*US07932031B2*
  US007932031B2                                 
(12)United States Patent(10)Patent No.: US 7,932,031 B2
 Lee et al. (45) Date of Patent:Apr.  26, 2011

(54)Methods for determining sensitivity to microtubule-stabilizing agents comprising ixabepilone by measuring the level of estrogen receptor 1 
    
(75)Inventors: Hyerim Lee,  Princeton, NJ (US); 
  Peter Shaw,  Yardley, PA (US); 
  Edwin A. Clark,  Pennington, NJ (US) 
(73)Assignee:Bristol-Myers Squibb Company,  Princeton, NJ (US), Type: US Company 
(*)Notice: Subject to any disclaimer, the term of this patent is extended or adjusted under 35 U.S.C. 154(b) by 0 days. 
(21)Appl. No.: 11/906,248 
(22)Filed: Oct.  1, 2007 
(65)Prior Publication Data 
 US 2008/0318228 A1 Dec.  25, 2008 
 Related U.S. Patent Documents 
(63) .
Continuation of application No. 11/289,102, filed on Nov.  29, 2005, now abandoned .
 
(60)Provisional application No. 60/631,993, filed on Nov.  30, 2004.
 
(51)Int. Cl. C12Q 001/68 (20060101)
(52)U.S. Cl. 435/6
(58)Field of Search  None

 
(56)References Cited
 
 U.S. PATENT DOCUMENTS
 5,569,588  A  10/1996    Ashby et al.     
 2002//0051978  A1  5/2002    Roth et al.     
 2003//0166223  A1  9/2003    Sims et al.     

 
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     * cited by examiner
 
     Primary Examiner —Laura B Goddard
     Art Unit — 1642
     Exemplary claim number — 1
 
(74)Attorney, Agent, or Firm — Paul D. Golian

(57)

Abstract

Biomarkers that are useful for identifying a mammal that will respond therapeutically or is responding therapeutically to a method of treating cancer that comprises administering a microtubule-stabilizing agent. In one aspect, the cancer is breast cancer, and the microtubule-stabilizing agent is an epothilone or analog or derivative thereof, or ixabepilone.
2 Claims, 10 Drawing Sheets, and 10 Figures


[0001] This application claims the benefit of U.S. Provisional Application No. 60/631,993 filed Nov. 30, 2004, and is a continuation of U.S. Non-Provisional application Ser. No. 11/289,102 filed Nov. 29, 2005 now abandoned, whose contents are hereby incorporated by reference in there entirety.

SEQUENCE LISTING

[0002] A compact disc labeled “Copy 1” contains the Sequence Listing as 10338 NP.ST25.txt. The Sequence Listing is 1686 KB in size and was recorded Nov. 29, 2005. The compact disk is 1 of 2 compact disks. A duplicate copy of the compact disc is labeled “Copy 2” and is 2 of 2 compact discs.
[0003] The compact disc and duplicate copy are identical and are hereby incorporated by reference into the present application.

FIELD OF THE INVENTION

[0004] The present invention relates generally to the field of pharmacogenomics, and more specifically to methods and procedures to determine drug sensitivity in patients to allow the identification of individualized genetic profiles which will aid in treating diseases and disorders.

BACKGROUND OF THE INVENTION

[0005] Cancer is a disease with extensive histoclinical heterogeneity. Although conventional histological and clinical features have been correlated to prognosis, the same apparent prognostic type of tumors varies widely in its responsiveness to therapy and consequent survival of the patient.
[0006] New prognostic and predictive markers, which would facilitate an individualization of therapy for each patient, are needed to accurately predict patient response to treatments, such as small molecule or biological molecule drugs, in the clinic. The problem may be solved by the identification of new parameters that could better predict the patient's sensitivity to treatment. The classification of patient samples is a crucial aspect of cancer diagnosis and treatment. The association of a patient's response to a treatment with molecular and genetic markers can open up new opportunities for treatment development in non-responding patients, or distinguish a treatment's indication among other treatment choices because of higher confidence in the efficacy. Further, the pre-selection of patients who are likely to respond well to a medicine, drug, or combination therapy may reduce the number of patients needed in a clinical study or accelerate the time needed to complete a clinical development program (M. Cockett et al., Current Opinion in Biotechnology, 11:602-609 (2000)).
[0007] The ability to predict drug sensitivity in patients is particularly challenging because drug responses reflect not only properties intrinsic to the target cells, but also a host's metabolic properties. Efforts to use genetic information to predict drug sensitivity have primarily focused on individual genes that have broad effects, such as the multidrug resistance genes, mdr1 and mrp1 (P. Sonneveld, J. Intern. Med., 247:521-534 (2000)).
[0008] The development of microarray technologies for large scale characterization of gene mRNA expression pattern has made it possible to systematically search for molecular markers and to categorize cancers into distinct subgroups not evident by traditional histopathological methods (J. Khan et al., Cancer Res., 58:5009-5013 (1998); A. A. Alizadeh et al., Nature, 403:503-511 (2000); M. Bittner et al., Nature, 406:536-540 (2000); J. Khan et al., Nature Medicine, 7(6):673-679 (2001); T. R. Golub et al., Science, 286:531-537 (1999); U. Alon et al., P. N. A. S. USA, 96:6745-6750 (1999)). Such technologies and molecular tools have made it possible to monitor the expression level of a large number of transcripts within a cell population at any given time (see, e.g., Schena et al., Science, 270:467-470 (1995); Lockhart et al., Nature Biotechnology, 14:1675-1680 (1996); Blanchard et al., Nature Biotechnology, 14:1649 (1996); U.S. Pat. No. 5,569,588 to Ashby et al.).
[0009] Recent studies demonstrate that gene expression information generated by microarray analysis of human tumors can predict clinical outcome (L. J. van't Veer et al., Nature, 415:530-536 (2002); T. Sorlie et al., P. N. A. S. USA, 98:10869-10874 (2001); M. Shipp et al., Nature Medicine, 8(1):68-74 (2002); G. Glinsky et al., The Journal of Clin. Invest., 113(6):913-923 (2004)). These findings bring hope that cancer treatment will be vastly improved by better predicting the response of individual tumors to therapy.
[0010] Needed are new and alternative methods and procedures to determine drug sensitivity in patients to allow the development of individualized genetic profiles which are necessary to treat diseases and disorders based on patient response at a molecular level.

SUMMARY OF THE INVENTION

[0011] The invention provides methods and procedures for determining patient sensitivity to one or more microtubule-stabilizing agents. The invention also provides methods of determining or predicting whether an individual requiring therapy for a disease state such as cancer will or will not respond to treatment, prior to administration of the treatment, wherein the treatment comprises administration of one or more microtubule-stabilizing agents.
[0012] In one aspect, the invention provides a method for identifying a mammal that will respond therapeutically to a method of treating cancer comprising administering a microtubule-stabilizing agent, wherein the method comprises: (a) measuring in the mammal the level of at least one biomarker selected from the biomarkers of Table 1 and Table 2; (b) exposing a biological sample from the mammal to the agent; (c) following the exposing in step (b), measuring in said biological sample the level of the at least one biomarker, wherein an increase in the level of the at least one biomarker measured in step (c) compared to the level of the at least one biomarker measured in step (a), predicts that the mammal will respond therapeutically to said method of treating cancer when said biomarker is from Table 1, and predicts that the mammal will not respond therapeutically to said method of treating cancer when said biomarker is from Table 2.
[0013] In another aspect, the invention provides a method for determining whether a mammal is responding therapeutically to a microtubule-stabilizing agent, comprising (a) exposing the mammal to the agent; and (b) following the exposing of step (a), measuring in the mammal the level of at least one biomarker selected from the biomarkers of Table 1 and Table 2, wherein an increase in the level of the at least one biomarker measured in step (b), compared to the level of the at least one biomarker in a mammal that has not been exposed to said agent, indicates that the mammal is responding to said agent when said biomarker is from Table 1, and indicates that the mammal is not responding to said agent when said biomarker is from Table 2.
[0014] In another aspect, the invention provides a method for predicting whether a mammal will respond therapeutically to a method of treating cancer comprising administering a microtubule-stabilizing agent, wherein the method comprises: (a) exposing a biological sample from the mammal to the microtubule-stabilizing agent; (b) following the exposing of step (a), measuring in said biological sample the level of at least one biomarker selected from the biomarkers of Table 1 or Table 2, wherein an increase in the level of the at least one biomarker measured in step (b), compared to the level of the at least one biomarker in a mammal that has not been exposed to said agent, predicts that the mammal will respond therapeutically to said method of treating cancer when said biomarker is from Table 1, and predicts that the mammal will not respond therapeutically to said method of treating cancer when said biomarker is from Table 2.
[0015] In another aspect, the invention provides a method for determining whether an agent stabilizes microtubules and has cytotoxic activity against rapidly proliferating cells, such as, tumor cells or other hyperproliferative cellular disease in a mammal, comprising: (a) exposing the mammal to the agent; and (b) following the exposing of step (a), measuring in the mammal the level of at least one biomarker selected from the biomarkers of Table 1 and Table 2, wherein an increase in the level of said at least one biomarker measured in step (b), compared to the level of the biomarker in a mammal that has not been exposed to said agent, indicates that the agent stabilizes microtubules and has cytotoxic activity against rapidly proliferating cells when said biomarker is from Table 1, and indicates that the agent does not stabilize microtubules and does not have cytotoxic activity against rapidly proliferating cells when said biomarker is from Table 2.
[0016] As used herein, respond therapeutically refers to the alleviation or abrogation of the cancer. This means that the life expectancy of an individual affected with the cancer will be increased or that one or more of the symptoms of the cancer will be reduced or ameliorated. The term encompasses a reduction in cancerous cell growth or tumor volume. Whether a mammal responds therapeutically can be measured by many methods well known in the art, such as PET imaging.
[0017] The amount of increase in the level of the at least one biomarker measured in the practice of the invention can be readily determined by one skilled in the art. In one aspect, the increase in the level of a biomarker is at least a two-fold difference, at least a three-fold difference, or at least a four-fold difference in the level of the biomarker.
[0018] The mammal can be, for example, a human, rat, mouse, dog, rabbit, pig sheep, cow, horse, cat, primate, or monkey.
[0019] The method of the invention can be, for example, an in vitro method wherein the step of measuring in the mammal the level of at least one biomarker comprises taking a biological sample from the mammal and then measuring the level of the biomarker(s) in the biological sample. The biological sample can comprise, for example, at least one of whole fresh blood, peripheral blood mononuclear cells, frozen whole blood, fresh plasma, frozen plasma, urine, saliva, skin, hair follicle, bone marrow, or tumor tissue.
[0020] The level of the at least one biomarker can be, for example, the level of protein and/or mRNA transcript of the biomarker(s).
[0021] The invention also provides an isolated biomarker selected from the biomarkers of Table 1 and Table 2. The biomarkers of the invention comprise sequences selected from the nucleotide and amino acid sequences provided in Table 1 and Table 2 and the Sequence Listing, as well as fragments and variants thereof.
[0022] The invention also provides a biomarker set comprising two or more biomarkers selected from the biomarkers of Table 1 and Table 2.
[0023] The invention also provides kits for determining or predicting whether a patient would be susceptible or resistant to a treatment that comprises one or more microtubule-stabilizing agents. The patient may have a cancer or tumor such as, for example, a breast cancer or tumor.
[0024] In one aspect, the kit comprises a suitable container that comprises one or more specialized microarrays of the invention, one or more microtubule-stabilizing agents for use in testing cells from patient tissue specimens or patient samples, and instructions for use. The kit may further comprise reagents or materials for monitoring the expression of a biomarker set at the level of mRNA or protein.
[0025] In another aspect, the invention provides a kit comprising two or more biomarkers selected from the biomarkers of Table 1 and Table 2.
[0026] In yet another aspect, the invention provides a kit comprising at least one of an antibody and a nucleic acid for detecting the presence of at least one of the biomarkers selected from the biomarkers of Table 1 and Table 2. In one aspect, the kit further comprises instructions for determining whether or not a mammal will respond therapeutically to a method of treating cancer comprising administering a microtubule-stabilizing agent. In another aspect, the instructions comprise the steps of (a) measuring in the mammal the level of at least one biomarker selected from the biomarkers of Table 1 and Table 2, (b) exposing the mammal to the microtubule-stabilizing agent, (c) following the exposing of step (b), measuring in the mammal the level of the at least one biomarker, wherein a difference in the level of the at least one biomarker measured in step (c) compared to the level of the at least one biomarker measured in step (a) predicts that the mammal will respond therapeutically to said method of treating cancer when said biomarker is from Table 1, and predicts that the mammal will not respond therapeutically to said method of treating cancer when said biomarker is from Table 2.
[0027] The invention also provides screening assays for determining if a patient will be susceptible or resistant to treatment with one or more microtubule-stabilizing agents.
[0028] The invention also provides a method of monitoring the treatment of a patient having a disease, wherein said disease is treated by a method comprising administering one or more microtubule-stabilizing agents.
[0029] The invention also provides individualized genetic profiles which are necessary to treat diseases and disorders based on patient response at a molecular level.
[0030] The invention also provides specialized microarrays, e.g., oligonucleotide microarrays or cDNA microarrays, comprising one or more biomarkers having expression profiles that correlate with either sensitivity or resistance to one or more microtubule-stabilizing agents.
[0031] The invention also provides antibodies, including polyclonal or monoclonal, directed against one or more biomarkers of the invention.
[0032] The invention will be better understood upon a reading of the detailed description of the invention when considered in connection with the accompanying figures.

BRIEF DESCRIPTION OF THE FIGURES

[0033] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.
[0034] FIG. 1 illustrates classification of breast cancer cell lines based on IC50 values measured against ixabepilone treatment. The cell lines were normalized based on a mean of the log(IC50) values (a dotted line) and divided into two groups defined as sensitive or resistant. The cell lines on the left are defined as sensitive since they are below the mean of the log(IC50)s, and those on the right are defined as resistant since they are above the mean.
[0035] FIG. 2 illustrates the error rates generated from each classifier identified. From the GeneCluster analysis, classifiers containing up to 250 genes were determined. Each predictor identified was evaluated with the error rate calculated through the leave-one out cross validation. As shown in FIG. 2, error rates for classifiers (8-250 genes) remain to be the minimum at zero.
[0036] FIG. 3 illustrates the error rate calculation on random permutation. Error rates generated from several examples of random classification (random 1-11) using the software GeneCluster are plotted against the predictor sets containing up to 250 genes. As shown in the plot, the error rate calculated based on the IC50-based classification is significantly lower than the error rates based on the random classification.
[0037] FIG. 4 illustrates the top 50 genes correlated with sensitivity for ixabepilone. The red and blue matrix represents the normalized expression patterns for each gene across the cell lines (brightest red indicates highest relative expression, darkest blue indicates lowest relative expression).
[0038] FIG. 5 illustrates gene expression level of microtubule associated protein, tau and estrogen receptor (also referred to herein as estrogen receptor 1, ER, and ER1). In order to explore the relationship between Tau and ER, gene expression levels of the two are plotted for each cell line. Tau expression levels are shown as a bar, and ER as a line.
[0039] FIG. 6 illustrates gene expression patterns of top 50 genes in 175 primary breast tumors. The red and green matrix represents the normalized expression patterns for each gene across the 175 primary breast tumors (brightest red indicates highest relative expression, green indicates lowest relative expression). A subset of tumors in a blue box shows relative high expression of the top 25 markers which expressed highly in the sensitive cell lines. On the other hand, the tumors in a magenta box show relatively high expression of the 25 genes expressed at elevated levels in the resistant cell lines.
[0040] FIG. 7 illustrates MAP tau and ER expression in breast tumors. In order to see the expression patterns of Tau and ER within tumors, their expression levels are plotted together. Tau is shown as a line, and ER as a bar. Tumors are arranged in an increasing order of Tau expression level. There seems to be a subset of tumors that express these genes highly, which suggest these are non-responders for ixabepilone.
[0041] FIG. 8 illustrates the biological networks implied in the mechanism of resistance to ixabepilone as determined by Ingenuity® pathway analysis using 200 preclinical candidate markers. There are nine major networks indicated above with their significance scores. As shown, ER pathway is the most implicated network.
[0042] FIG. 9 illustrates the most implicated ER network and shows the functional connectivity between ER and Tau.
[0043] FIG. 10 illustrates molecular intrinsic profiles of 134 patient tumors. A hierarchical clustering analysis was performed using about 9800 genes after eliminating low expressed and low variance genes. Row, gene; column, tumor. The tumors were classified into two major clusters and, interestingly, the majority of the samples from three Russian sites were clustered in the first cluster.

DETAILED DESCRIPTION OF THE INVENTION

[0044] The invention provides biomarkers that correlate with microtubule-stabilization agent sensitivity or resistance. These biomarkers can be employed for predicting response to one or more microtubule-stabilization agents. In one aspect, the biomarkers of the invention are those provided in Table 1, Table 2, and the Sequence Listing, including both polynucleotide and polypeptide sequences.
[0045] The biomarkers provided in Table 1 include the nucleotide sequences of SEQ ID NOS:1-100 and the amino acid sequences of SEQ ID NOS:201-299.
[0046] 
[00001] [TABLE-US-00001]
  TABLE 1
 
  BIOMARKERS (SENSITIVE)
      Affymetrix  
  Unigene title and     Probe   Gene
  SEQ ID NO:   Affymetrix Description   Set   Ontology
 
