Patent Publication Number: US-2013251710-A1

Title: Methods to Expand the Eligible Patient Population for HER2-Directed Targeted Therapies

Description:
CROSS-REFERENCE TO RELATED APPLICATION 
     The present application is a continuation-in-part application of U.S. patent application Ser. No. 13/093,563, which was filed on Apr. 25, 2011, which claims the benefit of U.S. Provisional Patent Application Ser. No. 61/327,460, which was filed on Apr. 23, 2010, both of which are incorporated herein by reference in their entirety. 
    
    
     STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT 
     Not Applicable 
     THE NAMES OF THE PARTIES TO A JOINT RESEARCH AGREEMENT 
     Not Applicable 
     INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ON A COMPACT DISC 
     Not Applicable 
     BACKGROUND OF THE INVENTION 
     Currently HER2-targeted therapies such as trastuzumab or lapatinib are only used in the treatment patients diagnosed with HER2 positive breast cancer, which comprise only 15% to 20% of all breast cancer patients. HER2 positivity is defined by either overexpression of HER2 protein, which is determined by immunohistochemical staining (3+ staining score by FDA approved Herceptest assay), or by amplification of the HER2 (ERBB2) gene, which is determined by fluorescence in situ hybridization assay (HER2/CEP17 ratio over 2 using FDA approved PathVysion assay). The current cut-offs for these assays were determined from clinical trials of patients diagnosed with metastatic or advanced breast cancer. 
     However, in a trial that tested the worth of addition of trastuzumab to adjuvant chemotherapy in the treatment of stage 2 or 3 breast cancer patients (NSABP trial B-31), even patients diagnosed with breast cancer that was classified as HER2 negative using currently used clinical HER2 assays (IHC and FISH) gained significant benefit from trastuzumab (Paik, et al.,  N Engl. J. Med.  358:1409-1411, 2008). In this study, degree of HER2 gene amplification or protein expression did not have any correlation with the degree of benefit from trastuzumab, directly challenging the use of currently used HER2 clinical assays (IHC and FISH) for selection of patients for adjuvant trastuzumab or other HER2 targeted therapies. 
     Therefore improved predictive tests for HER2-targeted therapies are clearly required. 
     BRIEF SUMMARY OF THE INVENTION 
     In order to develop better predictive test for HER2 targeted therapies, whole genome (transcriptome) gene expression profiling was performed on tumor specimens collected from patients enrolled in NSABP trial B-31 using microarrays (Agilent and Affymetrix platforms). As a result of this gene expression profiling effort, it was determined that mRNA expression levels of HER2 (ERBB2) itself is a predictor of the degree of benefit from trastuzumab in NSABP trial B-31. In addition, based on findings from NSABP trial B-31, is was determined that a large number of patients diagnosed with breast cancer that are classified as HER2 negative using current generation HER2 assays (IHC and FISH) are expected to derive benefit from trastuzumab or other HER2 targeted therapies. Therefore, the present disclosure details HER2 assays (based on measurement of HER2 mRNA) that provide a significant improvement over currently used HER2 assays (FISH and IHC) as a predictor of the degree of benefit from HER2 targeted therapies in the treatment of breast cancer in an adjuvant setting (stage 2 or 3 breast cancer). 
     Currently, breast cancer samples are assayed for HER2 protein levels or HER2 gene copy number, and based on this analysis the breast cancer samples are classified as “HER2 positive” or “HER2 negative.” Breast cancers that are classified as “HER2 positive” are candidates for treatment with a HER2-targeted therapy, such as trastuzumab, while those that are classified as “HER2 negative” are not candidates for HER2-targeted therapy. However, the inventors have determined that many breast cancers that are currently classified as “HER2 negative” still receive a therapeutic benefit from HER2-targeted therapies, such as trastuzumab. Therefore, the present disclosure provides improved assays that are more accurate in predicting the benefit from addition of a HER2-targeted therapy to chemotherapy. Breast cancer samples that were classified as “HER2 negative” by the assays previously described and used in the clinic are often classified as “HER2 positive” using the presently described HER2 mRNA assays. Therefore, numerous breast cancer patients that would not have been candidates for treatment with a HER2-targeted therapy based on the assays previously described and used in the clinic can be correctly identified as candidates for treatment with HER2-targeted therapies, such as trastuzumab, thus improving breast cancer patient care. 
     The present disclosure provides methods of identifying a cancer patient, for example a breast cancer patient, that has an increased benefit from the addition of a HER2-targeted therapy to chemotherapy, comprising assaying a tumor tissue sample from said patient for expression of HER2 mRNA, wherein a normalized HER2 mRNA expression level of about 6.0 or greater is indicative of a cancer patient that has a increased benefit from the addition of a HER2-targeted therapy to chemotherapy. In certain embodiments, normalized HER2 mRNA expression levels of about 6.0 to about 10.5 are indicative of a cancer patient that has an increased benefit from the addition of a HER2-targeted therapy to chemotherapy. In still other embodiments, normalized HER2 mRNA expression levels that are below the levels previously classified as “HER2 positive” are indicative of a cancer patient that has an increased benefit from the addition of a HER2-targeted therapy to chemotherapy. In particular aspects, normalized HER2 mRNA expression levels of about 6.0, about 6.5, about 7.0, about 7.5, about 8.0, about 8.5, about 9.0, about 9.5, about 10.0, or about 10.5 or greater are indicative of a cancer patient that has a increased benefit from the addition of a HER2-targeted therapy to chemotherapy. 
     In certain aspects of the present disclosure, the HER2-targeted therapy is trastuzumab, while in other aspects of the present disclosure the HER2-targeted therapy is lapatinib. In particular aspects of the present disclosure, the HER2-targeted therapy is combination of trastuzumab and lapatinib. It will be understood to the skilled artisan that other HER2-targeted therapies, either alone or in combination, could be used in conjunction with the teachings of the present disclosure. 
     The present disclosure also provides a method of identifying a cancer patient that has an increased benefit from the addition of a HER2-targeted therapy to a standard chemotherapy regimen, comprising assaying a tumor tissue sample from said patient for expression of HER2 or a HER2-related mRNA and estrogen receptor or an estrogen receptor-related mRNA, wherein a value outside of a range of a combined normalized HER2 mRNA expression level between about 11.0 and about 15.0 and a normalized estrogen receptor mRNA expression level of about 10.0 and about 12.0 is indicative of a cancer patient that has an increased benefit from the addition of a HER2-targeted therapy to a chemotherapy regimen. In certain embodiments the HER2-related mRNA is a c17orf37 or GRB7 mRNA. In other embodiments the estrogen receptor-related mRNA is a NAT1, GATA3, CA12 or IGF1R mRNA. 
     Thus, the present disclosure additionally provides methods of treating breast cancer in a patient in need of such treatment, comprising assaying a breast cancer or tumor tissue sample from said patient for expression of HER2 mRNA, and treating the patient with a HER2-targeted therapy and chemotherapy if the results of the assay indicate a normalized HER2 mRNA expression level of about 6.0 or greater. 
     The present disclosure further provides a method of treating breast cancer in a patient in need of such treatment, comprising assaying a tumor tissue sample from said patient for expression of HER2 or a HER2-related mRNA and estrogen receptor or an estrogen receptor-related mRNA, and treating the patient with a HER2-targeted therapy and a chemotherapy regimen if the results of the assay indicate a value outside of a range of a combined normalized HER2 or HER2-related mRNA expression level between about 11.0 and about 15.0 and a normalized estrogen receptor or estrogen receptor-related mRNA expression level of about 10.0 and about 12.0. In particular embodiments the HER2-related mRNA is a c17orf37 or GRB7 mRNA. In additional embodiments the estrogen receptor-related mRNA is a NAT1, GATA3, CA12 or IGF1R mRNA. 
    
    
     
       BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS 
         FIG. 1 . A plot of the log hazard ratio of trastuzumab in B-31 patients in relation to expression levels of HER2 mRNA. 
         FIG. 2 . A plot of the mRNA levels of samples classified as HER2 negative and HER2 positive from the B-31 and B-28 studies. 
         FIG. 3 . A plot showing the correlation between HER2 mRNA expression levels measured by the Nanostring method (nCounter assay) and the QuantigenePlex method. 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     Based on findings from NSABP trial B-31, a large number of patients diagnosed with breast cancer that are classified as HER2 negative using current generation HER2 assays (IHC and FISH) derived benefit from trastuzumab, a HER2-targeted therapy. Therefore, the present disclosure details HER2 assays (based on measurement of HER2 mRNA) that provide a significant improvement over currently used HER2 assays (FISH and IHC) as a predictor of the degree of benefit from HER2-targeted therapies in the treatment of breast cancer in an adjuvant setting (stage 2 or 3 breast cancer). In order to develop better predictive test for HER2 targeted therapies, whole genome (transcriptome) gene expression profiling was performed on tumor specimens collected from patients enrolled in NSABP trial B-31 using microarrays (Agilent and Affymetrix platforms). As a result of this gene expression profiling effort, it was determined that mRNA expression levels of HER2 (ERBB2) were a more accurate predictor of the degree of benefit from trastuzumab. 
     Although specific techniques for the quantitation of HER2 mRNA levels are discussed in the Example below, it will be understood by the skilled artisan that any technique currently used for quantitation of mRNA levels can be used in the practice of the present invention. 
     Therapeutic formulations are provided as pharmaceutical preparations for local administration to patients or subjects. The term “patient” or “subject” as used herein refers to human or animal subjects (animals being particularly useful as models for clinical efficacy of a particular composition). Selection of a suitable pharmaceutical preparation depends upon the method of administration chosen, and may be made according to protocols well-known to medicinal chemists. 
     The term “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutically active substances is well-known in the art. Except insofar as any conventional media or agent is incompatible with the platinum-based therapeutic agents, its use in the therapeutic compositions is contemplated. Supplementary active ingredients or therapeutic agents can also be used with the platinum-based therapeutic agents. 
     As used herein, “pharmaceutically-acceptable salts” refer to derivatives of the disclosed compounds wherein one or more components of the disclosed compounds are modified by making acid or base salts thereof. Examples of pharmaceutically-acceptable salts include, but are not limited to: mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. Thus, the term “acid addition salt” refers to the corresponding salt derivative of a component that has been prepared by the addition of an acid. The pharmaceutically-acceptable salts include the conventional salts or the quaternary ammonium salts of the component formed, for example, from inorganic or organic acids. For example, such conventional salts include, but are not limited to: those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, palmoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, and the like. Certain acidic or basic compounds may exist as zwitterions. All forms of the active agents, including free acid, free base, and zwitterions, are contemplated to be within the scope of the present disclosure. 
     A protein or antibody can be formulated into a composition in a neutral or salt form. Pharmaceutically acceptable salts include the acid addition salts (formed with the free amino groups of the protein), and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like. 
     In addition, the disclosed compositions or components thereof can be complexed with polyethylene glycol (PEG), metal ions, or incorporated into polymeric compounds such as polylactic acid, polyglycolic acid, hydrogels, dextran, and the like. Such compositions will influence the physical state, solubility, stability, rate of in vivo release, and rate of in vivo clearance, and are thus chosen according to the intended application. 
     The dosage unit forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form must be sterile and must be suitably fluid. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion, and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin. 
     Sterile injectable solutions are prepared by incorporating the disclosed compounds in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized ingredients into a sterile vehicle that contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum-drying and freeze-drying techniques, which yield a powder of the dosage unit plus any additional desired ingredient from a previously sterile-filtered solution thereof. 
     In certain aspects the present disclosure encompasses methods of treating or managing cancer, which comprise administering to a patient in need of such treatment or management a therapeutically effective amount of a disclosed composition or dosage unit thereof. In certain embodiments, such a compound or dosage unit is referred to as an active agent. Use of the disclosed compositions in the manufacture of a medicament for treating or preventing a disease or disorder is also contemplated. The present disclosure also encompasses compositions comprising a biologically or therapeutically effective amount of one or more of the disclosed compounds for use in the preparation of a medicament for use in treatment of cancer. 
     As used herein, and unless otherwise indicated, the terms “treat,” “treating,” and “treatment” contemplate an action that occurs while a patient is suffering from cancer, which reduces the severity of one or more symptoms or effects of cancer, or a related disease or disorder. As used herein, and unless otherwise indicated, the terms “manage,” “managing,” and “management” encompass preventing, delaying, or reducing the severity of a recurrence of cancer in a patient who has already suffered from the cancer. The terms encompass modulating the threshold, development, and/or duration of the cancer, or changing the way that a patient responds to the cancer. 
     As used herein, and unless otherwise specified, a “therapeutically effective amount” of a compound is an amount sufficient to provide any therapeutic benefit in the treatment or management of cancer, or to delay or minimize one or more symptoms associated with cancer. A therapeutically effective amount of a compound means an amount of the compound, alone or in combination with one or more other therapy and/or therapeutic agent, which provides any therapeutic benefit in the treatment or management of cancer, or related diseases or disorders. The term “therapeutically effective amount” can encompass an amount that cures cancer, improves or reduces cancer, reduces or avoids symptoms or causes of cancer, improves overall therapy, or enhances the therapeutic efficacy of another therapeutic agent. 
     Toxicity and therapeutic efficacy of the described compounds and compositions can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index, expressed as the ratio LD50/ED50. Compounds that exhibit large therapeutic indices are preferred. Compounds that exhibit toxic side effects may be used in certain embodiments, however, care should usually be taken to design delivery systems that target such compounds preferentially to the site of affected tissue, in order to minimize potential damage to uninfected cells and, thereby, reduce side effects. 
     Data obtained from cell culture assays and animal studies can be used in formulating a range of dosages for use in humans. In certain aspects of the present disclosure, the dosages of such compounds lie within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage may vary within this range depending on the dosage form employed and the route of administration utilized. For any compound used in the disclosed methods, the therapeutically effective dose can be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentration of the test compound that achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Plasma levels may be measured, for example, by high performance liquid chromatography. 
     When therapeutic treatment is contemplated, the appropriate dosage may also be determined using animal studies to determine the maximal tolerable dose, or MTD, of a bioactive agent per kilogram weight of the test subject. In general, at least one animal species tested is mammalian. Those skilled in the art regularly extrapolate doses for efficacy and avoiding toxicity to other species, including human. Before human studies of efficacy are undertaken, Phase I clinical studies help establish safe doses. Additionally, the bioactive agent may be complexed with a variety of well established compounds or structures that, for instance, enhance the stability of the bioactive agent, or otherwise enhance its pharmacological properties (e.g., increase in vivo half-life, reduce toxicity, etc.). 
     In certain embodiments of the present disclosure, the effective dose of the composition or dosage unit can be in the range of about 10 mg/kg to about 0.01 mg/kg, about 10 mg/kg to about 0.025 mg/kg, about 10 mg/kg to about 0.05 mg/kg, about 10 mg/kg to about 0.1 mg/kg, about 10 mg/kg to about 0.25 mg/kg, about 10 mg/kg to about 0.5 mg/kg, about 10 mg/kg to about 1 mg/kg, about 10 mg/kg to about 2.5 mg/kg, about 10 mg/kg to about 5 mg/kg, about 5 mg/kg to about 0.01 mg/kg, about 2.5 mg/kg to about 0.01 mg/kg, about 1 mg/kg to about 0.01 mg/kg, about 0.5 mg/kg to about 0.01 mg/kg, about 0.25 mg/kg to about 0.01 mg/kg, about 0.1 mg/kg to about 0.01 mg/kg, about 0.05 mg/kg to about 0.01 mg/kg, about 0.025 mg/kg to about 0.01 mg/kg, about 5 mg/kg to about 0.025 mg/kg, about 2.5 mg/kg to about 0.05 mg/kg, about 1 mg/kg to about 0.1 mg/kg, about 0.5 mg/kg to about 0.25 mg/kg, or about 3 mg/kg to about 0.1 mg/kg, or so. Thus, in particular embodiments, the effective dose of the composition or dosage unit is about 0.01 mg/kg, about 0.025 mg/kg, about 0.05 mg/kg, about 0.075 mg/kg, about 0.1 mg/kg, about 0.25 mg/kg, about 0.5 mg/kg, about 0.75 mg/kg, about 1 mg/kg, about 2.5 mg/kg, about 3 mg/kg, about 5 mg/kg, about 7.5 mg/kg, or about 10 mg/kg, or so. 
     The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventors to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention. The present invention is not to be limited in scope by the specific embodiments described herein, which are intended as single illustrations of individual aspects of the invention, and functionally equivalent methods and components are within the scope of the invention. Indeed, various modifications of the invention, in addition to those shown and described herein, will become apparent to those skilled in the art from the foregoing description. Such modifications are intended to fall within the scope of the appended claims. 
     Example 1 
     In The National Surgical Adjuvant Breast and Bowel Project (“NSABP”) clinical trial B31 cohort, the HER2 assays currently used in routine clinical practice to select patients for HER2 targeted therapies (namely IHC and FISH assays) failed to predict the degree of benefit from trastuzumab, and surprisingly, as shown in Table 1, even patients diagnosed with HER2 negative tumors gained the same degree of benefit as those with HER2 positive breast cancer defined by current HER2 assays (IHC and FISH) (Paik, et al.,  N. Engl. J. Med.  358:1409-1411, 2008). This data underscores the need to develop a new predictive test that can be used to predict the degree of benefit from HER2 targeted therapies in adjuvant setting. 
     
       
         
           
               
               
               
             
               
                   
                 TABLE 1 
               
             
            
               
                   
                   
               
               
                   
                 Treatment (events/total events) 
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 Central 
                   
                 Chemo plus 
                 RR 
                   
                 Interaction 
               
               
                 End Point 
                 HER2 Assay 
                 Chemo 
                 Trastuzumab 
                 (95% CI) 
                 p-value 
                 p-value 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 DFS 
                 Positive 
                 163/875 
                 85/804 
                 0.47 
                 &lt;0.001 
                 0.47 
               
               
                   
                   
                   
                   
                 (0.37-0.62) 
               
               
                   
                 Negative 
                 20/92 
                 7/82 
                 0.34 
                 0.014 
               
               
                   
                   
                   
                   
                 (0.14-0.80) 
               
               
                 Overall 
                 Positive 
                  55/875 
                 38/804 
                 0.66 
                 0.047 
                 0.08 
               
               
                 Survival 
                   
                   
                   
                 (0.43-0.99) 
               
               
                   
                 Negative 
                 10/92 
                 1/82 
                 0.08 
                 0.17 
               
               
                   
                   
                   
                   
                 (0.01-0.64) 
               
               
                   
               
            
           
         
       
     
     In Table 1, the end points were disease-free survival (“DFS”) or overall survival. The central HER2 assay results were defined as negative if they were negative by both fluorescence in situ hybridization (PathVysion™, Vysis) and immunohistochemical analysis (Herceptest™, Dako), and were defined as positive if either test was positive. Chemotherapy denotes 4 cycles of doxorubicin plus cyclophosphamide followed by 4 cycles of paclitaxel. The 95% confidence intervals (“CI”) and p-values were adjusted according to the number of positive nodes and estrogen-receptor status from the univariate Cox proportional-hazards model for each subgroup in the NSABP B-31 trial. 
     In order to develop a predictive test for the degree of benefit from trastuzumab or other HER2-targeted therapies, whole genome (trasnscriptome) gene expression profiling was performed on formalin fixed paraffin embedded tumor blocks collected from NSABP trial B-31, which tested the value of adding trastuzumab to standard adjuvant chemotherapy in the treatment of stage 2 or stage 3 breast cancer. The B-31 trial was largely enriched for HER2 positive breast cancer (90%), but also included HER2 negative breast cancer (10%). 
     The available tumor blocks from NSABP B-31 were divided into two randomly selected cohorts of discovery and validation sets. Microarray gene expression profiling was performed using both Agilent and Affymetrix arrays, and formal statistical tests (in Cox proportional hazard models controlling for clinical variables such as estrogen receptor status, tumor size, age, and number of metastatic axillary lymph nodes) were performed to test the interaction between gene expression and trastuzumab benefit. Since HER2 is a known target for trastuzumab, the initial a priori hypothesis was that HER2 (ERBB2) mRNA expression level is a linear predictor of the degree of benefit from trastuzumab, and improves upon the current generation of IHC- or FISH-based HER2 assays as a predictor of the degree of benefit from trastuzumab. 
     There are two independent oligonucleotide probes that hybridize to HER2 (ERBB2) mRNA in the Agilent microarray and three probes in the Affymetrix microarray. All five probes showed statistically significant interaction with trastuzumab as shown in Table 2, with interaction p-values ranging from 0.0075 to 00036. 
     
