop-Lox- KRAS_G12D gene.
  • adenoviral Cre Upon administration of adenoviral Cre by nasal inhalation, these mice develop lung cancers with high penetrance, leading to rapid death from the disease.
  • This mouse model therefore represents the most stringent and optimal model of KRAS-mutant human lung cancer and was therefore chosen for in vivo experiments; see Figure 20.
  • Mice received 20 mg/kg/d of 17-DMAG, a geldanamycin HSP90 inhibitor with almost identical structure as 17- AAG.
  • 17-DMAG a geldanamycin HSP90 inhibitor with almost identical structure as 17- AAG.
  • In vitro data confirmed that the biological effects seen with 17-AAG in the cell lines were identical to those seen with 17- DMAG (data not shown).
  • mice After only one week of treatment 2 of 3 mice showed dramatic regression of tumors as measured by MRI imaging; see Figure 21.
  • the third mice showed a slight but insignificant reduction of tumor burden, comparable to stable disease.
  • untreated mice invariably show rapid tumor progression and die quickly from disease.
  • the present invention refers to the following nucleotide sequences:
  • the present invention also provides techniques and methods wherein homologous sequences, and also genetic allelic variants and the like of the concise sequences provided herein are used. Preferably, such "variants' " are genetic variants.
  • SEQ lD No. 1 Nucleotide sequence of Homo sapiens v-abl Abelson murine leukemia viral oncogene homolog 2 (arg, Abelson-related gene) (ABL2), transcript variant c, mRKA (>gijl53266777MNM_001100108.11).
  • SEQ ID No. 2 Nucleotide sequence of Homo sapiens B lymphoid tyrosine kinase (BLK) 5 mRNA (>gi
  • EPH receptor A5 EPH receptor A5
  • transcript variant 1 mRNA (>gi
  • Nucleotide sequence of Homo sapiens v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog (avian) (SRC), transcript variant 1, mRNA (>g ⁇ 38202215
  • SEQ ID No. 9 Nucleotide sequence of Homo sapiens v-yes-1 Yamaguchi sarcoma viral oncogene homolog 1 (YESl), mRNA (>gi
  • SEQ ID No. 10 Nucleotide sequence of Homo sapiens v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS), transcript variant b, mRNA (>gi
  • KRAS Kirsten rat sarcoma viral oncogene homolog
  • transcript variant b mRNA (>gi
  • Basso AD et aLAkt forms an intracellular complex with heat shock protein 90 (Hsp90) and Cdc37 and is destabilized by inhibitors of Hsp90 function. J Biol C hem 277: 39858-66 (2002).
  • Cappuzzo F et al. Epidermal growth factor receptor gene and protein and gefitinib sensitivity in non-small-cell lung cancer. J Nail Cancer Inst 97: 643-55 (2005). Chen CF et al, Molecular genetic evidence supporting a novel human hepatocellular carcinoma tumor suppressor locus at 13ql2.11. Genes Chromosomes Cancer 44: 320-8 (2005).
  • Greshock J et al. Cancer cell lines as genetic models of their parent histology: analyses based on array comparative genomic hybridization. Cancer Res 67: 3594-600 (2007).
  • Greulich H et al. Oncogenic Transformation by Inhibitor-Sensitive and -Resistant EGFR Mutants.
  • McDermott U et al. Identification of genotype -correlated sensitivity to selective kinase inhibitors by using high-throughput tumor cell line profiling. Proc Natl Acad Sci U S A 104: 19936-41 (2007).
  • Neve RM et al. A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes. Cancer Cell 10: 515-27 (2006). Nevins JR et al.. Mining gene expression profiles: expression signatures as cancer phenotypes. Nat Rev Genet 8: 601-9 (2007).
  • EGF receptor gene mutations are common in lung cancers from "never smokers” and are associated with sensitivity of tumors to gefitinib and erlotinib. Proc Natl Acad Sci US A U)U 13306-1 1 (2004).
  • Non-small-cell lung cancer and Ba/F3 transformed cells harboring the ERBB2 G77 ⁇ insV_G/C mutation are sensitive to the dual-specific epidermal growth factor receptor and ERBB2 inhibitor HKI-272. Cancer Res 66: 6487-91 (2006).