  C6orf145:   “Consensus includes gb: AK024828.1   212923_s_at  
  chromosome 6/DEF = Homo sapiens cDNA: FLJ21175
  open reading   fis, clone CAS11071. /FEA = mRNA
  frame 145   /DB_XREF = gi: 10437233
  (LOC221749)   /UG = Hs.69388 hypothetical protein
  SEQ ID NOS: 1   FLJ20505”
  (DNA) and 201
  (amino acid)
  RTCD1: RNA“gb: NM_003729.1 /DEF = Homo sapiens   203594_at   assembly
  terminal phosphate   RNA 3-terminal phosphate cyclase     of
  cyclase domain 1   (RPC), mRNA. /FEA = mRNA     spliceosomal
  (LOC8634)   /GEN = RPC /PROD = RNA 3-terminal     tri-
  SEQ ID NOS: 2   phosphate cyclase     snRNP
  (DNA) and 202   /DB_XREF = gi: 4506588 /UG = Hs.27076
  (amino acid)   RNA 3-terminal phosphate cyclase
    /FL = gb: NM_003729.1”
  FXYD5: FXYD“gb: NM_014164.2 /DEF = Homo sapiens   218084_x_at   negative
  domain containing   FXYD domain-containing ion transport     regulation
  ion transport   regulator 5 (FXYD5), mRNA.     of
  regulator 5   /FEA = mRNA /GEN = FXYD5     calcium-
  (LOC53827)   /PROD = related to ion channel     dependent
  SEQ ID NOS: 3   /DB_XREF = gi: 11612664     cell-cell
  (DNA) and 203   /UG = Hs.294135 FXYD domain-     adhesion
  (amino acid)   containing ion transport regulator 5
    /FL = gb: NM_014164.2 gb: AF161462.1”
  G3BP: Ras-   Consensus includes gb: BG500067   201503_at   protein-
  GTPase-activating   /FEA = EST /DB_XREF = gi: 13461584     nucleus
  protein SH3-   /DB_XREF = est: 602545874F1     import
  domain-binding   /CLONE = IMAGE: 4668234
  protein   /UG = Hs.220689 Ras-GTPase-activating
  (LOC10146)   protein SH3-domain-binding protein
  SEQ ID NOS: 4   /FL = gb: U32519.1 gb: NM_005754.1
  (DNA) and 204
  (amino acid)
  FKBP1A: FK506“gb: NM_000801.1 /DEF = Homo sapiens   200709_at   protein
  binding protein   FK506-binding protein 1A (12 kD)     folding
  1A, 12 kDa   (FKBP1A), mRNA. /FEA = mRNA
  (LOC2280)   /GEN = FKBP1A /PROD = FK506-binding
  SEQ ID NOS: 5   protein 1A (12 kD)
  (DNA) and 205   /DB_XREF = gi: 4503724 /UG = Hs.752
  (amino acid)   FK506-binding protein 1A (12 kD)
    /FL = gb: BC001925.1 gb: M34539.1
    gb: NM_000801.1”
  VARS2: valyl-“gb: NM_006295.1 /DEF = Homo sapiens   201797_s_at   translational
  tRNA synthetase 2   valyl-tRNA synthetase 2 (VARS2),     elongation
  (LOC7407)   mRNA. /FEA = mRNA /GEN = VARS2
  SEQ ID NOS: 6   /PROD = valyl-tRNA synthetase 2
  (DNA) and 206   /DB_XREF = gi: 5454157
  (amino acid)   /UG = Hs.159637 valyl-tRNA synthetase
    2 /FL = gb: NM_006295.1”
  FKBP1A: FK506   Consensus includes gb: AI936769   214119_s_at   protein
  binding protein   /FEA = EST /DB_XREF = gi: 5675639     folding
  1A, 12 kDa   /DB_XREF = est: wp69c11.x1
  (LOC2280)   /CLONE = IMAGE: 2467028 /UG = Hs.752
  SEQ ID NOS: 7   FK506-binding protein 1A (12 kD)
  (DNA) and 207
  (amino acid)
  MTMR2:   “Consensus includes gb: AK027038.1   203211_s_at   protein
  myotubularin/DEF = Homo sapiens cDNA: FLJ23385     amino
  related protein 2   fis, clone HEP16802. /FEA = mRNA     acid
  (LOC8898)   /DB_XREF = gi: 10440053     dephosphorylation
  SEQ ID NOS: 8   /UG = Hs.181326 KIAA1073 protein
  (DNA) and 208   /FL = gb: AB028996.1 gb: NM_016156.1”
  (amino acid)
  PSMB8:   “gb: U17496.1 /DEF = Human proteasome   209040_s_at   proteolysis
  proteasome   subunit LMP7 (allele LMP7B) mRNA,     and
  (prosome,   complete cds. /FEA = mRNA     peptidolysis,
  macropain)   /GEN = LMP7 /PROD = proteasome     ubiquitin-
  subunit, beta type,   subunit LMP7 /DB_XREF = gi: 596139     dependent
  8 (large   /UG = Hs.180062 proteasome (prosome,     protein
  multifunctional   macropain) subunit, beta type, 8 (large     catabolism
  protease 7)   multifunctional protease 7)
  (LOC5696)   /FL = gb: U17496.1 gb: U17497.1”
  SEQ ID NOS: 9
  (DNA) and 209
  (amino acid)
  CTSC: cathepsin C“gb: NM_001814.1 /DEF = Homo sapiens   201487_at   proteolysis
  (LOC1075)   cathepsin C (CTSC), mRNA.     and
  SEQ ID NOS: 10   /FEA = mRNA /GEN = CTSC     peptidolysis
  (DNA) and 210   /PROD = cathepsin C
  (amino acid)   /DB_XREF = gi: 4503140 /UG = Hs.10029
    cathepsin C /FL = gb: NM_001814.1”
  ST5: suppression“gb: NM_005418.1 /DEF = Homo sapiens   202440_s_at
  of tumorigenicity 5   suppression of tumorigenicity 5 (ST5),
  (LOC6764)   mRNA. /FEA = mRNA /GEN = ST5
  SEQ ID NOS: 11   /PROD = suppression of tumorigenicity 5
  (DNA) and 211   /DB_XREF = gi: 4885612 /UG = Hs.79265
  (amino acid)   suppression of tumorigenicity 5
    /FL = gb: U15131.1 gb: U15779.1
    gb: NM_005418.1”
  MTMR2:   “Consensus includes gb: U58033.1   214649_s_at   protein
  myotubularin/DEF = Homo sapiens myotubularin     amino
  related protein 2   related protein 2 (MTMR2) mRNA,     acid
  (LOC8898)   partial cds. /FEA = mRNA     dephosphorylation
  SEQ ID NOS: 12   /GEN = MTMR2 /PROD = myotubularin
  (DNA) and 212   related protein 2 /DB_XREF = gi: 3912941
  (amino acid)   /UG = Hs.278491 myotubularin related
    protein 2 /FL = gb: NM_003912.1”
  PRNP: prion“gb: NM_000311.1 /DEF = Homo sapiens   201300_s_at
  protein (p27-30)   prion protein (p27-30) (Creutzfeld-Jakob
  (Creutzfeld-Jakob   disease, Gerstmann-Strausler-Scheinker
  disease,   syndrome, fatal familial insomnia)
  Gerstmann-   (PRNP), mRNA. /FEA = mRNA
  Strausler-   /GEN = PRNP /PROD = prion protein
  Scheinker   /DB_XREF = gi: 4506112 /UG = Hs.74621
  syndrome, fatal   prion protein (p27-30) (Creutzfeld-Jakob
  familial insomnia)   disease, Gerstmann-Strausler-Scheinker
  (LOC5621)   syndrome, fatal familial insomnia)
  SEQ ID NOS: 13   /FL = gb: AY008282.1 gb: M13899.1
  (DNA) and 213   gb: NM_000311.1”
  (amino acid)
  MET: met proto-   “gb: U19348.1 /DEF = Human (tpr-met   211599_x_at   signal
  oncogene   fusion) oncogene mRNA, complete cds.     transduction
  (hepatocyte   /FEA = mRNA /GEN = tprmet fusion
  growth factor   /PROD = tpr-met fusion protein
  receptor)   /DB_XREF = gi: 625085
  (LOC4233)   /FL = gb: U19348.1”
  SEQ ID NOS: 14
  (DNA) and 214
  (amino acid)
  MIRAB13:“Cluster Incl. W46406: zc31c10.s1 Homo   55081_at   vesicle-
  moleculesapiens cDNA, 3 end /clone = IMAGE-     mediated
  interacting with   323922 /clone_end = 3′ /gb = W46406     transport
  Rab13   /gi = 1331036 /ug = Hs.8535 /len = 568”
  (LOC85377)
  SEQ ID NOS: 15
  (DNA) and 215
  (amino acid)
  AKAP2: A kinase   Consensus includes gb: BE879367   202759_s_at
  (PRKA) anchor   /FEA = EST /DB_XREF = gi: 10328143
  protein 2   /DB_XREF = est: 601484628F1
  (LOC11217)   /CLONE = IMAGE: 3887262
  SEQ ID NOS: 16   /UG = Hs.42322 A kinase (PRKA) anchor
  (DNA) and 216   protein 2 /FL = gb: AB023137.1
  (amino acid)   gb: NM_007203.1
  MET: met proto-   Consensus includes gb: BG170541   203510_at   signal
  oncogene   /FEA = EST /DB_XREF = gi: 12677244     transduction
  (hepatocyte   /DB_XREF = est: 602322942F1
  growth factor   /CLONE = IMAGE: 4425947
  receptor)   /UG = Hs.285754 met proto-oncogene
  (LOC4233)   (hepatocyte growth factor receptor)
  SEQ ID NOS: 17   /FL = gb: J02958.1 gb: NM_000245.1
  (DNA) and 217
  (amino acid)
  STAC: SH3 and“gb: NM_003149.1 /DEF = Homo sapiens   205743_at   intracellular
  cysteine rich   src homology three (SH3) and cysteine     signaling
  domain   rich domain (STAC), mRNA.     cascade
  (LOC6769)   /FEA = mRNA /GEN = STAC /PROD = src
  SEQ ID NOS: 18   homology three (SH3) and cysteine
  (DNA) and 218   richdomain /DB_XREF = gi: 4507246
  (amino acid)   /UG = Hs.56045 src homology three
    (SH3) and cysteine rich domain
    /FL = gb: D86640.1 gb: NM_003149.1”
  CALU: calumenin   Consensus includes gb: BF939365   200755_s_at
  (LOC813)   /FEA = EST /DB_XREF = gi: 12356685
  SEQ ID NOS: 19   /DB_XREF = est: nad87h04.x1
  (DNA) and 219   /CLONE = IMAGE: 3410551
  (amino acid)   /UG = Hs.7753 calumenin
    /FL = gb: U67280.1 gb: AF013759.1
    gb: NM_001219.2
  MSN: moesin“gb: NM_002444.1 /DEF = Homo sapiens   200600_at   cell
  (LOC4478)   moesin (MSN), mRNA. /FEA = mRNA     motility
  SEQ ID NOS: 20   /GEN = MSN /PROD = moesin
  (DNA) and 220   /DB_XREF = gi: 4505256
  (amino acid)   /UG = Hs.170328 moesin
    /FL = gb: M69066.1 gb: NM_002444.1”
  ANXA1: annexin“gb: NM_000700.1 /DEF = Homo sapiens   201012_at   inflammatory
  A1 (LOC301)   annexin A1 (ANXA1), mRNA.     response,
  SEQ ID NOS: 21   /FEA = mRNA /GEN = ANXA1     cell
  (DNA) and 221   /PROD = annexin I     motility
  (amino acid)   /DB_XREF = gi: 4502100 /UG = Hs.78225
    annexin A1 /FL = gb: BC001275.1
    gb: NM_000700.1”
  CGI-100: CGI-100“gb: NM_016040.1 /DEF = Homo sapiens   202195_s_at   intracellular
  protein   CGI-100 protein (LOC50999), mRNA.     protein
  (LOC50999)   /FEA = mRNA /GEN = LOC50999     transport
  SEQ ID NOS: 22   /PROD = CGI-100 protein
  (DNA) and 222   /DB_XREF = gi: 7705583
  (amino acid)   /UG = Hs.296155 CGI-100 protein
    /FL = gb: AF151858.1 gb: NM_016040.1”
  CALU: calumenin“gb: NM_001219.2 /DEF = Homo sapiens   200757_s_at
  (LOC813)   calumenin (CALU), mRNA.
  SEQ ID NOS: 23   /FEA = mRNA /GEN = CALU
  (DNA) and 223   /PROD = calumenin precursor
  (amino acid)   /DB_XREF = gi: 6005991 /UG = Hs.7753
    calumenin /FL = gb: U67280.1
    gb: AF013759.1 gb: NM_001219.2”
  EGFR: epidermal   “gb: U95089.1 /DEF = Human truncated   210984_x_at   EGF
  growth factor   epidermal growth factor receptor-like     receptor
  receptor   protein precursor mRNA, complete cds.     signaling
  (erythroblastic   /FEA = mRNA /PROD = truncated     pathway
  leukemia viral (v-   epidermal growth factor receptor-
  erb-b) oncogene   likeprotein precursor
  homolog, avian)   /DB_XREF = gi: 2051984 /UG = Hs.77432
  (LOC1956)   epidermal growth factor receptor (avian
  SEQ ID NOS: 24   erythroblastic leukemia viral (v-erb-b)
  (DNA) and 224   oncogene homolog) /FL = gb: U95089.1”
  (amino acid)
  CASP4: caspase 4,   “gb: U25804.1 /DEF = Human Ich-2   209310_s_at   apoptosis
  apoptosis-related   cysteine protease mRNA, complete cds.
  cysteine protease   /FEA = mRNA /PROD = Ich-2
  (LOC837)   /DB_XREF = gi: 886049 /UG = Hs.74122
  SEQ ID NOS: 25   caspase 4, apoptosis-related cysteine
  (DNA) and 225   protease /FL = gb: U28976.1 gb: U28977.1
  (amino acid)   gb: U28978.1 gb: NM_001225.1
    gb: U25804.1 gb: U28014.1”
  DKFZP566E144:“gb: NM_015523.1 /DEF = Homo sapiens   218194_at   nucleotide
  small fragment   small fragment nuclease     metabolism
  nuclease   (DKFZP566E144), mRNA.
  (LOC25996)   /FEA = mRNA /GEN = DKFZP566E144
  SEQ ID NOS: 26   /PROD = small fragment nuclease
  (DNA) and 226   /DB_XREF = gi: 7661645 /UG = Hs.7527
  (amino acid)   small fragment nuclease
    /FL = gb: AF151872.1 gb: AL110239.1
    gb: NM_015523.1”
  AKR1B1: aldo-“gb: NM_001628.1 /DEF = Homo sapiens   201272_at   carbohydrate
  keto reductase   aldo-keto reductase family 1, member B1     metabolism
  family 1, member   (aldose reductase) (AKR1B1), mRNA.
  B1 (aldose   /FEA = mRNA /GEN = AKR1B1
  reductase)   /PROD = aldo-keto reductase family 1,
  (LOC231)   member B1 (aldosereductase)
  SEQ ID NOS: 27   /DB_XREF = gi: 4502048 /UG = Hs.75313
  (DNA) and 227   aldo-keto reductase family 1, member B1
  (amino acid)   (aldose reductase) /FL = gb: BC000260.1
    gb: BC005387.1 gb: J04795.1 gb: J05017.1
    gb: J05474.1 gb: M34720.1
    gb: NM_001628.1”
  CAV1: caveolin 1,   Consensus includes gb: AU147399   212097_at
  caveolae protein,   /FEA = EST /DB_XREF = gi: 11008920
  22 kDa (LOC857)   /DB_XREF = est: AU147399
  SEQ ID NOS: 28   /CLONE = MAMMA1000563
  (DNA) and 228/UG = Hs.74034 Homo sapiens clone
  (amino acid)   24651 mRNA sequence
  MBNL2:   Consensus includes gb: BE328496   203640_at
  muscleblind-like 2   /FEA = EST /DB_XREF = gi: 9202272
(Drosophila)   /DB_XREF = est: hs98f09.x1
  (LOC10150)   /CLONE = IMAGE: 3145289
  SEQ ID NOS: 29   /UG = Hs.283609 hypothetical protein
  (DNA) and 229   PRO2032 /FL = gb: AF116683.1
  (amino acid)   gb: NM_018615.1
  CRSP6: cofactor“gb: AF105421.1 /DEF = Homo sapiens   221517_s_at   regulation
  required for Sp1   vitamin D3 receptor interacting protein     of
  transcriptional   (DRIP80) mRNA, complete cds.     transcription,
  activation, subunit   /FEA = mRNA /GEN = DRIP80     DNA-
  6, 77 kDa   /PROD = vitamin D3 receptor interacting     dependent
  (LOC9440)   protein /DB_XREF = gi: 4838128
  SEQ ID NOS: 30   /UG = Hs.22630 cofactor required for Sp1
  (DNA) and 230   transcriptional activation, subunit 6
  (amino acid)   (77 kD) /FL = gb: AF105421.1”
  CALU: calumenin“gb: U67280.1 /DEF = Homo sapiens   200756_x_at
  (LOC813)   calumenin mRNA, complete cds.
  SEQ ID NOS: 31   /FEA = mRNA /PROD = calumenin
  (DNA) and 231   /DB_XREF = gi: 2809323 /UG = Hs.7753
  (amino acid)   calumenin /FL = gb: U67280.1
    gb: AF013759.1 gb: NM_001219.2”
  PTRF: polymerase   “Consensus includes gb: BC004295.1   208789_at
  I and transcript/DEF = Homo sapiens, clone
  release factor   IMAGE: 3622356, mRNA, partial cds.
  (LOC284119)   /FEA = mRNA /PROD = Unknown
  SEQ ID NOS: 32   (protein for IMAGE: 3622356)
  (DNA) and 232   /DB_XREF = gi: 13279151
  (amino acid)   /UG = Hs.29759 RNA POLYMERASE I
    AND TRANSCRIPT RELEASE
    FACTOR /FL = gb: AF312393.1”
  NUP155:“gb: NM_004298.1 /DEF = Homo sapiens   206550_s_at   nucleocytoplasmic
  nucleoporin   nucleoporin 155 kD (NUP155), mRNA.     transport
  155 kDa   /FEA = mRNA /GEN = NUP155
  (LOC9631)   /PROD = nucleoporin 155 kD
  SEQ ID NOS: 33   /DB_XREF = gi: 4758843 /UG = Hs.23255
  (DNA) and 233   nucleoporin 155 kD /FL = gb: AB018334.1
  (amino acid)   gb: NM_004298.1”
  DONSON:“gb: AF232674.1 /DEF = Homo sapiens   221677_s_at
  downstream   B17 mRNA, complete cds. /FEA = mRNA
  neighbor of SON   /PROD = B17 /DB_XREF = gi: 8118230
  (LOC29980)   /UG = Hs.17834 downstream neighbor of
  SEQ ID NOS: 34   SON /FL = gb: AF232674.1”
  (DNA) and 234
  (amino acid)
  CALU: calumenin   “Consensus includes gb: AF257659.1   214845_s_at
  (LOC813)/DEF = Homo sapiens crocalbin-like
  SEQ ID NOS: 35   protein mRNA, partial cds.
  (DNA) and 235   /FEA = mRNA /PROD = crocalbin-like
  (amino acid)   protein /DB_XREF = gi: 8515717
  /UG = Hs.302073 Homo sapiens
    crocalbin-like protein mRNA, partial
    cds”
  FAD104: FAD104“gb: NM_022763.1 /DEF = Homo sapiens   218618_s_at
  (LOC64778)   hypothetical protein FLJ23399
  SEQ ID NOS: 36   (FLJ23399), mRNA. /FEA = mRNA
  (DNA) and 236   /GEN = FLJ23399 /PROD = hypothetical
  (amino acid)   protein FLJ23399
    /DB_XREF = gi: 12232434
    /UG = Hs.299883 hypothetical protein
    FLJ23399 /FL = gb: NM_022763.1”
  EPHA2: EphA2“gb: NM_004431.1 /DEF = Homo sapiens   203499_at
  (LOC1969)   EphA2 (EPHA2), mRNA. /FEA = mRNA
  SEQ ID NOS: 37   /GEN = EPHA2 /PROD = EphA2
  (DNA) and 237   /DB_XREF = gi: 4758277
  (amino acid)   /UG = Hs.171596 EphA2
    /FL = gb: M59371.1 gb: NM_004431.1”
  PAK1IP1: PAK1“gb: NM_017906.1 /DEF = Homo sapiens   218886_at
  interacting protein   hypothetical protein FLJ20624
  1 (LOC55003)   (FLJ20624), mRNA. /FEA = mRNA
  SEQ ID NOS: 38   /GEN = FLJ20624 /PROD = hypothetical
  (DNA) and 238   protein FLJ20624
  (amino acid)   /DB_XREF = gi: 8923576 /UG = Hs.52256
    hypothetical protein FLJ20624
    /FL = gb: NM_017906.1”
  CTPS: CTP“gb: NM_001905.1 /DEF = Homo sapiens   202613_at   pyrimidine
  synthase   CTP synthase (CTPS), mRNA.     nucleotide
  (LOC1503)   /FEA = mRNA /GEN = CTPS     biosynthesis
  SEQ ID NOS: 39   /PROD = CTP synthase
  (DNA) and 239   /DB_XREF = gi: 4503132
  (amino acid)   /UG = Hs.251871 CTP synthase
    /FL = gb: NM_001905.1”
  CD44: CD44“gb: BC004372.1 /DEF = Homo sapiens,   209835_x_at   cell-cell
  antigen (homing   Similar to CD44 antigen (homing     adhesion
  function and   function and Indian blood group system),
  Indian blood group   clone MGC: 10468, mRNA, complete
  system) (LOC960)   cds. /FEA = mRNA /PROD = Similar to
  SEQ ID NOS: 40   CD44 antigen (homing function
  (DNA) and 240   andIndian blood group system)
  (amino acid)   /DB_XREF = gi: 13325117
    /UG = Hs.169610 CD44 antigen (homing
    function and Indian blood group system)
    /FL = gb: BC004372.1”
  CD97: CD97“gb: NM_001784.1 /DEF = Homo sapiens   202910_s_at   G-protein
  antigen (LOC976)   CD97 antigen (CD97), mRNA.     coupled
  SEQ ID NOS: 41   /FEA = mRNA /GEN = CD97     receptor
  (DNA) and 241   /PROD = CD97 antigen     protein
  (amino acid)   /DB_XREF = gi: 4502690./UG = Hs.3107     signaling
    CD97 antigen /FL = gb: NM_001784.1”     pathway
  SPTBN1: spectrin,   “Consensus includes gb: BE968833   212071_s_at
  beta, non-   /FEA = EST /DB_XREF = gi: 10579538
  erythrocytic 1   /DB_XREF = est: 601649861F1
  (LOC6711)   /CLONE = IMAGE: 3933782
  SEQ ID NOS: 42/UG = Hs.324648 Homo sapiens cDNA
  (DNA) and 242   FLJ13700 fis, clone PLACE2000216,
  (amino acid)   highly similar to SPECTRIN BETA
    CHAIN, BRAIN”
  SH3GLB1: SH3-“gb: AF263293.1 /DEF = Homo sapiens   209091_s_at
  domain GRB2-like   endophilin B1 mRNA, complete cds.
  endophilin B1   /FEA = mRNA /PROD = endophilin B1
  (LOC51100)   /DB_XREF = gi: 8118529
  SEQ ID NOS: 43   /UG = Hs.136309 SH3-containing protein
  (DNA) and 243   SH3GLB1 /FL = gb: AF263293.1”
  (amino acid)
  PGM1:“gb: NM_002633.1 /DEF = Homo sapiens   201968_s_at   glucose
  phosphoglucomutase   phosphoglucomutase 1 (PGM1), mRNA.     metabolism
  1 (LOC5236)   /FEA = mRNA /GEN = PGM1
  SEQ ID NOS: 44   /PROD = phosphoglucomutase 1
  (DNA) and 244   /DB_XREF = gi: 4505764 /UG = Hs.1869
  (amino acid)   phosphoglucomutase 1
    /FL = gb: BC001756.1 gb: M83088.1
    gb: NM_002633.1”
  SH3GLB1: SH3-“gb: AF257318.1 /DEF = Homo sapiens   210101_x_at
  domain GRB2-like   SH3-containing protein SH3GLB1
  endophilin B1   mRNA, complete cds. /FEA = mRNA
  (LOC51100)   /PROD = SH3-containing protein
  SEQ ID NOS: 45   SH3GLB1 /DB_XREF = gi: 8896091
  (DNA) and 245   /UG = Hs.136309 SH3-containing protein
  (amino acid)   SH3GLB1 /FL = gb: AF350371.1
    gb: AF151819.1 gb: NM_016009.1
    gb: AF257318.1”
  GBP1: guanylate“gb: BC002666.1 /DEF = Homo sapiens,   202269_x_at   immune
  binding protein 1,   guanylate binding protein 1, interferon-     response
  interferon-   inducible, 67 kD, clone MGC: 3949,
  inducible, 67 kDa   mRNA, complete cds. /FEA = mRNA
  (LOC2633)   /PROD = guanylate binding protein
  SEQ ID NOS: 46   1, interferon-inducible, 67 kD
  (DNA) and 246   /DB_XREF = gi: 12803662
  (amino acid)   /UG = Hs.62661 guanylate binding protein
    1, interferon-inducible, 67 kD
    /FL = gb: BC002666.1 gb: M55542.1
    gb: NM_002053.1”
  ADORA2B:“gb: NM_000676.1 /DEF = Homo sapiens   205891_at   adenylate
  adenosine A2b   adenosine A2b receptor (ADORA2B),     cyclase
  receptor (LOC136)   mRNA. /FEA = mRNA     activation
  SEQ ID NOS: 47   /GEN = ADORA2B /PROD = adenosine
  (DNA) and 247   A2b receptor /DB_XREF = gi: 4501950
  (amino acid)   /UG = Hs.45743 adenosine A2b receptor
    /FL = gb: M97759.1 gb: NM_000676.1”
  PLS3: plastin 3 (T“gb: NM_005032.2 /DEF = Homo sapiens   201215_at
  isoform)   plastin 3 (T isoform) (PLS3), mRNA.
  (LOC5358)   /FEA = mRNA /GEN = PLS3
  SEQ ID NOS: 48   /PROD = plastin 3 precursor
  (DNA) and 248   /DB_XREF = gi: 7549808 /UG = Hs.4114
  (amino acid)   plastin 3 (T isoform) /FL = gb: M22299.1
    gb: NM_005032.2”
  PDGFC: platelet“gb: NM_016205.1 /DEF = Homo sapiens   218718_at
  derived growth   platelet derived growth factor C
  factor C   (PDGFC), mRNA. /FEA = mRNA
  (LOC56034)   /GEN = PDGFC /PROD = secretory growth
  SEQ ID NOS: 49   factor-like protein fallotein
  (DNA) and 249   /DB_XREF = gi: 9994186 /UG = Hs.43080
  (amino acid)   platelet derived growth factor C
    /FL = gb: AF091434.1 gb: AF244813.1
    gb: AB033831.1 gb: NM_016205.1”
  MID1: midline 1“gb: NM_000381.1 /DEF = Homo sapiens   203637_s_at   microtubule
  (Opitz/BBB   midline 1 (OpitzBBB syndrome)     cytoskeleton
  syndrome)   (MID1), mRNA. /FEA = mRNA     organization
  (LOC4281)   /GEN = MID1 /PROD = midline 1     and
  SEQ ID NOS: 50   /DB_XREF = gi: 4557752 /UG = Hs.27695     biogenesis
  (DNA) and 250   midline 1 (OpitzBBB syndrome)
  (amino acid)   /FL = gb: AF269101.1 gb: AF230976.1
    gb: AF035360.1 gb: NM_000381.1”
  MET: met proto-   Consensus includes gb: BE870509   213807_x_at   signal
  oncogene   /FEA = EST /DB_XREF = gi: 10319285     transduction