       
         
           
               
               
               
               
             
               
                   
                 TABLE 2 
               
               
                   
                   
               
               
                   
                 Microarray Platform 
                 Probe 
                 Interaction p-value 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
            
               
                   
                 Agilent 
                 a_24_p284420 
                 0.00092 
               
               
                   
                 Agilent 
                 a_23_p89249 
                 0.00063 
               
               
                   
                 Affymetrix 
                 234354_x_at 
                 0.0013 
               
               
                   
                 Affymetrix 
                 216836_s_at 
                 0.00036 
               
               
                   
                 Affymetrix 
                 210930_s_at 
                 0.0075 
               
               
                   
                   
               
            
           
         
       
     
     Based on these findings, a new HER2 mRNA assay was developed using nanostring platform (Geiss, et al.,  Nat. Biotechnol.  26:317-325, 2008). The test is based on a commercially available technical platform from Nanostring but with custom designed probe sets including a specific set of reference genes (ACTB, RPLP0, H2ASY, SNRP70) to normalize the expression value of HER2 mRNA. This proprietary set of reference genes were selected from data mining of microarray data from NSABP trial B-27. 
     All available tumor blocks from the B-31 trial were examined, and formal statistical tests for interaction between HER2 mRNA and trastuzumab were performed. Nanostring-based HER2 mRNA was strongly predictive of the degree of benefit from trastuzumab in B-31. To illustrate this, log hazard of trastuzumab in B-31 patients is plotted in relation to expression levels of HER2 mRNA ( FIG. 1 ).  FIG. 1  shows a linear prediction of the degree of benefit from trastuzumab added to chemotherapy by the level of expression of HER2 mRNA in the treatment of breast cancer. Values above zero on the Y-axis means no benefit, and negative values on the Y-axis mean benefit from trastuzumab. HER2 mRNA levels in  FIG. 1  are based on nanostring assays, but other methods of measurement showed similar plots. 
     With increasing amounts of HER2 mRNA expression in the tumor tissue, there is an increasing degree of benefit from trastuzumab added to chemotherapy in B-31. The cut-off of trastuzumab benefit can be determined from  FIG. 1  with confidence intervals. The cut off based on B-31 data is 8.5 normalized HER2 mRNA expression level with a confidence interval of 6 to 10.5. 
     When this cut-off was applied to all breast cancer (B-31 study and B-28 study, which also compares 4 cycles of arimycin (doxorubicin) plus cyclophosphamide versus 4 cycles of AC followed by four cycles of TAXOL® (paclitaxel)), it became evident that a significant proportion of HER2 negative patients would benefit from trastuzumab ( FIG. 2 ).  FIG. 2  shows the identification of breast cancer patients who may benefit from trastuzumab in adjuvant setting (stage 2 or stage 3) based on HER2 mRNA measurement. The cut off derived from the nanostring HER2 mRNA assay is applied to a scattergram of tumors that are classified as either HER2 positive or HER2 negative by current clinical HER2 assays (IHC or FISH). The dotted line is the cut-off. It is clear that most breast cancers express HER2 mRNA at levels above the dotted line, suggesting that a significant proportion of patients with breast cancer are expected to benefit from trastuzumab. 
     Since HER2 mRNA expression levels linearly correlate with the degree of benefit from trastuzumab, this assay can be utilized to estimate the degree of benefit from trastuzumab before starting the treatment, and this information will help clinicians and patients decide whether to use HER2-targeted therapies, as well as considering other therapies. While the data in this Example is based on HER2 mRNA expression levels measured using either Agilent or Affymetrix arrays, or nanostring platform, the results are applicable broadly to any measure of HER2 mRNA, since a close correlation was demonstrated between HER2 mRNA measured by nanostring and other methods such as Quantigene Plex assay that were performed in a subset of B-31 samples ( FIG. 3 ).  FIG. 3  shows the correlation between HER2 mRNA expression levels measured by Nanostring method (nCounter assay) and QuantigenePlex method. 
     Example 2 
     NSABP trial B-31 suggested the efficacy of adjuvant trastuzumab for both HER2-positive and negative breast cancer. In order to develop a predictive model for trastuzumab benefit, gene expression profiling of archived tumor blocks from B-31 was performed. Cases with tumor blocks were randomly divided into a candidate discovery and a confirmation set. A predictive model was built from the candidate discovery cohort (N=588) through gene expression profiling with a custom designed nCounter assay that included candidate predictive and prognostic genes identified from microarray gene expression profiling. Pre-defined cut-points for the predictive model were tested in the confirmation set of 991 patients. Eight predictive genes associated with HER2 (ERBB2, c17orf37, GRB7) or ER (ESR1, NAT1, GATA3, CA12, IGF1R) were selected for the model building. Three dimensional subset treatment effect pattern plot using two principal components of these genes identified a subset with no benefit from trastuzumab, characterized by intermediate-level ERBB2 and high-level ESR1 mRNA expression. In the confirmation set (N=991), the predefined cut-points for this model classified patients into three subsets with differential benefit from trastuzumab with hazard ratios of 1.58 (95% CI: 0.67-3.69, N=100, p=0.29), 0.60 (95% CI: 0.41-0.89, p=0.011, N=449), and 0.28 (95% CI: 0.20-0.41, p&lt;0.0001, N=442). P-value for interaction between the model and trastuzumab was 0.0002. A gene expression based algorithm that predicts the degree of benefit from adjuvant trastuzumab has thus been developed. 
     Trastuzumab is a monoclonal antibody which is directed against HER2 protein overexpressed in approximately 20% of breast cancer patients with proven efficacy for both macro disease (metastatic and neo-adjuvant setting; Slamon, et al.,  N. Engl. J. Med.  344:783-792, 2001; Untch, et al.,  J. Clin. Oncol.  28:2024-2031, 2010) and micro-metastatic disease (adjuvant setting; Piccart-Gebhart, et al.,  N. Engl. J. Med.  353:1659-1672, 2005; Romond, et al.,  N. Engl. J. Med.  353:1673-1684, 2005). HER2 positive tumors showed a high rate of pathologic complete response to neo-adjuvant chemotherapy and complete responders tend to have favorable prognosis even without trastuzumab (Carey, et al.,  Clin. Cancer Res.  13:2329-2334, 2007). In the adjuvant setting, where many patients may have already derived significant benefit from surgery and chemo-endocrine therapy, benefit from addition of trastuzumab could be determined through a complex interaction between HER2 and other confounding variables. In addition, more robust tumor cell response to trastuzumab in adjuvant setting could be expected based on easier trastuzumab access to micro-metastatic tumor cells (Barok, et al.,  Mol. Cancer. Ther.  6:2065-2072, 2007), less compromised immune system favoring antibody dependent cell mediated cyto-toxicity through trastuzumab (Clynes, et al.,  Nat. Med.  6:443-446, 2000), and potential dependency of cancer stem cells on HER2 signaling pathway in the absence of HER2 over-expression (Nakanishi, et al.,  Br. J. Cancer  102:815-826, 2010). 
     NSABP trial B-31 demonstrated the efficacy of adjuvant trastuzumab added to chemo-endocrine therapy for HER2-positive breast cancer and also suggested a potential efficacy for HER2-negative breast cancer (Romond, et al., supra; Paik, et al.,  N. Engl. J. Med.  358:1409-1411, 2008). In order to develop a predictive model for the degree of benefit from adjuvant trastuzumab beyond clinical HER2 status, gene expression profiling of archived formalin fixed paraffin embedded tumor blocks (FFPET) from B-31 was performed using nCounter platform (Geiss, et al.,  Nat. Biotechnol.  26:317-325, 2008). nCounter platform allows multiplexed measurement of gene expression based on direct hybridization without involving enzyme reaction and is suited for profiling degraded RNA extracted from routinely processed FFPET. 
     Study Design and Patient Cohort 
     Developing a predictive algorithm using archived FFPET from a finished clinical trial is technically difficult due to degradation of RNA. For predictive model development, the following strategy was used. Among patients who participated in B-31 (N=2043), 1734 patients signed informed consent forms approved by a local Human Investigations Committee in accordance with an assurance filed with and approved by the Department of Health and Human Services to permit use of banked tissue for future studies for cancer and clinical follow up data, available estrogen receptor status, and number of positive nodes. Tumor blocks from 743 patients from the candidate discovery cohort of 800 randomly selected cases were subjected to microarray gene expression profiling to identify candidate predictive genes and prognostic genes, as 57 cases did not yield good RNA amplification product. While biologically relevant genes can be derived using the latter method, previous studies indicated that only about 30% of the genes identified using microarray platform when applied to FFPET could be validated using other technical platforms such as nCounter assay. 
     Therefore in order to minimize the risk when designing nCounter assay (462 genes) that has a potential to be developed into a clinical assay, not only were genes selected from microarray experiments included, but also other biologically or clinically interesting genes (see below). Since nCounter assay was designed based on follow-up data at the time of unblinding of the trial results, and eventual data analysis was based on 7 year follow up with twice the number of events, many predictive genes were no longer relevant, while other genes that were originally selected based on biology became candidate predictive genes. Because of these circumstances, only nCounter assay results are shown ignoring microarray results. 
     From the original 743 cases of candidate discovery cohort, after microarray experiments enough RNA was left to perform nCounter assay in 588 cases. Based on analysis of nCounter assay data from 588 cases from the candidate discovery cohort, a single predictive algorithm was committed to and cut-points for each of the categories with varying degrees of expected benefit from trastuzumab. Then these pre-specified cut-points in the remaining 991 cases (confirmation cohort) who were not used for the selection of genes for the predictive algorithm were assessed. There were 57 cases from the discovery cohort that were not subjected to microarray analyses that were included among 991 cases. 
     nCounter Assay 
     The nCounter assay was designed with 462 probes to include candidate prognostic and predictive genes from microarray data from the discovery cohort (198 predictive genes and 289 prognostic genes with overlap between the two), 42 prognostic genes from microarray data from NSABP trial B-27 (Bear, et al.,  J. Clin. Oncol.  24:2019-2027, 2006), PAM 50 genes (Parker, et al.,  J. Clin. Oncol.  27:1160-1167, 2009), Oncotype Dx genes (Paik, et al.,  N. Engl. J. Med  351:2817-2826, 2004), and 28 internal reference genes. One hundred nanograms of total RNA were used for the assay. The data for each tumor were normalized for technical variability with the sum of the positive controls inherent to nCounter assay and within sample reference normalized with the geometric mean of 4 internal reference genes (ACTB, RPLP0, SNRP70, H2AFY) which was selected from the microarray data analyses. 
     Statistical Analysis 
     Follow-up information was included up to October 2010. Patients from the control arm who crossed over to receive trastuzumab were censored at the time of cross over. The definition of the primary endpoint for this analysis (disease-free survival [DFS]) was previously described (Romond, et al., supra). Gene expression values were categorized into quartiles for screening possible predictive genes since many genes showed non-linearity of their association with treatment effect upon initial review of the data. The gene-by-treatment interaction was tested in the Cox proportional hazard models using the cross-product term of indicator variables for trastuzumab treatment and each marker status with adjustment for nodal status. For single markers other than estrogen receptor, analyses were adjusted for estrogen receptor and nodal status. Correlations between variables were assessed with Spearman&#39;s correlation coefficient (r). 
     The principal component analysis was performed on the final set of selected genes to determine the first two components that would capture most of the variation in the data. Once the two principal components has been chosen, interactions between treatments and the first two principal components (PC1 and PC2) of the candidate predictive genes from nCounter assay were evaluated by the Cox model as well as by means of the non-parametric sub-population treatment effect pattern plot (STEPP; Bonetti and Gelber,  Biostatistics  5:465-481, 2004), which is extended for three dimensions (3-D). (See below for detailed methods and code). The 3-D surface plot was drawn with spline interpolation to smooth the plot using S-PLUS ver.8.1 (TIBCO Software Inc., Palo Alto, Calif.). All statistical analyses were done with SAS ver.9.2 (SAS Institute Inc., Cary, N.C.). 
     STEPP methodology is an exploratory tool for treatment×covariate interaction. Originally, this approach only focused on one covariate, so it was extended for exploring two interaction effects simultaneously because it was believed the treatment effect would be affected by both HER2 associated genes and ER associated genes. For 3-D STEPP analysis, each subsequent subpopulation of 100 patients was formed by removing 50 patients with the lowest Covariate 1 (in this study, PC1) values from the current sub-population and replacing them with the next 50 patients in the ordered list, while fixing 400 sub-population based on the ordered Covariate2 (in this study, PC2) values. Once the moving process based on Covariate 1 values were done, the next subpopulation based on Covariate 2 values were defined by removing 100 patients with the lowest Covariate 2 values from the current subpopulation and replacing them with the next 100 patients in the ordered list. These processes continued until all patients were included in at least one subpopulation. After the overlapping subpopulations were identified, the treatment effect was estimated within each subpopulation using the COX regression models adjusting for nodal status. Furthermore, this calculation was done again exchanging subpopulation setting Covariate 1 for Covariate2 (thus, 400 patients were fixed based on Covariate2 values for consecutive 100 patients subpopulations based on Covariate2 values.) 3-D STEPP analysis results are then shown graphically. All computational processes are provided as an SAS macro program. 
     The SAS TDSTEPPplot Macro 
     % TDSTEPPplot is a SAS macro that visually examines the interaction effect of two continuous variables and treatment on failure time with 3D plots, applying COX proportional hazard model. This method is an extension of STEPP analysis, which was originally proposed by Bonetti and Gelber ( Stat. Med.  19:2595-2609, 2000). 
     Invocation and Details 
     In order to run this macro, the following may need to be included in the SAS program where the file 3dstepp.sas is saved such as: % include “c: \program file\mysasfiles\tdsteppmacro.sas”. Then execute the macro TDSTEPPplot. An example macro call is: options nonotes; % TDSTEPPplot(ds=data1, var1=var1, var2=var2, outds=outsm, rr1=300, rr2=400, r1=50, r2=100, cov=age, trt=treatment, time=surv, cens=censor, cind=1, maxhr=1.5); quit; options notes. 
     Definition of Macro Variables: 
     &lt;Parameters for the dataset&gt; DSN: name of the SAS data set containing survival times, status, and covariates. 
     &lt;Parameters for the variables&gt; Var1: continuous variable name of interest; Var2: another continuous variable name of interest time: survival time; cens: event status indicator variable; icens: censoring status indicator variable value (ex. 1); COVS: list of covariates, separated by blanks. Covariates must be continuous or dummy variables. 
     &lt;Parameters for STEPP analysis&gt; Rr1: the largest number of subjects in common among consecutive subpopulations for variable 1. Rr2: the number of subjects in each subpopulation for variable 1. (rr2&gt;rr1). R1: the largest number of subjects in common among consecutive subpopulations for variable 2. R2: the number of subjects in each subpopulation for variable 2. (r2&gt;r1) 
     &lt;Parameters for the outputs&gt; Outds: name of the SAS dataset to create a new output dataset for 3D plot. Maxhr: maximum value of Hazard ratio (Z axis) for the 3-D plot. 
     The Macro Program is shown in Table 3. 
     
       
         
           
               
             
               
                 TABLE 3 
               
               
                   
               
             
            
               
                 %macro stepp(r1=, r2=, ds=, var=, cov=, trt=, time=, cens=, cind= ); 
               
               
                 %let window=%eval(&amp;r2−&amp;r1); 
               
               
                 proc means data=&amp;ds; 
               
            
           
           
               
               
            
               
                   
                 var &amp;var; 
               
               
                   
                 output out=outds n=n; 
               
            
           
           
               
            
               
                 run; 
               
               
                 data outds;set outds; 
               
            
           
           
               
               
            
               
                   
                 k=int(n/&amp;window); 
               
               
                   
                 call symput(“k”,trim(put(k,best.))); 
               
               
                   
                 call symput(“obsn”,trim(put(n,best.))); 
               
            
           
           
               
            
               
                 run; 
               
               
                 proc rank data=&amp;ds out=&amp;ds; 
               
            
           
           
               
               
            
               
                   
                 var &amp;var; 
               
               
                   
                 ranks rank; 
               
            
           
           
               
            
               
                 run; 
               
               
                 %do i=1 %to &amp;k; 
               
            
           
           
               
               
            
               
                   
                 %let f=%eval(1+&amp;window*(&amp;i.−1)); 
               
               
                   
                 %let l=%eval(&amp;f+&amp;r2); 
               
               
                   
                 %if &amp;i&lt;&amp;k %then %do; 
               
            
           
           
               
               
            
               
                   
                 data data&amp;i; set &amp;ds; 
               
            
           
           
               
               
            
               
                   
                 if &amp;f=&lt; rank&lt;=&amp;l; 
               
            
           
           
               
            
               
                 %end; 
               
               
                 run; 
               
            
           
           
               
               
            
               
                   
                 %if &amp;i=&amp;k %then %do; 
               
            
           
           
               
               
            
               
                   
                 data data&amp;i; set &amp;ds; 
               
            
           
           
               
               
            
               
                   
                 if &amp;f=&lt; rank; 
               
            
           
           
               
            
               
                 %end; 
               
               
                 run; 
               
            
           
           
               
               
            
               
                   
                 proc means data=data&amp;i; 
               
            
           
           
               
               
            
               
                   
                 var &amp;var; 
               
               
                   
                 output out=out&amp;i median=med; 
               
            
           
           
               
               
            
               
                   
                 run; 
               
               
                   
                 data out&amp;i; set out&amp;i; 
               
            
           
           
               
               
            
               
                   
                 call symput(“median”,trim(put(med,best.))); 
               
            
           
           
               
               
            
               
                   
                 run; 
               
               
                   
                 proc phreg data=data&amp;i; 
               
            
           
           
               
               
            
               
                   
                 model &amp;time*&amp;cens(&amp;cind)=&amp;TRT &amp;cov /rl; 
               
               
                   
                 Hazardratio &amp;TRT; 
               
               
                   
                 ods output HazardRatios =hr&amp;i; 
               
            
           
           
               
               
            
               
                   
                 run; 
               
            
           
           
               
               
            
               
                   
                 data hr&amp;i; set 
               
               
                   
                 hr&amp;i; i=&amp;i; 
               
               
                   
                 median=&amp;median; 
               
            
           
           
               
               
            
               
                   
                 run; 
               
            
           
           
               
            
               
                 %end; 
               
               
                 data hr&amp;var; set %do s=1 %to &amp;k; hr&amp;s %end;; run; 
               
               
                 %mend; 
               
            
           
           
               
               
            
               
                   
                 %macro TDSTEPP(ds=, var2=, var1=, rr1=, rr2=, r1=, r2=, cov=, 
               
               
                   
                 trt=, time=, cens=, cind= ); 
               
            
           
           
               
            
               
                 data &amp;ds;set &amp;ds; drop rank:; run; 
               
               
                 %let window1=%eval(&amp;rr2−&amp;rr1); 
               
               
                 proc means data=&amp;ds; 
               
            
           
           
               
               
            
               
                   
                 var &amp;var1; 
               
               
                   
                 output out=outds1 n=n; 
               
            
           
           
               
            
               
                 run; 
               
               
                 data outds1;set outds1; 
               
            
           
           
               
               
            
               
                   
                 kk=int(n/&amp;window1); 
               
               
                   
                 call symput(“kk”,trim(put(kk,best.))); 
               
               
                   
                 call symput(“nall”,trim(put(n,best.))); 
               
            
           
           
               
            
               
                 run; 
               
               
                 proc rank data=&amp;ds out=&amp;ds; 
               
            
           
           
               
               
            
               
                   
                 var &amp;var1; 
               
               
                   
                 ranks rank1; 
               
            
           
           
               
            
               
                 run; 
               
               
                 %do q=1 %to &amp;kk; 
               
            
           
           
               
               
            
               
                   
                 %let f1=%eval(1+&amp;window1.*(&amp;q.−1)); 
               
               
                   
                 %let l1=%eval(&amp;f1+&amp;rr2.); 
               
               
                   
                 %if &amp;q&lt;&amp;kk %then %do; 
               
            
           
           
               
               
            
               
                   
                 data d&amp;q; set &amp;ds; if &amp;f1=&lt; rank1&lt;=&amp;l1; run; 
               
            
           
           
               
               
            
               
                   
                 %end; 
               
               
                   
                 %if &amp;q=&amp;kk %then %do; 
               
            
           
           
               
               
            
               
                   
                 data d&amp;q; set &amp;ds; if &amp;f1=&lt; rank1; run; 
               
            
           
           
               
               
            
               
                   
                 %end; 
               
               
                   
                 proc means data=d&amp;q; 
               
            
           
           
               
               
            
               
                   
                 var &amp;var1; 
               
               
                   
                 output out=out1_&amp;q median=med; 
               
            
           
           
               
               
            
               
                   
                 run; 
               
               
                   
                 data out1_&amp;q; set out1_&amp;q; 
               
            
           
           
               
               
            
               
                   
                 call symput(“median1”,trim(put(med,best.))); 
               
            
           
           
               
               
            
               
                   
                 run; 
               
            
           
           
               
               
            
               
                   
                 %stepp(r1=&amp;r1, r2=&amp;r2, ds=d&amp;q, var=&amp;var2, cov=&amp;cov, trt=&amp;trt, 
               
               
                   
                 time=&amp;time, cens=&amp;cens, cind=&amp;cind ); 
               
            
           
           
               
               
            
               
                   
                 data hrr&amp;q; set hr&amp;var2; 
               
            
           
           
               
               
            
               
                   
                 q=&amp;q; 
               
               
                   
                 &amp;var1=&amp;median1; 
               
               
                   
                 rename median=&amp;var2; 
               
            
           
           
               
               
            
               
                   
                 run; 
               
            
           
           
               
            
               
                 %end; 
               
               
                 data hrall&amp;var1; set %do t=1 %to &amp;kk; hrr&amp;t %end;; run; 
               
               
                 %mend; 
               
            
           
           
               
               
            
               
                   
                 %macro TDSTEPPplot(ds=, var1=, var2=, outds=, rr1=, rr2=, 
               
               
                   
                 r1=, r2=, cov=, trt=, time=, cens=, cind= , maxhr= ); 
               
            
           
           
               
            
               
                 ods listing close; 
               
               
                 %TDSTEPP(ds=&amp;ds, var2=&amp;var2, var1=&amp;var1, rr1=&amp;rr1, rr2=&amp;rr2, r1=&amp;r1, r2=&amp;r2, 
               
            
           
           
               
               
            
               
                   
                 cov=&amp;cov, trt=&amp;trt, time=&amp;time, cens=&amp;cens, cind=&amp;cind ); 
               
