WO2009106372A1 - Procédé de diagnostic - Google Patents

Procédé de diagnostic Download PDF

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WO2009106372A1
WO2009106372A1 PCT/EP2009/001915 EP2009001915W WO2009106372A1 WO 2009106372 A1 WO2009106372 A1 WO 2009106372A1 EP 2009001915 W EP2009001915 W EP 2009001915W WO 2009106372 A1 WO2009106372 A1 WO 2009106372A1
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jakl
protein
acute lymphoblastic
amino acid
cancer
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PCT/EP2009/001915
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English (en)
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Roberto Foa'
Marco Tartaglia
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Istituto Superiore Di Sanatà
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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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/118—Prognosis of disease development

Definitions

  • the present invention relates to a method of detecting the presence of, or a predisposition towards, cancer in a patient; a method of assaying a product for efficacy in the treatment or prophylaxis of cancer and to a method of treatment or prophylaxis of cancer in a patient.
  • the invention also relates to protein, polypeptide and nucleic acid sequences.
  • Acute lymphoblastic leukemia comprises a biologically heterogeneous group of clonal disorders that originate from the uncontrolled proliferation and expansion of immature lymphoblastic cells and are characterized by an extremely variable clinical outcome (1, 2).
  • substantial progress has been made towards understanding the molecular events contributing to malignant transformation. This has permitted the recognition of relevant prognostic factors and risk stratification, and has favored the implementation of therapeutic approaches based on cytogenetic and molecular lesions (2-5).
  • long-term survival in adults with ALL remains largely unsatisfactory, making the design of novel anti-leukemic drugs tailored to specific biological targets a current priority (4, 6).
  • JAK proteins associate constitutively with a variety of cytokine receptors lacking intrinsic kinase activity, and promote signal flow by phosphorylating tyrosyl residues of activated receptors to allow the recruitment and activation of STAT proteins. They share a complex multidomain structure characterized by a tyrosine kinase domain at the C-terminus, which is flanked by a catalytically inactive pseudokinase domain with regulatory function.
  • JAK family members preferentially associate with a diverse subset of cytokine receptors, each differentially expressed by individual cell lineages and tissues, facilitating specificity in function (9).
  • JAKl plays an essential and non-redundant role in mediating biological responses induced by a specific subgroup of cytokines controlling lymphoid cell precursor development (10).
  • Jakl '1' mouse pups exhibit a thymus markedly reduced in size, which is associated with a severe decrease in cellularity. Jakl loss of function is also associated with profound abnormalities in the B- cell compartment due to a block in differentiation at the pro-B/pre-B cell transition step, resulting in a deficit in the production of mature B lymphocytes (10).
  • the present invention is based on the finding that somatic activating JAKl mutations occur among adults with T-cell precursors ALL, and are associated with poor response to therapy and overall prognosis. This supports the view that upregulation of JAKl signaling contributes to malignancies of the lymphoid lineage.
  • a method of detecting the presence of, or a predisposition towards, cancer in a patient comprising detecting the presence of a mutant variant of the JAKl gene in a sample obtained from the patient, wherein the presence of the mutant variant is indicative of the presence of, or a predisposition towards, cancer in the patient.
  • the sample may be a sample of blood, tissue, bronchial lavage fluid or the like.
  • the mutant variant of the JAKl gene may be detected directly, such as by probing for the presence of DNA having the mutant variant, or may be detected indirectly by examining the RNA in the sample transcribed from the gene or the expressed protein itself.
  • the presence of a mutant variant of the protein can be detected by using antibodies specific for the mutant variant which have reduced affinity for the wild-type protein.
  • kits for detecting cancer, or a predisposition towards cancer, in a sample from a patient comprising means to detect the presence of a mutant variant of the JAKl gene in the sample.
  • the kit is used as described above.
  • the means may comprise, for example: opportunely selected primer pairs to amplify the JAKl coding sequence allowing these amplified genomic fragments to be sequenced or analyzed with other screening methods to identify disease-associated mutations; a nucleic acid probe capable of hybridising with the mutant variant of the JAKl gene or an antibody capable of binding to the protein encoded by the mutant variant.
  • the kit may also comprise a label attached to the probe or antibody and, optionally, instructions for use of the kit.
  • the JAKl gene comprises a nucleic acid sequence encoding a protein having at least 80% identity to SEQ. ID NO: 2. Preferably there is at least 90%, 95%, 99% or, most preferably 100% identity to SEQ ID NO:2.