  (hepatocyte   /DB_XREF = est: 601447096F1
  growth factor   /CLONE = IMAGE: 3851374
  receptor)   /UG = Hs.285754 met proto-oncogene
  (LOC4233)   (hepatocyte growth factor receptor)
  SEQ ID NOS: 51
  (DNA) and 251
  (amino acid)
  CHST6:“gb: NM_021615.1 /DEF = Homo sapiens   221059_s_at   proteoglycan
  carbohydrate (N-   carbohydrate (N-acetylglucosamine 6-O)     sulfate
  acetylglucosamine   sulfotransferase 6 (CHST6), mRNA.     transfer
  6-O)   /FEA = mRNA /GEN = CHST6
  sulfotransferase 6   /PROD = carbohydrate (N-
  (LOC4166)   acetylglucosamine 6-O)sulfotransferase 6
  SEQ ID NOS: 52   /DB_XREF = gi: 11055975
  (DNA) and 252   /UG = Hs.157439 carbohydrate (N-
  (amino acid)   acetylglucosamine 6-O) sulfotransferase
    6 /FL = gb: AF219990.1
    gb: NM_021615.1”
  MEIS2: Meis1,“gb: NM_020149.1 /DEF = Homo sapiens   207480_s_at   negative
  myeloid ecotropic   TALE homeobox protein Meis2e     regulation
  viral integration   (LOC56908), mRNA. /FEA = mRNA     of
  site 1 homolog 2   /GEN = LOC56908 /PROD = TALE     transcription
  (mouse)   homeobox protein Meis2e     from
  (LOC4212)   /DB_XREF = gi: 9910355     Pol II
  SEQ ID NOS: 53   /UG = Hs.283312 TALE homeobox     promoter
  (DNA) and 253   protein Meis2e /FL = gb: AF179899.1
  (amino acid)   gb: NM_020149.1”
  UPP1: uridine“gb: NM_003364.1 /DEF = Homo sapiens   203234_at   nucleoside
  phosphorylase 1   uridine phosphorylase (UP), mRNA.     metabolism
  (LOC7378)   /FEA = mRNA /GEN = UP /PROD = uridine
  SEQ ID NOS: 54   phosphorylase /DB_XREF = gi: 4507838
  (DNA) and 254   /UG = Hs.77573 uridine phosphorylase
  (amino acid)   /FL = gb: BC001405.1 gb: NM_003364.1”
  CD44: CD44   Consensus includes gb: AI493245   212014_x_at   cell-cell
  antigen (homing   /FEA = EST /DB_XREF = gi: 4394248     adhesion
  function and   /DB_XREF = est: ti30d08.x1
  Indian blood group   /CLONE = IMAGE: 2131983
  system) (LOC960)   /UG = Hs.169610 CD44 antigen (homing
  SEQ ID NOS: 55   function and Indian blood group system)
  (DNA) and 255
  (amino acid)
  BTG3: BTG   “Consensus includes gb: AI765445   213134_x_at   regulation
  family, member 3   /FEA = EST /DB_XREF = gi: 5231954     of cell
  (LOC10950)   /DB_XREF = est: wi80b08.x1     cycle
  SEQ ID NOS: 56   /CLONE = IMAGE: 2399607
  (DNA) and 256   /UG = Hs.77311 BTG family, member 3”
  (amino acid)
  FKBP1A: FK506“gb: BC005147.1 /DEF = Homo sapiens,   210186_s_at   protein
  binding protein   FK506-binding protein 1A (12 kD), clone     folding
  1A, 12 kDa   MGC: 2167, mRNA, complete cds.
  (LOC2280)   /FEA = mRNA /PROD = FK506-binding
  SEQ ID NOS: 57   protein 1A (12 kD)
  (DNA) and 257   /DB_XREF = gi: 13477342 /UG = Hs.752
  (amino acid)   FK506-binding protein 1A (12 kD)
    /FL = gb: BC005147.1”
  IFI16: interferon,“gb: NM_005531.1 /DEF = Homo sapiens   206332_s_at
  gamma-inducible   interferon, gamma-inducible protein 16
  protein 16   (IFI16), mRNA. /FEA = mRNA
  (LOC3428)   /GEN = IFI16 /PROD = interferon, gamma-
  SEQ ID NOS: 58   inducible protein 16
  (DNA) and 258   /DB_XREF = gi: 5031778
  (amino acid)   /UG = Hs.155530 interferon, gamma-
    inducible protein 16 /FL = gb: M63838.1
    gb: NM_005531.1”
  CD44: CD44   Consensus includes gb: BE903880   212063_at   cell-cell
  antigen (homing   /FEA = EST /DB_XREF = gi: 10395551     adhesion
  function and   /DB_XREF = est: 601494678F1
  Indian blood group   /CLONE = IMAGE: 3896970
  system) (LOC960)   /UG = Hs.323950 zinc finger protein 6
  SEQ ID NOS: 59   (CMPX1)
  (DNA) and 259
  (amino acid)
  IFI16: interferon,“gb: AF208043.1 /DEF = Homo sapiens   208966_x_at
  gamma-inducible   IFI16b (IFI16b) mRNA, complete cds.
  protein 16   /FEA = mRNA /GEN = IFI16b
  (LOC3428)   /PROD = IFI16b /DB_XREF = gi: 6644296
  SEQ ID NOS: 60   /UG = Hs.155530 interferon, gamma-
  (DNA) and 260   inducible protein 16
  (amino acid)   /FL = gb: AF208043.1”
  GNG12: guanine   Consensus includes gb: BG111761   212294_at   signal
  nucleotide binding   /FEA = EST /DB_XREF = gi: 12605267     transduction
  protein (G   /DB_XREF = est: 602285343F1
  protein), gamma   /CLONE = IMAGE: 4372619
  12 (LOC55970)/UG = Hs.8107 Homo sapiens mRNA;
  SEQ ID NOS: 61   cDNA DKFZp586B0918 (from clone
  (DNA) and 261   DKFZp586B0918)
  (amino acid)
  GSTP1:“gb: NM_000852.2 /DEF = Homo sapiens   200824_at   metabolism
  glutathione S-   glutathione S-transferase pi (GSTP1),
  transferase pi   mRNA. /FEA = mRNA /GEN = GSTP1
  (LOC2950)   /PROD = glutathione transferase
  SEQ ID NOS: 62   /DB_XREF = gi: 6552334
  (DNA) and 262   /UG = Hs.226795 glutathione S-
  (amino acid)   transferase pi /FL = gb: U62589.1
    gb: U30897.1 gb: NM_000852.2”
  MCAM:“gb: BC006329.1 /DEF = Homo sapiens,   211042_x_at
  melanoma cell   Similar to melanoma adhesion molecule,
  adhesion molecule   clone MGC: 12808, mRNA, complete
  (LOC4162)   cds. /FEA = mRNA /PROD = Similar to
  SEQ ID NOS: 63   melanoma adhesion molecule
  (DNA) and 263   /DB_XREF = gi: 13623456
  (amino acid)   /FL = gb: BC006329.1”
  MIRAB13:   “Consensus includes gb: BC001090.1   221779_at   vesicle-
  molecule/DEF = Homo sapiens, clone     mediated
  interacting with   IMAGE: 3504989, mRNA, partial cds.     transport
  Rab13   /FEA = mRNA /PROD = Unknown
  (LOC85377)   (protein for IMAGE: 3504989)
  SEQ ID NOS: 64   /DB_XREF = gi: 12654518 /UG = Hs.8535
  (DNA) and 264   hypothetical protein bA395L14.2”
  (amino acid)
  IFI16: interferon,   “Consensus includes gb: BG256677   208965_s_at
  gamma-inducible   /FEA = EST /DB_XREF = gi: 12766493
  protein 16   /DB_XREF = est: 602370865F1
  (LOC3428)   /CLONE = IMAGE: 4478872
  SEQ ID NOS: 65   /UG = Hs.155530 interferon, gamma-
  (DNA) and 265   inducible protein 16
  (amino acid)   /FL = gb: AF208043.1”
  DKFZp667G2110:   “Consensus includes gb: BE501352   214030_at
  hypothetical   /FEA = EST /DB_XREF = gi: 9703760
  protein   /DB_XREF = est: 7a41e05.x1
  DKFZp667G2110   /CLONE = IMAGE: 3221312
  (LOC131544)   /UG = Hs.23294 ESTs, Weakly similar to
  SEQ ID NOS: 66   T15138 hypothetical protein T28F2.4-
  (DNA)Caenorhabditis elegans C. elegans
  C1GALT1: core 1“gb: NM_020156.1 /DEF = Homo sapiens   219439_at
  UDP-galactose:N-   core1 UDP-galactose:N-
  acetylgalactosamine-   acetylgalactosamine-alpha-R beta 1,3-
  alpha-R beta 1,3-   galactosyltransferase (C1GALT1),
  galactosyltransferase   mRNA. /FEA = mRNA
  (LOC56913)   /GEN = C1GALT1 /PROD = core1UDP-
  SEQ ID NOS: 67   galactose:N-acetylgalactosamine-alpha-R
  (DNA) and 266   beta1,3-galactosyltransferase
  (amino acid)   /DB_XREF = gi: 9910143 /UG = Hs.46744
    core1 UDP-galactose:N-
    acetylgalactosamine-alpha-R beta 1,3-
    galactosyltransferase
    /FL = gb: AF155582.1 gb: NM_020156.1”
  RANGNRF: RAN“gb: NM_014185.1 /DEF = Homo sapiens   218526_s_at
  guanine nucleotide   HSPC165 protein (HSPC165), mRNA.
  release factor   /FEA = mRNA /GEN = HSPC165
  (LOC29098)   /PROD = HSPC165 protein
  SEQ ID NOS: 68   /DB_XREF = gi: 7661825 /UG = Hs.13605
  (DNA) and 267   HSPC165 protein /FL = gb: AF161514.1
  (amino acid)   gb: AF151070.1 gb: NM_014185.1
    gb: NM_016492.1 gb: AF168714.1
    gb: AF265206.1”
  ELL2: elongation   “Consensus includes gb: NM_012081.1   214446_at   regulation
  factor, RNA/DEF = Homo sapiens ELL-RELATED     of
  polymerase II, 2   RNA POLYMERASE II,     transcription,
  (LOC22936)   ELONGATION FACTOR (ELL2),     DNA-
  SEQ ID NOS: 69   mRNA. /FEA = CDS /GEN = ELL2     dependent
  (DNA) and 268   /PROD = ELL-RELATED RNA
  (amino acid)   POLYMERASE II,
    ELONGATIONFACTOR
    /DB_XREF = gi: 6912353
    /UG = Hs.173334 ELL-RELATED RNA
    POLYMERASE II, ELONGATION
    FACTOR /FL = gb: NM_012081.1”
  BIN1: bridging   “Consensus includes gb: AF043899.1   214439_x_at   synaptic
  integrator 1/DEF = Homo sapiens amphiphysin IIc1     transmission
  (LOC274)   mRNA, complete cds. /FEA = CDS
  SEQ ID NOS: 70   /PROD = amphiphysin IIc1
  (DNA) and 269   /DB_XREF = gi: 3064256
  (amino acid)   /UG = Hs.193163 bridging integrator 1
    /FL = gb: AF043899.1”
  M-RIP: myosin   “Consensus includes gb: AK025604.1   214771_x_at
  phosphatase-Rho/DEF = Homo sapiens cDNA: FLJ21951
  interacting protein   fis, clone HEP04968. /FEA = mRNA
  (LOC23164)   /DB_XREF = gi: 10438172
  SEQ ID NOS: 71   /UG = Hs.84883 KIAA0864 protein”
  (DNA) and 270
  (amino acid)
  MGC5306:“gb: BC001972.1 /DEF = Homo sapiens,   221580_s_at
  hypothetical   clone MGC: 5306, mRNA, complete cds.
  protein MGC5306   /FEA = mRNA /PROD = Unknown
  (LOC79101)   (protein for MGC: 5306)
  SEQ ID NOS: 72   /DB_XREF = gi: 12805036
  (DNA) and 271   /UG = Hs.301732 hypothetical protein
  (amino acid)   MGC5306 /FL = gb: BC001972.1”
  BTN3A3:“gb: NM_006994.2 /DEF = Homo sapiens   204820_s_at
  butyrophilin,   butyrophilin, subfamily 3, member A3
  subfamily 3,   (BTN3A3), mRNA. /FEA = mRNA
  member A3   /GEN = BTN3A3 /PROD = butyrophilin,
  (LOC10384)   subfamily 3, member A3
  SEQ ID NOS: 73   /DB_XREF = gi: 6325463
  (DNA) and 272   /UG = Hs.167741 butyrophilin, subfamily
  (amino acid)   3, member A3 /FL = gb: U90548.1
    gb: NM_006994.2”
  CAV2: caveolin 2“gb: NM_001233.1 /DEF = Homo sapiens   203324_s_at
  (LOC858)   caveolin 2 (CAV2), mRNA.
  SEQ ID NOS: 74   /FEA = mRNA /GEN = CAV2
  (DNA) and 273   /PROD = caveolin 2
  (amino acid)   /DB_XREF = gi: 4557412
    /UG = Hs.139851 caveolin 2
    /FL = gb: BC005256.1 gb: AF035752.1
    gb: NM_001233.1”
  IFNGR1:“gb: NM_000416.1 /DEF = Homo sapiens   202727_s_at   signal
  interferon gamma   interferon gamma receptor 1 (IFNGR1),     transduction
  receptor 1   mRNA. /FEA = mRNA /GEN = IFNGR1
  (LOC3459)   /PROD = interferon gamma receptor 1
  SEQ ID NOS: 75   /DB_XREF = gi: 4557879
  (DNA) and 274   /UG = Hs.180866 interferon gamma
  (amino acid)   receptor 1 /FL = gb: BC005333.1
    gb: J03143.1 gb: NM_000416.1”
  MGC5297:“gb: NM_024091.1 /DEF = Homo sapiens   219200_at
  hypothetical   hypothetical protein MGC5297
  protein MGC5297   (MGC5297), mRNA. /FEA = mRNA
  (LOC79072)   /GEN = MGC5297 /PROD = hypothetical
  SEQ ID NOS: 76   protein MGC5297
  (DNA) and 275   /DB_XREF = gi: 13129089
  (amino acid)   /UG = Hs.23856 hypothetical protein
    MGC5297 /FL = gb: BC001295.1
    gb: NM_024091.1”
  TGFBI:“gb: NM_000358.1 /DEF = Homo sapiens   201506_at   cell
  transforming   transforming growth factor, beta-     adhesion
  growth factor,   induced, 68 kD (TGFBI), mRNA.
  beta-induced,   /FEA = mRNA /GEN = TGFBI
  68 kDa (LOC7045)   /PROD = transforming growth factor,
  SEQ ID NOS: 77   beta-induced, 68 kD
  (DNA) and 276   /DB_XREF = gi: 4507466
  (amino acid)   /UG = Hs.118787 transforming growth
    factor, beta-induced, 68 kD
    /FL = gb: BC000097.1 gb: BC004972.1
    gb: M77349.1 gb: NM_000358.1”
  AKAP2: A kinase“gb: NM_007203.1 /DEF = Homo sapiens   202760_s_at
  (PRKA) anchor   A kinase (PRKA) anchor protein 2
  protein 2   (AKAP2), mRNA. /FEA = mRNA
  (LOC11217)   /GEN = AKAP2 /PROD = A kinase
  SEQ ID NOS: 78   (PRKA) anchor protein 2
  (DNA) and 277   /DB_XREF = gi: 6005708 /UG = Hs.42322
  (amino acid)   A kinase (PRKA) anchor protein 2
    /FL = gb: AB023137.1 gb: NM_007203.1”
  QKI: quaking   Consensus includes gb: AI114716   212263_at   signal
  homolog, KH   /FEA = EST /DB_XREF = gi: 6360061     transduction
  domain RNA   /DB_XREF = est: HA1315 /UG = Hs.15020
  binding (mouse)   homolog of mouse quaking QKI (KH
  (LOC9444)   domain RNA binding protein)
  SEQ ID NOS: 79   /FL = gb: AF142419.1 gb: AF142422.1
  (DNA) and 278
  (amino acid)
  PRNP: prion   “Consensus includes gb: AV725328   215707_s_at
  protein (p27-30)   /FEA = EST /DB_XREF = gi: 10830606
  (Creutzfeld-Jakob   /DB_XREF = est: AV725328
  disease,   /CLONE = HTCAVD03 /UG = Hs.74621
  Gerstmann-   prion protein (p27-30) (Creutzfeld-Jakob
  Strausler-   disease, Gerstmann-Strausler-Scheinker
  Scheinker   syndrome, fatal familial insomnia)”
  syndrome, fatal
  familial insomnia)
  (LOC5621)
  SEQ ID NOS: 80
  (DNA) and 279
  (amino acid)
  HIC: I-mfa“gb: AF054589.1 /DEF = Homo sapiens   211675_s_at
  domain-containing   HIC protein isoform p40 and HIC protein
  protein   isoform p32 mRNAs, complete cds.
  (LOC29969)   /FEA = mRNA /PROD = HIC protein
  SEQ ID NOS: 81   isoform p32; HIC protein isoform p40
  (DNA) and 280   /DB_XREF = gi: 3426297
  (amino acid)   /FL = gb: AF054589.1”
  POPDC3: popeye“gb: NM_022361.1 /DEF = Homo sapiens   219926_at
  domain containing   popeye protein 3 (POP3), mRNA.
  3 (LOC64208)   /FEA = mRNA /GEN = POP3
  SEQ ID NOS: 82   /PROD = popeye protein 3
  (DNA) and 281   /DB_XREF = gi: 11641280
  (amino acid)   /UG = Hs.303154 popeye protein 3
    /FL = gb: AF204171.1 gb: NM_022361.1”
  FLJ10315:“gb: NM_018056.1 /DEF = Homo sapiens   218770_s_at
  hypothetical   hypothetical protein FLJ10315
  protein FLJ10315   (FLJ10315), mRNA. /FEA = mRNA
  (LOC55116)   /GEN = FLJ10315 /PROD = hypothetical
  SEQ ID NOS: 83   protein FLJ10315
  (DNA) and 282   /DB_XREF = gi: 8922347 /UG = Hs.25544
  (amino acid)   hypothetical protein FLJ10315
    /FL = gb: AL136695.1 gb: NM_018056.1”
  PSMB9:“gb: NM_002800.1 /DEF = Homo sapiens   204279_at   proteolysis
  proteasome   proteasome (prosome, macropain)     and
  (prosome,   subunit, beta type, 9 (large     peptidolysis,
  macropain)   multifunctional protease 2) (PSMB9),     ubiquitin-
  subunit, beta type,   mRNA. /FEA = mRNA /GEN = PSMB9     dependent
  9 (large   /PROD = proteasome (prosome,     protein
  multifunctional   macropain) subunit, betatype, 9 (large     catabolism
  protease 2)   multifunctional protease 2)
  (LOC5698)   /DB_XREF = gi: 4506204 /UG = Hs.9280
  SEQ ID NOS: 84   proteasome (prosome, macropain)
  (DNA) and 283   subunit, beta type, 9 (large
  (amino acid)   multifunctional protease 2)
    /FL = gb: U01025.1 gb: NM_002800.1”
  DEPDC1: DEP“gb: NM_017779.1 /DEF = Homo sapiens   220295_x_at
  domain containing   hypothetical protein FLJ20354
  1 (LOC55635)   (FLJ20354), mRNA. /FEA = mRNA
  SEQ ID NOS: 85   /GEN = FLJ20354 /PROD = hypothetical
  (DNA) and 284   protein FLJ20354
  (amino acid)   /DB_XREF = gi: 8923327
    /UG = Hs.133260 hypothetical protein
    FLJ20354 /FL = gb: NM_017779.1”
  EGFR: epidermal   Consensus includes gb: AW157070   201983_s_at   EGF
  growth factor   /FEA = EST /DB_XREF = gi: 6228471     receptor
  receptor   /DB_XREF = est: au91e07.x1     signaling
  (erythroblastic   /CLONE = IMAGE: 2783652     pathway
  leukemia viral (v-   /UG = Hs.77432 epidermal growth factor
  erb-b) oncogene   receptor (avian erythroblastic leukemia
  homolog, avian)   viral (v-erb-b) oncogene homolog)
  (LOC1956)   /FL = gb: NM_005228.1
  SEQ ID NOS: 86
  (DNA) and 285
  (amino acid)
  AMPD2:   Consensus includes gb: AI916249   212360_at   purine
  adenosine   /FEA = EST /DB_XREF = gi: 5636104     nucleotide
  monophosphate   /DB_XREF = est: wg99c01.x1     metabolism
  deaminase 2   /CLONE = IMAGE: 2379360
  (isoform L)   /UG = Hs.82927 adenosine
  (LOC271)   monophosphate deaminase 2 (isoform L)
  SEQ ID NOS: 87   /FL = gb: NM_004037.2
  (DNA) and 286
  (amino acid)
  GLS: glutaminase“gb: AF158555.1 /DEF = Homo sapiens   221510_s_at   glutamine
  (LOC2744)   glutaminase C mRNA, complete cds.     catabolism
  SEQ ID NOS: 88   /FEA = mRNA /PROD = glutaminase C
  (DNA) and 287   /DB_XREF = gi: 5690371
  (amino acid)   /UG = Hs.239189 glutaminase
    /FL = gb: AF158555.1 gb: AF097492.1”
  EBNA1BP2:“gb: NM_006824.1 /DEF = Homo sapiens   201323_at   ribosome
  EBNA1 binding   nucleolar protein p40; homolog of yeast     biogenesis
  protein 2   EBNA1-binding protein (P40), mRNA.
  (LOC10969)   /FEA = mRNA /GEN = P40
  SEQ ID NOS: 89   /PROD = nucleolar protein p40; homolog
  (DNA) and 288   of yeastEBNA1-binding protein
  (amino acid)   /DB_XREF = gi: 5803110 /UG = Hs.74407
    nucleolar protein p40; homolog of yeast
    EBNA1-binding protein
    /FL = gb: U86602.1 gb: NM_006824.1”
  VIM: vimentin   Consensus includes gb: AI922599   201426_s_at
  (LOC7431)   /FEA = EST /DB_XREF = gi: 5658563
  SEQ ID NOS: 90   /DB_XREF = est: wm90b11.x1
  (DNA) and 289   /CLONE = IMAGE: 2443197
  (amino acid)   /UG = Hs.297753 vimentin
    /FL = gb: BC000163.2 gb: NM_003380.1
  ZNF258: zinc“gb: NM_007167.1 /DEF = Homo sapiens   219924_s_at   development
  finger protein 258   zinc finger protein 258 (ZNF258),
  (LOC9204)   mRNA. /FEA = mRNA /GEN = ZNF258
  SEQ ID NOS: 91   /PROD = zinc finger protein 258
  (DNA) and 290   /DB_XREF = gi: 6005977
  (amino acid)   /UG = Hs.301637 zinc finger protein 258
    /FL = gb: AF055470.1 gb: NM_007167.1”
  SGCE:“gb: NM_003919.1 /DEF = Homo sapiens   204688_at   muscle
  sarcoglycan,   sarcoglycan, epsilon (SGCE), mRNA.     development
  epsilon   /FEA = mRNA /GEN = SGCE
  (LOC8910)   /PROD = sarcoglycan, epsilon
  SEQ ID NOS: 92   /DB_XREF = gi: 10835046
  (DNA) and 291   /UG = Hs.110708 sarcoglycan, epsilon
  (amino acid)   /FL = gb: NM_003919.1 gb: AF036364.1”
  CD44: CD44   “gb: M24915.1 /DEF = Human CDw44   204490_s_at   cell-cell
  antigen (homing   antigen, complete cds. /FEA = mRNA     adhesion
  function and   /DB_XREF = gi: 180196 /UG = Hs.169610
  Indian blood group   CD44 antigen (homing function and
  system) (LOC960)   Indian blood group system)
  SEQ ID NOS: 93   /FL = gb: NM_000610.1 gb: U40373.1
  (DNA) and 292   gb: M59040.1 gb: M24915.1”
  (amino acid)
  SHCBP1: likely“gb: NM_024745.1 /DEF = Homo sapiens   219493_at
  ortholog of mouse   hypothetical protein FLJ22009
  Shc SH2-domain   (FLJ22009), mRNA. /FEA = mRNA
  binding protein 1   /GEN = FLJ22009 /PROD = hypothetical
  (LOC79801)   protein FLJ22009
  SEQ ID NOS: 94   /DB_XREF = gi: 13376069
  (DNA) and 293   /UG = Hs.123253 hypothetical protein
  (amino acid)   FLJ22009 /FL = gb: NM_024745.1”
  IMP-3: IGF-II“gb: NM_006547.1 /DEF = Homo sapiens   203820_s_at   protein
  mRNA-binding   IGF-II mRNA-binding protein 3     biosynthesis
  protein 3   (KOC1), mRNA. /FEA = mRNA
  (LOC10643)   /GEN = KOC1 /PROD = IGF-II mRNA-
  SEQ ID NOS: 95   binding protein 3
  (DNA) and 294   /DB_XREF = gi: 5729900 /UG = Hs.79440
  (amino acid)   IGF-II mRNA-binding protein 3
    /FL = gb: U97188.1 gb: U76705.1
    gb: AF117108.1 gb: NM_006547.1”
  BTG3: BTG“gb: NM_006806.1 /DEF = Homo sapiens   205548_s_at   regulation
  family, member 3   BTG family, member 3 (BTG3), mRNA.     of cell
  (LOC10950)   /FEA = mRNA /GEN = BTG3     cycle
  SEQ ID NOS: 96   /PROD = BTG family, member 3
  (DNA) and 295   /DB_XREF = gi: 5802989 /UG = Hs.77311
  (amino acid)   BTG family, member 3
    /FL = gb: D64110.1 gb: NM_006806.1”
  RAI14: retinoic“gb: NM_015577.1 /DEF = Homo sapiens   202052_s_at
  acid induced 14   novel retinal pigment epithelial gene
  (LOC26064)   (NORPEG), mRNA. /FEA = mRNA
  SEQ ID NOS: 97   /GEN = NORPEG
  (DNA) and 296   /PROD = DKFZP564G013 protein
  (amino acid)   /DB_XREF = gi: 13470085
    /UG = Hs.15165 novel retinal pigment
    epithelial gene /FL = gb: NM_015577.1
    gb: AF155135.1”
  QKI: quaking   Consensus includes gb: AA149639   212262_at   signal
  homolog, KH   /FEA = EST /DB_XREF = gi: 1720440     transduction
  domain RNA   /DB_XREF = est: zl39c06.s1
  binding (mouse)   /CLONE = IMAGE: 504298
  (LOC9444)   /UG = Hs.15020 homolog of mouse
  SEQ ID NOS: 98   quaking QKI (KH domain RNA binding
  (DNA) and 297   protein) /FL = gb: AF142419.1
  (amino acid)   gb: AF142422.1
  CGI-100: CGI-100   “Consensus includes gb: AL117354   202194_at   intracellular
  protein   /DEF = Human DNA sequence from clone     protein
  (LOC50999)   RP5-976O13 on chromosome 1p21.2-22.2     transport
  SEQ ID NOS: 99   Contains part of the gene for CGI-
  (DNA) and 298   100 protein, 3 isoforms of the gene for
  (amino acid)   M96 protein, ESTs, STSs, GSSs and a
    CpG Island /FEA = mRNA_1
    /DB_XREF = gi: 6822199
    /UG = Hs.296155 CGI-100 protein
    /FL = gb: AF151858.1 gb: NM_016040.1”
  CTSZ: cathepsin Z“gb: AF073890.1 /DEF = Homo sapiens   210042_s_at   proteolysis
  (LOC1522)   cathepsin X precursor, mRNA, complete     and
  SEQ ID NOS: 100   cds. /FEA = mRNA /PROD = cathepsin X     peptidolysis
  (DNA) and 299   precursor /DB_XREF = gi: 3650497
  (amino acid)   /UG = Hs.252549 cathepsin Z
    /FL = gb: AF032906.1 gb: AF073890.1
    gb: NM_001336.1 gb: AF136273.1”
 