            
           
           
               
            
               
                 quit; 
               
            
           
           
               
               
            
               
                   
                 %TDSTEPP(ds=&amp;ds, var2=&amp;var1, var1=&amp;var2, rr1=&amp;rr1, rr2=&amp;rr2, 
               
               
                   
                 r1=&amp;r1, r2=&amp;r2, cov=&amp;cov, trt=&amp;trt, time=&amp;time, cens=&amp;cens, 
               
               
                   
                 cind=&amp;cind ); 
               
            
           
           
               
            
               
                 ods listing; 
               
               
                 data hrall; set hrall&amp;var1 hrall&amp;var2;run; 
               
               
                 proc means data=hrall; 
               
            
           
           
               
               
            
               
                   
                 var &amp;var1; 
               
               
                   
                 output out=out1 max=max1 min=min1; 
               
            
           
           
               
            
               
                 run; 
               
               
                 data out1; 
               
            
           
           
               
               
            
               
                   
                 set out1; 
               
               
                   
                 call symput(“max1”,trim(put(max1,best.))); 
               
               
                   
                 call symput(“min1”,trim(put(min1,best.))); 
               
            
           
           
               
            
               
                 run; 
               
               
                 proc means data=hrall; 
               
            
           
           
               
               
            
               
                   
                 var &amp;var2; 
               
               
                   
                 output out=out2 max=max2 min=min2; 
               
            
           
           
               
            
               
                 run; 
               
               
                 data out2; set out2; 
               
            
           
           
               
               
            
               
                   
                 call symput(“max2”,trim(put(max2,best.))); 
               
            
           
           
               
            
               
                 call symput(“min2”,trim(put(min2,best.))); 
               
               
                 run; 
               
               
                 proc g3grid data=hrall out=&amp;outds; 
               
            
           
           
               
               
            
               
                   
                 grid &amp;var1*&amp;var2=HazardRatio / spline smooth=.2 
               
            
           
           
               
               
            
               
                   
                 axis1=&amp;min1. to &amp;max1. by 0.5 
               
               
                   
                 axis2=&amp;min2. to &amp;max2. by 0.5; 
               
            
           
           
               
            
               
                 run; 
               
               
                 goptions reset=all border ; 
               
               
                 axis3 order=(0 to &amp;maxhr by 0.1) label=none; 
               
               
                 proc g3d data=&amp;outds; 
               
            
           
           
               
               
            
               
                   
                 plot &amp;var1*&amp;var2-HazardRatio / rotate=60 grid zaxis=axis3 zticknum=14 
               
            
           
           
               
               
            
               
                   
                 zmin=0 zmax=1.5; 
               
            
           
           
               
            
               
                 run; 
               
               
                 quit; 
               
               
                 %mend; 
               
               
                   
               
            
           
         
       
     
     Results 
     Results of nCounter Assay in the Candidate Discovery Cohort (N=588) and Development of a Prediction Model 
     Although microarray gene expression analyses of 743 tumors from the discovery cohort were performed, the genes discovered from the microarray experiments could only be partially technically validated using other platforms such as nCounter assay. Therefore other biologically and clinically relevant genes were included in the design of the nCounter assay. nCounter assay is ideal for multiplexed quantification of relative gene expression levels using RNA extracted from FFPET samples since it uses short hybridization sequences and does not depend on enzymatic reaction. 
     In order to develop a predictive algorithm, it was first tried to identify reproducibly predictive genes by performing ten-fold jack-knifing process. The results of statistical tests for gene-by-trastuzumab interaction terms in Cox models adjusting for the number of positive nodes are shown in Table 4. 
     
       
         
           
               
               
               
               
               
             
               
                 TABLE 4 
               
               
                   
               
               
                 Number 
                   
                   
                   
                   
               
               
                 of times 
               
               
                 significant 
                 mean 
                 maximum 
                 minimum 
               
               
                 during 
                 p-value 
                 p-value 
                 p-value 
               
               
                 1-fold Jack 
                 from 10- 
                 from 10 
                 from 10 
               
               
                 Knifing 
                 fold jack 
                 fold jack 
                 fold jack 
               
               
                 process 
                 knifing 
                 knifing 
                 knifing 
                 gene symbol 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 10 
                 0.0025 
                 0.0054 
                 0.0002 
                 FLOT2 
               
               
                 10 
                 0.0049 
                 0.01 
                 0.0008 
                 UNC119 
               
               
                 10 
                 0.0051 
                 0.0136 
                 0.0008 
                 TUBB2C 
               
               
                 10 
                 0.0054 
                 0.0131 
                 0.0016 
                 XYLT1 
               
               
                 10 
                 0.0057 
                 0.0151 
                 0.0018 
                 SLC39A14 
               
               
                 10 
                 0.0059 
                 0.0269 
                 0.0007 
                 CA12 
               
               
                 10 
                 0.007 
                 0.0154 
                 0.001 
                 GATA3 
               
               
                 9 
                 0.0078 
                 0.0509 
                 0.0003 
                 GTF3C2 
               
               
                 10 
                 0.0088 
                 0.0223 
                 0.0014 
                 SLC39A14 
               
               
                 10 
                 0.0095 
                 0.025 
                 0.0013 
                 CA12 
               
               
                 10 
                 0.0145 
                 0.0347 
                 0.0024 
                 FTH1 
               
               
                 10 
                 0.0155 
                 0.0385 
                 0.0013 
                 SUPT6H 
               
               
                 10 
                 0.0156 
                 0.0349 
                 0.0041 
                 ACVR1B 
               
               
                 9 
                 0.0166 
                 0.0533 
                 0.005 
                 DKFZP434A0131 
               
               
                 10 
                 0.0181 
                 0.0357 
                 0.0014 
                 RPL23A 
               
               
                 9 
                 0.0188 
                 0.0825 
                 0.0012 
                 ILF2 
               
               
                 9 
                 0.0194 
                 0.0591 
                 0.0056 
                 DNAJC4 
               
               
                 10 
                 0.02 
                 0.0477 
                 0.002 
                 ABHD2 
               
               
                 10 
                 0.0214 
                 0.0476 
                 0.0093 
                 ZACN 
               
               
                 9 
                 0.0239 
                 0.0976 
                 0.0041 
                 TPBG 
               
               
                 9 
                 0.0241 
                 0.053 
                 0.0052 
                 DNAJC4 
               
               
                 10 
                 0.0242 
                 0.0396 
                 0.0034 
                 FAM84B 
               
               
                 9 
                 0.0243 
                 0.0562 
                 0.0042 
                 SPDEF 
               
               
                 8 
                 0.0277 
                 0.0808 
                 0.0074 
                 DAD1 
               
               
                 8 
                 0.0297 
                 0.1148 
                 0.0039 
                 CASC3 
               
               
                 9 
                 0.03 
                 0.0535 
                 0.0044 
                 MYADM 
               
               
                 9 
                 0.0316 
                 0.1292 
                 0.0079 
                 PTTG1 
               
               
                 8 
                 0.0329 
                 0.0827 
                 0.0059 
                 UHMK1 
               
               
                 6 
                 0.0346 
                 0.0666 
                 0.0059 
                 TMBIM6 
               
               
                 8 
                 0.0348 
                 0.0911 
                 0.006 
                 THOP1 
               
               
                 9 
                 0.0364 
                 0.0863 
                 0.0058 
                 ANGPTL2 
               
               
                 8 
                 0.0366 
                 0.139 
                 0.005 
                 ISOC1 
               
               
                 9 
                 0.0379 
                 0.086 
                 0.0131 
                 TMSB10 
               
               
                 9 
                 0.0388 
                 0.2252 
                 0.0056 
                 PIK3CA 
               
               
                 7 
                 0.0401 
                 0.107 
                 0.0097 
                 SLC7A2 
               
               
                 6 
                 0.0407 
                 0.1022 
                 0.0088 
                 ORC6L 
               
               
                 6 
                 0.0408 
                 0.0607 
                 0.0116 
                 SPP1 
               
               
                 6 
                 0.0411 
                 0.0881 
                 0.0083 
                 CD9 
               
               
                 7 
                 0.0426 
                 0.095 
                 0.009 
                 PCK2 
               
               
                 7 
                 0.0433 
                 0.097 
                 0.0125 
                 CEACAM1 
               
               
                 6 
                 0.0437 
                 0.0896 
                 0.0159 
                 RPL21 
               
               
                 7 
                 0.0442 
                 0.1008 
                 0.0084 
                 C17orf37 
               
               
                 7 
                 0.0458 
                 0.1119 
                 0.016 
                 KHSRP 
               
               
                 7 
                 0.0462 
                 0.1588 
                 0.0111 
                 RASSF7 
               
               
                 5 
                 0.0466 
                 0.073 
                 0.0196 
                 RPL21 
               
               
                 7 
                 0.0477 
                 0.1475 
                 0.0127 
                 RPL34 
               
               
                 6 
                 0.0485 
                 0.1114 
                 0.0064 
                 ERBB2 
               
               
                 6 
                 0.0489 
                 0.1281 
                 0.0116 
                 RPL23A 
               
               
                 6 
                 0.0497 
                 0.1363 
                 0.0083 
                 NUF2 
               
               
                 5 
                 0.0516 
                 0.0997 
                 0.0122 
                 EGFR 
               
               
                 6 
                 0.0525 
                 0.1375 
                 0.0126 
                 ENPP1 
               
               
                 7 
                 0.0528 
                 0.0949 
                 0.0138 
                 ZNF609 
               
               
                 6 
                 0.0542 
                 0.1148 
                 0.007 
                 NLK 
               
               
                 6 
                 0.0574 
                 0.1421 
                 0.0096 
                 ERBB2 
               
               
                 3 
                 0.0593 
                 0.0954 
                 0.0112 
                 IGF1R 
               
               
                 8 
                 0.0603 
                 0.2704 
                 0.0089 
                 L3MBTL2 
               
               
                 5 
                 0.0612 
                 0.1314 
                 0.0336 
                 LOXL3 
               
               
                 5 
                 0.0617 
                 0.1868 
                 0.0046 
                 TPBG 
               
               
                 6 
                 0.0623 
                 0.1546 
                 0.0151 
                 ACVR1B 
               
               
                 4 
                 0.0631 
                 0.1314 
                 0.0217 
                 PTP4A2 
               
               
                 3 
                 0.0636 
                 0.116 
                 0.0202 
                 GATA3 
               
               
                 6 
                 0.0648 
                 0.1645 
                 0.0038 
                 PRR3 
               
               
                 5 
                 0.0656 
                 0.2032 
                 0.0131 
                 SLC39A14 
               
               
                 4 
                 0.0657 
                 0.1155 
                 0.0106 
                 C9orf58 
               
               
                 5 
                 0.0665 
                 0.1483 
                 0.026 
                 B4GALT1 
               
               
                 6 
                 0.0676 
                 0.2062 
                 0.0203 
                 TBX21 
               
               
                 5 
                 0.0682 
                 0.1752 
                 0.014 
                 FBXW11 
               
               
                 5 
                 0.0687 
                 0.1844 
                 0.0097 
                 MTCH2 
               
               
                 4 
                 0.0701 
                 0.2389 
                 0.0297 
                 ZNF124 
               
               
                 4 
                 0.0705 
                 0.151 
                 0.0154 
                 XYLT1 
               
               
                 5 
                 0.0714 
                 0.1418 
                 0.024 
                 KRT7 
               
               
                 3 
                 0.079 
                 0.1425 
                 0.0191 
                 PADI2 
               
               
                 4 
                 0.0797 
                 0.174 
                 0.0259 
                 CA12 
               
               
                 2 
                 0.0875 
                 0.162 
                 0.0256 
                 KRT7 
               
               
                 4 
                 0.088 
                 0.2208 
                 0.0259 
                 PTP4A2 
               
               
                 3 
                 0.0889 
                 0.3082 
                 0.0289 
                 EHMT1 
               
               
                 1 
                 0.0908 
                 0.1753 
                 0.0179 
                 ANGPTL4 
               
               
                 3 
                 0.0912 
                 0.2083 
                 0.0217 
                 LASS6 
               
               
                 1 
                 0.0914 
                 0.1663 
                 0.0157 
                 IGKV1-5 
               
               
                 3 
                 0.0914 
                 0.1889 
                 0.0359 
                 MTCH2 
               
               
                 2 
                 0.0925 
                 0.1579 
                 0.0336 
                 KIF2C 
               
               
                 4 
                 0.0926 
                 0.233 
                 0.024 
                 ASPHD2 
               
               
                 4 
                 0.0949 
                 0.2731 
                 0.0235 
                 KLHL25 
               
               
                 4 
                 0.0952 
                 0.2981 
                 0.0222 
                 GRB7 
               
               
                 2 
                 0.0952 
                 0.23 
                 0.0335 
                 MED13L 
               
               
                 4 
                 0.096 
                 0.1809 
                 0.0187 
                 FAM127A 
               
               
                 4 
                 0.0966 
                 0.2073 
                 0.0305 
                 FAM148A 
               
               
                 2 
                 0.0975 
                 0.2084 
                 0.011 
                 MYB 
               
               
                 2 
                 0.0978 
                 0.2047 
                 0.03 
                 SNX5 
               
               
                 3 
                 0.0987 
                 0.2458 
                 0.0268 
                 ZC3H15 
               
               
                 7 
                 0.0993 
                 0.3792 
                 0.0168 
                 ELN 
               
               
                 2 
                 0.1 
                 0.3233 
                 0.0221 
                 PTP4A2 
               
               
                 1 
                 0.1013 
                 0.1945 
                 0.0263 
                 MYADM 
               
               
                 3 
                 0.1027 
                 0.2017 
                 0.037 
                 C1orf93 
               
               
                 2 
                 0.1038 
                 0.2977 
                 0.0315 
                 B4GALT1 
               
               
                 3 
                 0.1039 
                 0.2121 
                 0.023 
                 ESR1 
               
               
                 3 
                 0.1044 
                 0.2061 
                 0.027 
                 UBE2W 
               
               
                 3 
                 0.1052 
                 0.3319 
                 0.0388 
                 UBE2C 
               
               
                 0 
                 0.1056 
                 0.1868 
                 0.0637 
                 SOX4 
               
               
                 4 
                 0.1065 
                 0.3086 
                 0.0148 
                 LOC442270 
               
               
                 4 
                 0.1066 
                 0.3086 
                 0.0158 
                 TMBIM6 
               
               
                 4 
                 0.1075 
                 0.2562 
                 0.0207 
                 PGRMC2 
               
               
                 2 
                 0.1077 
                 0.2182 
                 0.0332 
                 IGF1R 
               
               
                 2 
                 0.1077 
                 0.2508 
                 0.0213 
                 SSBP2 
               
               
                 3 
                 0.1079 
                 0.2653 
                 0.0374 
                 ZC3HAV1L 
               
               
                 2 
                 0.1085 
                 0.2962 
                 0.0098 
                 MGC70870 
               
               
                 1 
                 0.1094 
                 0.212 
                 0.0432 
                 MYADM 
               
               
                 5 
                 0.1117 
                 0.3441 
                 0.0161 
                 TMBIM6 
               
               
                 1 
                 0.1118 
                 0.2436 
                 0.0399 
                 ACAD9 
               
               
                 2 
                 0.1126 
                 0.3157 
                 0.0165 
                 ESR1 
               
               
                 0 
                 0.1147 
                 0.3465 
                 0.0518 
                 NDC80 
               
               
                 3 
                 0.1148 
                 0.2837 
                 0.0251 
                 KCNE1 
               
               
                 0 
                 0.115 
                 0.2392 
                 0.0533 
                 THOP1 
               
               
                 4 
                 0.1153 
                 0.3512 
                 0.0352 
                 ABHD2 
               
               
                 3 
                 0.1153 
                 0.4057 
                 0.0161 
                 MGC70870 
               
               
                 1 
                 0.1164 
                 0.2129 
                 0.0337 
                 RPL21 
               
               
                 1 
                 0.1164 
                 0.4675 
                 0.03 
                 CLIC1 
               
               
                 1 
                 0.1168 
                 0.2156 
                 0.0485 
                 TMBIM1 
               
               
                 1 
                 0.1188 
                 0.2397 
                 0.0306 
                 MIA 
               
               
                 4 
                 0.119 
                 0.3063 
                 0.0135 
                 PSMD3 
               
               
                 2 
                 0.1225 
                 0.3202 
                 0.0164 
                 KLHL25 
               
               
                 4 
                 0.1228 
                 0.3382 
                 0.0353 
                 AURKA 
               
               
                 2 
                 0.1236 
                 0.2737 
                 0.0325 
                 KRT18 
               
               
                 1 
                 0.124 
                 0.2089 
                 0.0441 
                 POLR2L 
               
               
                 1 
                 0.1247 
                 0.2142 
                 0.0416 
                 BLVRA 
               
               
                 2 
                 0.1259 
                 0.2619 
                 0.0266 
                 PRPF40A 
               
               
                 1 
                 0.1264 
                 0.2285 
                 0.0147 
                 TBXAS1 
               
               
                 3 
                 0.1268 
                 0.4372 
                 0.0173 
                 KCNE1 
               
               
                 1 
                 0.127 
                 0.4015 
                 0.0486 
                 LSM14A 
               
               
                 2 
                 0.1273 
                 0.3058 
                 0.0308 
                 FURIN 
               
               
                 1 
                 0.1275 
                 0.2569 
                 0.0403 
                 ADFP 
               
               
                 4 
                 0.1278 
                 0.3995 
                 0.0283 
                 L3MBTL2 
               
               
                 2 
                 0.1286 
                 0.4374 
                 0.0383 
                 FOXA1 
               
               
                 1 
                 0.1289 
                 0.4577 
                 0.0392 
                 LOC442270 
               
               
                 2 
                 0.1301 
                 0.2629 
                 0.0402 
                 Kua-UEV 
               
               
                 1 
                 0.1301 
                 0.2505 
                 0.0402 
                 TBX10 
               
               
                 1 
                 0.1306 
                 0.2214 
                 0.0389 
                 SREBF2 
               
               
                 0 
                 0.1318 
                 0.2826 
                 0.061 
                 C17orf37 
               
               
                 1 
                 0.1321 
                 0.2319 
                 0.0381 
                 UBTD1 
               
               
                 3 
                 0.1326 
                 0.3082 
                 0.0319 
                 NAT1 
               
               
                 1 
                 0.1326 
                 0.2468 
                 0.0374 
                 RPL34 
               
               
                 0 
                 0.1326 
                 0.2183 
                 0.0542 
                 UHMK1 
               
               
                 1 
                 0.133 
                 0.2916 
                 0.0287 
                 SPP1 
               
               
                 0 
                 0.1335 
                 0.3095 
                 0.0751 
                 RBM14 
               
               
                 0 
                 0.1336 
                 0.311 
                 0.0622 
                 HSPBP1 
               
               
                 1 
                 0.1336 
                 0.2821 
                 0.0335 
                 TYMS 
               
               
                 0 
                 0.1341 
                 0.2537 
                 0.0501 
                 ANLN 
               
               
                 1 
                 0.1346 
                 0.2968 
                 0.0488 
                 KRT81 
               
               
                 2 
                 0.1349 
                 0.2318 
                 0.0339 
                 CUGBP1 
               
               
                 1 
                 0.1351 
                 0.2631 
                 0.0451 
                 PPP2R2D 
               
               
                 1 
                 0.1354 
                 0.2027 
                 0.0454 
                 BBC3 
               
               
                 1 
                 0.1356 
                 0.207 
                 0.0375 
                 KRT81 
               
               
                 1 
                 0.1363 
                 0.3257 
                 0.0262 
                 LOXL3 
               
               
                 3 
                 0.1383 
                 0.3505 
                 0.0219 
                 ORMDL3 
               
               
                 3 
                 0.1385 
                 0.4207 
                 0.0208 
                 CCL21 
               
               
                 0 
                 0.1391 
                 0.2146 
                 0.0549 
                 HSPBP1 
               
               
                 0 
                 0.1391 
                 0.3087 
                 0.0597 
                 LOXL3 
               
               
                 1 
                 0.1397 
                 0.2978 
                 0.0203 
                 FKBP3 
               
               
                 0 
                 0.1402 
                 0.2267 
                 0.0605 
                 UGCG 
               
               
                 1 
                 0.1405 
                 0.2634 
                 0.0396 
                 MYB 
               
               
                 1 
                 0.1405 
                 0.3013 
                 0.0494 
                 ORC6L 
               
               
                 0 
                 0.1428 
                 0.2423 
                 0.0658 
                 POLR2L 
               
               
                 1 
                 0.1432 
                 0.2928 
                 0.0424 
                 FBXO15 
               
               
                 0 
                 0.1435 
                 0.2405 
                 0.0832 
                 CRTC2 
               
               
                 1 
                 0.1441 
                 0.4018 
                 0.0405 
                 TBX10 
               
               
                 2 
                 0.1449 
                 0.2684 
                 0.0376 
                 GUSBL2 
               
               
                 0 
                 0.1452 
                 0.3304 
                 0.0655 
                 UNC119 
               
               
                 1 
                 0.1452 
                 0.321 
                 0.045 
                 CYBRD1 
               
               
                 0 
                 0.1476 
                 0.3627 
                 0.0568 
                 PTTG1 
               
               
                 1 
                 0.1477 
                 0.3087 
                 0.0474 
                 ESR1 
               
               
                 1 
                 0.1504 
                 0.3412 
                 0.0392 
                 ACVR1B 
               
               
                 0 
                 0.1512 
                 0.2638 
                 0.054 
                 TYMS 
               
               
                 2 
                 0.1519 
                 0.2818 
                 0.0492 
                 FURIN 
               
               
                 0 
                 0.1519 
                 0.3033 
                 0.0643 
                 POM121L9P 
               
               
                 1 
                 0.1546 
                 0.3953 
                 0.0426 
                 FTH1 
               
               
                 1 
                 0.1568 
                 0.2943 
                 0.0162 
                 FBXO15 
               
               
                 0 
                 0.1587 
                 0.3133 
                 0.0543 
                 CFLP1 
               
               
                 1 
                 0.1588 
                 0.2832 
                 0.027 
                 SFRP1 
               
               
                 1 
                 0.1597 
                 0.2758 
                 0.0486 
                 FLJ22795 
               
               
                 1 
                 0.1612 
                 0.4295 
                 0.0377 
                 PIK3CA 
               
               
                 1 
                 0.1631 
                 0.3609 
                 0.049 
                 CIAPIN1 
               
               
                 0 
                 0.1642 
                 0.2968 
                 0.0794 
                 URM1 
               
               
                 0 
                 0.1648 
                 0.2875 
                 0.0641 
                 NEBL 
               
               
                 2 
                 0.1654 
                 0.3332 
                 0.0344 
                 PGR 
               
               
                 0 
                 0.1665 
                 0.3038 
                 0.0658 
                 SPTAN1 
               
               
                 1 
                 0.1668 
                 0.5187 
                 0.0318 
                 TPBG 
               
               
                 1 
                 0.1671 
                 0.4113 
                 0.0406 
                 BRAF 
               
               
                 0 
                 0.1676 
                 0.2907 
                 0.0648 
                 CCDC24 
               
               
                 1 
                 0.1679 
                 0.2974 
                 0.0463 
                 SNHG5 
               
               
                 2 
                 0.1699 
                 0.3429 
                 0.0247 
                 REPS2 
               
               
                 0 
                 0.1704 
                 0.335 
                 0.0583 
                 FURIN 
               
               
                 1 
                 0.1704 
                 0.3224 
                 0.0487 
                 AKT1 
               
               
                 1 
                 0.1708 
                 0.3422 
                 0.0481 
                 ANGPTL2 
               
               
                 0 
                 0.1708 
                 0.3428 
                 0.0518 
                 KIAA0652 
               
               
                 1 
                 0.1728 
                 0.263 
                 0.0425 
                 PSMD3 
               
               
                 0 
                 0.1733 
                 0.2574 
                 0.0904 
                 TAPBP 
               
               
                 1 
                 0.1737 
                 0.2782 
                 0.0484 
                 HERC2P4 
               
               
                 1 
                 0.1737 
                 0.5989 
                 0.0482 
                 C16orf14 
               