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Abstract

La présente invention concerne un procédé permettant de sélectionner (a) une ou des cellules, (a) un ou des tissus ou (a) une ou des cultures cellulaires présentant une susceptibilité au dasatinib. L'invention concerne également une méthode permettant de déterminer la sensibilité d'une cellule tumorale mammifère ou d'une cellule cancéreuse au traitement par dasatinib, En outre, l'invention concerne un procédé in vitro permettant d'identifier un sujet répondant ou un patient sensible au dasatinib, ainsi que les utilisations d'un oligo- ou polynucléotide capable de détecter (an) le status d'amplification d'au moins un gène sélectionné dans le groupe comprenant les gènes SRC, EPHA3, FRK, EPHA5, EPHA8, YES, ABL2, LCK et BLK. La présente invention concerne également une méthode permettant de diagnostiquer un cancer des poumons à grandes cellules ainsi qu'un procédé permettant de contrôler l'efficacité d'un traitement de ce cancer. En outre, l'invention concerne une méthode permettant de prédire l'efficacité d'un traitement du cancer, en particulier, d'un cancer du poumon à grandes cellules. L'invention concerne également, l'utilisation d'un animal non humain (transgénique) ou d'une cellule transgénique présentant au moins un gène marqueur amplifié tel qu'il est défini dans la description, afin de cribler et/ou de valider un médicament pour le traitement de ce cancer, ainsi qu'une trousse permettant de mettre en oeuvre les méthodes susmentionnées.
PCT/EP2009/060641 2008-08-18 2009-08-17 Susceptibilité au dasatinib Ceased WO2010020619A2 (fr)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2562265A1 (fr) 2011-08-22 2013-02-27 Lead Discovery Center GmbH Sensibilité à des inhibiteurs sélectifs de CDK9
WO2013026874A1 (fr) 2011-08-22 2013-02-28 Lead Discovery Center Gmbh Inhibiteurs de cdk9 pour traiter le carcinome de la ligne médiane
EP2600150A3 (fr) * 2011-12-01 2014-03-05 Sysmex Corporation Procédé de détermination de la sensibilité de cellules tumorales au dasatinib et programme informatique
WO2014043628A1 (fr) * 2012-09-14 2014-03-20 Memorial Sloan-Kettering Cancer Center Gènes associés à une sensibilité au dasatinib
WO2021043953A1 (fr) * 2019-09-05 2021-03-11 Pamgene Bv Signatures d'activité kinase pour prédire la réponse de patients atteints d'un carcinome pulmonaire non à petites cellules à un inhibiteur de point de contrôle immunitaire pd-1 ou pd-l1

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US20060019284A1 (en) * 2004-06-30 2006-01-26 Fei Huang Identification of polynucleotides for predicting activity of compounds that interact with and/or modulate protein tyrosine kinases and/or protein tyrosine kinase pathways in lung cancer cells
US20070154915A1 (en) * 2005-11-04 2007-07-05 Johji Inazawa Method for detecting cancer and a method for suppressing cancer
US7908091B2 (en) * 2006-03-17 2011-03-15 Prometheus Laboratories Inc. Methods of predicting and monitoring tyrosine kinase inhibitor therapy

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2562265A1 (fr) 2011-08-22 2013-02-27 Lead Discovery Center GmbH Sensibilité à des inhibiteurs sélectifs de CDK9
WO2013026890A1 (fr) 2011-08-22 2013-02-28 Lead Discovery Center Gmbh Susceptibilité à des inhibiteurs sélectifs de cdk9
WO2013026874A1 (fr) 2011-08-22 2013-02-28 Lead Discovery Center Gmbh Inhibiteurs de cdk9 pour traiter le carcinome de la ligne médiane
EP2600150A3 (fr) * 2011-12-01 2014-03-05 Sysmex Corporation Procédé de détermination de la sensibilité de cellules tumorales au dasatinib et programme informatique
WO2014043628A1 (fr) * 2012-09-14 2014-03-20 Memorial Sloan-Kettering Cancer Center Gènes associés à une sensibilité au dasatinib
US10113200B2 (en) 2012-09-14 2018-10-30 Memorial Sloan-Kettering Cancer Center Genes associated with dasatinib sensitivity
WO2021043953A1 (fr) * 2019-09-05 2021-03-11 Pamgene Bv Signatures d'activité kinase pour prédire la réponse de patients atteints d'un carcinome pulmonaire non à petites cellules à un inhibiteur de point de contrôle immunitaire pd-1 ou pd-l1

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