[0047] The biomarkers provided in Table 2 include the nucleotide sequences of SEQ ID NOS:101-200 and the amino acid sequences of SEQ ID NOS:300-395.
[0048] 
[00002] [TABLE-US-00002]
  TABLE 2
 
  BIOMARKERS (RESISTANT)
      Affymetrix  
  Unigene title and     Probe   Gene
  SEQ ID NO:   Affymetrix Description   Set   Ontology
 
  GATA3: GATA“gb: BC003070.1 /DEF = Homo sapiens,   209604_s_at   proteolysis
  binding protein 3   GATA-binding protein 3, clone     and
  (LOC2625)   MGC: 2346, mRNA, complete cds.     peptidolysis
  SEQ ID NOS: 101   /FEA = mRNA /PROD = GATA-binding
  (DNA) and 300   protein 3 /DB_XREF = gi: 13111765
  (amino acid)   /UG = Hs.169946 GATA-binding protein
    3 /FL = gb: BC003070.1 gb: M69106.1
    gb: NM_002051.1”
  TFF1: trefoil“gb: NM_003225.1 /DEF = Homo sapiens   205009_at   carbohydrate
  factor 1 (breast   trefoil factor 1 (breast cancer, estrogen-     metabolism,
  cancer, estrogen-   inducible sequence expressed in) (TFF1),     cell
  inducible sequence   mRNA. /FEA = mRNA /GEN = TFF1     growth
  expressed in)   /PROD = trefoil factor 1 (breast     and/or
  (LOC7031)   cancer, estrogen-inducible sequence     maintenance
  SEQ ID NOS: 102   expressed in) /DB_XREF = gi: 4507450
  (DNA) and 301   /UG = Hs.1406 trefoil factor 1 (breast
  (amino acid)   cancer, estrogen-inducible sequence
    expressed in) /FL = gb: NM_003225.1”
  ZFYVE21: zinc“gb: NM_024071.1 /DEF = Homo sapiens   219929_s_at
  finger, FYVE   hypothetical protein MGC2550
  domain containing   (MGC2550), mRNA. /FEA = mRNA
  21 (LOC79038)   /GEN = MGC2550 /PROD = hypothetical
  SEQ ID NOS: 103   protein MGC2550
  (DNA) and 302   /DB_XREF = gi: 13129053
  (amino acid)   /UG = Hs.318498 hypothetical protein
    MGC2550 /FL = gb: BC001130.1
    gb: NM_024071.1”
  ATP5G2: ATP“gb: D13119.1 /DEF = Homo sapiens P2   208764_s_at   proton
  synthase, H+   mRNA for ATP synthase subunit c,     transport
  transporting,   complete cds. /FEA = mRNA /GEN = P2
  mitochondrial F0   /PROD = ATP synthase subunit c
  complex, subunit c   precursor /DB_XREF = gi: 285909
  (subunit 9),   /UG = Hs.89399 ATP synthase, H+
  isoform 2   transporting, mitochondrial F0 complex,
  (LOC517)   subunit c (subunit 9), isoform 2
  SEQ ID NOS: 104   /FL = gb: D13119.1”
  (DNA) and 303
  (amino acid)
  EGFL5: EGF-like-   “Consensus includes gb: W68084   212830_at   metabolism
  domain, multiple 5   /FEA = EST /DB_XREF = gi: 1376954
  (LOC1955)   /DB_XREF = est: zd42f12.s1
  SEQ ID NOS: 105   /CLONE = IMAGE: 343343 /UG = Hs.5599
  (DNA) and 304   EGF-like-domain, multiple 5”
  (amino acid)
  MCCC2:“gb: AB050049.1 /DEF = Homo sapiens   209624_s_at   leucine
  methylcrotonoyl-   mccb mRNA for non-biotin containing     catabolism
  Coenzyme A   subunit of 3-methylcrotonyl-CoA
  carboxylase 2   carboxylase, complete cds.
  (beta)   /FEA = mRNA /GEN = mccb /PROD = non-
  (LOC64087)   biotin containing subunit of3-
  SEQ ID NOS: 106   methylcrotonyl-CoA carboxylase
  (DNA) and 305   /DB_XREF = gi: 10934058
  (amino acid)   /UG = Hs.167531 methylcrotonoyl-
    Coenzyme A carboxylase 2 (beta)
    /FL = gb: AB050049.1 gb: AF310971.1
    gb: AF301000.1 gb: NM_022132.2”
  ABAT: 4-“gb: AF237813.1 /DEF = Homo sapiens   209460_at
  aminobutyrate   NPD009 mRNA, complete cds.
  aminotransferase   /FEA = mRNA /PROD = NPD009
  (LOC18)   /DB_XREF = gi: 9963907
  SEQ ID NOS: 107   /UG = Hs.283675 NPD009 protein
  (DNA) and 306   /FL = gb: NM_020686.1 gb: AF237813.1”
  (amino acid)
  —: Clone   “Consensus includes gb: AV700224   208774_at   signal
  IMAGE: 3869896,   /FEA = EST /DB_XREF = gi: 10302195     transduction
  mRNA   /DB_XREF = est: AV700224
  (LOC388434)   /CLONE = GKCARG01 /UG = Hs.75852
  SEQ ID NOS: 108   casein kinase 1, delta
  (DNA)   /FL = gb: BC003558.1”
  FEM1B: fem-1   Consensus includes gb: AI799061   212367_at   induction
homolog b (C. elegans)   /FEA = EST /DB_XREF = gi: 5364533     of
  (LOC10116)   /DB_XREF = est: we98a10.x1     apoptosis
  SEQ ID NOS: 109   /CLONE = IMAGE: 2349114
  (DNA) and 307/UG = Hs.6048 FEM-1 (C. elegans)
  (amino acid)   homolog b /FL = gb: AF178632.1
    gb: NM_015322.1 gb: AF204883.1
  SLC35A1: solute“gb: NM_006416.1 /DEF = Homo sapiens   203306_s_at
  carrier family 35   solute carrier family 35 (CMP-sialic acid
  (CMP-sialic acid   transporter), member 1 (SLC35A1),
  transporter),   mRNA. /FEA = mRNA /GEN = SLC35A1
  member A1   /PROD = solute carrier family 35 (CMP-
  (LOC10559)   sialic acidtransporter), member 1
  SEQ ID NOS: 110   /DB_XREF = gi: 5453620 /UG = Hs.82921
  (DNA) and 308   solute carrier family 35 (CMP-sialic acid
  (amino acid)   transporter), member 1 /FL = gb: D87969.1
    gb: NM_006416.1”
  ZNF607: zinc   Consensus includes gb: AL560017   200658_s_at   DNA
  finger protein 607   /FEA = EST /DB_XREF = gi: 12906073     metabolism
  (LOC84775)   /DB_XREF = est: AL560017
  SEQ ID NOS: 111   /CLONE = CS0DG004YD08 (5 prime)
  (DNA) and 309   /UG = Hs.75323 prohibitin
  (amino acid)   /FL = gb: NM_002634.2
  FLJ11164:“gb: NM_018346.1 /DEF = Homo sapiens   218307_at
  hypothetical   hypothetical protein FLJ11164
  protein FLJ11164   (FLJ11164), mRNA. /FEA = mRNA
  (LOC55316)   /GEN = FLJ11164 /PROD = hypothetical
  SEQ ID NOS: 112   protein FLJ11164
  (DNA) and 310   /DB_XREF = gi: 8922910 /UG = Hs.8033
  (amino acid)   hypothetical protein FLJ11164
    /FL = gb: NM_018346.1”
  ABAT: 4-“gb: NM_000663.1 /DEF = Homo sapiens   206527_at   aminobutyrate
  aminobutyrate   4-aminobutyrate aminotransferase     metabolism
  aminotransferase   (ABAT), nuclear gene encoding
  (LOC18)   mitochondrial protein, mRNA.
  SEQ ID NOS: 113   /FEA = mRNA /GEN = ABAT /PROD = 4-
  (DNA) and 311   aminobutyrate aminotransferase
  (amino acid)   precursor /DB_XREF = gi: 4501846
    /UG = Hs.1588 4-aminobutyrate
    aminotransferase /FL = gb: NM_000663.1
    gb: L32961.1”
  SLC19A2: solute“gb: AF153330.1 /DEF = Homo sapiens   209681_at   small
  carrier family 19   thiamine carrier 1 (TC1) mRNA,     molecule
  (thiamine   complete cds. /FEA = mRNA /GEN = TC1     transport
  transporter),   /PROD = thiamine carrier 1
  member 2   /DB_XREF = gi: 5453325 /UG = Hs.30246
  (LOC10560)   solute carrier family 19 (thiamine
  SEQ ID NOS: 114   transporter), member 2
  (DNA) and 312   /FL = gb: AF153330.1 gb: AF135488.1
  (amino acid)   gb: AF160812.1”
  SLC9A3R1: solute“gb: NM_004252.1 /DEF = Homo sapiens   201349_at   intracellular
  carrier family 9   solute carrier family 9 (sodiumhydrogen     signaling
  (sodium/hydrogen   exchanger), isoform 3 regulatory factor 1     cascade
  exchanger),   (SLC9A3R1), mRNA. /FEA = mRNA
  isoform 3 regulator   /GEN = SLC9A3R1 /PROD = solute carrier
  1 (LOC9368)   family 9 (sodiumhydrogenexchanger),
  SEQ ID NOS: 115   isoform 3 regulatory factor 1
  (DNA) and 313   /DB_XREF = gi: 4759139
  (amino acid)   /UG = Hs.184276 solute carrier family 9
    (sodiumhydrogen exchanger), isoform 3
    regulatory factor 1 /FL = gb: BC001443.1
    gb: BC003361.1 gb: AF036241.1
    gb: AF015926.1 gb: NM_004252.1”
  ICA1: islet cell   “gb: L21181.1 /DEF = Human autoantigen   210547_x_at
  autoantigen 1,   p69 mRNA, complete cds. /FEA = mRNA
  69 kDa (LOC3382)   /PROD = autoantigen p69
  SEQ ID NOS: 116   /DB_XREF = gi: 437366 /UG = Hs.167927
  (DNA) and 314   islet cell autoantigen 1 (69 kD)
  (amino acid)   /FL = gb: L21181.1”
  CIRBP: cold“gb: NM_001280.1 /DEF = Homo sapiens   200811_at   response
  inducible RNA   cold inducible RNA-binding protein     to cold
  binding protein   (CIRBP), mRNA. /FEA = mRNA
  (LOC1153)   /GEN = CIRBP /PROD = cold inducible
  SEQ ID NOS: 117   RNA-binding protein
  (DNA) and 315   /DB_XREF = gi: 4502846
  (amino acid)   /UG = Hs.119475 cold inducible RNA-
    binding protein /FL = gb: D78134.1
    gb: BC000403.1 gb: BC000901.1
    gb: AF021336.1 gb: NM_001280.1”
  C14orf114:“gb: NM_018199.1 /DEF = Homo sapiens   218363_at
  chromosome 14   hypothetical protein FLJ10738
  open reading   (FLJ10738), mRNA. /FEA = mRNA
  frame 114   /GEN = FLJ10738 /PROD = hypothetical
  (LOC55218)   protein FLJ10738
  SEQ ID NOS: 118   /DB_XREF = gi: 8922630 /UG = Hs.5457
  (DNA) and 316   hypothetical protein FLJ10738
  (amino acid)   /FL = gb: BC001962.1 gb: NM_018199.1”
  GREB1: GREB1“gb: NM_014668.1 /DEF = Homo sapiens   205862_at
  protein   KIAA0575 gene product (KIAA0575),
  (LOC9687)   mRNA. /FEA = mRNA /GEN = KIAA0575
  SEQ ID NOS: 119   /PROD = KIAA0575 gene product
  (DNA) and 317   /DB_XREF = gi: 7662187
  (amino acid)   /UG = Hs.193914 KIAA0575 gene
    product /FL = gb: AB011147.1
    gb: NM_014668.1”
  ESR1: estrogen“gb: NM_000125.1 /DEF = Homo sapiens   205225_at   nuclear
  receptor 1   estrogen receptor 1 (ESR1), mRNA.     hormone
  (LOC2099)   /FEA = mRNA /GEN = ESR1     receptor,
  SEQ ID NOS: 120   /PROD = estrogen receptor 1     cellular
  (DNA) and 318   /DB_XREF = gi: 4503602 /UG = Hs.1657     proliferation
  (amino acid)   estrogen receptor 1     and
    /FL = gb: NM_000125.1”     differentiation
  ABAT: 4-“gb: AF237813.1 /DEF = Homo sapiens   209459_s_at
  aminobutyrate   NPD009 mRNA, complete cds.
  aminotransferase   /FEA = mRNA /PROD = NPD009
  (LOC18)   /DB_XREF = gi: 9963907
  SEQ ID NOS: 121   /UG = Hs.283675 NPD009 protein
  (DNA) and 319   /FL = gb: NM_020686.1 gb: AF237813.1”
  (amino acid)
  ABCG1: ATP-“gb: NM_004915.2 /DEF = Homo sapiens   204567_s_at   small
  binding cassette,   ATP-binding cassette, sub-family G     molecule
  sub-family G   (WHITE), member 1 (ABCG1),     transport
  (WHITE), member   transcript variant 1, mRNA.
  1 (LOC9619)   /FEA = mRNA /GEN = ABCG1
  SEQ ID NOS: 122   /PROD = ATP-binding cassette sub-
  (DNA) and 320   family G member 1isoform a
  (amino acid)   /DB_XREF = gi: 8051574 /UG = Hs.10237
    ATP-binding cassette, sub-family G
    (WHITE), member 1
    /FL = gb: NM_004915.2”
  SLC35B1: solute“gb: NM_005827.1 /DEF = Homo sapiens   202433_at   transport
  carrier family 35,   UDP-galactose transporter related
  member B1   (UGTREL1), mRNA. /FEA = mRNA
  (LOC10237)   /GEN = UGTREL1 /PROD = UDP-
  SEQ ID NOS: 123   galactose transporter related
  (DNA) and 321   /DB_XREF = gi: 5032212
  (amino acid)   /UG = Hs.154073 UDP-galactose
    transporter related /FL = gb: D87989.1
    gb: NM_005827.1”
  TOB1: transducer   “Consensus includes gb: AA675892   202704_at   negative
  of ERBB2, 1   /FEA = EST /DB_XREF = gi: 2775239     regulation
  (LOC10140)   /DB_XREF = est: b03503s     of cell
  SEQ ID NOS: 124   /CLONE = b03503 /UG = Hs.178137     proliferation
  (DNA) and 322   transducer of ERBB2, 1
  (amino acid)   /FL = gb: D38305.1 gb: NM_005749.1”
  FOXA1: forkhead“gb: NM_004496.1 /DEF = Homo sapiens   204667_at   regulation
  box A1   hepatocyte nuclear factor 3, alpha     of
  (LOC3169)   (HNF3A), mRNA. /FEA = mRNA     transcription,
  SEQ ID NOS: 125   /GEN = HNF3A /PROD = hepatocyte     DNA-
  (DNA) and 323   nuclear factor 3, alpha     dependent
  (amino acid)   /DB_XREF = gi: 4758533
    /UG = Hs.299867 hepatocyte nuclear
    factor 3, alpha /FL = gb: U39840.1
    gb: NM_004496.1”
  LARGE: like-   “Consensus includes gb: AB011181.2   215543_s_at   muscle
  glycosyltransferase/DEF = Homo sapiens mRNA for     maintenance,
  (LOC9215)   KIAA0609 protein, partial cds.     glycosphingolipid
  SEQ ID NOS: 126   /FEA = mRNA /GEN = KIAA0609     biosynthesis
  (DNA) and 324   /PROD = KIAA0609 protein
  (amino acid)   /DB_XREF = gi: 6683718 /UG = Hs.25220
    like-glycosyltransferase”
  AKT1: v-akt“gb: NM_005163.1 /DEF = Homo sapiens   207163_s_at   signal
  murine thymoma   v-akt murine thymoma viral oncogene     transduction
  viral oncogene   homolog 1 (AKT1), mRNA.
  homolog 1   /FEA = mRNA /GEN = AKT1
  (LOC207)   /PROD = serinethreonine protein kinase
  SEQ ID NOS: 127   /DB_XREF = gi: 4885060 /UG = Hs.71816
  (DNA) and 325   v-akt murine thymoma viral oncogene
  (amino acid)   homolog 1 /FL = gb: M63167.1
    gb: NM_005163.1”
  CTPS2: CTP“gb: NM_019857.1 /DEF = Homo sapiens   219080_s_at
  synthase II   CTP synthase II (CTPS2), mRNA.
  (LOC56474)   /FEA = mRNA /GEN = CTPS2
  SEQ ID NOS: 128   /PROD = CTP synthase II
  (DNA) and 326   /DB_XREF = gi: 9789918 /UG = Hs.58553
  (amino acid)   CTP synthase II /FL = gb: AF226667.1
    gb: NM_019857.1”
  RBM8A: RNA   Consensus includes gb: AI738479   214113_s_at   nuclear
  binding motif   /FEA = EST /DB_XREF = gi: 5100460     mRNA
  protein 8A   /DB_XREF = est: wi32d06.x1     splicing,
  (LOC9939)   /CLONE = IMAGE: 2391947     via
  SEQ ID NOS: 129   /UG = Hs.65648 RNA binding motif     spliceosome
  (DNA) and 327   protein 8A
  (amino acid)
  SIAH2: seven in   “gb: U76248.1 /DEF = Human hSIAH2   209339_at   small
  absentia homolog   mRNA, complete cds. /FEA = mRNA     GTPase
2 (Drosophila)   /PROD = hSIAH2     mediated
  (LOC6478)   /DB_XREF = gi: 2673967 /UG = Hs.20191     signal
  SEQ ID NOS: 130seven in absentia (Drosophila) homolog     transduction
  (DNA) and 328   2 /FL = gb: U76248.1 gb: NM_005067.1”
  (amino acid)
  FLJ13855:“gb: NM_023079.1 /DEF = Homo sapiens   217750_s_at   ubiquitin
  hypothetical   hypothetical protein FLJ13855     cycle
  protein FLJ13855   (FLJ13855), mRNA. /FEA = mRNA
  (LOC65264)   /GEN = FLJ13855 /PROD = hypothetical
  SEQ ID NOS: 131   protein FLJ13855
  (DNA) and 329   /DB_XREF = gi: 12751494
  (amino acid)   /UG = Hs.168232 hypothetical protein
    FLJ13855 /FL = gb: NM_023079.1”
  ITPK1: inositol“gb: AF279372.1 /DEF = Homo sapiens   210740_s_at   signal
  1,3,4-triphosphate   inositol 1,3,4-trisphosphate 56-kinase     transduction
  5/6 kinase   mRNA, complete cds. /FEA = mRNA
  (LOC3705)   /PROD = inositol 1,3,4-trisphosphate 56-
  SEQ ID NOS: 132   kinase /DB_XREF = gi: 12006345
  (DNA) and 330   /UG = Hs.6453 inositol 1,3,4-triphosphate
  (amino acid)   56 kinase /FL = gb: AF279372.1”
  SEPX1:“gb: NM_016332.1 /DEF = Homo sapiens   217977_at
  selenoprotein X, 1   selenoprotein X, 1 (SEPX1), mRNA.
  (LOC51734)   /FEA = mRNA /GEN = SEPX1
  SEQ ID NOS: 133   /PROD = selenoprotein X, 1
  (DNA) and 331   /DB_XREF = gi: 7706510
  (amino acid)   /UG = Hs.279623 selenoprotein X, 1
    /FL = gb: AF187272.1 gb: BC003127.1
    gb: AF166124.1 gb: NM_016332.1”
  IRX5: iroquois   “gb: U90304.1 /DEF = Human iroquois-   210239_at   regulation
  homeobox protein   class homeodomain protein IRX-2a     of
  5 (LOC10265)   mRNA, complete cds. /FEA = mRNA     transcription,
  SEQ ID NOS: 134   /PROD = iroquois-class homeodomain     DNA-
  (DNA) and 332   protein IRX-2a /DB_XREF = gi: 1899219     dependent
  (amino acid)   /UG = Hs.25351 iroquois homeobox
    protein 5 /FL = gb: U90304.1
    gb: NM_005853.1”
  PTEN:“gb: BC005821.1 /DEF = Homo sapiens,   211711_s_at   regulation
  phosphatase and   phosphatase and tensin homolog     of CDK
  tensin homolog   (mutated in multiple advanced cancers     activity
  (mutated in   1), clone MGC: 11227, mRNA, complete
  multiple advanced   cds. /FEA = mRNA /PROD = phosphatase
  cancers 1)   and tensin homolog (mutated inmultiple
  (LOC5728)   advanced cancers 1)
  SEQ ID NOS: 135   /DB_XREF = gi: 13543309
  (DNA) and 333   /FL = gb: BC005821.1”
  (amino acid)
  DP1: polyposis“gb: BC000232.1 /DEF = Homo sapiens,   208873_s_at
  locus protein 1   Similar to deleted in polyposis 1, clone
  (LOC7905)   MGC: 2267, mRNA, complete cds.
  SEQ ID NOS: 136   /FEA = mRNA /PROD = Similar to deleted
  (DNA) and 334   in polyposis 1 /DB_XREF = gi: 12652946
  (amino acid)   /UG = Hs.178112 DNA segment, single
    copy probe LNS-CAILNS-CAII (deleted
    in polyposis /FL = gb: BC000232.1”
  KIAA1002:“gb: NM_014925.1 /DEF = Homo sapiens   203831_at
  KIAA1002 protein   KIAA1002 protein (KIAA1002), mRNA.
  (LOC22864)   /FEA = mRNA /GEN = KIAA1002
  SEQ ID NOS: 137   /PROD = KIAA1002 protein
  (DNA) and 335   /DB_XREF = gi: 7662441
  (amino acid)   /UG = Hs.102483 KIAA1002 protein
    /FL = gb: AB023219.1 gb: AF113695.1
    gb: NM_014925.1”
  PDCD4:   “Consensus includes gb: N92498   212593_s_at   apoptosis
  programmed cell   /FEA = EST /DB_XREF = gi: 1264807
  death 4 (neoplastic   /DB_XREF = est: zb28a04.s1
  transformation   /CLONE = IMAGE: 304878
  inhibitor)/UG = Hs.326248 Homo sapiens cDNA:
  (LOC27250)   FLJ22071 fis, clone HEP11691”
  SEQ ID NOS: 138
  (DNA) and 336
  (amino acid)
  APPBP2: amyloid   Consensus includes gb: AV681579   202629_at   intracellular
  beta precursor   /FEA = EST /DB_XREF = gi: 10283442     protein
  protein   /DB_XREF = est: AV681579     transport
  (cytoplasmic tail)   /CLONE = GKBAFE05 /UG = Hs.84084
  binding protein 2   amyloid beta precursor protein
  (LOC10513)   (cytoplasmic tail)-binding protein 2
  SEQ ID NOS: 139   /FL = gb: AF017782.1 gb: NM_006380.1
  (DNA) and 337
  (amino acid)
  ACVR1B: activin   Consensus includes gb: AL117643.1   213198_at   transmembrane
  A receptor, type IB/DEF = Homo sapiens mRNA; cDNA     receptor
  (LOC91)   DKFZp434M245 (from clone     protein
  SEQ ID NOS: 140   DKFZp434M245). /FEA = mRNA     serine/threonine
  (DNA) and 338   /DB_XREF = gi: 5912233 /UG = Hs.5288     kinase
  (amino acid)Homo sapiens mRNA; cDNA     signaling
    DKFZp434M245 (from clone     pathway
    DKFZp434M245)
  TLE3: transducin-   Consensus includes gb: AW873621   212770_at   regulation
  like enhancer of   /FEA = EST /DB_XREF = gi: 8007674     of
  split 3 (E(sp1)   /DB_XREF = est: ho64d03.x1     transcription,
  homolog,   /CLONE = IMAGE: 3042149     DNA-
Drosophila)   /UG = Hs.31305 KIAA1547 protein     dependent
  (LOC7090)
  SEQ ID NOS: 141
  (DNA) and 339
  (amino acid)
  CIRBP: cold“gb: NM_001280.1 /DEF = Homo sapiens   200810_s_at   response
  inducible RNA   cold inducible RNA-binding protein     to cold
  binding protein   (CIRBP), mRNA. /FEA = mRNA
  (LOC1153)   /GEN = CIRBP /PROD = cold inducible
  SEQ ID NOS: 142   RNA-binding protein
  (DNA) and 340   /DB_XREF = gi: 4502846
  (amino acid)   /UG = Hs.119475 cold inducible RNA-
    binding protein /FL = gb: D78134.1
    gb: BC000403.1 gb: BC000901.1
    gb: AF021336.1 gb: NM_001280.1”
  ABCA3: ATP-“gb: NM_001089.1 /DEF = Homo sapiens   204343_at   drug
  binding cassette,   ATP-binding cassette, sub-family A     resistance
  sub-family A   (ABC1), member 3 (ABCA3), mRNA.
  (ABC1), member   /FEA = mRNA /GEN = ABCA3
  3 (LOC21)   /PROD = ATP-binding cassette, sub-
  SEQ ID NOS: 143   family A member 3
  (DNA) and 341   /DB_XREF = gi: 4501848 /UG = Hs.26630
  (amino acid)   ATP-binding cassette, sub-family A
    (ABC1), member 3 /FL = gb: U78735.1
    gb: NM_001089.1”
  MTSS1:“gb: NM_014751.1 /DEF = Homo sapiens   203037_s_at
  metastasis   KIAA0429 gene product (KIAA0429),
  suppressor 1   mRNA. /FEA = mRNA /GEN = KIAA0429
  (LOC9788)   /PROD = KIAA0429 gene product
  SEQ ID NOS: 144   /DB_XREF = gi: 7662113 /UG = Hs.77694
  (DNA) and 342   KIAA0429 gene product
  (amino acid)   /FL = gb: AB007889.1 gb: NM_014751.1”
  CA12: carbonic“gb: NM_001218.2 /DEF = Homo sapiens   203963_at   one-
  anhydrase XII   carbonic anhydrase XII (CA12), mRNA.     carbon
  (LOC771)   /FEA = mRNA /GEN = CA12     compound
  SEQ ID NOS: 145   /PROD = carbonic anhydrase XII     metabolism
  (DNA) and 343   precursor /DB_XREF = gi: 9951924
  (amino acid)   /UG = Hs.5338 carbonic anhydrase XII
    /FL = gb: AF037335.1 gb: AF051882.1
    gb: NM_001218.2”
  FRAT2: frequently“gb: AB045118.1 /DEF = Homo sapiens   209864_at
  rearranged in   FRAT2 mRNA, complete cds.
  advanced T-cell   /FEA = mRNA /GEN = FRAT2
  lymphomas 2   /PROD = FRAT2
  (LOC23401)   /DB_XREF = gi: 13365650
  SEQ ID NOS: 146   /UG = Hs.140720 GSK-3 binding protein
  (DNA) and 344   FRAT2 /FL = gb: AB045118.1”
  (amino acid)
  SUPT4H1:“gb: NM_003168.1 /DEF = Homo sapiens   201484_at   chromatin
  suppressor of Ty 4suppressor of Ty (S. cerevisiae) 4     modeling
homolog 1 (S. cerevisiae)   homolog 1 (SUPT4H1), mRNA.
  (LOC6827)   /FEA = mRNA /GEN = SUPT4H1
  SEQ ID NOS: 147/PROD = suppressor of Ty (S. cerevisiae) 4
  (DNA) and 345   homolog 1 /DB_XREF = gi: 4507310
  (amino acid)   /UG = Hs.79058 suppressor of Ty
  (S. cerevisiae) 4 homolog 1
    /FL = gb: BC002802.1 gb: U43923.1
    gb: U38818.1 gb: U38817.1
    gb: NM_003168.1”
  UBPH: similar to“gb: NM_019116.1 /DEF = Homo sapiens   205687_at
  ubiquitin binding   similar to ubiquitin binding protein
  protein   (UBPH), mRNA. /FEA = mRNA