               
                 0 
                 0.1747 
                 0.4094 
                 0.076 
                 SLC7A2 
               
               
                 1 
                 0.1748 
                 0.2931 
                 0.0435 
                 ABCF2 
               
               
                 1 
                 0.1754 
                 0.3594 
                 0.0213 
                 NAT1 
               
               
                 1 
                 0.1754 
                 0.3153 
                 0.0395 
                 SSBP2 
               
               
                 0 
                 0.1771 
                 0.4647 
                 0.0501 
                 CIAPIN1 
               
               
                 0 
                 0.1782 
                 0.3452 
                 0.0946 
                 ILF2 
               
               
                 0 
                 0.1785 
                 0.3608 
                 0.0722 
                 TMEM174 
               
               
                 1 
                 0.1808 
                 0.5008 
                 0.0497 
                 NECAB3 
               
               
                 0 
                 0.1819 
                 0.3369 
                 0.0685 
                 YWHAZ 
               
               
                 2 
                 0.1823 
                 0.3831 
                 0.0104 
                 CD9 
               
               
                 1 
                 0.1828 
                 0.3374 
                 0.0272 
                 LCE3E 
               
               
                 0 
                 0.1834 
                 0.2813 
                 0.0974 
                 THSD4 
               
               
                 0 
                 0.1844 
                 0.245 
                 0.1379 
                 ACTB 
               
               
                 1 
                 0.1848 
                 0.2965 
                 0.0409 
                 IGHV1-69 
               
               
                 0 
                 0.1863 
                 0.4299 
                 0.073 
                 C20orf144 
               
               
                 0 
                 0.1869 
                 0.4663 
                 0.0743 
                 PIK3CA 
               
               
                 1 
                 0.1873 
                 0.3484 
                 0.033 
                 NAT1 
               
               
                 1 
                 0.1879 
                 0.3093 
                 0.0366 
                 CTSL2 
               
               
                 0 
                 0.1881 
                 0.3848 
                 0.0554 
                 GTF3C2 
               
               
                 0 
                 0.1882 
                 0.3169 
                 0.0591 
                 TFRC 
               
               
                 0 
                 0.1884 
                 0.3586 
                 0.0856 
                 KRT81 
               
               
                 0 
                 0.1892 
                 0.3056 
                 0.0958 
                 REPS2 
               
               
                 0 
                 0.1895 
                 0.378 
                 0.0625 
                 GRB7 
               
               
                 0 
                 0.1917 
                 0.3347 
                 0.068 
                 ATAD3A 
               
               
                 1 
                 0.192 
                 0.3626 
                 0.0271 
                 HPS6 
               
               
                 0 
                 0.1923 
                 0.3721 
                 0.0738 
                 CEP55 
               
               
                 0 
                 0.1934 
                 0.4 
                 0.0528 
                 GTF3C2 
               
               
                 0 
                 0.194 
                 0.4355 
                 0.0746 
                 GCGR 
               
               
                 0 
                 0.1949 
                 0.5025 
                 0.052 
                 CD9 
               
               
                 0 
                 0.1954 
                 0.4391 
                 0.0603 
                 ZFP36L1 
               
               
                 0 
                 0.1958 
                 0.3983 
                 0.1003 
                 IGH@ 
               
               
                 0 
                 0.196 
                 0.552 
                 0.0925 
                 ZFP36L1 
               
               
                 1 
                 0.1962 
                 0.3786 
                 0.0328 
                 hCG_1642354 
               
               
                 0 
                 0.1971 
                 0.5528 
                 0.0552 
                 CXorf56 
               
               
                 0 
                 0.1982 
                 0.2783 
                 0.0534 
                 CASC3 
               
               
                 0 
                 0.1986 
                 0.3211 
                 0.0519 
                 UBE2N 
               
               
                 0 
                 0.1999 
                 0.4872 
                 0.0861 
                 DKFZP434A0131 
               
               
                 0 
                 0.2 
                 0.5259 
                 0.0536 
                 MAPT 
               
               
                 0 
                 0.2013 
                 0.4253 
                 0.0667 
                 IL6ST 
               
               
                 0 
                 0.2024 
                 0.2755 
                 0.1174 
                 ASPHD2 
               
               
                 0 
                 0.2028 
                 0.4818 
                 0.0768 
                 GCGR 
               
               
                 0 
                 0.2033 
                 0.4887 
                 0.0667 
                 KRTAP6-3 
               
               
                 0 
                 0.2039 
                 0.4344 
                 0.1259 
                 PPIA 
               
               
                 1 
                 0.2051 
                 0.6008 
                 0.0316 
                 HRH2 
               
               
                 0 
                 0.2052 
                 0.4033 
                 0.0771 
                 SFRP1 
               
               
                 0 
                 0.2056 
                 0.3406 
                 0.0645 
                 POLDIP2 
               
               
                 0 
                 0.2064 
                 0.3995 
                 0.0858 
                 IDUA 
               
               
                 0 
                 0.2074 
                 0.4174 
                 0.0627 
                 MELK 
               
               
                 0 
                 0.209 
                 0.5488 
                 0.0754 
                 LAYN 
               
               
                 0 
                 0.209 
                 0.4421 
                 0.1147 
                 ZC3H15 
               
               
                 0 
                 0.2108 
                 0.2828 
                 0.1321 
                 SIAH2 
               
               
                 0 
                 0.2109 
                 0.3212 
                 0.0948 
                 PADI2 
               
               
                 1 
                 0.2117 
                 0.5989 
                 0.0417 
                 RAB27B 
               
               
                 0 
                 0.2129 
                 0.4565 
                 0.1031 
                 ENO1 
               
               
                 0 
                 0.2136 
                 0.3619 
                 0.0562 
                 CD24 
               
               
                 0 
                 0.2147 
                 0.3855 
                 0.099 
                 SLC25A5 
               
               
                 0 
                 0.2155 
                 0.4141 
                 0.0593 
                 CLIC1 
               
               
                 0 
                 0.2161 
                 0.3202 
                 0.1025 
                 RAB27B 
               
               
                 1 
                 0.2161 
                 0.4247 
                 0.0206 
                 CCL21 
               
               
                 0 
                 0.2179 
                 0.3817 
                 0.1189 
                 MYB 
               
               
                 0 
                 0.2182 
                 0.3441 
                 0.0967 
                 C1orf212 
               
               
                 0 
                 0.22 
                 0.4717 
                 0.0563 
                 MRPS36 
               
               
                 0 
                 0.22 
                 0.4899 
                 0.1048 
                 PCBD2 
               
               
                 0 
                 0.2217 
                 0.405 
                 0.1032 
                 KRT14 
               
               
                 0 
                 0.2227 
                 0.3292 
                 0.1048 
                 THSD4 
               
               
                 0 
                 0.224 
                 0.4659 
                 0.0945 
                 UGCG 
               
               
                 0 
                 0.2244 
                 0.4271 
                 0.0562 
                 ARL17 
               
               
                 0 
                 0.225 
                 0.4044 
                 0.0935 
                 CSNK1D 
               
               
                 0 
                 0.2251 
                 0.45 
                 0.0864 
                 GSTM1 
               
               
                 0 
                 0.2251 
                 0.5311 
                 0.0705 
                 RPLP0 
               
               
                 0 
                 0.2257 
                 0.5385 
                 0.0566 
                 KRTAP6-3 
               
               
                 0 
                 0.2268 
                 0.4222 
                 0.0921 
                 PHACTR4 
               
               
                 0 
                 0.2281 
                 0.5222 
                 0.0595 
                 C20orf67 
               
               
                 0 
                 0.229 
                 0.6194 
                 0.1066 
                 PSMC5 
               
               
                 0 
                 0.2295 
                 0.4129 
                 0.0914 
                 PCTK2 
               
               
                 0 
                 0.23 
                 0.4047 
                 0.06 
                 hCG_1642354 
               
               
                 0 
                 0.2303 
                 0.423 
                 0.0704 
                 HIBCH 
               
               
                 0 
                 0.2308 
                 0.4319 
                 0.079 
                 CHD6 
               
               
                 1 
                 0.2326 
                 0.4362 
                 0.0331 
                 CASC3 
               
               
                 0 
                 0.233 
                 0.3518 
                 0.1259 
                 ANLN 
               
               
                 0 
                 0.233 
                 0.5514 
                 0.0531 
                 LOC730275 
               
               
                 0 
                 0.2331 
                 0.4251 
                 0.1088 
                 ETS2 
               
               
                 0 
                 0.2331 
                 0.3735 
                 0.1297 
                 IMPAD1 
               
               
                 1 
                 0.2335 
                 0.3654 
                 0.0476 
                 RASSF7 
               
               
                 1 
                 0.2335 
                 0.4652 
                 0.0481 
                 RPS28 
               
               
                 0 
                 0.2352 
                 0.7334 
                 0.0923 
                 SMCP 
               
               
                 0 
                 0.2353 
                 0.4502 
                 0.1493 
                 AK1 
               
               
                 1 
                 0.2355 
                 0.3806 
                 0.0327 
                 POLR3H 
               
               
                 0 
                 0.237 
                 0.3771 
                 0.122 
                 NME3 
               
               
                 0 
                 0.2371 
                 0.3776 
                 0.1038 
                 FBXW11 
               
               
                 1 
                 0.2372 
                 0.5584 
                 0.0377 
                 HIST1H2AA 
               
               
                 0 
                 0.2376 
                 0.3633 
                 0.0901 
                 BIRC5 
               
               
                 0 
                 0.2377 
                 0.4972 
                 0.0864 
                 SLC39A6 
               
               
                 0 
                 0.2386 
                 0.5099 
                 0.0784 
                 POLD4 
               
               
                 1 
                 0.2393 
                 0.4379 
                 0.0434 
                 TRIB3 
               
               
                 0 
                 0.2401 
                 0.3576 
                 0.1003 
                 C9orf58 
               
               
                 0 
                 0.2402 
                 0.5139 
                 0.0824 
                 TMSB10 
               
               
                 0 
                 0.2405 
                 0.576 
                 0.0778 
                 NEBL 
               
               
                 0 
                 0.2405 
                 0.6161 
                 0.0885 
                 ST6GALNAC4 
               
               
                 0 
                 0.241 
                 0.4734 
                 0.0764 
                 POLDIP2 
               
               
                 0 
                 0.2434 
                 0.4973 
                 0.0992 
                 BLVRA 
               
               
                 1 
                 0.2439 
                 0.5358 
                 0.0238 
                 HPS6 
               
               
                 0 
                 0.244 
                 0.5325 
                 0.0631 
                 RAF1 
               
               
                 0 
                 0.2456 
                 0.5851 
                 0.0711 
                 GGA2 
               
               
                 0 
                 0.2464 
                 0.4561 
                 0.1281 
                 SREBF2 
               
               
                 0 
                 0.2467 
                 0.516 
                 0.1311 
                 PITPNC1 
               
               
                 0 
                 0.2475 
                 0.5387 
                 0.0732 
                 LOC346887 
               
               
                 0 
                 0.2476 
                 0.5254 
                 0.063 
                 CCNB1 
               
               
                 0 
                 0.2481 
                 0.4521 
                 0.1128 
                 SIAH2 
               
               
                 0 
                 0.2488 
                 0.4233 
                 0.1136 
                 C8orf73 
               
               
                 0 
                 0.2499 
                 0.4664 
                 0.0668 
                 IL6ST 
               
               
                 0 
                 0.2503 
                 0.608 
                 0.0958 
                 ABHD2 
               
               
                 0 
                 0.2506 
                 0.4927 
                 0.0903 
                 BAG1 
               
               
                 0 
                 0.251 
                 0.4168 
                 0.075 
                 LOC346887 
               
               
                 1 
                 0.2517 
                 0.6055 
                 0.0282 
                 KRTAP6-3 
               
               
                 0 
                 0.2521 
                 0.5378 
                 0.0596 
                 HN1 
               
               
                 1 
                 0.2528 
                 0.4936 
                 0.0453 
                 ADAMTSL5 
               
               
                 0 
                 0.2529 
                 0.6362 
                 0.0702 
                 HIST2H2BE 
               
               
                 0 
                 0.2531 
                 0.5166 
                 0.1086 
                 BRAF 
               
               
                 0 
                 0.2539 
                 0.7009 
                 0.0587 
                 FOXA1 
               
               
                 0 
                 0.2556 
                 0.5933 
                 0.1421 
                 FLJ35390 
               
               
                 0 
                 0.2557 
                 0.5763 
                 0.0907 
                 GUSB 
               
               
                 1 
                 0.2589 
                 0.5904 
                 0.0293 
                 CTSL2 
               
               
                 1 
                 0.2595 
                 0.488 
                 0.0483 
                 HERC2P4 
               
               
                 0 
                 0.2615 
                 0.4635 
                 0.1195 
                 ZNF124 
               
               
                 0 
                 0.2629 
                 0.4835 
                 0.0752 
                 EPN2 
               
               
                 0 
                 0.263 
                 0.4065 
                 0.145 
                 CEACAM1 
               
               
                 0 
                 0.2641 
                 0.5647 
                 0.0611 
                 VPS45A 
               
               
                 0 
                 0.2642 
                 0.523 
                 0.0875 
                 TUBB2C 
               
               
                 1 
                 0.2653 
                 0.6265 
                 0.0399 
                 GPR160 
               
               
                 0 
                 0.2654 
                 0.591 
                 0.0825 
                 MAPT 
               
               
                 0 
                 0.2661 
                 0.5232 
                 0.1261 
                 BCAS1 
               
               
                 0 
                 0.2673 
                 0.4542 
                 0.1233 
                 NME3 
               
               
                 0 
                 0.2678 
                 0.5888 
                 0.0602 
                 MDM2 
               
               
                 1 
                 0.2686 
                 0.5369 
                 0.0419 
                 THRAP1 
               
               
                 0 
                 0.2695 
                 0.5733 
                 0.1591 
                 CCDC74A 
               
               
                 0 
                 0.2713 
                 0.6686 
                 0.146 
                 SNHG5 
               
               
                 0 
                 0.2718 
                 0.5621 
                 0.0842 
                 HDAC4 
               
               
                 0 
                 0.272 
                 0.5353 
                 0.1183 
                 SF3B3 
               
               
                 0 
                 0.2721 
                 0.5958 
                 0.1201 
                 C19orf28 
               
               
                 0 
                 0.2724 
                 0.4892 
                 0.1002 
                 ADFP 
               
               
                 0 
                 0.2732 
                 0.6091 
                 0.1586 
                 HSPA8 
               
               
                 0 
                 0.2741 
                 0.3818 
                 0.1661 
                 MED13L 
               
               
                 0 
                 0.2755 
                 0.6604 
                 0.1028 
                 ANGPTL4 
               
               
                 1 
                 0.277 
                 0.8196 
                 0.0452 
                 BTG2 
               
               
                 0 
                 0.2799 
                 0.7136 
                 0.0581 
                 KCNE1 
               
               
                 0 
                 0.2801 
                 0.7793 
                 0.0869 
                 LOC100128062 
               
               
                 0 
                 0.2808 
                 0.429 
                 0.1566 
                 C8orf73 
               
               
                 0 
                 0.2809 
                 0.4603 
                 0.082 
                 H2AFY 
               
               
                 0 
                 0.2816 
                 0.4557 
                 0.0651 
                 AK1 
               
               
                 0 
                 0.282 
                 0.6567 
                 0.0946 
                 HERC2P4 
               
               
                 0 
                 0.2823 
                 0.4587 
                 0.1823 
                 MYBL2 
               
               
                 0 
                 0.2823 
                 0.556 
                 0.1192 
                 BCL2 
               
               
                 0 
                 0.2828 
                 0.4205 
                 0.1837 
                 FAM83E 
               
               
                 0 
                 0.2828 
                 0.5267 
                 0.0706 
                 RAB22A 
               
               
                 0 
                 0.283 
                 0.5934 
                 0.0936 
                 EPN2 
               
               
                 0 
                 0.2834 
                 0.6302 
                 0.1097 
                 KRT5 
               
               
                 0 
                 0.2837 
                 0.4656 
                 0.0873 
                 FKBP3 
               
               
                 0 
                 0.2837 
                 0.4049 
                 0.1193 
                 PLD3 
               
               
                 0 
                 0.2862 
                 0.4718 
                 0.1611 
                 RAF1 
               
               
                 0 
                 0.2862 
                 0.9851 
                 0.1052 
                 CD24 
               
               
                 0 
                 0.2865 
                 0.4496 
                 0.1157 
                 CYB561 
               
               
                 0 
                 0.2867 
                 0.5223 
                 0.1209 
                 RASD2 
               
               
                 0 
                 0.2871 
                 0.4746 
                 0.1387 
                 MYBL2 
               
               
                 0 
                 0.288 
                 0.3936 
                 0.2119 
                 URM1 
               
               
                 0 
                 0.2889 
                 0.5035 
                 0.0531 
                 NDUFB9 
               
               
                 1 
                 0.2896 
                 0.6973 
                 0.0225 
                 PGM5 
               
               
                 0 
                 0.2905 
                 0.5741 
                 0.059 
                 MBNL1 
               
               
                 0 
                 0.2908 
                 0.5972 
                 0.1081 
                 SOX4 
               
               
                 0 
                 0.2912 
                 0.4792 
                 0.1038 
                 FLJ22659 
               
               
                 0 
                 0.2927 
                 0.4588 
                 0.143 
                 RRM2 
               
               
                 0 
                 0.2934 
                 0.548 
                 0.0809 
                 BIRC5 
               
               
                 0 
                 0.2936 
                 0.6343 
                 0.0742 
                 LSM14A 
               
               
                 0 
                 0.2938 
                 0.5409 
                 0.1466 
                 CDH3 
               
               
                 0 
                 0.2942 
                 0.5172 
                 0.0861 
                 SUPT6H 
               
               
                 0 
                 0.2946 
                 0.5327 
                 0.0864 
                 SCNN1D 
               
               
                 0 
                 0.2952 
                 0.4817 
                 0.1476 
                 VEGFA 
               
               
                 0 
                 0.2961 
                 0.5157 
                 0.1302 
                 PXN 
               
               
                 0 
                 0.2982 
                 0.9336 
                 0.0857 
                 KLHL25 
               
               
                 0 
                 0.2987 
                 0.4708 
                 0.1476 
                 MTOR 
               
               
                 0 
                 0.2995 
                 0.5097 
                 0.1354 
                 C9orf58 
               