  • the mutant variant of the JAKl gene comprises a nucleic acid sequence encoding a protein having at least 80% identity to SEQ. ID NO: 2 and in which at least one of the amino acid residues at the following positions is substituted with an alternative amino acid: 204, 512, 634, 653, 724, and 879. More preferably there is at least 90%, 95%, 99% or, most preferably 100% identity to SEQ ID NO:2 (aside from the substituted amino acid).
  • the mutant variant of the JAKl gene comprises a nucleic acid sequence encoding a protein having at least 80% identity to SEQ. ID NO: 2 and in which the protein has at least one amino acid substitution selected from the group consisting of: Lys204Met, Ser512Leu, Ala634Asp, Leu653Phe, Arg724His, Arg879Ser, Arg879Cys, Arg879His and combinations thereof. More preferably there is at least 90%, 95%, 99% or, most preferably 100% identity to SEQ ID NO:2 (aside from the substituted amino acid).
  • the patient is an adult (i.e. over the age of 16 years).
  • the cancer is leukaemia such as acute lymphoblastic leukaemia, in particular, B-cell precursor acute lymphoblastic leukaemia or, more frequently, T-cell precursor acute lymphoblastic leukaemia.
  • leukaemia such as acute lymphoblastic leukaemia, in particular, B-cell precursor acute lymphoblastic leukaemia or, more frequently, T-cell precursor acute lymphoblastic leukaemia.
  • a method of assaying a product for efficacy in the treatment or prophylaxis of cancer comprising determining the kinase activity of the mutant variant of the JAKl protein, or activation level of signalling pathways modulated by JAKl protein, in the presence and absence of the product wherein a reduced kinase activity of the mutant variant, or a reduced activation of signalling pathways modulated by the JAKl protein, in the presence of the product as compared with kinase activity in the absence of the product, or with the activation level of signalling pathways modulated by the JAKl protein, is indicative that the product is efficacious in the treatment or prophylaxis of cancer.
  • determining the kinase activity, or more generally JAKl functional upregulation comprises determining at least one of: the level of STATl, AKT and ERK phosphorylation.
  • the JAKl protein is present in a cell during the assay process.
  • the JAKl protein comprises a sequence having at least 80% identity to SEQ. ID NO: 2. Preferably there is at least 90%, 95%, 99% or, most preferably 100% identity to SEQ ID NO:2.
  • the mutant variant of the JAKl protein comprises a sequence having at least 80% identity to SEQ. ID NO: 2 and in which at least one of the amino acid residues at the following positions is substituted with an alternative amino acid: 204, 512, 634, 653, 724, and 879. More preferably there is at least 90%, 95%, 99% or, most preferably 100% identity to SEQ ID NO:2 (aside from the substituted amino acid).
  • the mutant variant of the JAKl protein comprises a sequence having at least 80% identity to SEQ. ID NO: 2 and in which the protein has at least one amino acid substitution selected from the group consisting of: Lys204Met, Ser512Leu, Ala634Asp, Leu653Phe, Arg724His, Arg879Ser, Arg879Cys, Arg879His and combinations thereof. More preferably there is at least 90%, 95%, 99% or, most preferably 100% identity to SEQ ID NO:2 (aside from the substituted amino acid).
  • the cancer is acute lymphoblastic leukaemia, in particular, T-cell precursor acute lymphoblastic leukaemia or B-cell precursor acute lymphoblastic leukaemia.
  • a method of treatment or prophylaxis of cancer in a patient comprising administering to the patient a therapeutically effective quantity of a product that inhibits the activity of the JAKl protein.
  • the JAKl protein comprises a sequence having at least 80% identity to SEQ. ID NO: 2. Preferably there is at least 90%, 95%, 99% or, most preferably 100% identity to SEQ ID NO:2.
  • the product inhibits the signalling activity of the JAKl protein.
  • the product inhibits the kinase activity of the JAKl protein.
  • the product inhibits the expression of the JAKl protein.
  • the cancer is acute lymphoblastic leukaemia, in particular, T-cell precursor acute lymphoblastic leukaemia or B-cell precursor acute lymphoblastic leukaemia.
  • An exemplary product that inhibits the activity of the JAKl protein is a molecule that mimics a JAKl substrate and binds to the active site blocking the catalytic activity of the protein, or is an antibody or an antigen-binding fragment thereof that binds specifically to the protein. It is particularly preferred that the antibody is specific for an epitope of the protein that incorporates the substituted amino acid, thereby being specific for the mutant variant of the JAKl protein.