  (LOC56061)   /GEN = UBPH /PROD = similar to
  SEQ ID NOS: 148   ubiquitin binding protein
  (DNA) and 346   /DB_XREF = gi: 9507222
  (amino acid)   /UG = Hs.288620 similar to ubiquitin
    binding protein /FL = gb: NM_019116.1”
  MGC50853:   “Consensus includes gb: AL043266   212400_at
  hypothetical   /FEA = EST /DB_XREF = gi: 5935844
  protein   /DB_XREF = est: DKFZp434L1423_s1
  MGC50853   /CLONE = DKFZp434L1423
  (LOC399665)   /UG = Hs.111334 ferritin, light
  SEQ ID NOS: 149   polypeptide”
  (DNA) and 347
  (amino acid)
  TBL1X:   Consensus includes gb: AV753028   213400_s_at   signal
  transducin (beta)-   /FEA = EST /DB_XREF = gi: 10910876     transduction
  like 1X-linked   /DB_XREF = est: AV753028
  (LOC6907)   /CLONE = NPDBCD07 /UG = Hs.76536
  SEQ ID NOS: 150   transducin (beta)-like 1
  (DNA) and 348
  (amino acid)
  FLJ11280:   Consensus includes gb: AL561943   221856_s_at
  hypothetical   /FEA = EST /DB_XREF = gi: 12909874
  protein FLJ11280   /DB_XREF = est: AL561943
  (LOC55793)   /CLONE = CS0DB002YO04 (3 prime)
  SEQ ID NOS: 151   /UG = Hs.3346 hypothetical protein
  (DNA) and 349   FLJ11280
  (amino acid)
  RHOB: ras   “Consensus includes gb: AI263909   212099_at   Rho
  homolog gene   /FEA = EST /DB_XREF = gi: 3872112     protein
  family, member B   /DB_XREF = est: qi08f09.x1     signal
  (LOC388)   /CLONE = IMAGE: 1855913     transduction
  SEQ ID NOS: 152   /UG = Hs.204354 ras homolog gene
  (DNA) and 350   family, member B
  (amino acid)   /FL = gb: NM_004040.1”
  LASS6: LAG1   “Consensus includes gb: BG289001   212442_s_at
  longevity   /FEA = EST /DB_XREF = gi: 13044404
  assurance homolog   /DB_XREF = est: 602381262F1
6 (S. cerevisiae)   /CLONE = IMAGE: 4499078
  (LOC253782)/UG = Hs.101282 Homo sapiens cDNA:
  SEQ ID NOS: 153   FLJ21238 fis, clone COL01115”
  (DNA) and 351
  (amino acid)
  KIAA0515:   “Consensus includes gb: AB011087.1   212068_s_at
  KIAA0515/DEF = Homo sapiens mRNA for
  (LOC84726)   KIAA0515 protein, partial cds.
  SEQ ID NOS: 154   /FEA = mRNA /GEN = KIAA0515
  (DNA)   /PROD = KIAA0515 protein
    /DB_XREF = gi: 3043553
    /UG = Hs.108945 KIAA0515 protein”
  MCCC2:   Consensus includes gb: AW439494   209623_at
  methylcrotonoyl-   /FEA = EST /DB_XREF = gi: 6974800
  Coenzyme A   /DB_XREF = est: xt19c01.x1
  carboxylase 2   /CLONE = IMAGE: 2779584
  (beta)   /UG = Hs.167531 methylcrotonoyl-
  (LOC64087)   Coenzyme A carboxylase 2 (beta)
  SEQ ID NOS: 155   /FL = gb: AB050049.1 gb: AF310971.1
  (DNA) and 352   gb: AF301000.1 gb: NM_022132.2
  (amino acid)
  TFF3: trefoil“gb: NM_003226.1 /DEF = Homo sapiens   204623_at   phosphoenolpyruvate-
  factor 3 (intestinal)   trefoil factor 3 (intestinal) (TFF3),     dependent
  (LOC7033)   mRNA. /FEA = mRNA /GEN = TFF3     sugar
  SEQ ID NOS: 156   /PROD = trefoil factor 3 (intestinal)     phosphotransferase
  (DNA) and 353   /DB_XREF = gi: 4507452 /UG = Hs.82961     system
  (amino acid)   trefoil factor 3 (intestinal)
    /FL = gb: L08044.1 gb: L15203.1
    gb: NM_003226.1”
  GATA3: GATA   Consensus includes gb: AI796169   209603_at   defense
  binding protein 3   /FEA = EST /DB_XREF = gi: 5361632     response
  (LOC2625)   /DB_XREF = est: wh43d10.x1
  SEQ ID NOS: 157   /CLONE = IMAGE: 2383507
  (DNA) and 354   /UG = Hs.169946 GATA-binding protein
  (amino acid)   3 /FL = gb: BC003070.1 gb: M69106.1
    gb: NM_002051.1
  CEBPA:“gb: NM_004364.1 /DEF = Homo sapiens   204039_at
  CCAAT/enhancer   CCAATenhancer binding protein
  binding protein   (CEBP), alpha (CEBPA), mRNA.
  (C/EBP), alpha   /FEA = mRNA /GEN = CEBPA
  (LOC1050)   /PROD = CCAATenhancer binding
  SEQ ID NOS: 158   protein (CEBP), alpha
  (DNA) and 355   /DB_XREF = gi: 4757971 /UG = Hs.76171
  (amino acid)   CCAATenhancer binding protein
    (CEBP), alpha /FL = gb: NM_004364.1”
  LOC92482:   “Consensus includes gb: AK025724.1   213224_s_at
  hypothetical/DEF = Homo sapiens cDNA: FLJ22071
  protein LOC92482   fis, clone HEP11691. /FEA = mRNA
  (LOC92482)   /DB_XREF = gi: 10438333
  SEQ ID NOS: 159/UG = Hs.326248 Homo sapiens cDNA:
  (DNA)   FLJ22071 fis, clone HEP11691”
  FLJ13910:   Consensus includes gb: BF671894   212482_at
  hypothetical   /FEA = EST /DB_XREF = gi: 11945789
  protein FLJ13910   /DB_XREF = est: 602151796F1
  (LOC64795)   /CLONE = IMAGE: 4292999
  SEQ ID NOS: 160   /UG = Hs.75277 hypothetical protein
  (DNA) and 356   FLJ13910
  (amino acid)
  C14orf130:“gb: NM_018108.1 /DEF = Homo sapiens   218108_at
  chromosome 14   hypothetical protein FLJ10483
  open reading   (FLJ10483), mRNA. /FEA = mRNA
  frame 130   /GEN = FLJ10483 /PROD = hypothetical
  (LOC55148)   protein FLJ10483
  SEQ ID NOS: 161   /DB_XREF = gi: 8922451 /UG = Hs.6877
  (DNA) and 357   hypothetical protein FLJ10483
  (amino acid)   /FL = gb: NM_018108.1”
  CDKN1B: cyclin-“gb: BC001971.1 /DEF = Homo sapiens,   209112_at   regulation
  dependent kinase   Similar to cyclin-dependent kinase     of CDK
  inhibitor 1B (p27,   inhibitor 1B (p27, Kip1), clone     activity
  Kip1) (LOC1027)   MGC: 5304, mRNA, complete cds.
  SEQ ID NOS: 162   /FEA = mRNA /PROD = Similar to cyclin-
  (DNA) and 358   dependent kinase inhibitor 1B (p27, Kip1)
  (amino acid)   /DB_XREF = gi: 12805034
    /UG = Hs.238990 cyclin-dependent kinase
    inhibitor 1B (p27, Kip1)
    /FL = gb: BC001971.1 gb: NM_004064.1
    gb: U10906.1 gb: AF247551.1
    gb: AY004255.1”
  APPBP2: amyloid“gb: NM_006380.1 /DEF = Homo sapiens   202631_s_at   intracellular
  beta precursor   amyloid beta precursor protein     protein
  protein   (cytoplasmic tail)-binding protein 2     transport
  (cytoplasmic tail)   (APPBP2), mRNA. /FEA = mRNA
  binding protein 2   /GEN = APPBP2 /PROD = amyloid beta
  (LOC10513)   precursor protein (cytoplasmictail)-
  SEQ ID NOS: 163   binding protein 2
  (DNA) and 359   /DB_XREF = gi: 5453552 /UG = Hs.84084
  (amino acid)   amyloid beta precursor protein
    (cytoplasmic tail)-binding protein 2
    /FL = gb: AF017782.1 gb: NM_006380.1”
  LOC81558:“gb: NM_030802.1 /DEF = Homo sapiens   221249_s_at
  C/EBP-induced   CEBP-induced protein (LOC81558),
  protein   mRNA. /FEA = mRNA /GEN = LOC81558
  (LOC81558)   /PROD = CEBP-induced protein
  SEQ ID NOS: 164   /DB_XREF = gi: 13540589
  (DNA) and 360   /FL = gb: NM_030802.1”
  (amino acid)
  FLJ20274:   Consensus includes gb: AL134904   213025_at
  hypothetical   /FEA = EST /DB_XREF = gi: 6603091
  protein FLJ20274   /DB_XREF = est: DKFZp762M0710_s1
  (LOC55623)   /CLONE = DKFZp762M0710
  SEQ ID NOS: 165   /UG = Hs.268371 hypothetical protein
  (DNA) and 361   FLJ20274
  (amino acid)
  RAB11A:“gb: NM_004663.1 /DEF = Homo sapiens   200864_s_at   intracellular
  RAB11A, member   RAB11A, member RAS oncogene family     protein
  RAS oncogene   (RAB11A), mRNA. /FEA = mRNA     transport
  family (LOC8766)   /GEN = RAB11A /PROD = RAB11A,
  SEQ ID NOS: 166   member RAS oncogene family
  (DNA) and 362   /DB_XREF = gi: 4758983 /UG = Hs.75618
  (amino acid)   RAB11A, member RAS oncogene family
    /FL = gb: AF000231.1 gb: NM_004663.1”
  —: MRNA; cDNA   Consensus includes gb: BE967207   212114_at
  DKFZp313P052   /FEA = EST /DB_XREF = gi: 11773627
  (from clone   /DB_XREF = est: 601661094R1
  DKFZp313P052)   /CLONE = IMAGE: 3916174
  (LOC387869)   /UG = Hs.165590 ribosomal protein S13
  SEQ ID NOS: 167
  (DNA) and 363
  (amino acid)
  NPEPPS:   “Consensus includes gb: AJ132583.1   201455_s_at   proteolysis
  aminopeptidase/DEF = Homo sapiens mRNA for     and
  puromycin   puromycin sensitive aminopeptidase,     peptidolysis
  sensitive   partial. /FEA = mRNA
  (LOC9520)   /PROD = puromycin sensitive
  SEQ ID NOS: 168   aminopeptidase /DB_XREF = gi: 4210725
  (DNA) and 364   /UG = Hs.293007 aminopeptidase
  (amino acid)   puromycin sensitive
    /FL = gb: NM_006310.1”
  UBL3: ubiquitin-“gb: AF044221.1 /DEF = Homo sapiens   201534_s_at
  like 3 (LOC5412)   HCG-1 protein (HCG-1) mRNA,
  SEQ ID NOS: 169   complete cds. /FEA = mRNA
  (DNA) and 365   /GEN = HCG-1 /PROD = HCG-1 protein
  (amino acid)   /DB_XREF = gi: 4105251
    /UG = Hs.173091 ubiquitin-like 3
    /FL = gb: AF044221.1 gb: AL080177.1
    gb: NM_007106.1”
  BAMBI: BMP and“gb: NM_012342.1 /DEF = Homo sapiens   203304_at
  activin membrane-   putative transmembrane protein (NMA),
  bound inhibitor   mRNA. /FEA = mRNA /GEN = NMA
homolog (Xenopus   /PROD = putative transmembrane protein
laevis)   /DB_XREF = gi: 6912533 /UG = Hs.78776
  (LOC25805)   putative transmembrane protein
  SEQ ID NOS: 170   /FL = gb: U23070.1 gb: NM_012342.1”
  (DNA) and 366
  (amino acid)
  GABPB2: GA“gb: NM_005254.2 /DEF = Homo sapiens   204618_s_at   regulation
  binding protein   GA-binding protein transcription factor,     of
  transcription   beta subunit 1 (53 kD) (GABPB1),     transcription,
  factor, beta subunit   transcript variant beta, mRNA.     DNA-
  2, 47 kDa   /FEA = mRNA /GEN = GABPB1     dependent
  (LOC2553)   /PROD = GA-binding protein
  SEQ ID NOS: 171   transcription factor, betasubunit 1
  (DNA) and 367   (53 kD), isoform beta 1
  (amino acid)   /DB_XREF = gi: 8051592 /UG = Hs.78915
    GA-binding protein transcription factor,
    beta subunit 1 (53 kD) /FL = gb: U13045.1
    gb: NM_005254.2”
  MAPT:   “gb: J03778.1 /DEF = Human microtubule-   206401_s_at   microtubule
  microtubule-   associated protein tau mRNA, complete     cytoskeleton
  associated protein   cds. /FEA = mRNA /GEN = MTBT1     organization
  tau (LOC4137)   /DB_XREF = gi: 338684 /UG = Hs.101174     and
  SEQ ID NOS: 172   microtubule-associated protein tau     biogenesis
  (DNA) and 368   /FL = gb: BC000558.1 gb: J03778.1
  (amino acid)   gb: NM_016841.1”
  WBSCR21:   Consensus includes gb: AI923458   221927_s_at
  Williams Beuren   /FEA = EST /DB_XREF = gi: 5659422
  syndrome   /DB_XREF = est: wn85h04.x1
  chromosome   /CLONE = IMAGE: 2452663
  region 21/UG = Hs.182476 Homo sapiens clone
  (LOC83451)   PP1226 unknown mRNA
  SEQ ID NOS: 173
  (DNA) and 369
  (amino acid)
  ZNF278: zinc“gb: AF242522.1 /DEF = Homo sapiens   211392_s_at
  finger protein 278   krueppel-related zinc finger protein
  (LOC23598)   SBZF5 mRNA, complete cds.
  SEQ ID NOS: 174   /FEA = mRNA /PROD = krueppel-related
  (DNA) and 370   zinc finger protein SBZF5
  (amino acid)   /DB_XREF = gi: 9802041 /UG = Hs.27801
    zinc finger protein 278
    /FL = gb: AF242522.1”
  SUPT4H1:“gb: BC002802.1 /DEF = Homo sapiens,   201483_s_at   chromatin
  suppressor of Ty 4suppressor of Ty (S. cerevisiae) 4     modeling
homolog 1 (S. cerevisiae)   homolog 1, clone MGC: 3864, mRNA,
  (LOC6827)   complete cds. /FEA = mRNA
  SEQ ID NOS: 175/PROD = suppressor of Ty (S. cerevisiae) 4
  (DNA) and 371   homolog 1 /DB_XREF = gi: 12803910
  (amino acid)   /UG = Hs.79058 suppressor of Ty
  (S. cerevisiae) 4 homolog 1
    /FL = gb: BC002802.1 gb: U43923.1
    gb: U38818.1 gb: U38817.1
    gb: NM_003168.1”
  RAB4B: RAB4B,“gb: NM_016154.1 /DEF = Homo sapiens   219807_x_at
  member RAS   ras-related GTP-binding protein 4b
  oncogene family   (RAB4B), mRNA. /FEA = mRNA
  (LOC53916)   /GEN = RAB4B /PROD = ras-related GTP-
  SEQ ID NOS: 176   binding protein 4b
  (DNA) and 372   /DB_XREF = gi: 7706672
  (amino acid)/UG = Hs.279771 Homo sapiens
    TR00071289_m (RAB4B), mRNA
    /FL = gb: AF165522.1 gb: NM_016154.1”
  PEX11B:“gb: NM_003846.1 /DEF = Homo sapiens   202658_at   peroxisome
  peroxisomal   peroxisomal biogenesis factor 11B     organization
  biogenesis factor   (PEX11B), mRNA. /FEA = mRNA     and
  11B (LOC8799)   /GEN = PEX11B /PROD = peroxisomal     biogenesis
  SEQ ID NOS: 177   biogenesis factor 11B
  (DNA) and 373   /DB_XREF = gi: 4505718 /UG = Hs.83023
  (amino acid)   peroxisomal biogenesis factor 11B
    /FL = gb: AF093670.1 gb: AB018080.1
    gb: NM_003846.1”
  LASS6: LAG1   “Consensus includes gb: AI658534   212446_s_at
  longevity   /FEA = EST /DB_XREF = gi: 4762104
  assurance homolog   /DB_XREF = est: tu17g01.x1
6 (S. cerevisiae)   /CLONE = IMAGE: 2251344
  (LOC253782)/UG = Hs.101282 Homo sapiens cDNA:
  SEQ ID NOS: 178   FLJ21238 fis, clone COL01115”
  (DNA) and 374
  (amino acid)
  C10orf86:“gb: BC005212.1 /DEF = Homo sapiens,   211376_s_at
  chromosome 10   Similar to hypothetical protein
  open reading   FLJ20003, clone MGC: 12228, mRNA,
  frame 86   complete cds. /FEA = mRNA
  (LOC54780)   /PROD = Similar to hypothetical protein
  SEQ ID NOS: 179   FLJ20003 /DB_XREF = gi: 13528824
  (DNA) and 375   /UG = Hs.258798 hypothetical protein
  (amino acid)   FLJ20003 /FL = gb: BC005212.1”
  PLEKHF2:“gb: NM_024613.1 /DEF = Homo sapiens   218640_s_at
  pleckstrin   hypothetical protein FLJ13187
  homology domain   (FLJ13187), mRNA. /FEA = mRNA
  containing, family   /GEN = FLJ13187 /PROD = hypothetical
  F (with FYVE   protein FLJ13187
  domain) member 2   /DB_XREF = gi: 13375826
  (LOC79666)   /UG = Hs.29724 hypothetical protein
  SEQ ID NOS: 180   FLJ13187 /FL = gb: NM_024613.1”
  (DNA) and 376
  (amino acid)
  KIAA0261:   “Consensus includes gb: D87450.1   212267_at
  KIAA0261   /DEF = Human mRNA for KIAA0261
  (LOC23063)   gene, partial cds. /FEA = mRNA
  SEQ ID NOS: 181   /GEN = KIAA0261
  (DNA) and 377   /DB_XREF = gi: 1665788
  (amino acid)   /UG = Hs.154978 KIAA0261 protein”
  TIP120A: TBP-gb: AL136810.1 /DEF = Homo sapiens   208839_s_at
  interacting protein   mRNA; cDNA DKFZp434G0222 (from
  (LOC55832)   clone DKFZp434G0222); complete cds.
  SEQ ID NOS: 182   /FEA = mRNA /GEN = DKFZp434G0222
  (DNA) and 378   /PROD = hypothetical protein
  (amino acid)   /DB_XREF = gi: 12053130
    /UG = Hs.184786 TBP-interacting protein
    /FL = gb: AL136810.1
  GATA3: GATA   Consensus includes gb: AI796169   209602_s_at   defense
  binding protein 3   /FEA = EST /DB_XREF = gi: 5361632     response
  (LOC2625)   /DB_XREF = est: wh43d10.x1
  SEQ ID NOS: 183   /CLONE = IMAGE: 2383507
  (DNA) and 379   /UG = Hs.169946 GATA-binding protein
  (amino acid)   3 /FL = gb: BC003070.1 gb: M69106.1
    gb: NM_002051.1
  CGI-85: CGI-85“gb: NM_017635.1 /DEF = Homo sapiens   218242_s_at
  protein   hypothetical protein FLJ20039
  (LOC51111)   (FLJ20039), mRNA. /FEA = mRNA
  SEQ ID NOS: 184   /GEN = FLJ20039 /PROD = hypothetical
  (DNA) and 380   protein FLJ20039
  (amino acid)   /DB_XREF = gi: 8923045
    /UG = Hs.267448 hypothetical protein
    FLJ20039 /FL = gb: NM_017635.1”
  C20orf11:“gb: NM_017896.1 /DEF = Homo sapiens   218448_at
  chromosome 20   hypothetical protein FLJ20602
  open reading   (FLJ20602), mRNA. /FEA = mRNA
  frame 11   /GEN = FLJ20602 /PROD = hypothetical
  (LOC54994)   protein FLJ20602
  SEQ ID NOS: 185   /DB_XREF = gi: 8923556
  (DNA) and 381   /UG = Hs.103808 hypothetical protein
  (amino acid)   FLJ20602 /FL = gb: NM_017896.1”
  IGF1R: insulin-   Consensus includes gb: H05812   203628_at   signal
  like growth factor   /FEA = EST /DB_XREF = gi: 869364     transduction
  1 receptor   /DB_XREF = est: yl77f04.s1
  (LOC3480)   /CLONE = IMAGE: 44149
  SEQ ID NOS: 186   /UG = Hs.239176 insulin-like growth
  (DNA) and 382   factor 1 receptor /FL = gb: NM_000875.2
  (amino acid)
  LOC51315:“gb: NM_016618.1 /DEF = Homo sapiens   218303_x_at
  hypothetical   hypothetical protein (LOC51315),
  protein LOC51315   mRNA. /FEA = mRNA /GEN = LOC51315
  (LOC51315)   /PROD = hypothetical protein
  SEQ ID NOS: 187   /DB_XREF = gi: 7706155 /UG = Hs.5721
  (DNA) and 383   hypothetical protein /FL = gb: AF208845.1
  (amino acid)   gb: AF217520.1 gb: NM_016618.1”
  PBP: prostatic“gb: NM_002567.1 /DEF = Homo sapiens   205353_s_at
  binding protein   prostatic binding protein (PBP), mRNA.
  (LOC5037)   /FEA = mRNA /GEN = PBP
  SEQ ID NOS: 188   /PROD = prostatic binding protein
  (DNA) and 384   /DB_XREF = gi: 4505620 /UG = Hs.80423
  (amino acid)   prostatic binding protein
    /FL = gb: D16111.1 gb: NM_002567.1”
  KIAA0602:“Cluster Incl. AB011174: Homo sapiens   34406_at
  KIAA0602 protein   mRNA for KIAA0602 protein, partial
  (LOC23241)   cds /cds = (0,2889) /gb = AB011174
  SEQ ID NOS: 189   /gi = 3043727 /ug = Hs.37656 /len = 3428”
  (DNA)
  MYST2: MYST“gb: NM_007067.1 /DEF = Homo sapiens   200049_at   regulation
  histone   histone acetyltransferase (HBOA),     of
  acetyltransferase 2   mRNA. /FEA = mRNA /GEN = HBOA     transcription,
  (LOC11143)   /PROD = histone acetyltransferase     DNA-
  SEQ ID NOS: 190   /DB_XREF = gi: 5901961 /UG = Hs.21907     dependent
  (DNA) and 385   histone acetyltransferase
  (amino acid)   /FL = gb: AF074606.1 gb: AF140360.1
    gb: NM_007067.1”
  C6orf211:“gb: NM_024573.1 /DEF = Homo sapiens   218195_at
  chromosome 6   hypothetical protein FLJ12910
  open reading   (FLJ12910), mRNA. /FEA = mRNA
  frame 211   /GEN = FLJ12910 /PROD = hypothetical
  (LOC79624)   protein FLJ12910
  SEQ ID NOS: 191   /DB_XREF = gi: 13375745
  (DNA) and 386   /UG = Hs.15929 hypothetical protein
  (amino acid)   FLJ12910 /FL = gb: NM_024573.1”
  C20orf149:“gb: NM_024299.1 /DEF = Homo sapiens   218010_x_at
  chromosome 20   hypothetical protein MGC2479
  open reading   (MGC2479), mRNA. /FEA = mRNA
  frame 149   /GEN = MGC2479 /PROD = hypothetical
  (LOC79144)   protein MGC2479
  SEQ ID NOS: 192   /DB_XREF = gi: 13236523
  (DNA) and 387   /UG = Hs.79625 hypothetical protein
  (amino acid)   MGC2479 /FL = gb: BC002531.1
    gb: NM_024299.1”
  LLGL2: lethal“gb: NM_004524.1 /DEF = Homo sapiens   203713_s_at
  giant larvaelethal giant larvae (Drosophila) homolog
  homolog 2   2 (LLGL2), mRNA. /FEA = mRNA
(Drosophila)   /GEN = LLGL2 /PROD = lethal giant
  (LOC3993)larvae (Drosophila) homolog 2
  SEQ ID NOS: 193   /DB_XREF = gi: 4758679 /UG = Hs.3123
  (DNA) and 388lethal giant larvae (Drosophila) homolog
  (amino acid)   2 /FL = gb: NM_004524.1”
  KIAA0882:   Consensus includes gb: AI348094   212956_at
  KIAA0882 protein   /FEA = EST /DB_XREF = gi: 4085300
  (LOC23158)   /DB_XREF = est: qp61g12.x1
  SEQ ID NOS: 194   /CLONE = IMAGE: 1927558
  (DNA) and 389   /UG = Hs.90419 KIAA0882 protein
  (amino acid)
  CA12: carbonic“gb: BC001012.1 /DEF = Homo sapiens,   204508_s_at
  anhydrase XII   hypothetical protein FLJ20151, clone
  (LOC771)   MGC: 1073, mRNA, complete cds.
  SEQ ID NOS: 195   /FEA = mRNA /PROD = hypothetical
  (DNA) and 390   protein FLJ20151
  (amino acid)   /DB_XREF = gi: 12654376
    /UG = Hs.279916 hypothetical protein
    FLJ20151 /FL = gb: BC001012.1
    gb: NM_017689.1”
  SLC2A10: solute“gb: NM_030777.1 /DEF = Homo sapiens   221024_s_at   glucose
  carrier family 2   solute carrier family 2 (facilitated     transport
  (facilitated glucose   glucose transporter), member 10
  transporter),   (SLC2A10), mRNA. /FEA = mRNA
  member 10   /GEN = SLC2A10 /PROD = solute carrier
  (LOC81031)   family 2 (facilitated glucosetransporter),
  SEQ ID NOS: 196   member 10 /DB_XREF = gi: 13540546
  (DNA) and 391   /FL = gb: NM_030777.1”
  (amino acid)
  TRIM37: tripartite   “Consensus includes gb: AK022701.1   213009_s_at
  motif-containing/DEF = Homo sapiens cDNA FLJ12639
  37 (LOC4591)   fis, clone NT2RM4001938, highly
  SEQ ID NOS: 197similar to Homo sapiens mRNA for
  (DNA) and 392   KIAA0898 protein. /FEA = mRNA
  (amino acid)   /DB_XREF = gi: 10434250 /UG = Hs.8164
    Mulibrey nanism”
  AP1G1: adaptor-   “Consensus includes gb: AL050025.1   215867_x_at   endocytosis
  related protein/DEF = Homo sapiens mRNA; cDNA
  complex 1, gamma   DKFZp564D066 (from clone
  1 subunit   DKFZp564D066); partial cds.
  (LOC164)   /FEA = mRNA /GEN = DKFZp564D066
  SEQ ID NOS: 198   /PROD = hypothetical protein
  (DNA) and 393   /DB_XREF = gi: 4884095 /UG = Hs.5344
  (amino acid)   adaptor-related protein complex 1,
    gamma 1 subunit”
  UBL3: ubiquitin-“gb: NM_007106.1 /DEF = Homo sapiens   201535_at
  like 3 (LOC5412)   ubiquitin-like 3 (UBL3), mRNA.
  SEQ ID NOS: 199   /FEA = mRNA /GEN = UBL3
  (DNA) and 394   /PROD = ubiquitin-like 3
  (amino acid)   /DB_XREF = gi: 6005927
    /UG = Hs.173091 ubiquitin-like 3
    /FL = gb: AF044221.1 gb: AL080177.1
    gb: NM_007106.1”
  CYB561:   “Consensus includes gb: U06715.1   217200_x_at   secretory
  cytochrome b-561   /DEF = Human cytochrome B561,     vesicle-
  (LOC1534)   HCYTO B561, mRNA, partial cds.     specific
  SEQ ID NOS: 200   /FEA = mRNA /GEN = B561     electron
  (DNA) and 395   /PROD = HCYTO B561     transport
  (amino acid)   /DB_XREF = gi: 476590 /UG = Hs.153028     protein
    cytochrome b-561”
 