               
                 0 
                 0.2996 
                 0.5883 
                 0.1308 
                 GPRIN1 
               
               
                 0 
                 0.3012 
                 0.5351 
                 0.1026 
                 ANGPTL4 
               
               
                 0 
                 0.3025 
                 0.6279 
                 0.1452 
                 LSM14A 
               
               
                 0 
                 0.3031 
                 0.5252 
                 0.147 
                 CSNK1D 
               
               
                 0 
                 0.3045 
                 0.6755 
                 0.1408 
                 ADNP 
               
               
                 0 
                 0.3051 
                 0.5116 
                 0.192 
                 ACBD6 
               
               
                 0 
                 0.3056 
                 0.8967 
                 0.1003 
                 ABCF2 
               
               
                 0 
                 0.3064 
                 0.4939 
                 0.1184 
                 IDUA 
               
               
                 0 
                 0.3084 
                 0.5233 
                 0.1024 
                 TBX21 
               
               
                 0 
                 0.3093 
                 0.4011 
                 0.1673 
                 MARVELD2 
               
               
                 0 
                 0.3108 
                 0.7302 
                 0.1545 
                 RPS25 
               
               
                 0 
                 0.3111 
                 0.5627 
                 0.1679 
                 SNRP70 
               
               
                 0 
                 0.3119 
                 0.4873 
                 0.1966 
                 PKP3 
               
               
                 0 
                 0.3141 
                 0.6169 
                 0.1701 
                 TMEM97 
               
               
                 0 
                 0.3144 
                 0.5136 
                 0.0961 
                 CFLP1 
               
               
                 0 
                 0.3169 
                 0.3966 
                 0.0784 
                 BBC3 
               
               
                 0 
                 0.3183 
                 0.5439 
                 0.1563 
                 TMBIM1 
               
               
                 0 
                 0.3186 
                 0.5027 
                 0.1206 
                 SLC25A29 
               
               
                 0 
                 0.319 
                 0.5045 
                 0.226 
                 NXPH3 
               
               
                 0 
                 0.3196 
                 0.6268 
                 0.1121 
                 MMP11 
               
               
                 0 
                 0.3208 
                 0.9706 
                 0.0888 
                 ZNF225 
               
               
                 0 
                 0.3221 
                 0.6674 
                 0.1763 
                 ANLN 
               
               
                 0 
                 0.323 
                 0.5941 
                 0.1247 
                 TMCO1 
               
               
                 0 
                 0.3231 
                 0.4697 
                 0.1157 
                 ZFP36L1 
               
               
                 0 
                 0.3234 
                 0.7005 
                 0.0918 
                 LENG8 
               
               
                 0 
                 0.3241 
                 0.6112 
                 0.1165 
                 ORMDL3 
               
               
                 0 
                 0.3294 
                 0.4768 
                 0.262 
                 IGH@ 
               
               
                 1 
                 0.3308 
                 0.899 
                 0.0292 
                 UBE2N 
               
               
                 0 
                 0.3313 
                 0.7257 
                 0.1573 
                 IGKV2-24 
               
               
                 0 
                 0.3323 
                 0.5303 
                 0.1746 
                 EXO1 
               
               
                 0 
                 0.3323 
                 0.8389 
                 0.1382 
                 PPP2R2D 
               
               
                 0 
                 0.3338 
                 0.6559 
                 0.1059 
                 NDC80 
               
               
                 0 
                 0.3348 
                 0.5634 
                 0.1276 
                 ELAVL4 
               
               
                 0 
                 0.3348 
                 0.7748 
                 0.1789 
                 ACBD6 
               
               
                 0 
                 0.3352 
                 0.5109 
                 0.2166 
                 ENO1 
               
               
                 0 
                 0.3352 
                 0.5497 
                 0.171 
                 PCBD2 
               
               
                 0 
                 0.3362 
                 0.8215 
                 0.119 
                 MAFK 
               
               
                 0 
                 0.3363 
                 0.5947 
                 0.1581 
                 ZNF124 
               
               
                 0 
                 0.3382 
                 0.739 
                 0.1145 
                 SFRS1 
               
               
                 0 
                 0.3399 
                 0.653 
                 0.1391 
                 GINS2 
               
               
                 0 
                 0.3413 
                 0.8706 
                 0.1732 
                 DDX42 
               
               
                 0 
                 0.3426 
                 0.5415 
                 0.1592 
                 RPL34 
               
               
                 0 
                 0.3429 
                 0.7113 
                 0.106 
                 EHD2 
               
               
                 0 
                 0.3436 
                 0.5471 
                 0.1454 
                 LOC643159 
               
               
                 0 
                 0.3439 
                 0.6676 
                 0.0879 
                 C1orf212 
               
               
                 0 
                 0.3441 
                 0.7145 
                 0.091 
                 MPDU1 
               
               
                 0 
                 0.3442 
                 0.7819 
                 0.1394 
                 PKP3 
               
               
                 0 
                 0.3454 
                 0.466 
                 0.2367 
                 FBXW11 
               
               
                 0 
                 0.3461 
                 0.5657 
                 0.1716 
                 PDZK1IP1 
               
               
                 0 
                 0.3465 
                 0.4862 
                 0.1421 
                 LOC285830 
               
               
                 0 
                 0.3473 
                 0.622 
                 0.1932 
                 NDUFB9 
               
               
                 0 
                 0.3477 
                 0.5942 
                 0.0997 
                 TMEM174 
               
               
                 0 
                 0.3491 
                 0.7214 
                 0.116 
                 POLR3H 
               
               
                 0 
                 0.3508 
                 0.7058 
                 0.0731 
                 RPS21 
               
               
                 0 
                 0.3513 
                 0.6841 
                 0.2525 
                 FTH1 
               
               
                 0 
                 0.3516 
                 0.5412 
                 0.1676 
                 LOC442260 
               
               
                 0 
                 0.3517 
                 0.9903 
                 0.205 
                 FNDC4 
               
               
                 0 
                 0.3524 
                 0.6925 
                 0.0946 
                 HN1 
               
               
                 0 
                 0.3527 
                 0.6956 
                 0.2332 
                 UBTD1 
               
               
                 0 
                 0.3532 
                 0.6078 
                 0.1287 
                 IGF1R 
               
               
                 1 
                 0.3536 
                 0.8895 
                 0.0234 
                 C16orf42 
               
               
                 0 
                 0.3537 
                 0.6313 
                 0.1657 
                 ROMO1 
               
               
                 0 
                 0.3545 
                 0.6306 
                 0.1157 
                 TMEM45B 
               
               
                 0 
                 0.3554 
                 0.5963 
                 0.0961 
                 CLPB 
               
               
                 0 
                 0.3573 
                 0.6657 
                 0.1533 
                 EPOR 
               
               
                 0 
                 0.3574 
                 0.6869 
                 0.1213 
                 HEATR3 
               
               
                 0 
                 0.3601 
                 0.5999 
                 0.1576 
                 HDGFRP3 
               
               
                 0 
                 0.3636 
                 0.76 
                 0.1202 
                 TBC1D10B 
               
               
                 0 
                 0.3649 
                 0.6348 
                 0.1448 
                 PGR 
               
               
                 0 
                 0.3662 
                 0.751 
                 0.1596 
                 PYCR1 
               
               
                 0 
                 0.3671 
                 0.5514 
                 0.2303 
                 C1orf104 
               
               
                 0 
                 0.3672 
                 0.6094 
                 0.2087 
                 FAM148A 
               
               
                 0 
                 0.3672 
                 0.6682 
                 0.1416 
                 TAF2 
               
               
                 0 
                 0.3678 
                 0.8589 
                 0.1345 
                 CCL21 
               
               
                 0 
                 0.3682 
                 0.611 
                 0.1873 
                 LARS 
               
               
                 0 
                 0.3704 
                 0.5236 
                 0.1487 
                 PRELID1 
               
               
                 0 
                 0.3708 
                 0.7718 
                 0.1132 
                 PLEKHF2 
               
               
                 0 
                 0.3721 
                 0.9015 
                 0.1233 
                 MDM2 
               
               
                 0 
                 0.3737 
                 0.6432 
                 0.0805 
                 HN1 
               
               
                 0 
                 0.3752 
                 0.7665 
                 0.1833 
                 GATA3 
               
               
                 0 
                 0.3753 
                 0.5781 
                 0.1844 
                 RND3 
               
               
                 0 
                 0.3754 
                 0.4618 
                 0.2416 
                 RAF1 
               
               
                 0 
                 0.3758 
                 0.9211 
                 0.1171 
                 DRD2 
               
               
                 0 
                 0.3766 
                 0.6449 
                 0.1948 
                 LOC442270 
               
               
                 0 
                 0.3773 
                 0.5883 
                 0.1959 
                 ADCY7 
               
               
                 0 
                 0.3795 
                 0.7988 
                 0.1846 
                 BAG1 
               
               
                 0 
                 0.3805 
                 0.6565 
                 0.1112 
                 FBXO15 
               
               
                 0 
                 0.3805 
                 0.711 
                 0.2165 
                 MLPH 
               
               
                 0 
                 0.3806 
                 0.7101 
                 0.1911 
                 FLAD1 
               
               
                 0 
                 0.3807 
                 0.7426 
                 0.192 
                 LOC730275 
               
               
                 0 
                 0.3813 
                 0.614 
                 0.1423 
                 KIAA0310 
               
               
                 0 
                 0.3814 
                 0.6025 
                 0.1592 
                 LASS6 
               
               
                 0 
                 0.3816 
                 0.6542 
                 0.1053 
                 LOC442260 
               
               
                 0 
                 0.3817 
                 0.57 
                 0.1464 
                 IGJ 
               
               
                 0 
                 0.3823 
                 0.7164 
                 0.1995 
                 C15orf52 
               
               
                 0 
                 0.3828 
                 0.8011 
                 0.2048 
                 CAPS 
               
               
                 0 
                 0.3853 
                 0.7792 
                 0.1397 
                 GPR160 
               
               
                 0 
                 0.3855 
                 0.6771 
                 0.2797 
                 RPLP0 
               
               
                 0 
                 0.3861 
                 0.6249 
                 0.1425 
                 B4GALT1 
               
               
                 0 
                 0.3861 
                 0.6816 
                 0.1504 
                 SLC39A6 
               
               
                 0 
                 0.3865 
                 0.9027 
                 0.0602 
                 MKI67 
               
               
                 0 
                 0.3868 
                 0.6311 
                 0.2163 
                 SNRP70 
               
               
                 0 
                 0.3895 
                 0.6042 
                 0.2299 
                 UBE2T 
               
               
                 0 
                 0.3896 
                 0.8027 
                 0.2385 
                 CCDC25 
               
               
                 0 
                 0.3905 
                 0.5502 
                 0.0701 
                 KIF2C 
               
               
                 0 
                 0.3924 
                 0.7239 
                 0.1798 
                 PYCR1 
               
               
                 0 
                 0.3936 
                 0.6936 
                 0.1433 
                 CSNK1D 
               
               
                 0 
                 0.3945 
                 0.5928 
                 0.2017 
                 HYPK 
               
               
                 0 
                 0.3964 
                 0.8984 
                 0.1292 
                 ISOC1 
               
               
                 0 
                 0.3979 
                 0.8908 
                 0.0577 
                 HNRPAB 
               
               
                 0 
                 0.3984 
                 0.5033 
                 0.317 
                 MBOAT2 
               
               
                 0 
                 0.3991 
                 0.8319 
                 0.121 
                 PGM5 
               
               
                 0 
                 0.4004 
                 0.555 
                 0.1985 
                 MMP11 
               
               
                 0 
                 0.4004 
                 0.6644 
                 0.1864 
                 TMSB10 
               
               
                 0 
                 0.4007 
                 0.7334 
                 0.1492 
                 ADNP 
               
               
                 0 
                 0.4011 
                 0.9234 
                 0.1192 
                 GAMT 
               
               
                 0 
                 0.4015 
                 0.575 
                 0.1961 
                 MBOAT2 
               
               
                 0 
                 0.4016 
                 0.8532 
                 0.1565 
                 ETS2 
               
               
                 0 
                 0.4016 
                 0.9722 
                 0.1626 
                 PRPF40A 
               
               
                 0 
                 0.4024 
                 0.619 
                 0.2141 
                 PADI2 
               
               
                 0 
                 0.4029 
                 0.5256 
                 0.2863 
                 MIA 
               
               
                 0 
                 0.4046 
                 0.7492 
                 0.1653 
                 MRPS12 
               
               
                 0 
                 0.405 
                 0.789 
                 0.2283 
                 LOC400590 
               
               
                 0 
                 0.4062 
                 0.8739 
                 0.2331 
                 GALNT2 
               
               
                 0 
                 0.407 
                 0.6108 
                 0.1169 
                 MTOR 
               
               
                 0 
                 0.4086 
                 0.751 
                 0.2249 
                 CYBRD1 
               
               
                 0 
                 0.4088 
                 0.8071 
                 0.1578 
                 RASD2 
               
               
                 0 
                 0.411 
                 0.7435 
                 0.1713 
                 LOC401397 
               
               
                 0 
                 0.4112 
                 0.8497 
                 0.1897 
                 SLC6A19 
               
               
                 0 
                 0.4123 
                 0.6129 
                 0.2875 
                 UBTD1 
               
               
                 0 
                 0.4123 
                 0.7862 
                 0.171 
                 CLIP3 
               
               
                 0 
                 0.413 
                 0.6012 
                 0.1462 
                 MGST3 
               
               
                 0 
                 0.4138 
                 0.7788 
                 0.1071 
                 AGPS 
               
               
                 0 
                 0.4138 
                 0.9223 
                 0.1197 
                 CDC20 
               
               
                 0 
                 0.4166 
                 0.7074 
                 0.2572 
                 GSTM1 
               
               
                 0 
                 0.4177 
                 0.693 
                 0.2366 
                 THRAP1 
               
               
                 0 
                 0.4188 
                 0.7162 
                 0.1654 
                 GINS2 
               
               
                 0 
                 0.4188 
                 0.7825 
                 0.0888 
                 RPL23A 
               
               
                 0 
                 0.42 
                 0.6814 
                 0.2875 
                 MGST3 
               
               
                 0 
                 0.4201 
                 0.6393 
                 0.191 
                 VPRBP 
               
               
                 0 
                 0.4206 
                 0.8924 
                 0.0643 
                 ARL8A 
               
               
                 0 
                 0.421 
                 0.5552 
                 0.2112 
                 RPS25 
               
               
                 0 
                 0.4218 
                 0.8356 
                 0.1252 
                 KRTAP5-9 
               
               
                 0 
                 0.4218 
                 0.8082 
                 0.2512 
                 C16orf42 
               
               
                 0 
                 0.4234 
                 0.7464 
                 0.1311 
                 PSMC5 
               
               
                 0 
                 0.4241 
                 0.9274 
                 0.1459 
                 CTSL2 
               
               
                 0 
                 0.4243 
                 0.7231 
                 0.2217 
                 DHPS 
               
               
                 0 
                 0.4256 
                 0.8829 
                 0.2063 
                 ADORA3 
               
               
                 0 
                 0.4261 
                 0.83 
                 0.2359 
                 MFSD1 
               
               
                 0 
                 0.4273 
                 0.7555 
                 0.1193 
                 VPS18 
               
               
                 0 
                 0.4284 
                 0.7952 
                 0.1388 
                 SLC25A31 
               
               
                 0 
                 0.4284 
                 0.8508 
                 0.1151 
                 UBFD1 
               
               
                 0 
                 0.4286 
                 0.8975 
                 0.1875 
                 C1QL2 
               
               
                 0 
                 0.4287 
                 0.5217 
                 0.3514 
                 IDUA 
               
               
                 0 
                 0.4291 
                 0.7443 
                 0.1741 
                 STK11IP 
               
               
                 0 
                 0.4295 
                 0.8537 
                 0.1405 
                 ARL17 
               
               
                 0 
                 0.4299 
                 0.7508 
                 0.1631 
                 FLAD1 
               
               
                 0 
                 0.433 
                 0.9939 
                 0.2011 
                 NDC80 
               
               
                 0 
                 0.434 
                 0.7919 
                 0.1512 
                 KRTAP2-4 
               
               
                 0 
                 0.4342 
                 0.671 
                 0.1749 
                 KRT18 
               
               
                 0 
                 0.4343 
                 0.7493 
                 0.118 
                 PGRMC2 
               
               
                 0 
                 0.4354 
                 0.9375 
                 0.1572 
                 MSI2 
               
               
                 0 
                 0.4359 
                 0.6397 
                 0.2313 
                 TRABD 
               
               
                 0 
                 0.4362 
                 0.7742 
                 0.1324 
                 ROMO1 
               
               
                 0 
                 0.4366 
                 0.7986 
                 0.1156 
                 PCBD2 
               
               
                 0 
                 0.4367 
                 0.9087 
                 0.2691 
                 BCAS1 
               
               
                 0 
                 0.4368 
                 0.7495 
                 0.186 
                 BRD2 
               
               
                 0 
                 0.4369 
                 0.6463 
                 0.1931 
                 IGHA1 
               
               
                 0 
                 0.4371 
                 0.8285 
                 0.1683 
                 MAZ 
               
               
                 0 
                 0.4385 
                 0.7279 
                 0.2212 
                 FGFR4 
               
               
                 0 
                 0.4386 
                 0.5905 
                 0.202 
                 CD24 
               
               
                 0 
                 0.4388 
                 0.7507 
                 0.1749 
                 AK1 
               
               
                 0 
                 0.439 
                 0.7462 
                 0.1921 
                 TBX10 
               
               
                 0 
                 0.4395 
                 0.8645 
                 0.1521 
                 GAMT 
               
               
                 0 
                 0.4401 
                 0.8157 
                 0.2912 
                 LOC442260 
               
               
                 0 
                 0.4402 
                 0.7576 
                 0.1101 
                 UBE2N 
               
               
                 0 
                 0.4411 
                 0.9307 
                 0.1845 
                 FNDC4 
               
               
                 0 
                 0.4411 
                 0.688 
                 0.2201 
                 SUPT6H 
               
               
                 0 
                 0.4412 
                 0.8623 
                 0.091 
                 CCDC74A 
               
               
                 0 
                 0.4438 
                 0.8942 
                 0.1259 
                 SFRS1 
               
               
                 0 
                 0.4462 
                 0.7729 
                 0.1764 
                 RPS14 
               
               
                 0 
                 0.4462 
                 0.8168 
                 0.125 
                 C16orf14 
               
               
                 0 
                 0.4463 
                 0.9565 
                 0.0946 
                 KRTAP2-4 
               
               
                 0 
                 0.4466 
                 0.6565 
                 0.1693 
                 BBC3 
               
               
                 0 
                 0.4469 
                 0.803 
                 0.1084 
                 FAM84B 
               
               
                 0 
                 0.447 
                 0.845 
                 0.2457 
                 ZACN 
               
               
                 0 
                 0.4507 
                 0.7808 
                 0.1479 
                 ZNF704 
               
               
                 0 
                 0.4508 
                 0.9147 
                 0.2458 
                 VDAC1 
               
               
                 0 
                 0.4515 
                 0.7983 
                 0.1594 
                 MRPS36 
               
               
                 0 
                 0.452 
                 0.7063 
                 0.2498 
                 SSBP2 
               
               
                 0 
                 0.4527 
                 0.6211 
                 0.1544 
                 TBC1D9 
               
               
                 0 
                 0.4531 
                 0.9055 
                 0.2287 
                 RPAP1 
               
               
                 0 
                 0.4539 
                 0.8174 
                 0.1172 
                 C15orf52 
               
               
                 0 
                 0.4548 
                 0.9576 
                 0.1074 
                 ORC6L 
               
               
                 0 
                 0.4569 
                 0.8157 
                 0.1044 
                 ADNP 
               
               
                 0 
                 0.4595 
                 0.7648 
                 0.1369 
                 EPOR 
               
               
                 0 
                 0.4611 
                 0.9802 
                 0.1373 
                 KGFLP1 
               
               
                 0 
                 0.4612 
                 0.9553 
                 0.1917 
                 ARL8A 
               
               
                 0 
                 0.4612 
                 0.7023 
                 0.2153 
                 DDX42 
               
               
                 0 
                 0.4613 
                 0.6274 
                 0.3089 
                 KRTAP19-1 
               
               
                 0 
                 0.4618 
                 0.9914 
                 0.1633 
                 FLOT2 
               
               
                 0 
                 0.4619 
                 0.6638 
                 0.2464 
                 NEDD8 
               
               
                 0 
                 0.4641 
                 0.7182 
                 0.2446 
                 C20orf67 
               