  • the product that inhibits the activity of the JAKl protein may comprise part of a pharmaceutical composition which also comprises a pharmaceutically acceptable carrier, diluent or excipient (see Remington's Pharmaceutical Sciences in US Pharmacopoeia, 1984, Mack Publishing Company, Easton, PA, USA).
  • a pharmaceutically acceptable carrier diluent or excipient
  • the dose required for a patient may be determined using methods known in the art, for example, by dose-response experiments.
  • the product may be administered by a range of routes, for example, orally or parenterally.
  • an isolated protein comprising a sequence having at least 80% identity to SEQ. ID NO. 2 and wherein at least one of the amino acid residues at the following positions is substituted with an alternative amino acid: 204, 512, 634, 653, 724, and 879. More preferably there is at least 90%, 95%, 99% or, most preferably 100% identity to SEQ ID NO:2 (aside from the substituted amino acid).
  • the protein comprises an amino acid substitution selected from the group consisting of: Lys204Met, Ser512Leu, Ala634Asp, Leu653Phe, Arg724His, Arg879Ser, Arg879Cys, Arg879His and combinations thereof.
  • an isolated polypeptide comprising a fragment of the isolated protein of the invention, wherein the fragment is at least 8 amino acids long and includes the substituted amino acid residue. It is preferred that the fragment is at least 9, 10, 1 1, 12, 14, 16 or 20 amino acids long.
  • an isolated nucleic acid sequence comprising a sequence encoding the isolated protein or the isolated polypeptide of the invention.
  • ALL means "acute lymphoblastic leukaemia”.
  • CM conditional medium
  • Detecting the presence of cancer, or a predisposition towards cancer includes: providing a diagnosis of cancer; assessing the progression of the condition after initial diagnosis; monitoring the response of the condition to a treatment; and establishing the extent of a patient's condition (i.e. staging). For example, by making a quanitative assessment of the proportion of cells in a patient's sample that contain a mutant variant of the JAKl gene, the relative advancement of the cancer may be determined. The term also includes detecting one step of the molecular events causing malignancy.
  • DHPLC means "denaturing high performance liquid chromatography”.
  • ERK means "extracellular signal-regulated kinase”.
  • the percentage "identity" between two sequences is determined using the BLASTP algorithm version 2.2.2 (Altschul, Stephen F., Thomas L. Madden, Alejandro A.
  • “Mutant variant” when referring to a protein or nucleic acid sequence means a protein or nucleic acid sequence having at least one amino acid or nucleotide deletion, insertion or substitution as compared with a wild type sequence.
  • “Prophylaxis” includes lowering the incidence of a condition as well as complete prevention thereof.
  • Treatment includes remission or reduction of symptoms and alleviation of the condition as well as complete eradication of the condition.
  • Figure IA depicts representative electropherograms showing the occurrence of somatically acquired JAKl mutations in subjects with T-ALL. In all cases, mutations were observed at diagnosis (above), but were undetectable during remission (below).
  • Figure IB shows JAKl domain structure and location of affected residues.
  • the predicted amino acid substitutions resulting from the JAKl mutations are positioned below the cartoon of the protein with its functional domains indicated (left) and shown in JAKl three-dimensional modeled structure (right).
  • Figure 1 C depicts electropherograms showing the occurrence of mutations in a fraction of leukemic cells of two individuals with T-ALL.
  • the mutant allele constituted only a portion of the amplified fragment from BM obtained at diagnosis (blasts >70% of total cells) (above).
  • the heterozygous status of each subject for an intragenic polymorphic site (below) is shown for comparison.
  • Figure 2 A shows STATl phosphorylation assays. Basal and IFN- ⁇ -stimulated endogenous STATl phosphorylation in JAKl -defective U4C cells transiently transfected with wild type (WT) JAKl or selected mutants was studied. Blots are representative of three experiments performed.
  • FIG. 2B shows the results of STATl activation assays. Basal and IFN- ⁇ -stimulated endogenous STATl transcriptional activity in JAKl -defective U4C cells transiently cotransfected with p-GAS-Luc and phRL-TK constructs, and wild type (WT) JAKl or a mutant allele was studied.
  • STATl -induced luciferase gene expression levels were determined by measuring the luciferase activity (counts per second, CPS) normalized to the activity of the Renilla luciferase, using a dual luciferase reporter assay system. Activity ratios are expressed as averages of three replicates ⁇ SD.