[0049] Certain biomarkers were of particular interest. Microtubule-associated protein tau was identified as one of the resistance markers, and has been shown to bind at the close site of microtubule where Taxol® binds to. It is believed that Taxol® interferes microtubule and Tau interaction, but Tau's interaction seems more resistant than Taxol® (R. Dye et al., J. Biological Chem., 268, 6847-6850 (1993)). Therefore, this further validates the observation that Tau expressing cells are more resistant to ixabepilone treatment as ixabepilone binds at the same site of Taxol® in tubulin. Another interesting resistance biomarker is estrogen receptor. In general, estrogen-receptor status is predictive of response to hormonal treatments. (J. C. Chang et al., Lancet, 362, 362-369 (2003)). However, it was interesting to observe estrogen receptor as a strong marker for the resistance to ixabepilone. ER has not been previously suggested as a predictive marker of a patient's response to chemotherapy. More interestingly, microtubule associated protein tau is estrogen induced (M. West et al., P. N. A. S. USA, 98, 11462-11467 (2001)). ER and Tau were also found as resistance markers in an analysis of Taxol® (data not provided), and this suggests that Tau and ER both are likely to be the resistance markers for microtubule-stabilizing agents such as ixabepilone and Taxol®.
[0050] Several other genes appear promising as potential markers including transporter genes (ATP-binding cassette, sub-family G (WHITE), member 1 and ATP-binding cassette, sub-family A (ABC1), member 3), Midline 1 (C. Berti et al., BMC Cell Biol., February 29; 5(1):9 (2004)), LMP7 and etc. The differential expression patterns of these biomarkers were distinct between the two phenotypes of the cell lines (sensitive and resistant). In addition, their biological functions are involved in drug resistance mechanism or related with microtubule functions. Furthermore, their differential expression patterns observed within tumors support their potential as response markers.