               
                 0 
                 0.4647 
                 0.8964 
                 0.1143 
                 LOC642852 
               
               
                 0 
                 0.4663 
                 0.8471 
                 0.2224 
                 HYPK 
               
               
                 0 
                 0.4672 
                 0.7489 
                 0.1967 
                 LAYN 
               
               
                 0 
                 0.469 
                 0.8757 
                 0.2289 
                 CCDC25 
               
               
                 0 
                 0.4693 
                 0.6757 
                 0.1736 
                 OGFR 
               
               
                 0 
                 0.4712 
                 0.7487 
                 0.2692 
                 RPS14 
               
               
                 0 
                 0.4715 
                 0.9546 
                 0.1425 
                 MTOR 
               
               
                 0 
                 0.472 
                 0.8737 
                 0.1708 
                 FLOT2 
               
               
                 0 
                 0.4738 
                 0.9688 
                 0.1734 
                 PXN 
               
               
                 0 
                 0.4742 
                 0.9384 
                 0.0861 
                 SLC25A5 
               
               
                 0 
                 0.4747 
                 0.7689 
                 0.2258 
                 PLD3 
               
               
                 0 
                 0.4753 
                 0.7034 
                 0.2917 
                 TMEM45B 
               
               
                 0 
                 0.4754 
                 0.6429 
                 0.2767 
                 CUGBP1 
               
               
                 0 
                 0.4763 
                 0.7979 
                 0.2544 
                 MRPS36 
               
               
                 0 
                 0.4763 
                 0.9343 
                 0.1583 
                 ZNF704 
               
               
                 0 
                 0.4766 
                 0.9207 
                 0.2153 
                 CCDC24 
               
               
                 0 
                 0.4767 
                 0.864 
                 0.2005 
                 SMG1 
               
               
                 0 
                 0.477 
                 0.9732 
                 0.1225 
                 UBFD1 
               
               
                 0 
                 0.4783 
                 0.6856 
                 0.1954 
                 ADORA3 
               
               
                 0 
                 0.4799 
                 0.6935 
                 0.2601 
                 SLC25A28 
               
               
                 0 
                 0.4812 
                 0.9412 
                 0.1861 
                 NME3 
               
               
                 0 
                 0.4813 
                 0.839 
                 0.1565 
                 HNRNPA1L2 
               
               
                 0 
                 0.4825 
                 0.6575 
                 0.1904 
                 KCNE4 
               
               
                 0 
                 0.4843 
                 0.9812 
                 0.1786 
                 LCE3E 
               
               
                 0 
                 0.4846 
                 0.7526 
                 0.2009 
                 SLC25A29 
               
               
                 0 
                 0.4847 
                 0.7087 
                 0.3026 
                 LOC649178 
               
               
                 0 
                 0.4851 
                 0.6757 
                 0.3091 
                 GPR160 
               
               
                 0 
                 0.4855 
                 0.7326 
                 0.1245 
                 PRR3 
               
               
                 0 
                 0.4858 
                 0.9316 
                 0.3019 
                 LOC285830 
               
               
                 0 
                 0.4861 
                 0.7334 
                 0.1708 
                 IGKV1-5 
               
               
                 0 
                 0.4862 
                 0.7291 
                 0.2521 
                 C19orf28 
               
               
                 0 
                 0.4874 
                 0.8667 
                 0.284 
                 CSNK1A1 
               
               
                 0 
                 0.4877 
                 0.7902 
                 0.2337 
                 UBE2W 
               
               
                 0 
                 0.4883 
                 0.8346 
                 0.2097 
                 POGZ 
               
               
                 0 
                 0.4888 
                 0.8615 
                 0.1317 
                 DPY19L4 
               
               
                 0 
                 0.4901 
                 0.9288 
                 0.2347 
                 CXXC5 
               
               
                 0 
                 0.4915 
                 0.8431 
                 0.198 
                 MGC24125 
               
               
                 0 
                 0.4933 
                 0.8563 
                 0.3301 
                 KGFLP1 
               
               
                 0 
                 0.4934 
                 0.7928 
                 0.225 
                 TP53 
               
               
                 0 
                 0.4936 
                 0.8377 
                 0.1802 
                 PTK2 
               
               
                 0 
                 0.4944 
                 0.8622 
                 0.2002 
                 GPR22 
               
               
                 0 
                 0.4952 
                 0.9651 
                 0.1877 
                 EXO1 
               
               
                 0 
                 0.496 
                 0.9096 
                 0.2492 
                 KRT17 
               
               
                 0 
                 0.4965 
                 0.8691 
                 0.2708 
                 DDX42 
               
               
                 0 
                 0.4971 
                 0.956 
                 0.2376 
                 RPS28 
               
               
                 0 
                 0.4973 
                 0.9344 
                 0.2066 
                 ERBB4 
               
               
                 0 
                 0.4978 
                 0.9956 
                 0.2911 
                 CLIP3 
               
               
                 0 
                 0.4987 
                 0.7218 
                 0.313 
                 PTK2 
               
               
                 0 
                 0.5001 
                 0.9207 
                 0.2011 
                 GSN 
               
               
                 0 
                 0.5004 
                 0.9599 
                 0.2139 
                 KIAA1815 
               
               
                 0 
                 0.5013 
                 0.8068 
                 0.3743 
                 STEAP3 
               
               
                 0 
                 0.5017 
                 0.8404 
                 0.2708 
                 KRT18P28 
               
               
                 0 
                 0.5039 
                 0.7031 
                 0.2445 
                 RASSF7 
               
               
                 0 
                 0.5042 
                 0.9162 
                 0.1926 
                 PLD4 
               
               
                 0 
                 0.5046 
                 0.9301 
                 0.2515 
                 ADCYAP1 
               
               
                 0 
                 0.5054 
                 0.9395 
                 0.2081 
                 LSMD1 
               
               
                 0 
                 0.5063 
                 0.7963 
                 0.204 
                 NXPH3 
               
               
                 0 
                 0.5064 
                 0.8855 
                 0.1803 
                 SLC6A19 
               
               
                 0 
                 0.5066 
                 0.717 
                 0.3463 
                 STEAP3 
               
               
                 0 
                 0.5086 
                 0.9977 
                 0.1348 
                 TBC1D10B 
               
               
                 0 
                 0.5093 
                 0.9829 
                 0.1077 
                 CDC6 
               
               
                 0 
                 0.51 
                 0.8542 
                 0.1738 
                 TAPBP 
               
               
                 0 
                 0.5101 
                 0.7863 
                 0.2278 
                 HNRNPA1L2 
               
               
                 0 
                 0.5109 
                 0.9359 
                 0.1779 
                 CCNB1 
               
               
                 0 
                 0.511 
                 0.9949 
                 0.2187 
                 AURKA 
               
               
                 0 
                 0.5118 
                 0.6619 
                 0.2239 
                 VPS37B 
               
               
                 0 
                 0.5119 
                 0.977 
                 0.1873 
                 DPY19L4 
               
               
                 0 
                 0.5128 
                 0.9219 
                 0.0507 
                 MGC4093 
               
               
                 0 
                 0.5132 
                 0.8587 
                 0.3225 
                 SLC30A10 
               
               
                 0 
                 0.5137 
                 0.6977 
                 0.3608 
                 EHMT1 
               
               
                 0 
                 0.5158 
                 0.8182 
                 0.3438 
                 TFRC 
               
               
                 0 
                 0.5159 
                 0.8333 
                 0.2781 
                 CCDC24 
               
               
                 0 
                 0.5164 
                 0.9912 
                 0.1939 
                 RCL1 
               
               
                 0 
                 0.5171 
                 0.8637 
                 0.1361 
                 KRT14 
               
               
                 0 
                 0.5187 
                 0.9362 
                 0.1708 
                 MTCH2 
               
               
                 0 
                 0.52 
                 0.8154 
                 0.2767 
                 SPTAN1 
               
               
                 0 
                 0.521 
                 0.9374 
                 0.1112 
                 BDH2 
               
               
                 0 
                 0.5211 
                 0.7856 
                 0.2716 
                 VPS37B 
               
               
                 0 
                 0.5222 
                 0.8935 
                 0.2842 
                 KRT18 
               
               
                 0 
                 0.5224 
                 0.9437 
                 0.2132 
                 DRD2 
               
               
                 0 
                 0.5233 
                 0.9794 
                 0.2427 
                 SLC6A19 
               
               
                 0 
                 0.524 
                 0.7061 
                 0.374 
                 ADFP 
               
               
                 0 
                 0.5259 
                 0.8753 
                 0.1395 
                 LOC346887 
               
               
                 0 
                 0.5264 
                 0.8196 
                 0.2214 
                 HIBCH 
               
               
                 0 
                 0.5264 
                 0.759 
                 0.2759 
                 TCEB2 
               
               
                 0 
                 0.5267 
                 0.8973 
                 0.3408 
                 MBNL1 
               
               
                 0 
                 0.5274 
                 0.9924 
                 0.2231 
                 HNRNPA1L2 
               
               
                 0 
                 0.5275 
                 0.6931 
                 0.4012 
                 PHGDH 
               
               
                 0 
                 0.5285 
                 0.8256 
                 0.2666 
                 KHSRP 
               
               
                 0 
                 0.5298 
                 0.8335 
                 0.4093 
                 ELN 
               
               
                 0 
                 0.5317 
                 0.9032 
                 0.3174 
                 CUGBP1 
               
               
                 0 
                 0.532 
                 0.7359 
                 0.2556 
                 PHB2 
               
               
                 0 
                 0.5322 
                 0.9112 
                 0.1902 
                 UGDH 
               
               
                 0 
                 0.5325 
                 0.8362 
                 0.1965 
                 MPDU1 
               
               
                 0 
                 0.5327 
                 0.8594 
                 0.2145 
                 CCNB1 
               
               
                 0 
                 0.5329 
                 0.9997 
                 0.2088 
                 TMEM121 
               
               
                 0 
                 0.5337 
                 0.7695 
                 0.2558 
                 SCUBE2 
               
               
                 0 
                 0.5338 
                 0.7466 
                 0.3408 
                 CCDC25 
               
               
                 0 
                 0.5339 
                 0.934 
                 0.0906 
                 MGC4093 
               
               
                 0 
                 0.5345 
                 0.7322 
                 0.2295 
                 TBC1D9 
               
               
                 0 
                 0.5347 
                 0.9482 
                 0.2669 
                 FABP5 
               
               
                 0 
                 0.5352 
                 0.833 
                 0.2314 
                 POM121L9P 
               
               
                 0 
                 0.536 
                 0.8139 
                 0.286 
                 SPDEF 
               
               
                 0 
                 0.5364 
                 0.6418 
                 0.3183 
                 SNX11 
               
               
                 0 
                 0.5365 
                 0.6782 
                 0.3338 
                 ARHGEF11 
               
               
                 0 
                 0.5366 
                 0.9142 
                 0.2159 
                 IFI27L1 
               
               
                 0 
                 0.5366 
                 0.8427 
                 0.2727 
                 KRTAP13-2 
               
               
                 0 
                 0.5377 
                 0.9448 
                 0.2555 
                 BLVRA 
               
               
                 0 
                 0.5379 
                 0.8465 
                 0.2358 
                 MAP3K13 
               
               
                 0 
                 0.5379 
                 0.9508 
                 0.1348 
                 DKFZP434A0131 
               
               
                 0 
                 0.5392 
                 0.9921 
                 0.0838 
                 RCL1 
               
               
                 0 
                 0.5393 
                 0.8098 
                 0.3437 
                 MSI2 
               
               
                 0 
                 0.5397 
                 0.8193 
                 0.2463 
                 NECAB3 
               
               
                 0 
                 0.5404 
                 0.904 
                 0.1486 
                 SLC16A8 
               
               
                 0 
                 0.5405 
                 0.7599 
                 0.1872 
                 FARP2 
               
               
                 0 
                 0.5407 
                 0.9228 
                 0.2023 
                 RPS3A 
               
               
                 0 
                 0.5416 
                 0.8974 
                 0.2815 
                 PGRMC2 
               
               
                 0 
                 0.5432 
                 0.7967 
                 0.2505 
                 KCNE4 
               
               
                 0 
                 0.5436 
                 0.9841 
                 0.1934 
                 IGHV1-69 
               
               
                 0 
                 0.5437 
                 0.9798 
                 0.0918 
                 SFRP1 
               
               
                 0 
                 0.5443 
                 0.9506 
                 0.2256 
                 SLC30A10 
               
               
                 0 
                 0.5446 
                 0.9415 
                 0.1843 
                 FOXC1 
               
               
                 0 
                 0.545 
                 0.9833 
                 0.2468 
                 CXorf56 
               
               
                 0 
                 0.5465 
                 0.9348 
                 0.3156 
                 SMS 
               
               
                 0 
                 0.548 
                 0.873 
                 0.259 
                 MIA 
               
               
                 0 
                 0.5503 
                 0.7081 
                 0.3715 
                 Kua-UEV 
               
               
                 0 
                 0.5509 
                 0.9427 
                 0.2898 
                 UGDH 
               
               
                 0 
                 0.552 
                 0.9927 
                 0.2002 
                 RPS3A 
               
               
                 0 
                 0.5524 
                 0.9194 
                 0.1359 
                 UBE2C 
               
               
                 0 
                 0.5525 
                 0.8963 
                 0.2301 
                 FAM110A 
               
               
                 0 
                 0.5527 
                 0.8168 
                 0.3731 
                 LARS 
               
               
                 0 
                 0.5531 
                 0.7951 
                 0.3719 
                 RND3 
               
               
                 0 
                 0.5532 
                 0.9411 
                 0.318 
                 SPP1 
               
               
                 0 
                 0.5542 
                 0.8005 
                 0.297 
                 C19orf28 
               
               
                 0 
                 0.5552 
                 0.9352 
                 0.2419 
                 FAM83E 
               
               
                 0 
                 0.5555 
                 0.9513 
                 0.3464 
                 PDZK1IP1 
               
               
                 0 
                 0.5555 
                 0.9528 
                 0.3324 
                 PHRF1 
               
               
                 0 
                 0.556 
                 0.9262 
                 0.3201 
                 PRPF40A 
               
               
                 0 
                 0.558 
                 0.8458 
                 0.25 
                 RAB22A 
               
               
                 0 
                 0.5586 
                 0.753 
                 0.2644 
                 FABP5 
               
               
                 0 
                 0.5592 
                 0.9235 
                 0.275 
                 RELB 
               
               
                 0 
                 0.5599 
                 0.9114 
                 0.3087 
                 HSPA8 
               
               
                 0 
                 0.56 
                 0.8345 
                 0.2174 
                 ATAD3A 
               
               
                 0 
                 0.5604 
                 0.9916 
                 0.2094 
                 YWHAZ 
               
               
                 0 
                 0.5604 
                 0.9009 
                 0.1947 
                 CLPB 
               
               
                 0 
                 0.5623 
                 0.9171 
                 0.3045 
                 TEX2 
               
               
                 0 
                 0.5632 
                 0.961 
                 0.3307 
                 DNAJC4 
               
               
                 0 
                 0.5635 
                 0.95 
                 0.2629 
                 SPTAN1 
               
               
                 0 
                 0.564 
                 0.9526 
                 0.1988 
                 SULT1A2 
               
               
                 0 
                 0.5641 
                 0.9053 
                 0.1616 
                 FNDC4 
               
               
                 0 
                 0.565 
                 0.9711 
                 0.1287 
                 BCAS1 
               
               
                 0 
                 0.5667 
                 0.7795 
                 0.2515 
                 PGR 
               
               
                 0 
                 0.5673 
                 0.9139 
                 0.2956 
                 MALAT1 
               
               
                 0 
                 0.5688 
                 0.8143 
                 0.3014 
                 LOC200810 
               
               
                 0 
                 0.5698 
                 0.9129 
                 0.3094 
                 KCNE4 
               
               
                 0 
                 0.5699 
                 0.8543 
                 0.2923 
                 TMEM97 
               
               
                 0 
                 0.5703 
                 0.9738 
                 0.2347 
                 IGKV1-5 
               
               
                 0 
                 0.5704 
                 0.9449 
                 0.3253 
                 UNC119 
               
               
                 0 
                 0.5714 
                 0.938 
                 0.2877 
                 DAD1 
               
               
                 0 
                 0.5724 
                 0.8395 
                 0.3078 
                 ZC3HAV1L 
               
               
                 0 
                 0.5727 
                 0.9818 
                 0.2937 
                 GPR22 
               
               
                 0 
                 0.573 
                 0.8676 
                 0.311 
                 MAZ 
               
               
                 0 
                 0.573 
                 0.8893 
                 0.2804 
                 PYCR1 
               
               
                 0 
                 0.5746 
                 0.9841 
                 0.2752 
                 TP53 
               
               
                 0 
                 0.5755 
                 0.7404 
                 0.0867 
                 MAD2L2 
               
               
                 0 
                 0.5759 
                 0.8088 
                 0.214 
                 METTL3 
               
               
                 0 
                 0.577 
                 0.9375 
                 0.294 
                 FRMD4A 
               
               
                 0 
                 0.5773 
                 0.961 
                 0.0755 
                 GALNT10 
               
               
                 0 
                 0.5775 
                 0.888 
                 0.3911 
                 HPS6 
               
               
                 0 
                 0.5781 
                 0.8532 
                 0.2051 
                 MFSD1 
               
               
                 0 
                 0.5801 
                 0.9834 
                 0.2924 
                 KIAA0146 
               
               
                 0 
                 0.5801 
                 0.9337 
                 0.3372 
                 PCK2 
               
               
                 0 
                 0.5804 
                 0.8573 
                 0.4032 
                 GPRIN1 
               
               
                 0 
                 0.5806 
                 0.8192 
                 0.2239 
                 IFI27L1 
               
               
                 0 
                 0.5809 
                 0.9849 
                 0.2686 
                 MAFK 
               
               
                 0 
                 0.5826 
                 0.9883 
                 0.0957 
                 RPL10 
               
               
                 0 
                 0.5832 
                 0.8703 
                 0.2143 
                 NECAB3 
               
               
                 0 
                 0.584 
                 0.8789 
                 0.2226 
                 BTG2 
               
               
                 0 
                 0.5841 
                 0.9721 
                 0.327 
                 KIAA0146 
               
               
                 0 
                 0.5842 
                 0.9272 
                 0.2407 
                 LOC652261 
               
               
                 0 
                 0.5853 
                 0.9545 
                 0.2107 
                 PHRF1 
               
               
                 0 
                 0.5854 
                 0.9175 
                 0.2564 
                 ST6GALNAC4 
               
               
                 0 
                 0.5856 
                 0.9715 
                 0.1879 
                 RPS21 
               
               
                 0 
                 0.5872 
                 0.849 
                 0.3134 
                 UBR2 
               
               
                 0 
                 0.5881 
                 0.9613 
                 0.2782 
                 KIAA0652 
               
               
                 0 
                 0.5881 
                 0.8298 
                 0.3702 
                 MVP 
               
               
                 0 
                 0.5885 
                 0.8047 
                 0.2628 
                 TCEB2 
               
               
                 0 
                 0.5885 
                 0.8458 
                 0.34 
                 CHD6 
               
               
                 0 
                 0.5886 
                 0.9733 
                 0.3825 
                 TMEM19 
               
               
                 0 
                 0.5902 
                 0.9988 
                 0.2531 
                 FRAG1 
               
               
                 0 
                 0.5903 
                 0.7583 
                 0.2754 
                 CAPS 
               
               
                 0 
                 0.5917 
                 0.9714 
                 0.3515 
                 CLPP 
               
               
                 0 
                 0.5918 
                 0.9612 
                 0.3871 
                 HIST2H2BE 
               
               
                 0 
                 0.5937 
                 0.8122 
                 0.1991 
                 KIAA1920 
               
               
                 0 
                 0.594 
                 0.8599 
                 0.3944 
                 NXPH3 
               
               
                 0 
                 0.596 
                 0.9099 
                 0.2733 
                 PKP3 
               
               
                 0 
                 0.596 
                 0.9531 
                 0.3264 
                 C14orf1 
               