  • FIG. 2C shows Ba/F3 survival assays. Wild type (WT) or mutant JAKl transduced Ba/F3 cells were grown in absence of IL-3 (left) or with 0.5% or 5% WEHI-3B cell conditional medium (CM) as source of IL-3 (right). Cell numbers (average of three replicates ⁇ SD) were counted at the indicated time points (left) or at day 3 of culture (right).
  • WT Wild type
  • CM cell conditional medium
  • FIG. 2D shows Stat5, Akt and Erk phosphorylation assays.
  • Endogenous Stat5 Tyr694, Akt Ser473 and Erk 1/2 Thr202/Tyr204 phosphorylation levels from lysates of Ba/F3 cells transduced with wild type (WT) Jakl or a mutant form and cultured without IL-3 (left) or with 2% WEHI-3B cell CM as the source of IL-3 (right) were studied.
  • Activation of Stat5 pStat5/Stat5
  • AKT pAkt/Akt
  • Erkl/2 pErk/Erk
  • FIG. 3 A shows supervised hierarchical clustering of gene expression profiles performed on blasts from 16 adult T-ALL patients, with or without a JAKl mutation.
  • Figure 3B depicts Kaplan-Meier estimates of disease-free survival (DFS) (above) and overall survival (OS) (below) in subjects with (gray) or without (black) a JAK mutation.
  • DFS disease-free survival
  • OS overall survival
  • Multivariate analysis confirmed the statistical significance of the reduced DFS and OV among JAKl mutation-posive patients, and excluded a significant contribution of the more advanced age of these subjects.
  • Figure 4 shows JAKl domain structure and location of JAKl, JAK2 and JAK3 residues reported to be mutated in myeloproliferative disorders and leukemias.
  • the predicted amino acid substitutions are positioned below the cartoon of the JAKl protein with its functional domains indicated above (JAKl, top; JAK2, middle; JAK3, bottom) (left) and in its three-dimensional modeled structure (JAKl, Ser512, Lys204, Arg724, Ala634 and Arg879; JAK2 Lys607, Leu ⁇ l l and Val617; JAK3 Prol32, Val722 and Ala572) (right).
  • Figure 5 shows gene expression profiles of JAKl mutation-positive and mutation- negative blasts from adult subjects with T-ALL. Unsupervised hierarchical clustering was performed on blasts from 16 adult T-ALL patients, with or without a JAKl mutation. Relative expression levels are shown according to the reported color scale diagram (below). Clustering based on 1345 probe sets grouped the expression profiles of the 5 JAKl mutation-positive patients (light gray at top of figure) into 2 distinct clusters.
  • Figure 6 shows GFP expression levels of purified Ba/F3 cells transduced with the pMX-JAKl -IRES-GFP bicistronic retroviral vector coding for wild type (WT) Jakl or one of the three generated mutants.
  • SEQ. ID NO: 1 is the wild type nucleotide sequence of the JAKl gene held under GenBank Accession no. NM_002227.2. The protein coding sequence is also shown.
  • SEQ. ID NO: 2 is the protein sequence encoded by the nucleotide sequence of SEQ. ID NO: 1.
  • SEQ. ID NO: 3 is another wild type protein sequence of JAKl, which has 99.4% identity to SEQ. ID NO: 2.
  • Table IV List of the oligonucleotide sequences used to amplify the entire coding sequence of the JAKl gene (exons 1 to 24).
  • Lysed samples were analyzed by 10% SDS-PAGE, transferred to a polyvinylidene difluoride membrane (Pierce) and probed with anti-phospho-Statl (9171, Cell Signaling), anti- Statl (9172, Cell Signaling) and anti-Jakl (3332, Cell Signaling) antibodies. Endogenous STATl transcriptional activity was assessed by luciferase transactivation assays in cells co-transfected with a p-GAS-Luc construct, switched to serum-starvation medium (8 h) and then stimulated with IFN- ⁇ (1000 units/ml, 16 h) or left unstimulated.
  • IFN- ⁇ 1000 units/ml, 16 h
  • STATl -induced luciferase expression was assessed and normalized using a dual luciferase reporter assay system (Promega) and a phRL-TK plasmid constitutively expressing the Renilla luciferase.
  • Each of the three leukemia-associated mutations was also introduced in the murine JAKl cDNA cloned in the bicistronic retroviral vector pMX-IRES-GFP. Constructs were transfected into Phoenix or BOSC packaging cells to produce retroviruses, and murine Ba/F3 (mantained in RPMI- 1640 medium containing 10% FCS, 1% L- glutamine and 10% WEHI-3B cell CM) cells were infected with retroviral supernatants. GFP-positive populations were purified by flow-cytometric sorting and then expanded. Equal GFP expression levels of transduced cells were confirmed by FACS analysis (Figure 6).