Microtubule-Stabilizing Agents

[0051] Agents that affect microtubule-stabilization are well known in the art. These agents have cytotoxic activity against rapidly proliferating cells, such as, tumor cells or other hyperproliferative cellular disease.
[0052] In one aspect, the microtubule-stabilizing agent is an epothilone, or analog or derivative thereof. The epothilones, including analogs and derivatives thereof, may be found to exert microtubule-stabilizing effects similar to paclitaxel (Taxol®) and, hence, cytotoxic activity against rapidly proliferating cells, such as, tumor cells or other hyperproliferative cellular disease.
[0053] Suitable microtubule-stabilizing agents are disclosed, for example, in the following PCT publications hereby incorporated by reference: WO93/10121; WO98/22461; WO99/02514; WO99/58534; WO00/39276; WO02/14323; WO02/72085; WO02/98868; WO03/070170; WO03/77903; WO03/78411; WO04/80458; WO04/56832; WO04/14919; WO03/92683; WO03/74053; WO03/57217; WO03/22844; WO03/103712; WO03/07924; WO02/74042; WO02/67941; WO01/81342; WO00/66589; WO00/58254; WO99/43320; WO99/42602; WO99/39694; WO99/16416; WO 99/07692; WO99/03848; WO99/01124; and WO 98/25929.
[0054] In another aspect, the microtubule-stabilizing agent is ixabepilone. Ixabepilone is a semi-synthetic analog of the natural product epothilone B that binds to tubulin in the same binding site as paclitaxel, but interacts with tubulin differently. (P. Giannakakou et al., P. N. A. S. USA, 97, 2904-2909 (2000)).
[0055] In another aspect, the microtubule-stabilizing agent is a taxane. The taxanes are well known in the art and include, for example, paclitaxel (Taxol®) and docetaxel (Taxotere®).

Biomarkers and Biomarker Sets

[0056] The invention includes individual biomarkers and biomarker sets having both diagnostic and prognostic value in disease areas in which microtubule-stabilization and/or cytotoxic activity against rapidly proliferating cells, such as, tumor cells or other hyperproliferative cellular disease is of importance, e.g., in cancers or tumors. The biomarker sets comprise a plurality of biomarkers such as, for example, a plurality of the biomarkers provided in Table 1 and Table 2, that highly correlate with resistance or sensitivity to one or more microtubule-stabilizing agents.
[0057] The biomarker sets of the invention enable one to predict or reasonably foretell the likely effect of one or more microtubule-stabilizing agents in different biological systems or for cellular responses. The biomarker sets can be used in in vitro assays of microtubule-stabilizing agent response by test cells to predict in vivo outcome. In accordance with the invention, the various biomarker sets described herein, or the combination of these biomarker sets with other biomarkers or markers, can be used, for example, to predict how patients with cancer might respond to therapeutic intervention with one or more microtubule-stabilizing agents.
[0058] A biomarker set of cellular gene expression patterns correlating with sensitivity or resistance of cells following exposure of the cells to one or more microtubule-stabilizing agents provides a useful tool for screening one or more tumor samples before treatment with the microtubule-stabilizing agent. The screening allows a prediction of cells of a tumor sample exposed to one or more microtubule-stabilizing agents, based on the expression results of the biomarker set, as to whether or not the tumor, and hence a patient harboring the tumor, will or will not respond to treatment with the microtubule-stabilizing agent.
[0059] The biomarker or biomarker set can also be used as described herein for monitoring the progress of disease treatment or therapy in those patients undergoing treatment for a disease involving a microtubule-stabilizing agent.
[0060] The biomarkers also serve as targets for the development of therapies for disease treatment. Such targets may be particularly applicable to treatment of breast cancers or tumors. Indeed, because these biomarkers are differentially expressed in sensitive and resistant cells, their expression patterns are correlated with relative intrinsic sensitivity of cells to treatment with microtubule-stabilizing agents. Accordingly, the biomarkers highly expressed in resistant cells may serve as targets for the development of new therapies for the tumors which are resistant to microtubule-stabilizing agents.
[0061] The level of biomarker protein and/or mRNA can be determined using methods well known to those skilled in the art. For example, quantification of protein can be carried out using methods such as ELISA, 2-dimensional SDS PAGE, Western blot, immunopreciptation, immunohistochemistry, fluorescence activated cell sorting (FACS), or flow cytometry. Quantification of mRNA can be carried out using methods such as PCR, array hybridization, Northern blot, in-situ hybridization, dot-blot, Taqman, or RNAse protection assay.

Microarrays

[0062] The invention also includes specialized microarrays, e.g., oligonucleotide microarrays or cDNA microarrays, comprising one or more biomarkers, showing expression profiles that correlate with either sensitivity or resistance to one or more microtubule-stabilizing agents. Such microarrays can be employed in in vitro assays for assessing the expression level of the biomarkers in the test cells from tumor biopsies, and determining whether these test cells are likely to be resistant or sensitive to microtubule-stabilizing agents. For example, a specialized microarray can be prepared using all the biomarkers, or subsets thereof, as described herein and shown in Table 1 and Table 2. Cells from a tissue or organ biopsy can be isolated and exposed to one or more of the microtubule-stabilizing agents. Following application of nucleic acids isolated from both untreated and treated cells to one or more of the specialized microarrays, the pattern of gene expression of the tested cells can be determined and compared with that of the biomarker pattern from the control panel of cells used to create the biomarker set on the microarray. Based upon the gene expression pattern results from the cells that underwent testing, it can be determined if the cells show a resistant or a sensitive profile of gene expression. Whether or not the tested cells from a tissue or organ biopsy will respond to one or more of the microtubule-stabilizing agents and the course of treatment or therapy can then be determined or evaluated based on the information gleaned from the results of the specialized microarray analysis.

Antibodies

[0063] The invention also includes antibodies, including polyclonal or monoclonal, directed against one or more of the polypeptide biomarkers. Such antibodies can be used in a variety of ways, for example, to purify, detect, and target the biomarkers of the invention, including both in vitro and in vivo diagnostic, detection, screening, and/or therapeutic methods.

Kits

[0064] The invention also includes kits for determining or predicting whether a patient would be susceptible or resistant to a treatment that comprises one or more microtubule-stabilizing agents. The patient may have a cancer or tumor such as, for example, a breast cancer or tumor. Such kits would be useful in a clinical setting for use in testing a patient's biopsied tumor or other cancer samples, for example, to determine or predict if the patient's tumor or cancer will be resistant or sensitive to a given treatment or therapy with a microtubule-stabilizing agent. The kit comprises a suitable container that comprises: one or more microarrays, e.g., oligonucleotide microarrays or cDNA microarrays, that comprise those biomarkers that correlate with resistance and sensitivity to microtubule-stabilizing agents; one or more microtubule-stabilizing agents for use in testing cells from patient tissue specimens or patient samples; and instructions for use. In addition, kits contemplated by the invention can further include, for example, reagents or materials for monitoring the expression of biomarkers of the invention at the level of mRNA or protein, using other techniques and systems practiced in the art such as, for example, RT-PCR assays, which employ primers designed on the basis of one or more of the biomarkers described herein, immunoassays, such as enzyme linked immunosorbent assays (ELISAs), immunoblotting, e.g., Western blots, or in situ hybridization, and the like, as further described herein.

Application of Biomarkers and Biomarker Sets

[0065] The biomarkers and biomarker sets may be used in different applications. Biomarker sets can be built from any combination of biomarkers listed in Table 1 and Table 2 to make predictions about the likely effect of any microtubule-stabilizing agent in different biological systems. The various biomarkers and biomarkers sets described herein can be used, for example, as diagnostic or prognostic indicators in disease management, to predict how patients with cancer might respond to therapeutic intervention with a microtubule-stabilizing agent, and to predict how patients might respond to therapeutic intervention that affects microtubule-stabilization and/or cytotoxic activity against rapidly proliferating cells, such as, tumor cells or other hyperproliferative cellular disease.
[0066] The biomarkers have both diagnostic and prognostic value in diseases areas in which microtubule-stabilization and/or cytotoxic activity against rapidly proliferating cells, such as, tumor cells or other hyperproliferative cellular disease is of importance.
[0067] In accordance with the invention, cells from a patient tissue sample, e.g., a tumor or cancer biopsy, can be assayed to determine the expression pattern of one or more biomarkers prior to treatment with one or more microtubule-stabilizing agents. In one aspect, the tumor or cancer is breast cancer. Success or failure of a treatment can be determined based on the biomarker expression pattern of the cells from the test tissue (test cells), e.g., tumor or cancer biopsy, as being relatively similar or different from the expression pattern of a control set of the one or more biomarkers. Thus, if the test cells show a biomarker expression profile which corresponds to that of the biomarkers in the control panel of cells which are sensitive to the microtubule-stabilizing agent, it is highly likely or predicted that the individual's cancer or tumor will respond favorably to treatment with the microtubule-stabilizing agent. By contrast, if the test cells show a biomarker expression pattern corresponding to that of the biomarkers of the control panel of cells which are resistant to the microtubule-stabilizing agent, it is highly likely or predicted that the individual's cancer or tumor will not respond to treatment with the microtubule-stabilizing agent.
[0068] The invention also provides a method of monitoring the treatment of a patient having a disease treatable by one or more microtubule-stabilizing agents. The isolated test cells from the patient's tissue sample, e.g., a tumor biopsy or tumor sample, can be assayed to determine the expression pattern of one or more biomarkers before and after exposure to a microtubule-stabilizing agent. The resulting biomarker expression profile of the test cells before and after treatment is compared with that of one or more biomarkers as described and shown herein to be highly expressed in the control panel of cells that are either resistant or sensitive to a microtubule-stabilizing agent. Thus, if a patient's response is sensitive to treatment by a microtubule-stabilizing agent, based on correlation of the expression profile of the one or biomarkers, the patient's treatment prognosis can be qualified as favorable and treatment can continue. Also, if, after treatment with a microtubule-stabilizing agent, the test cells don't show a change in the biomarker expression profile corresponding to the control panel of cells that are sensitive to the microtubule-stabilizing agent, it can serve as an indicator that the current treatment should be modified, changed, or even discontinued. This monitoring process can indicate success or failure of a patient's treatment with a microtubule-stabilizing agent and such monitoring processes can be repeated as necessary or desired.
[0069] The biomarkers of the invention can be used to predict an outcome prior to having any knowledge about a biological system. Essentially, a biomarker can be considered to be a statistical tool. Biomarkers are useful in predicting the phenotype that is used to classify the biological system.
[0070] Although the complete function of all of the biomarkers are not currently known, some of the biomarkers are likely to be directly or indirectly involved in microtubule-stabilization and/or cytotoxic activity against rapidly proliferating cells. In addition, some of the biomarkers may function in metabolic or other resistance pathways specific to the microtubule-stabilizing agents tested. Notwithstanding, knowledge about the function of the biomarkers is not a requisite for determining the accuracy of a biomarker according to the practice of the invention.

EXAMPLES

Example 1

Identification of Biomarkers

Methods

[0071] Cell Lines and Cytotoxicity Assay
[0072] 23 breast cancer cell lines were assayed for their sensitivity to ixabepilone. Each cell line was exposed to ixabepilone for 72 hours, and growth inhibition was assessed by the CellTiter 96® Aqueous Non-Radioactive Cell proliferation Assay (Promega) for IC50 measurements. Then, the concentration of the ixabepilone required for 50% growth inhibition was calculated as the IC50. For each experimental condition, at least triplicate measurements were carried out for each cell line. The 23 cell lines were assayed for their IC50 measurements twice, and these two separate IC50 data sets were used for the following analysis.
[0073] Training Set Selection
[0074] For analysis, training cell lines were chosen in the following manner. The 23 cell lines were assigned into the classes “sensitive” or “resistant” using IC50 values; log(IC50) values were normalized based on the mean and the standard deviation (SD) across the 23 cell lines for each IC50 data set. (J. E. Staunton et al., P. N. A. S. USA. 98, 10787-10792 (2001)) The cell lines with the normalized log(IC50) below the mean of log(IC50)s were classified as sensitive and above as resistant. Subsequently, classification of the cell lines were compared in two separate experiments and 18 cell lines that exhibited consistent IC50 and classification were chosen as a training set for subsequent marker analysis. Five cell lines with inconsistent IC50 and classification were considered to be intermediate and were eliminated from the analysis.
[0075] RNA Extraction and Gene Expression Data
[0076] The 23 breast cancer cell lines were grown to 50-70% confluent in RPMI media with FBS 10% at 37° C. and 5% CO2. RNA was isolated using the RNeasy Mini kit (Qiagen) according to the manufacturer's instructions. 10 ug of total RNA was used to prepare biotinylated cRNA targets as described in Affymetrix protocol. Targets were hybridized to Affymetrix high-density HU133 A and B set that consist of 44,000 probe sets containing ˜32,000 genes. The chips were washed and stained using recommended procedures for GeneChip®. Expression values were calculated and scaled to 1500 by using Affymetrix GeneChip® software.
[0077] k-Nearest Neighbors (KNN) Analysis
[0078] GeneCluster software was used to find a set of marker genes. First, genes with greater than 100 average difference were filtered. Then, genes were excluded if they varied by less than 2-fold and 1000 average difference change across 18 training cell lines. Subsequently, intensity units across the cell lines for each gene were normalized to the mean and variance. The genes were ranked according to the correlation between their expression level, and the sensitivity and resistance profile of the training cell lines. A marker gene selection process was carried out by KNN algorithm which fed only the genes with higher correlation with the target class. The KNN algorithm sets the class of the data point to the majority class appearing in the k closest training set samples. This marker selection is done by sorting the genes according to the signal-to-noise statistics, [μ1(g)−μ2(g)]/[σ1(g)+σ2(g)], described as the correlation function where [μ1(g), μ2(g)] and [σ1(g), σ2(g)] denote the means and SDs of the expression levels of gene g for the samples in class 1 and class 2, respectively. The magnitude of correlation values indicates the strength of the correlation between gene expression and class distinction.
[0079] Leave-One-Out Cross-Validation Analysis and Random Classification
[0080] Predictors with 1-250 genes were used for cross-validation of the training set. For each predictor, cross-validation was performed with the entire training set; one cell line was removed, the classifier was trained on the remaining cell lines and then tested for its ability to classify the withheld cell line. This procedure was repeated for each cell line in the training set. For random classification analysis, GeneCluster was used to generate random class vectors and calculate error rates.
[0081] Clustering and Tree View
[0082] Gene expression data were analyzed by the software Cluster and TreeView.
[0083] Breast Tumors and Gene Expression Data
[0084] RNAs extracted from 175 breast tumors resected at the surgery were obtained from the Karolinska Institute (Stockholm, Sweden). These RNA samples were profiled using Affymetrix Human U133 sets and their gene expression data were used for the analysis.