               
                 0 
                 0.5961 
                 0.8943 
                 0.3773 
                 FBXO25 
               
               
                 0 
                 0.5962 
                 0.9994 
                 0.3779 
                 GINS2 
               
               
                 0 
                 0.5963 
                 0.9871 
                 0.2586 
                 CYB561 
               
               
                 0 
                 0.5964 
                 0.7641 
                 0.3282 
                 KIAA1920 
               
               
                 0 
                 0.5971 
                 0.917 
                 0.2213 
                 YWHAZ 
               
               
                 0 
                 0.5974 
                 0.9979 
                 0.2618 
                 VEGFA 
               
               
                 0 
                 0.5976 
                 0.9969 
                 0.2048 
                 THOP1 
               
               
                 0 
                 0.5985 
                 0.8242 
                 0.2282 
                 MMP11 
               
               
                 0 
                 0.5987 
                 0.885 
                 0.2829 
                 EMP2 
               
               
                 0 
                 0.599 
                 0.981 
                 0.2009 
                 ERBB2IP 
               
               
                 0 
                 0.5991 
                 0.9446 
                 0.2839 
                 CHD6 
               
               
                 0 
                 0.6009 
                 0.923 
                 0.2616 
                 SCNN1D 
               
               
                 0 
                 0.6021 
                 0.9153 
                 0.1722 
                 MAD2L2 
               
               
                 0 
                 0.6025 
                 0.9562 
                 0.4342 
                 CNFN 
               
               
                 0 
                 0.6026 
                 0.8597 
                 0.3629 
                 EPN2 
               
               
                 0 
                 0.6027 
                 0.9924 
                 0.2293 
                 METTL3 
               
               
                 0 
                 0.6035 
                 0.9529 
                 0.3295 
                 ENPP1 
               
               
                 0 
                 0.6038 
                 0.9669 
                 0.2092 
                 CFLP1 
               
               
                 0 
                 0.6054 
                 0.9875 
                 0.3096 
                 SLAIN2 
               
               
                 0 
                 0.6056 
                 0.8544 
                 0.329 
                 DDX50 
               
               
                 0 
                 0.6083 
                 0.9704 
                 0.1802 
                 CDC6 
               
               
                 0 
                 0.6091 
                 0.9572 
                 0.355 
                 RBM14 
               
               
                 0 
                 0.6092 
                 0.9814 
                 0.2331 
                 POGZ 
               
               
                 0 
                 0.6097 
                 0.9903 
                 0.2645 
                 MMD 
               
               
                 0 
                 0.6097 
                 0.9724 
                 0.3251 
                 RPS3A 
               
               
                 0 
                 0.6097 
                 0.9948 
                 0.2198 
                 CXXC5 
               
               
                 0 
                 0.6119 
                 0.8747 
                 0.3061 
                 ZNF609 
               
               
                 0 
                 0.6121 
                 0.9526 
                 0.2149 
                 CLNS1A 
               
               
                 0 
                 0.6124 
                 0.9051 
                 0.2536 
                 AURKA 
               
               
                 0 
                 0.6125 
                 0.9951 
                 0.2624 
                 MAPT 
               
               
                 0 
                 0.6125 
                 0.9932 
                 0.1133 
                 BDH2 
               
               
                 0 
                 0.6125 
                 0.923 
                 0.2215 
                 ZC3H15 
               
               
                 0 
                 0.6126 
                 0.9195 
                 0.2967 
                 LOC442019 
               
               
                 0 
                 0.6141 
                 0.9381 
                 0.3814 
                 EGFR 
               
               
                 0 
                 0.6143 
                 0.9516 
                 0.1436 
                 GHR 
               
               
                 0 
                 0.6146 
                 0.9331 
                 0.4167 
                 BMP2K 
               
               
                 0 
                 0.6148 
                 0.8948 
                 0.1792 
                 SMARCD2 
               
               
                 0 
                 0.6159 
                 0.9354 
                 0.2584 
                 ACTR3B 
               
               
                 0 
                 0.6161 
                 0.9631 
                 0.2091 
                 IMPAD1 
               
               
                 0 
                 0.6162 
                 0.9736 
                 0.3626 
                 LOC401397 
               
               
                 0 
                 0.6165 
                 0.9373 
                 0.4053 
                 RBM14 
               
               
                 0 
                 0.6166 
                 0.9981 
                 0.3839 
                 TUBB2C 
               
               
                 0 
                 0.6169 
                 0.894 
                 0.3824 
                 PTEN 
               
               
                 0 
                 0.617 
                 0.8699 
                 0.412 
                 AKT1 
               
               
                 0 
                 0.6171 
                 0.921 
                 0.3615 
                 LOC285830 
               
               
                 0 
                 0.6173 
                 0.8837 
                 0.3683 
                 RHBDD1 
               
               
                 0 
                 0.6174 
                 0.9658 
                 0.2774 
                 TEX2 
               
               
                 0 
                 0.6176 
                 0.9403 
                 0.296 
                 RPS14 
               
               
                 0 
                 0.6186 
                 0.913 
                 0.3621 
                 MPDU1 
               
               
                 0 
                 0.6187 
                 0.8494 
                 0.3357 
                 TP53 
               
               
                 0 
                 0.6188 
                 0.8623 
                 0.1966 
                 LOC653391 
               
               
                 0 
                 0.6194 
                 0.997 
                 0.2664 
                 RAB27B 
               
               
                 0 
                 0.6196 
                 0.8972 
                 0.1835 
                 FAM84B 
               
               
                 0 
                 0.6205 
                 0.7705 
                 0.379 
                 PSMD3 
               
               
                 0 
                 0.6205 
                 0.946 
                 0.3166 
                 RPS28 
               
               
                 0 
                 0.621 
                 0.9781 
                 0.2499 
                 KGFLP1 
               
               
                 0 
                 0.6212 
                 0.9183 
                 0.2461 
                 ZC3HAV1L 
               
               
                 0 
                 0.6212 
                 0.8697 
                 0.3782 
                 CLIC1 
               
               
                 0 
                 0.6218 
                 0.8033 
                 0.3738 
                 CRTC2 
               
               
                 0 
                 0.6224 
                 0.9059 
                 0.2255 
                 MALAT1 
               
               
                 0 
                 0.6233 
                 0.9923 
                 0.3238 
                 MDM2 
               
               
                 0 
                 0.6234 
                 0.7724 
                 0.3386 
                 PTTG1 
               
               
                 0 
                 0.6237 
                 0.8941 
                 0.2905 
                 POM121L9P 
               
               
                 0 
                 0.6253 
                 0.89 
                 0.1562 
                 ABCF2 
               
               
                 0 
                 0.6255 
                 0.8224 
                 0.3288 
                 ELN 
               
               
                 0 
                 0.6259 
                 0.9521 
                 0.4245 
                 LENG8 
               
               
                 0 
                 0.6266 
                 0.9074 
                 0.4328 
                 LOC649178 
               
               
                 0 
                 0.6267 
                 0.9657 
                 0.3544 
                 FKBP3 
               
               
                 0 
                 0.627 
                 0.9617 
                 0.4084 
                 TCEB2 
               
               
                 0 
                 0.6272 
                 0.9282 
                 0.2686 
                 SLC39A6 
               
               
                 0 
                 0.6276 
                 0.9633 
                 0.4689 
                 ST6GALNAC4 
               
               
                 0 
                 0.628 
                 0.9829 
                 0.3149 
                 CLPP 
               
               
                 0 
                 0.628 
                 0.8813 
                 0.2685 
                 DDX50 
               
               
                 0 
                 0.6281 
                 0.9477 
                 0.3428 
                 XYLT1 
               
               
                 0 
                 0.6289 
                 0.9378 
                 0.3864 
                 GALNT2 
               
               
                 0 
                 0.6313 
                 0.9804 
                 0.2038 
                 KRT14 
               
               
                 0 
                 0.6314 
                 0.9661 
                 0.2993 
                 SOX4 
               
               
                 0 
                 0.633 
                 0.9948 
                 0.3844 
                 AKT1 
               
               
                 0 
                 0.6331 
                 0.9148 
                 0.4279 
                 C17orf37 
               
               
                 0 
                 0.634 
                 0.986 
                 0.2757 
                 PRELID1 
               
               
                 0 
                 0.6347 
                 0.862 
                 0.5175 
                 VPS37B 
               
               
                 0 
                 0.6349 
                 0.9252 
                 0.3608 
                 CENPF 
               
               
                 0 
                 0.6356 
                 0.7589 
                 0.4233 
                 SNRP70 
               
               
                 0 
                 0.6364 
                 0.8009 
                 0.4371 
                 SCNN1D 
               
               
                 0 
                 0.6367 
                 0.8482 
                 0.3367 
                 HDGFRP3 
               
               
                 0 
                 0.6367 
                 0.9998 
                 0.3151 
                 THRAP1 
               
               
                 0 
                 0.637 
                 0.9236 
                 0.3972 
                 PRKD3 
               
               
                 0 
                 0.6376 
                 0.9146 
                 0.4745 
                 SREBF2 
               
               
                 0 
                 0.6376 
                 0.9847 
                 0.2059 
                 C1QL2 
               
               
                 0 
                 0.6377 
                 0.8456 
                 0.3928 
                 ERBB2 
               
               
                 0 
                 0.6379 
                 0.9837 
                 0.2563 
                 MSN 
               
               
                 0 
                 0.6381 
                 0.9377 
                 0.3416 
                 SELO 
               
               
                 0 
                 0.6382 
                 0.8633 
                 0.3039 
                 CACNG7 
               
               
                 0 
                 0.6388 
                 0.9836 
                 0.1054 
                 VDAC1 
               
               
                 0 
                 0.6402 
                 0.9715 
                 0.2158 
                 TEX2 
               
               
                 0 
                 0.6404 
                 0.8321 
                 0.4685 
                 FBXO25 
               
               
                 0 
                 0.6409 
                 0.9168 
                 0.223 
                 RPL10 
               
               
                 0 
                 0.6409 
                 0.8981 
                 0.284 
                 DAD1 
               
               
                 0 
                 0.6412 
                 0.9677 
                 0.4101 
                 CAPS 
               
               
                 0 
                 0.642 
                 0.957 
                 0.2921 
                 H2AFY 
               
               
                 0 
                 0.642 
                 0.9992 
                 0.331 
                 ILF2 
               
               
                 0 
                 0.642 
                 0.9431 
                 0.0661 
                 RPS21 
               
               
                 0 
                 0.643 
                 0.9307 
                 0.4789 
                 TFRC 
               
               
                 0 
                 0.6433 
                 0.9527 
                 0.3386 
                 PTK2 
               
               
                 0 
                 0.6449 
                 0.956 
                 0.362 
                 BRD2 
               
               
                 0 
                 0.6458 
                 0.9681 
                 0.3661 
                 RPS2 
               
               
                 0 
                 0.6464 
                 0.8606 
                 0.265 
                 KRTAP2-4 
               
               
                 0 
                 0.6466 
                 0.9062 
                 0.4614 
                 UBFD1 
               
               
                 0 
                 0.6468 
                 0.9405 
                 0.4443 
                 IRGC 
               
               
                 0 
                 0.6474 
                 0.9228 
                 0.479 
                 IGHA1 
               
               
                 0 
                 0.648 
                 0.8942 
                 0.3385 
                 SPDEF 
               
               
                 0 
                 0.65 
                 0.8887 
                 0.255 
                 MYBL2 
               
               
                 0 
                 0.6508 
                 0.9669 
                 0.3298 
                 FGFR4 
               
               
                 0 
                 0.651 
                 0.9915 
                 0.1339 
                 PGM5 
               
               
                 0 
                 0.652 
                 0.9904 
                 0.4144 
                 HIST2H2BE 
               
               
                 0 
                 0.6523 
                 0.914 
                 0.3021 
                 SF3B3 
               
               
                 0 
                 0.6523 
                 0.917 
                 0.3656 
                 DHPS 
               
               
                 0 
                 0.6528 
                 0.9862 
                 0.351 
                 FRMD4A 
               
               
                 0 
                 0.6532 
                 0.8503 
                 0.3567 
                 TBC1D10B 
               
               
                 0 
                 0.6533 
                 0.9472 
                 0.3838 
                 GSN 
               
               
                 0 
                 0.6535 
                 0.9044 
                 0.3562 
                 NUF2 
               
               
                 0 
                 0.6538 
                 0.8938 
                 0.3959 
                 GAMT 
               
               
                 0 
                 0.6539 
                 0.9385 
                 0.3718 
                 PCK2 
               
               
                 0 
                 0.6544 
                 0.8728 
                 0.4005 
                 FRMD4A 
               
               
                 0 
                 0.6546 
                 0.9983 
                 0.1605 
                 IGJ 
               
               
                 0 
                 0.6552 
                 0.999 
                 0.4203 
                 RRM2 
               
               
                 0 
                 0.6554 
                 0.8896 
                 0.4308 
                 ANGPTL2 
               
               
                 0 
                 0.6554 
                 0.9756 
                 0.3819 
                 BDH2 
               
               
                 0 
                 0.6555 
                 0.9101 
                 0.2109 
                 AGPS 
               
               
                 0 
                 0.6555 
                 0.9767 
                 0.2594 
                 CIAPIN1 
               
               
                 0 
                 0.6556 
                 0.8917 
                 0.431 
                 ALG13 
               
               
                 0 
                 0.6559 
                 0.9564 
                 0.3499 
                 PRKD3 
               
               
                 0 
                 0.6561 
                 0.842 
                 0.3549 
                 GUSB 
               
               
                 0 
                 0.6565 
                 0.8903 
                 0.4051 
                 SLC16A8 
               
               
                 0 
                 0.6565 
                 0.9731 
                 0.4523 
                 ACTR3B 
               
               
                 0 
                 0.657 
                 0.8964 
                 0.3038 
                 PRR3 
               
               
                 0 
                 0.6579 
                 0.9279 
                 0.4306 
                 PTPRA 
               
               
                 0 
                 0.6595 
                 0.8248 
                 0.3348 
                 NME4 
               
               
                 0 
                 0.6601 
                 0.8765 
                 0.4759 
                 ACTR3B 
               
               
                 0 
                 0.6617 
                 0.9884 
                 0.2203 
                 ACAD9 
               
               
                 0 
                 0.6618 
                 0.8665 
                 0.4001 
                 FLJ22659 
               
               
                 0 
                 0.6624 
                 0.9275 
                 0.3648 
                 BCL2 
               
               
                 0 
                 0.664 
                 0.999 
                 0.3043 
                 ARL17 
               
               
                 0 
                 0.6651 
                 0.9212 
                 0.4236 
                 SNHG5 
               
               
                 0 
                 0.666 
                 0.9938 
                 0.2815 
                 MMD 
               
               
                 0 
                 0.6662 
                 0.9768 
                 0.2878 
                 JMJD1B 
               
               
                 0 
                 0.6663 
                 0.9424 
                 0.385 
                 HYPK 
               
               
                 0 
                 0.6673 
                 0.9537 
                 0.4065 
                 KRT18P28 
               
               
                 0 
                 0.6681 
                 0.9837 
                 0.4513 
                 LENG8 
               
               
                 0 
                 0.6681 
                 0.9342 
                 0.4758 
                 PHACTR4 
               
               
                 0 
                 0.6686 
                 0.9637 
                 0.2579 
                 GUSBL2 
               
               
                 0 
                 0.6687 
                 0.9857 
                 0.3898 
                 ACAD9 
               
               
                 0 
                 0.6695 
                 0.9823 
                 0.314 
                 ADORA3 
               
               
                 0 
                 0.6705 
                 0.9952 
                 0.3506 
                 MBNL1 
               
               
                 0 
                 0.6718 
                 0.9308 
                 0.3587 
                 C14orf1 
               
               
                 0 
                 0.6723 
                 0.9928 
                 0.2639 
                 GGA2 
               
               
                 0 
                 0.6727 
                 0.9366 
                 0.2944 
                 PCSK6 
               
               
                 0 
                 0.6727 
                 0.9055 
                 0.2316 
                 SLC25A5 
               
               
                 0 
                 0.6733 
                 0.9629 
                 0.2627 
                 C20orf144 
               
               
                 0 
                 0.6735 
                 0.9881 
                 0.3268 
                 HIBCH 
               
               
                 0 
                 0.6742 
                 0.9071 
                 0.507 
                 KRTAP13-2 
               
               
                 0 
                 0.6742 
                 0.9975 
                 0.4199 
                 RND3 
               
               
                 0 
                 0.6751 
                 0.9585 
                 0.3661 
                 MYC 
               
               
                 0 
                 0.6754 
                 0.9252 
                 0.3509 
                 PSMC5 
               
               
                 0 
                 0.6757 
                 0.9927 
                 0.3963 
                 ALG13 
               
               
                 0 
                 0.677 
                 0.9799 
                 0.472 
                 FRAG1 
               
               
                 0 
                 0.677 
                 0.9922 
                 0.4235 
                 VPRBP 
               
               
                 0 
                 0.6772 
                 0.9503 
                 0.3955 
                 HSPBP1 
               
               
                 0 
                 0.6776 
                 0.8631 
                 0.2532 
                 HNRPAB 
               
               
                 0 
                 0.6778 
                 0.8877 
                 0.4002 
                 CEACAM1 
               
               
                 0 
                 0.6785 
                 0.9998 
                 0.4248 
                 RPS2 
               
               
                 0 
                 0.6785 
                 0.9782 
                 0.3189 
                 CD63 
               
               
                 0 
                 0.6789 
                 0.9996 
                 0.2981 
                 TRABD 
               
               
                 0 
                 0.68 
                 0.8164 
                 0.5369 
                 C1orf212 
               
               
                 0 
                 0.6805 
                 0.9543 
                 0.4227 
                 MKI67 
               
               
                 0 
                 0.6806 
                 0.9661 
                 0.384 
                 PLD3 
               
               
                 0 
                 0.6809 
                 0.9294 
                 0.4219 
                 SMARCD2 
               
               
                 0 
                 0.6814 
                 0.9555 
                 0.4695 
                 PTEN 
               
               
                 0 
                 0.6822 
                 0.9686 
                 0.3218 
                 PIAS1 
               
               
                 0 
                 0.6824 
                 0.9467 
                 0.2595 
                 PRKD3 
               
               
                 0 
                 0.6826 
                 0.9225 
                 0.4275 
                 ELAVL4 
               
               
                 0 
                 0.683 
                 0.8928 
                 0.3574 
                 KIAA0310 
               
               
                 0 
                 0.6831 
                 0.9722 
                 0.4076 
                 GCGR 
               
               
                 0 
                 0.6833 
                 0.9901 
                 0.4466 
                 MMD 
               
               
                 0 
                 0.6841 
                 0.9864 
                 0.3263 
                 BTG2 
               
               
                 0 
                 0.6843 
                 0.9552 
                 0.4427 
                 GNPTG 
               
               
                 0 
                 0.6844 
                 0.9991 
                 0.3038 
                 SNX11 
               
               
                 0 
                 0.6844 
                 0.9304 
                 0.4359 
                 UBR2 
               
               
                 0 
                 0.6854 
                 0.9374 
                 0.2301 
                 SMCP 
               
               
                 0 
                 0.6855 
                 0.9261 
                 0.3341 
                 EMP2 
               
               
                 0 
                 0.6869 
                 0.9749 
                 0.3817 
                 ACTB 
               
               
                 0 
                 0.6885 
                 0.8771 
                 0.4549 
                 ENPP1 
               
               
                 0 
                 0.6887 
                 0.9139 
                 0.3565 
                 FAM148A 
               
               
                 0 
                 0.6887 
                 0.9374 
                 0.4136 
                 NUDCD3 
               
               
                 0 
                 0.6893 
                 0.9416 
                 0.3452 
                 RPL10 
               
               
                 0 
                 0.6895 
                 0.9063 
                 0.2827 
                 GGA2 
               
               
                 0 
                 0.6911 
                 0.9949 
                 0.2505 
                 CLPP 
               
               
                 0 
                 0.6934 
                 0.922 
                 0.4453 
                 LOC642852 
               
               
                 0 
                 0.6934 
                 0.9627 
                 0.4467 
                 CNFN 
               
               
                 0 
                 0.6937 
                 0.9749 
                 0.4044 
                 KIF2C 
               
               
                 0 
                 0.6939 
                 0.9652 
                 0.3715 
                 POLDIP2 
               
               
                 0 
                 0.6943 
                 0.9799 
                 0.3867 
                 C20orf20 
               
               
                 0 
                 0.6948 
                 0.9357 
                 0.4145 
                 LAYN 
               
               
                 0 
                 0.695 
                 0.9938 
                 0.4247 
                 MRPS12 
               
               
                 0 
                 0.695 
                 0.9715 
                 0.3227 
                 UBE2T 
               
               
                 0 
                 0.6962 
                 0.9383 
                 0.2122 
                 ZNF225 
               
               
                 0 
                 0.6969 
                 0.9185 
                 0.2386 
                 HIST1H2AA 
               
               
                 0 
                 0.697 
                 0.982 
                 0.3922 
                 SLC25A31 
               
               
                 0 
                 0.6973 
                 0.9732 
                 0.4419 
                 PHACTR4 
               
               
                 0 
                 0.6974 
                 0.9251 
                 0.5496 
                 UBE2C 
               
               
                 0 
                 0.6978 
                 0.8539 
                 0.4938 
                 KIAA2013 
               
               
                 0 
                 0.6988 
                 0.9634 
                 0.526 
                 CD63 
               
               
                 0 
                 0.6989 
                 0.976 
                 0.3421 
                 EHD2 
               
               
                 0 
                 0.699 
                 0.9151 
                 0.3437 
                 THSD4 
               
               
                 0 
                 0.6991 
                 0.9421 
                 0.2866 
                 CENPF 
               
               
                 0 
                 0.6993 
                 0.8774 
                 0.3055 
                 VEGFA 
               
               
                 0 
                 0.6995 
                 0.976 
                 0.4145 
                 PPIA 
               
               
                 0 
                 0.6997 
                 0.9205 
                 0.421 
                 KIAA1815 
               
               
                 0 
                 0.6997 
                 0.9916 
                 0.2685 
                 NME4 
               
               
                 0 
                 0.6998 
                 0.9845 
                 0.3316 
                 SNX11 
               
               
                 0 
                 0.7005 
                 0.9193 
                 0.4062 
                 TMEM19 
               
               
                 0 
                 0.7005 
                 0.9378 
                 0.4089 
                 CACNG7 
               
               
                 0 
                 0.7006 
                 0.9797 
                 0.2201 
                 C20orf67 
               
               
                 0 
                 0.7014 
                 0.937 
                 0.3042 
                 DHPS 
               
               
                 0 
                 0.7015 
                 0.9375 
                 0.4427 
                 TMEM45B 
               
               
                 0 
                 0.7018 
                 0.9624 
                 0.4117 
                 C16orf42 
               
               
                 0 
                 0.7021 
                 0.9396 
                 0.3035 
                 TMEM121 
               
               
                 0 
                 0.7026 
                 0.9773 
                 0.4756 
                 DRD2 
               
               
                 0 
                 0.7027 
                 0.9797 
                 0.2284 
                 FAM173B 
               
               
                 0 
                 0.7029 
                 0.9749 
                 0.4154 
                 TMEM121 
               
               
                 0 
                 0.7032 
                 0.9691 
                 0.2979 
                 KRT18P28 
               
               
                 0 
                 0.7034 
                 0.9682 
                 0.3619 
                 KRTAP13-2 
               
               
                 0 
                 0.704 
                 0.9634 
                 0.4285 
                 MGAT4B 
               
               
                 0 
                 0.7045 
                 0.896 
                 0.4424 
                 hCG_1642354 
               