  • Transduced Ba/F3 cells were cultured in the absence or presence of IL-3 (0.5% or 5% WEHI-3B cell CM) for assaying cytokine independence and response, and viable cells were counted by trypan-blue exclusion.
  • IL-3 0.5% or 5% WEHI-3B cell CM
  • viable cells were counted by trypan-blue exclusion.
  • Ba/F3 cells were starved in RPMI- 1640 medium containing 1% BSA (5 h) and then stimulated with 2% WEHI-3B cell CM (30 min) or left unstimulated.
  • kinase domain (residues 876-1 153) was modeled by homology to the crystallographic structures of the following kinase domains obtained from the Protein Data Bank (pdb): JAK2 (pdb code 2b7a, identity 54%), JAK3 (pdb code lyvj, identity 51%), FGFRl (pdb code lfgi, identity 35%) and FGFR2 (pdb code loec, identity 34%).
  • Residues 604-852 of the pseudokinase domain were modeled by homology to the structures of the kinase domains of RET (pdb code 2ivt, identity 26%) and ABL (pdb code lfpu, identity 24%).
  • the SH2 domain was modeled by homology to the structures of the following SH2 domains: GRBlO (pdb code lnrv, identity 23%), LCK (pdb code llkk, identity 21%), HCK (pdb code lqcf, identity 20%) and SHP-2 (pdb code 2shp, C-terminal SH2 domain, identity 16%).
  • the FERM domain (residues 34- 420) was modeled by homology to the structures of the FERM domains of the following proteins: ezrin (pdb code Ini2, identity 12%), radixin (pdb code Ij 19, identity 14%), moesin (pdb code Ie5w, identity 12%) and focal adhesion kinase (pdb code 2al6, identity 15%).
  • ezrin pdb code Ini2, identity 12%
  • radixin pdb code Ij 19, identity 14%)
  • moesin pdb code Ie5w, identity 12%)
  • focal adhesion kinase (pdb code 2al6, identity 15%).
  • a possible relative orientation of the four domains was obtained by superimposing them on a complete model of JAK2 (31).
  • RNA quality profiling was checked by agarose gel electrophoresis and spectrophotometry. HGU 133 Plus 2.0 gene chips (Affymetrix) were used to determine gene expression profiles. The detailed protocol for sample preparation and microarray processing is available from the.
  • Oligonucleotide microarray analysis and gene expression data were performed using the dChip software (www.dchip.org) (33), which utilizes an invariant set normalization method where the array with median overall intensity is chosen as the baseline for normalization. Model-based expressions were computed for each array and probe set using only perfect match probes. For unsupervised analysis, non-specific criteria included the requirement for individual gene expression levels to be higher than 100 in at least 20% of samples, and for the ratio of SD to the mean expression across samples to be included between 0.8 and 1000. Analysis of Variance (ANOVA) with .P-value ⁇ 0.001 was performed to compare profiles obtained from T-ALL patients with or without a JAKl mutation.
  • ANOVA Analysis of Variance
  • DPLC denaturing high performance liquid chromatography
  • Table I List of non-synonymous JAKl changes identified in subjects with acute lymphoblastic leukemia.
  • genotyping of genomic DNAs from BM obtained during remission demonstrated the somatic origin of the 1535OT (Ser512Leu), 1901C>A (Ala634Asp) and 2171G>A (Arg724His) changes in the leukemic clones (Fig. IA and Table I).
  • missense changes 184A>G (Ile62Val) and 1078C>T (Arg360Trp), were deemed non-pathogenic variants, as they were observed in non-leukemic cells of affected patients or in unaffected control subjects.
  • JAKl gene mutations occur in ALL and are more frequently observed among adult individuals with involvement of the T-cell lineage.
  • JAKl mutations To examine the effects of the identified mutations on protein function, wild type JAKl or a mutant form (A634D, R724H and R879C) was expressed transiently in JAKl- defective human fibrosarcoma U4C cells, and endogenous STATl phosphorylation was compared basally and following stimulation with IFN- ⁇ (Fig. 2A). Consistent with previous studies (10, 11), untransfected cells lacking functional JAKl did not exhibit STATl phosphorylation in response to IFN- ⁇ . All JAKl mutants promoted an enhanced response to the ligand compared to wild type JAKl .