Results

[0085] Drug sensitivity data (IC50) was used as a template for determining the phenotype of the cell lines as resistant or sensitive. Initially, two separate IC50 data sets were generated for 23 breast cancer cell lines. As a first step for the analysis, the log(IC50) value for each cell line was calculated and normalized using the mean of log(IC50)s and SD across the cell lines (J. E. Staunton et al., P. N. A. S. USA. 98, 10787-10792 (2001)) in each IC50 data panel. Then the cell lines were divided into two classes using the following method; the normalized log(IC50)s above the mean are defined as resistant and below as sensitive (FIG. 1). After comparing the classification of the cell lines in the two data sets, 18 cell lines that displayed the consistent classification and IC50 values in two separate experiments were selected and utilized for marker selection.
[0086] Subsequently, the gene expression data of the 18 cell lines were analyzed to identify genes that were highly correlated with observed phenotype defined as sensitive or resistant. From the GeneCluster analysis, classifiers that consisted of up to 250 correlated genes were selected and tested through leave-one out cross validation; by holding back one cell line, training on the remaining cell lines, predicting the class of the withheld cell line, and repeating this cycle for each cell line in the training set. Each gene was ranked according to the correlation in the training set between its expression level and the sensitivity-resistance class distinction. Each classifier identified from the analysis was evaluated with the error rate as shown in the FIG. 2. In order to assess whether or not the classifiers can be observed by chance, the error rates calculated from random classification were examined. Shown as an example in FIG. 3, error rates generated from random classifications were significantly higher than that from IC50-based classification.
[0087] From the GeneCluster analysis, 200 genes (Tables 1 and 2) were identified whose expression levels were highly correlated with the sensitivity-resistance class distinction based on the KNN analysis and the T-test. Among these genes, the 50 marker candidates most closely correlated with sensitivity-resistance class distinction (first 25 sensitive markers of Table 1 and first 25 resistant markers of Table 2) were selected for further analysis.
[0088] As shown in FIG. 4, these 50 genes showed distinct expression patterns between sensitive and resistant cell lines. For example, the top 25 markers correlated with sensitivity (one of which was Proteasome subunit, beta type 8 (LMP7)) as shown in FIG. 4 were highly expressed (shown in red) in sensitive cell lines, but at a lower level (shown in blue) in the resistant cell lines. In contrast, the top 25 markers correlated with resistance showed the opposite expression pattern as these genes were highly expressed in the resistant cell lines, but at a lower level in the sensitive cell lines.
[0089] Among 200 genes identified, it is interesting to find estrogen receptor (ER) as one of the resistance markers. As shown in FIG. 4, ER expression levels were highly correlated with the resistance to ixabepilone. ER was highly expressed in the resistant cell lines, but its expression was very low in the sensitive cell lines. Although the resistance mechanism of ER is not obvious, the data suggests that ER might be involved in drug resistance associated with microtubule stabilizing agents. In fact, ER was also found as a resistance marker in our analysis of paclitaxel (data not shown). In addition to ER, a microtubule-associated protein, tau (Tau), was also identified as one of the resistance markers. This protein has been proposed to bind close to the Taxol® binding site on β-tubulin and stabilize microtubules in a similar way to Taxol® (S. Kar et al., EMBO J., 22, 70-77. (2003)). Therefore, Tau is likely to affect the Taxol® bound to microtubules and, presumably, ixabepilone in a similar way since ixabepilone binds at the same site as paclitaxel on β-tubulin. Interestingly, Tau is estrogen induced (M. West et al., P. N. A. S. USA, 98, 11462-11467 (2001)). Evidently, they seem to be co-regulated as shown in a plot of Tau expression level against ER (FIG. 5). Their correlation coefficient value is 0.7.
[0090] Among the sensitivity markers identified, LMP7 is particularly interesting because it appears to be connected to Tau's function. In general, the proteasome is a multicatalytic proteinase complex responsible for the degradation of most intracellular proteins, including proteins crucial to cell cycle regulation and programmed cell death, or apoptosis (P. Voorhees et al., Clin. Cancer Res., 9, 6316-6325 (2003)). Among many proteins processed by the proteasome, Tau is degraded by the 20S proteasome in vitro in an ubiquitin-independent manner (D. David et al., J. of Neurochemistry, 83, 176-185 (2002)). This supports LMP7 as one of the sensitivity markers because LMP7 presumably facilitates interaction between ixabepilone and microtubules by degrading Tau and making ixabepilone more accessible to the microtubules.
[0091] One type of drug resistance mechanism is based on the function of a group of transporter proteins, able to prevent the intracellular accumulation of anticancer drugs by an efflux mechanisms (F. Leonessa et al., Endocr. Relat. Cancer., 10, 43-73 (2003)). Several transporter genes were identified as potential resistance markers as they were highly expressed in the resistant cell lines. These genes include ATP-binding cassette, sub-family G (WHITE), member 1 and ATP-binding cassette, sub-family A (ABC1), member 3. They are ATP dependent transporters which may be involved in lipid transport, and act as an efflux pump for chemotherapeutics drugs respectively (M. Gottesman et al., Nat. Rev. Cancer., January; 2(1):48-58 (2002)).
[0092] In addition, genes implied in microtubule functions are particularly interesting since ixabepilone is a microtubule-stabilizing agents. Microtubules are essential components of the cytoskeleton and involved in cell motility and transport, and maintenance of cell shape. The dynamic nature of a microtubule whose ability to polymerize and depolymerize, is essential for the segregation of chromosomes during mitosis (C. Bode et al., Biochemistry, 41, 3870-3874 (2002)). Therefore, marker genes such as midline 1 (C. Berti et al., BMC Cell Biol., February 29; 5(1):9 (2004)) and annexin A1 (L. C. Alldridge et al., Exp. Cell Res., October 15; 290(1):93-107 (2003)) that are implied in those functions can be involved in the mechanism of drug resistance. The biomarkers are categorized by their biological functions in Tables 1 and 2.
[0093] To study the use of these genes as response prediction markers in vivo, the expression pattern of these 50 genes in 175 breast cancer biopsies obtained from the Karolinska Institute was examined. The 50 genes were used to cluster the expression patterns of tumors. As shown in FIG. 6, these tumors were found to show patterns of expression that allowed sub-classification of these tumors into distinct groups as seen in the cell line study. Among many genes, Tau and ER were examined in tumors which were identified as resistance markers from cell lines. As shown in the FIG. 7, there is a trend in which Tau and ER seem to correlate as seen in the cell line study. In fact, both genes are highly expressed in a subset of tumors. These tumors are presumed to be non-responders for ixabepilone treatment.

Example 2

Further Evaluation of ER and Tau Biomarkers

[0094] Estrogen receptor (ER) and tau (Tau) were identified as biomarkers since their expression patterns were highly correlated with resistance to ixabepilone. In addition, it was found that the ER pathway was the most implicated biological network for resistance to ixabepilone based on the pathway analysis using preclinical candidate markers (FIGS. 9 and 10). Interestingly, Tau was recently identified as the gene most correlated with pathological complete response for T/FAC neoadjuvant treatment in breast cancer patients (M. Ayers et al, J. Clin. Oncol., 22(12):2284-93 (2004); R. Rouzier et al., P.N.A.S., June 7; 102(23):8315-20 (2005)). Following this report, our preclinical study on paclitaxel supported the clinical findings of Tau as a novel mediator of paclitaxel sensitivity (P. Wagner et al., Cell Cycle, September; 4(9):1149-52 (2005)). ER and Tau were evaluated for their predictability of response to ixabepilone in CA163-080 trial.

Methods

CA163-080 Study

[0095] CA163-080 is an exploratory genomic phase II study that was conducted in breast cancer patients who received ixabepilone as a neoadjuvant treatment. The primary objective of this study was to identify predictive markers of response to ixabepilone through gene expression profiling of pre-treatment breast cancer biopsies. Patients with invasive stage IIA-IIIB breast adenocarcinoma (tumor size≧3 cm diameter) received 40 mg/m2 ixabepilone as a 3-hour infusion on Day 1 for up to four 21-day cycles, followed by surgery within 3-4 weeks of completion of chemotherapy. A total of 164 patients were enrolled in this study. Biopsies for gene expression analysis were obtained both pre- and post-treatment. Upon isolation of biopsies from the patients, samples were either snap frozen in liquid nitrogen or placed into RNAlater solution overnight, followed by removal from the RNAlater solution. All samples were kept at −70° C. until use.

Evaluation of Pathological Response

[0096] Pathological response was assessed using the Sataloff classification system (D. Sataloff et al., J. Am. Coll. Surg., 180(3):297-306 (1995)) and used as an end point for the pharmacogenomic analysis. The pathologic response was evaluated in the primary tumor site at the end of treatment and prior to surgery by assessing histologic changes compared with baseline as following: At the primary tumor site, cellular modifications were evaluated in both the infiltrating tumoral component and in the possible ductal component, to determine viable residual infiltrating component (% of total tumoral mass); residual ductal component (% of total tumoral mass); the mitotic index. Pathologic Complete Response (pCR) in the breast only was defined as T-A, Total or near total therapeutic effect in primary site. Based on this criteria, responders included patients with pCR while non-responders included patients who failed to demonstrate pCR. The response rate was defined as the number of responders divided by the number of treated patients.

Gene Expression Profiling

[0097] Total RNA was isolated using the RNeasy Mini kit (Qiagen) according to the manufacturer's instructions by Karolinska Institute (Stockholm, Sweden). A total of 134 patients with more than 1 μg of total RNA with good quality were included in the data set for the final genomic analysis. Samples were profiled in a randomized order by batches to minimize the experimental bias. Each batch consisted of about 15 subject samples and 2 experimental controls using RNA extracted from HeLa cells. The expression profiling was done following a complete randomization with an effort to balance the number of samples from two tissue collection procedures (RNAlater and liquid nitrogen), two mRNA preparation methods (standard and DNA supernatants), tissue collection sites, and time of RNA sample preparation within in each batch. The mRNA samples from each subject was processed with HG-U133A 2.0 GeneChip® arrays on the Affymetrix platform and quantitated with GeneChip® Operating Software (GCOS) V1.0 (Affymetrix). The HG-U133A 2.0 GeneChip® array consists of about 22,276 probe sets, each containing about 15 perfect match and corresponding mismatch 25 mer oligonucleotide probes from specific gene sequences.

Gene Expression Data Processing

[0098] The gene expression data were transformed using base two logarithm. The Robust Multichip Average (RMA) method (C. Clopper et al., Biometrika, 26:404-13 (1934)) was used to normalize the raw expression data. The gene expression measures of each gene were centered at zero and rescaled to have a 1-unit standard deviation.

Stromal Effects in Tumor Biopsies

[0099] A hierarchical clustering analysis was performed in order to examine molecular profiles of tumor biopsies (FIG. 10). Among the two highest level clusters, the tumor samples from three Russian sites (site numbers 16, 24 and 25) were mostly clustered in the first cluster (Fisher's exact test, p-value <0.01). It appeared that genes implied in lymphocyte functions such as MHC, CD antigen, and IgG were enriched in this cluster. Therefore, this led to the concern that the tumors from the three Russian sites contained more stroma in the biopsies than other sites, thus contaminating the RNA samples with respect to gene expression for tumor tissue alone. In addition, the mean tumor size from the three Russian sites (16, 24 and 25) was larger than that for the other sites (mean 5.43 vs. mean 4.55, t-test p-value=0.022). Moreover, the pCR response rate was lower than that of the other sites (pCR 19.8% v.s. 17.2%, chi-squared p-value=0.73). This suggested that tumors from the three Russian sites were distinct from the others. Thus, a subgroup analysis excluding the three Russian sites was also performed.

Statistical Analysis

[0100] Logistic regression (F. Hsieh et al., Stat. Med., 17(14):1623-34 (1998)) was used to explore the relationships between the expression of genes and response to ixabepilone. The following model was fitted for each gene separately:
[0101]  [see pdf for image]
where Y=1 represents a responder and X is the gene expression measure. For each gene, the probability from a two-tailed Score test of whether the estimate of b1=0 was used to rank the most interesting genes for further investigation. eb1 is the odds ratio of being a non-responder for a one unit increase in gene expression relative to the average expression for the sample of subjects. Odds ratios and 95% confidence limits were reported.
[0102] Subjects were randomly assigned to the equal sized training set (n=67) or the test set (n=67), for responders and non-responders groups separately. The gene expression of ER and Tau were considered as potential predictors for response. Single logistic regression (SLR) was used to build the predictive model based on the training set, and the model performance was assessed on the test set. The prediction error, sensitivity, specificity, PPV (positive predictive value), and NPV (negative predictive value) as well as their 95% confidence intervals of the SLR model were estimated.

Results

Estrogen Receptor (ER) and Tau

[0103] For ER, patients whose predicted probability of being responders was greater than 0.3 were classified as responders. The ER prognostic sensitivity, specificity, PPV, NPV, and their 95% confidence intervals of the SLR model are 0.64 (0.35, 0.85), 0.79 (0.66, 0.87), 0.37 (0.19, 0.59), and 0.92 (0.80, 0.97), respectively. For Tau, patients whose predicted probability of being responders was greater than 0.25 were classified as responders. The Tau prognostic sensitivity, specificity, PPV, NPV, and their 95% confidence intervals of the SLR model are 0.55 (0.28, 0.79), 0.73 (0.60, 0.83), 0.29 (0.14, 0.50), and 0.89 (0.77, 0.95), respectively.

Estrogen Receptor (ER) and Tau without Russian Sites 16, 24 and 25

[0104] For ER and Tau separately as SLR predictors, patients whose predicted probability of being responders was greater than 0.5 were classified as responders. The estrogen receptor 1 prognostic sensitivity, specificity, PPV, NPV and their 95% confidence intervals of the SLR model are 0.67 (0.35, 0.88), 0.83 (0.69, 0.92), 0.46 (0.23, 0.71), and 0.92 (0.79, 0.97), respectively. The Tau prognostic sensitivity, specificity, PPV, NPV, and their 95% confidence intervals of the SLR model are 0.44 (0.19, 0.73), 0.88 (0.75, 0.95), 0.44 (0.19, 0.73), and 0.88 (0.75, 0.95), respectively.

Conclusion/Discussion

[0105] A total of 164 patients were enrolled in CA163-080 study. The quality and quantity of RNA samples obtained from pre-treatment biopsies was fairly good as 134 patients (85%) had RNA samples with >1 μg good quality which did not require additional amplification for gene expression profiling. Stromal contamination in the tumor biopsies was raised as a potential problem for the analysis. It appeared that the three Russian sites 16, 24 and 25 might have more stromal tissues in the samples compared to others based on the hierarchical clustering analysis. In addition, the tumors from these three Russian sites were larger than others at baseline. Although further analysis is needed to confirm this hypothesis, it raised an important issue for analyzing clinical samples that are inherently heterogeneous.
[0106] Among preclinical candidate markers, ER and Tau were examined in CA163-080 for their predictability. In our preclinical work, ER and Tau had been identified as biomarkers since their expression patterns were highly correlated with resistance to ixabepilone. In CA163-080, ER predicted well for pCR whether or not the three Russian sites were included in the analysis. However, the highest PPV was obtained when these sites were excluded. In another study finding predictive markers of response to combination chemotherapy with paclitaxel, 5-Fluorouracil, adriamycin and cyclophosphamide, the ER regulated gene Tau was identified as the best predictor of response (M. Ayers et al., J. Clin. Oncol., 22(12):2284-93 (2004)). Work done by this group has also demonstrated in vitro that knocking down Tau levels using small interfering RNA (siRNA) increases the sensitivity of breast cancer cell lines to paclitaxel treatment (R. Rouzier et al., P.N.A.S., June 7; 102(23):8315-20 (2005)). The proposed mechanism is that high levels of Tau inhibit binding of paclitaxel to the taxane binding site on β-tubulin. Tau gene expression was therefore also examined for ability to predict response to ixabepilone. The PPV (0.44) with this gene was similar to that for ER (0.46) in the subset excluding the 3 Russian sites.
[0107] Thus, ER and Tau demonstrated their utility as a predictors for response to ixabepilone and can be used as biomarkers for identifying the pCR responders to ixabepilone.

Example 3

Production of Antibodies Against the Biomarkers

[0108] Antibodies against the biomarkers can be prepared by a variety of methods. For example, cells expressing a biomarker polypeptide can be administered to an animal to induce the production of sera containing polyclonal antibodies directed to the expressed polypeptides. In one aspect, the biomarker protein is prepared and isolated or otherwise purified to render it substantially free of natural contaminants, using techniques commonly practiced in the art. Such a preparation is then introduced into an animal in order to produce polyclonal antisera of greater specific activity for the expressed and isolated polypeptide.
[0109] In one aspect, the antibodies of the invention are monoclonal antibodies (or protein binding fragments thereof). Cells expressing the biomarker polypeptide can be cultured in any suitable tissue culture medium, however, it is preferable to culture cells in Earle's modified Eagle's medium supplemented to contain 10% fetal bovine serum (inactivated at about 56° C.), and supplemented to contain about 10 g/l nonessential amino acids, about 1,00 U/ml penicillin, and about 100 μg/ml streptomycin.
[0110] The splenocytes of immunized (and boosted) mice can be extracted and fused with a suitable myeloma cell line. Any suitable myeloma cell line can be employed in accordance with the invention, however, it is preferable to employ the parent myeloma cell line (SP2/0), available from the ATCC (Manassas, Va.). After fusion, the resulting hybridoma cells are selectively maintained in HAT medium, and then cloned by limiting dilution as described by Wands et al. (1981, Gastroenterology, 80:225-232). The hybridoma cells obtained through such a selection are then assayed to identify those cell clones that secrete antibodies capable of binding to the polypeptide immunogen, or a portion thereof.
[0111] Alternatively, additional antibodies capable of binding to the biomarker polypeptide can be produced in a two-step procedure using anti-idiotypic antibodies. Such a method makes use of the fact that antibodies are themselves antigens and, therefore, it is possible to obtain an antibody that binds to a second antibody. In accordance with this method, protein specific antibodies can be used to immunize an animal, preferably a mouse. The splenocytes of such an immunized animal are then used to produce hybridoma cells, and the hybridoma cells are screened to identify clones that produce an antibody whose ability to bind to the protein-specific antibody can be blocked by the polypeptide. Such antibodies comprise anti-idiotypic antibodies to the protein-specific antibody and can be used to immunize an animal to induce the formation of further protein-specific antibodies.

Example 4

Immunofluorescence Assays

[0112] The following immunofluorescence protocol may be used, for example, to verify biomarker protein expression on cells or, for example, to check for the presence of one or more antibodies that bind biomarkers expressed on the surface of cells. Briefly, Lab-Tek II chamber slides are coated overnight at 4° C. with 10 micrograms/milliliter (μg/ml) of bovine collagen Type II in DPBS containing calcium and magnesium (DPBS++). The slides are then washed twice with cold DPBS++ and seeded with 8000 CHO—CCR5 or CHO pC4 transfected cells in a total volume of 125 g and incubated at 37° C. in the presence of 95% oxygen/5% carbon dioxide.
[0113] The culture medium is gently removed by aspiration and the adherent cells are washed twice with DPBS++ at ambient temperature. The slides are blocked with DPBS++ containing 0.2% BSA (blocker) at 0-4° C. for one hour. The blocking solution is gently removed by aspiration, and 125 μl of antibody containing solution (an antibody containing solution may be, for example, a hybridoma culture supernatant which is usually used undiluted, or serum/plasma which is usually diluted, e.g., a dilution of about 1/100 dilution). The slides are incubated for 1 hour at 0-4° C. Antibody solutions are then gently removed by aspiration and the cells are washed five times with 400 μl of ice cold blocking solution. Next, 125 μl of 1 μg/ml rhodamine labeled secondary antibody (e.g., anti-human IgG) in blocker solution is added to the cells. Again, cells are incubated for 1 hour at 0-4° C.
[0114] The secondary antibody solution is then gently removed by aspiration and the cells are washed three times with 400 μl of ice cold blocking solution, and five times with cold DPBS++. The cells are then fixed with 125 μl of 3.7% formaldehyde in DPBS++ for 15 minutes at ambient temperature. Thereafter, the cells are washed five times with 400 μl of DPBS++ at ambient temperature. Finally, the cells are mounted in 50% aqueous glycerol and viewed in a fluorescence microscope using rhodamine filters.
[0115] Although the invention has been described in some detail by way of illustration and example for purposes of clarity and understanding, it will be apparent that certain changes and modifications may be practiced within the scope of the appended claims.
(57)

Claims

1. A method for treating breast cancer in a mammal in need thereof comprising:
(a) measuring an expression level of an RNA transcript or its expression product in a breast cancer tissue biological sample from the mammal wherein the RNA transcript is an estrogen receptor 1 transcript;
(b) determining a normalized expression level of the estrogen receptor 1 transcript or its expression product, wherein the normalized expression level of the estrogen receptor 1 transcript or its expression product negatively correlates with an increased likelihood of a beneficial response to said method of treatment; and
(c) administering a microtubule-stabilizing agent comprising ixabepilone to said mammal when said mammal has said increased likelihood of said beneficial response.
2. The method of claim 1 wherein the breast cancer is invasive breast carcinoma.
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