  • CM WEHI-3B cell conditional medium
  • Jakl mutants conferred IL-3 -independent growth to cells, whereas cells expressing wild type Jakl or the R879C Jakl mutant retained dependence on the cytokine for survival.
  • cells expressing each of the three mutants exhibited enhanced growth in response to IL-3.
  • Ba/F3 cells expressing the A634D Jakl mutant exhibited enhanced Stat5, Akt and Erk phosphorylation basally and following stimulation, while a higher phosphorylation level of these signal transducers in cells expressing the R724H Jakl protein was observed in cultures maintained in presence of IL-3 (Fig. 2D).
  • JAKl ' s SH2 domain does not function as phosphotyrosyl-binding domain but rather plays a structural role in stabilizing the conformation of the FERM domain (21), which mediates its association to cytokine receptors and exerts an as-yet- uncharacterized restraint on catalytic function (22, 23).
  • the molecular mechanism through which these mutations affect JAKl function remains to be explained. Structural and functional consequences were not obvious for the activating changes affecting residues Arg 724 and Arg ,879
  • ISG15, ISGF3G, IFI44L and IRFl were overrepresented in all the JAKl mutation- positive subjects, further supporting the gain of function role of the ALL-associated JAKl lesions.
  • JAK2 and JAK3 Functional upregulation of two members of the JAK family, JAK2 and JAK3, has recently been discovered in myeloproliferative disorders and other malignancies of the myeloid lineage (18-20, 24).
  • the JAK2 V617F amino acid change occurs in the majority of polycythemia vera cases and in approximately 50% of individuals with essential thrombocythemia or idiopathic myelofibrosis.
  • the available data support the view that this recurrent change, which affects the pseudokinase domain of the protein, induces constitutive activation of the kinase and hypersensitivity to cytokines.
  • JAKl is expressed widely and participates in intracellular signaling elicited by class II cytokine receptors and receptors that utilize the gpl30 or ⁇ c receptor subunit. While the hematopoietic defects in Jakl '1' mice were restricted to the lymphoid cell compartment as a result of an impaired response to IL-7 (10) and the present findings indicate a cell- context dependence of somatically acquired JAKl mutations' contribution to leukemogenesis, this kinase may be involved in other malignancies. A concomitant genetic event, including a mutation affecting other members of the JAK family, may synergize with the JAKl defect to promote aberrant cell proliferation and/or survival in a cell-specific context.
  • the experimental data disclosed herein provide the first evidence that JAKl gene defects are associated with a poor response to therapy, frequent relapse of the disease and reduced overall survival, identifying such mutations as a novel informative prognostic marker occurring in a sizable proportion of adult T-ALL.
  • the data also enable the development of therapeutic approaches tailored at interfering with JAKl signaling, expression and kinase activity.
  • JAK-STAT signaling from interferons to cytokines. J. Biol. Chem. 282:20059-20063.
  • Interleukin-9 is a major anti-apoptotic factor for thymic lymphomas. Blood 85:1300-1305.
  • JAK2 Janus kinase 2
  • J AK2 Janus kinase 2
  • Jakl SH2 domain does not fulfill a classical SH2 function in Jak/STAT signaling but plays a structural role for receptor interaction and up-regulation of receptor surface expression. J. Biol. Chem. 280:25760-25768.
  • the JAK2V617F activating mutation occurs in chronic myelomonocytic leukemia and acute myeloid leukemia, but not in acute lymphoblastic leukemia or chronic lymphocytic leukemia.

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Abstract

L'invention porte sur un procédé de détection de la présence d'un cancer ou d'une prédisposition envers un cancer chez un patient. Le procédé comprend la détection de la présence d'un variant mutant du gène JAK1 dans un échantillon obtenu à partir du patient. La présence du variant mutant est indicative de la présence du cancer ou d'une prédisposition envers un cancer chez le patient.
PCT/EP2009/001915 2008-02-29 2009-02-27 Procédé de diagnostic WO2009106372A1 (fr)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015023866A1 (fr) * 2013-08-15 2015-02-19 The Regents Of The University Of Michigan Méthodes et biomarqueurs pour la détection et le traitement de la leucémie à lymphocytes t matures
WO2015184061A3 (fr) * 2014-05-28 2016-01-21 Dana-Farber Cancer Institute, Inc. Activation de biomarqueurs de la jak prédictifs de réponse à un anti-corps inhibiteur de point de contrôle immunitaire

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