               
                 0 
                 0.7047 
                 0.9711 
                 0.4327 
                 EPOR 
               
               
                 0 
                 0.7049 
                 0.9221 
                 0.3743 
                 SELO 
               
               
                 0 
                 0.7051 
                 0.9618 
                 0.3537 
                 RPS25 
               
               
                 0 
                 0.7052 
                 0.8959 
                 0.3767 
                 RCL1 
               
               
                 0 
                 0.7057 
                 0.9958 
                 0.3935 
                 SLAIN2 
               
               
                 0 
                 0.706 
                 0.9118 
                 0.2027 
                 ARL8A 
               
               
                 0 
                 0.7071 
                 0.9575 
                 0.3423 
                 FAM110A 
               
               
                 0 
                 0.7071 
                 0.9793 
                 0.3858 
                 GPR22 
               
               
                 0 
                 0.7075 
                 0.9443 
                 0.2604 
                 IGKV2-24 
               
               
                 0 
                 0.7076 
                 0.9262 
                 0.4323 
                 GSN 
               
               
                 0 
                 0.708 
                 0.9902 
                 0.3368 
                 ERBB2IP 
               
               
                 0 
                 0.708 
                 0.8642 
                 0.436 
                 TMEM97 
               
               
                 0 
                 0.7086 
                 0.932 
                 0.3669 
                 MGST3 
               
               
                 0 
                 0.7087 
                 0.9934 
                 0.3936 
                 MELK 
               
               
                 0 
                 0.709 
                 0.9772 
                 0.4655 
                 SF3B3 
               
               
                 0 
                 0.7091 
                 0.942 
                 0.4158 
                 HDAC4 
               
               
                 0 
                 0.711 
                 0.983 
                 0.4759 
                 UBE2T 
               
               
                 0 
                 0.7122 
                 0.9998 
                 0.4827 
                 PCSK6 
               
               
                 0 
                 0.7129 
                 0.9708 
                 0.4504 
                 PITPNC1 
               
               
                 0 
                 0.7141 
                 0.9939 
                 0.469 
                 LOC401397 
               
               
                 0 
                 0.7144 
                 0.9806 
                 0.2118 
                 MKI67 
               
               
                 0 
                 0.715 
                 0.8717 
                 0.4644 
                 EGFR 
               
               
                 0 
                 0.7153 
                 0.8907 
                 0.4257 
                 LASS6 
               
               
                 0 
                 0.7153 
                 0.9665 
                 0.4881 
                 PCTK2 
               
               
                 0 
                 0.7155 
                 0.9625 
                 0.2872 
                 UQCR 
               
               
                 0 
                 0.7157 
                 0.9843 
                 0.4601 
                 TBC1D9 
               
               
                 0 
                 0.7162 
                 0.9271 
                 0.4327 
                 HSPA8 
               
               
                 0 
                 0.7163 
                 0.8728 
                 0.4736 
                 LSMD1 
               
               
                 0 
                 0.7167 
                 0.9993 
                 0.3291 
                 LOC730275 
               
               
                 0 
                 0.7169 
                 0.9736 
                 0.2485 
                 CDC6 
               
               
                 0 
                 0.7172 
                 0.9637 
                 0.4451 
                 KRT7 
               
               
                 0 
                 0.7175 
                 0.8705 
                 0.4871 
                 SIAH2 
               
               
                 0 
                 0.7181 
                 0.9807 
                 0.3871 
                 LOC653391 
               
               
                 0 
                 0.7185 
                 0.9998 
                 0.3324 
                 SLC30A10 
               
               
                 0 
                 0.7187 
                 0.91 
                 0.3928 
                 FBXO25 
               
               
                 0 
                 0.7193 
                 0.9777 
                 0.3884 
                 FAM127A 
               
               
                 0 
                 0.7199 
                 0.9324 
                 0.3211 
                 C16orf14 
               
               
                 0 
                 0.7199 
                 0.9397 
                 0.4944 
                 CSNK1A1 
               
               
                 0 
                 0.7204 
                 0.998 
                 0.3332 
                 ZNF225 
               
               
                 0 
                 0.721 
                 0.9495 
                 0.4755 
                 LOC100128062 
               
               
                 0 
                 0.7216 
                 0.993 
                 0.4682 
                 RAB22A 
               
               
                 0 
                 0.7226 
                 0.9953 
                 0.3791 
                 GALNT10 
               
               
                 0 
                 0.7226 
                 0.8765 
                 0.5133 
                 BMP2K 
               
               
                 0 
                 0.7227 
                 0.9492 
                 0.5545 
                 IGHA1 
               
               
                 0 
                 0.7227 
                 0.968 
                 0.5317 
                 LOC642852 
               
               
                 0 
                 0.7227 
                 0.9944 
                 0.5061 
                 SMARCD2 
               
               
                 0 
                 0.7233 
                 0.9504 
                 0.3534 
                 KIAA2013 
               
               
                 0 
                 0.7238 
                 0.9173 
                 0.4837 
                 HNRPA3 
               
               
                 0 
                 0.7238 
                 0.9597 
                 0.449 
                 SLC25A31 
               
               
                 0 
                 0.7239 
                 0.9859 
                 0.2638 
                 FLAD1 
               
               
                 0 
                 0.7241 
                 0.9939 
                 0.318 
                 MGC24125 
               
               
                 0 
                 0.7246 
                 0.9829 
                 0.4979 
                 MGC24125 
               
               
                 0 
                 0.725 
                 0.9312 
                 0.572 
                 KIAA0310 
               
               
                 0 
                 0.7251 
                 0.9792 
                 0.4393 
                 VDAC1 
               
               
                 0 
                 0.7252 
                 0.9648 
                 0.4113 
                 TMEM174 
               
               
                 0 
                 0.7256 
                 0.9579 
                 0.4541 
                 STEAP3 
               
               
                 0 
                 0.7265 
                 0.963 
                 0.366 
                 PCTK2 
               
               
                 0 
                 0.727 
                 0.9992 
                 0.4787 
                 RPLP0 
               
               
                 0 
                 0.7272 
                 0.9074 
                 0.4215 
                 H2AFY 
               
               
                 0 
                 0.7281 
                 0.9945 
                 0.4109 
                 GALNT2 
               
               
                 0 
                 0.7282 
                 0.9207 
                 0.411 
                 URM1 
               
               
                 0 
                 0.7282 
                 0.9215 
                 0.2106 
                 C1orf93 
               
               
                 0 
                 0.7283 
                 0.9244 
                 0.4316 
                 PTEN 
               
               
                 0 
                 0.7284 
                 0.9758 
                 0.4679 
                 C1QL2 
               
               
                 0 
                 0.7286 
                 0.9269 
                 0.4543 
                 HEATR3 
               
               
                 0 
                 0.7286 
                 0.9906 
                 0.4525 
                 PTPRA 
               
               
                 0 
                 0.7304 
                 0.9659 
                 0.4943 
                 KIAA1815 
               
               
                 0 
                 0.7305 
                 0.9643 
                 0.3026 
                 PITPNC1 
               
               
                 0 
                 0.7311 
                 0.9658 
                 0.531 
                 SMG1 
               
               
                 0 
                 0.7313 
                 0.887 
                 0.5312 
                 STK11IP 
               
               
                 0 
                 0.7314 
                 0.9265 
                 0.5148 
                 ISOC1 
               
               
                 0 
                 0.7321 
                 0.9483 
                 0.4796 
                 MRPS12 
               
               
                 0 
                 0.7324 
                 0.9566 
                 0.2202 
                 MALAT1 
               
               
                 0 
                 0.7328 
                 0.9623 
                 0.2481 
                 GNPTG 
               
               
                 0 
                 0.733 
                 0.9962 
                 0.4347 
                 TRIB3 
               
               
                 0 
                 0.7345 
                 0.928 
                 0.4988 
                 ACBD6 
               
               
                 0 
                 0.7349 
                 0.9197 
                 0.4493 
                 BRD2 
               
               
                 0 
                 0.7354 
                 0.9875 
                 0.4886 
                 KRT17 
               
               
                 0 
                 0.7361 
                 0.9868 
                 0.3632 
                 EXO1 
               
               
                 0 
                 0.7367 
                 0.9275 
                 0.4796 
                 IGHV1-69 
               
               
                 0 
                 0.7372 
                 0.9624 
                 0.4535 
                 C1orf104 
               
               
                 0 
                 0.7375 
                 0.9398 
                 0.4891 
                 LOC400590 
               
               
                 0 
                 0.7378 
                 0.9654 
                 0.4863 
                 IL6ST 
               
               
                 0 
                 0.7383 
                 0.9381 
                 0.3418 
                 UBR2 
               
               
                 0 
                 0.7394 
                 0.9405 
                 0.3804 
                 MGAT4B 
               
               
                 0 
                 0.7395 
                 0.9226 
                 0.5431 
                 UQCR 
               
               
                 0 
                 0.7401 
                 0.9929 
                 0.3967 
                 NEDD8 
               
               
                 0 
                 0.7412 
                 0.9511 
                 0.5199 
                 ARHGEF11 
               
               
                 0 
                 0.7417 
                 0.9628 
                 0.3511 
                 MCCD1 
               
               
                 0 
                 0.7417 
                 0.9515 
                 0.4353 
                 MSI2 
               
               
                 0 
                 0.7421 
                 0.9273 
                 0.5576 
                 EMP2 
               
               
                 0 
                 0.7422 
                 0.9362 
                 0.6205 
                 MARVELD2 
               
               
                 0 
                 0.7422 
                 0.9361 
                 0.5301 
                 MYO1F 
               
               
                 0 
                 0.7433 
                 0.9757 
                 0.453 
                 C1orf93 
               
               
                 0 
                 0.7434 
                 0.9692 
                 0.4942 
                 ORMDL3 
               
               
                 0 
                 0.7438 
                 0.9593 
                 0.4619 
                 UGCG 
               
               
                 0 
                 0.7441 
                 0.9964 
                 0.506 
                 MGC70870 
               
               
                 0 
                 0.7444 
                 0.9194 
                 0.3783 
                 ATXN2 
               
               
                 0 
                 0.7448 
                 0.967 
                 0.3434 
                 ADCY7 
               
               
                 0 
                 0.745 
                 0.9971 
                 0.5858 
                 CEP55 
               
               
                 0 
                 0.7455 
                 0.9163 
                 0.5755 
                 LSMD1 
               
               
                 0 
                 0.7455 
                 0.8844 
                 0.5856 
                 PXN 
               
               
                 0 
                 0.7462 
                 0.9987 
                 0.5026 
                 SMCP 
               
               
                 0 
                 0.7463 
                 0.9602 
                 0.5405 
                 SNX5 
               
               
                 0 
                 0.7476 
                 0.9991 
                 0.3192 
                 C20orf20 
               
               
                 0 
                 0.7482 
                 0.8923 
                 0.4533 
                 CYB561 
               
               
                 0 
                 0.7483 
                 0.9933 
                 0.2156 
                 VPS45A 
               
               
                 0 
                 0.7484 
                 0.9947 
                 0.4546 
                 LOC649178 
               
               
                 0 
                 0.7485 
                 0.9019 
                 0.4989 
                 MAP3K13 
               
               
                 0 
                 0.7492 
                 0.9644 
                 0.5197 
                 SLC16A8 
               
               
                 0 
                 0.7498 
                 0.9895 
                 0.2785 
                 TAPBP 
               
               
                 0 
                 0.75 
                 0.8713 
                 0.3743 
                 ZNF592 
               
               
                 0 
                 0.7501 
                 0.9806 
                 0.5118 
                 ARFGEF2 
               
               
                 0 
                 0.7503 
                 0.9745 
                 0.2555 
                 CIZ1 
               
               
                 0 
                 0.7507 
                 0.9536 
                 0.3272 
                 FLJ35390 
               
               
                 0 
                 0.7507 
                 0.9894 
                 0.5138 
                 ADAMTSL5 
               
               
                 0 
                 0.7508 
                 0.9461 
                 0.5709 
                 IGH@ 
               
               
                 0 
                 0.7508 
                 0.9937 
                 0.3778 
                 NLK 
               
               
                 0 
                 0.751 
                 0.9693 
                 0.4256 
                 MCCD1 
               
               
                 0 
                 0.7511 
                 0.9154 
                 0.5499 
                 LOC652261 
               
               
                 0 
                 0.7511 
                 0.9991 
                 0.371 
                 ATAD3A 
               
               
                 0 
                 0.7514 
                 0.987 
                 0.4115 
                 LOC442019 
               
               
                 0 
                 0.7514 
                 0.9624 
                 0.4926 
                 RHBDD1 
               
               
                 0 
                 0.752 
                 0.9436 
                 0.3224 
                 FKSG30 
               
               
                 0 
                 0.7521 
                 0.9559 
                 0.5952 
                 CXorf56 
               
               
                 0 
                 0.7523 
                 0.9819 
                 0.4708 
                 RPAP1 
               
               
                 0 
                 0.7525 
                 0.9973 
                 0.5524 
                 ELAVL4 
               
               
                 0 
                 0.7526 
                 0.9579 
                 0.4707 
                 CLNS1A 
               
               
                 0 
                 0.7528 
                 0.9443 
                 0.4462 
                 TYMS 
               
               
                 0 
                 0.7529 
                 0.9963 
                 0.5272 
                 C8orf73 
               
               
                 0 
                 0.7534 
                 0.953 
                 0.3571 
                 KHSRP 
               
               
                 0 
                 0.7536 
                 0.9868 
                 0.5323 
                 KRTAP5-9 
               
               
                 0 
                 0.7538 
                 0.9801 
                 0.3808 
                 FAM83E 
               
               
                 0 
                 0.7538 
                 0.941 
                 0.4053 
                 VPRBP 
               
               
                 0 
                 0.7543 
                 0.9004 
                 0.6412 
                 MBOAT2 
               
               
                 0 
                 0.7549 
                 0.9813 
                 0.2649 
                 CEP55 
               
               
                 0 
                 0.7551 
                 0.9893 
                 0.243 
                 LCE3E 
               
               
                 0 
                 0.7551 
                 0.9603 
                 0.4614 
                 MLPH 
               
               
                 0 
                 0.7556 
                 0.9967 
                 0.3562 
                 KIAA0146 
               
               
                 0 
                 0.7561 
                 0.961 
                 0.2536 
                 CACNG7 
               
               
                 0 
                 0.7562 
                 0.952 
                 0.3203 
                 GALNT10 
               
               
                 0 
                 0.7564 
                 0.9683 
                 0.2773 
                 EHD2 
               
               
                 0 
                 0.7569 
                 0.9906 
                 0.5289 
                 CXXC5 
               
               
                 0 
                 0.7571 
                 0.9337 
                 0.5152 
                 CD63 
               
               
                 0 
                 0.7573 
                 0.9824 
                 0.337 
                 JMJD1B 
               
               
                 0 
                 0.7576 
                 0.957 
                 0.4758 
                 FRAG1 
               
               
                 0 
                 0.7576 
                 0.9839 
                 0.5753 
                 TRABD 
               
               
                 0 
                 0.7581 
                 0.9954 
                 0.4265 
                 NAT10 
               
               
                 0 
                 0.7591 
                 0.9745 
                 0.4933 
                 MAFK 
               
               
                 0 
                 0.7607 
                 0.978 
                 0.3086 
                 POLR2L 
               
               
                 0 
                 0.761 
                 0.9127 
                 0.506 
                 HRH2 
               
               
                 0 
                 0.7622 
                 0.984 
                 0.4022 
                 KIAA0652 
               
               
                 0 
                 0.7622 
                 0.9742 
                 0.4243 
                 CLNS1A 
               
               
                 0 
                 0.7628 
                 0.9515 
                 0.5061 
                 REPS2 
               
               
                 0 
                 0.7635 
                 0.9124 
                 0.4856 
                 TBX21 
               
               
                 0 
                 0.7642 
                 0.9801 
                 0.4484 
                 MED13L 
               
               
                 0 
                 0.7649 
                 0.9628 
                 0.4702 
                 DDX50 
               
               
                 0 
                 0.765 
                 0.9625 
                 0.4859 
                 EHMT1 
               
               
                 0 
                 0.765 
                 0.9878 
                 0.3723 
                 MLPH 
               
               
                 0 
                 0.766 
                 0.9948 
                 0.4551 
                 POLR3H 
               
               
                 0 
                 0.7661 
                 0.9813 
                 0.4048 
                 EVL 
               
               
                 0 
                 0.7663 
                 0.9967 
                 0.389 
                 POGZ 
               
               
                 0 
                 0.7664 
                 0.9321 
                 0.5645 
                 KRTAP5-9 
               
               
                 0 
                 0.7666 
                 0.9956 
                 0.3713 
                 CDC20 
               
               
                 0 
                 0.7669 
                 0.9805 
                 0.4749 
                 HDGFRP3 
               
               
                 0 
                 0.7672 
                 0.9786 
                 0.3668 
                 DOT1L 
               
               
                 0 
                 0.7672 
                 0.9904 
                 0.3309 
                 SLAIN2 
               
               
                 0 
                 0.7673 
                 0.9873 
                 0.5417 
                 NUF2 
               
               
                 0 
                 0.7677 
                 0.9881 
                 0.5894 
                 SLC25A28 
               
               
                 0 
                 0.7685 
                 0.9939 
                 0.409 
                 KIAA2013 
               
               
                 0 
                 0.7685 
                 0.9925 
                 0.4487 
                 SMG1 
               
               
                 0 
                 0.7688 
                 0.9988 
                 0.4309 
                 PPIA 
               
               
                 0 
                 0.7692 
                 0.9851 
                 0.4793 
                 MSN 
               
               
                 0 
                 0.7706 
                 0.9768 
                 0.4737 
                 HNRPAB 
               
               
                 0 
                 0.7708 
                 0.9968 
                 0.505 
                 DOT1L 
               
               
                 0 
                 0.7708 
                 0.9438 
                 0.4485 
                 BRAF 
               
               
                 0 
                 0.7709 
                 0.9432 
                 0.5775 
                 ROMO1 
               
               
                 0 
                 0.7709 
                 0.9916 
                 0.4307 
                 ACTB 
               
               
                 0 
                 0.771 
                 0.9931 
                 0.3584 
                 MAZ 
               
               
                 0 
                 0.771 
                 0.996 
                 0.4555 
                 RHBDD1 
               
               
                 0 
                 0.7711 
                 0.9295 
                 0.605 
                 GSTM1 
               
               
                 0 
                 0.7715 
                 0.9934 
                 0.482 
                 BCL2 
               
               
                 0 
                 0.7716 
                 0.9675 
                 0.559 
                 NDUFB9 
               
               
                 0 
                 0.7718 
                 0.9952 
                 0.324 
                 UGDH 
               
               
                 0 
                 0.7723 
                 0.9955 
                 0.3569 
                 ZACN 
               
               
                 0 
                 0.7727 
                 0.9836 
                 0.5241 
                 TAF2 
               
               
                 0 
                 0.7735 
                 0.9948 
                 0.4459 
                 MELK 
               
               
                 0 
                 0.7737 
                 0.9772 
                 0.5689 
                 ERBB4 
               
               
                 0 
                 0.7744 
                 0.9909 
                 0.437 
                 NLK 
               
               
                 0 
                 0.7755 
                 0.9979 
                 0.4124 
                 RPS2 
               
               
                 0 
                 0.7755 
                 0.955 
                 0.498 
                 CDH3 
               
               
                 0 
                 0.7757 
                 0.9979 
                 0.3835 
                 ZNF704 
               
               
                 0 
                 0.7758 
                 0.987 
                 0.4859 
                 ZNF609 
               
               
                 0 
                 0.776 
                 0.948 
                 0.6177 
                 VPS45A 
               
               
                 0 
                 0.7763 
                 0.998 
                 0.3259 
                 C20orf144 
               
               
                 0 
                 0.7777 
                 0.9665 
                 0.2782 
                 C1orf104 
               
               
                 0 
                 0.7785 
                 0.9589 
                 0.4912 
                 RPAP1 
               
               
                 0 
                 0.7786 
                 0.9215 
                 0.4194 
                 IGKV2-24 
               
               
                 0 
                 0.7788 
                 0.9928 
                 0.5772 
                 JMJD1B 
               
               
                 0 
                 0.7789 
                 0.9847 
                 0.4949 
                 KRTAP19-1 
               
               
                 0 
                 0.7789 
                 0.9834 
                 0.4657 
                 MCCD1 
               
               
                 0 
                 0.7793 
                 0.9677 
                 0.6465 
                 UBE2W 
               
               
                 0 
                 0.7794 
                 0.9996 
                 0.4779 
                 SLC7A2 
               
               
                 0 
                 0.7798 
                 0.959 
                 0.4224 
                 CIZ1 
               
               
                 0 
                 0.7801 
                 0.9777 
                 0.5373 
                 CRTC2 
               
               
                 0 
                 0.7806 
                 0.9715 
                 0.3712 
                 FARP2 
               
               
                 0 
                 0.7809 
                 0.9938 
                 0.5772 
                 KRTAP19-1 
               
               
                 0 
                 0.7811 
                 0.9545 
                 0.5695 
                 FGFR4 
               
               
                 0 
                 0.7813 
                 0.9954 
                 0.4387 
                 CYBRD1 
               
               
                 0 
                 0.7814 
                 0.932 
                 0.6018 
                 LOC652261 
               
               
                 0 
                 0.7818 
                 0.9813 
                 0.5384 
                 TBXAS1 
               
               
                 0 
                 0.7822 
                 0.9972 
                 0.4964 
                 UHMK1 
               
               
                 0 
                 0.7824 
                 0.9583 
                 0.5898 
                 ASPHD2 
               
               
                 0 
                 0.7831 
                 0.9568 
                 0.6717 
                 KRT5 
               
               
                 0 
                 0.7832 
                 0.9533 
                 0.4823 
                 IRGC 
               
               
                 0 
                 0.7835 
                 0.9207 
                 0.5815 
                 EVL 
               
               
                 0 
                 0.7835 
                 0.9866 
                 0.344 
                 FOXA1 
               
               
                 0 
                 0.7835 
                 0.9765 
                 0.5165 
                 ZNF592 
               
               
                 0 
                 0.7852 
                 0.9288 
                 0.4759 
                 CDH3 
               
               
                 0 
                 0.7854 
                 0.9934 
                 0.5758 
                 GPRIN1 
               
               
                 0 
                 0.7857 
                 0.9679 
                 0.5555 
                 NUDCD3 
               
               
                 0 
                 0.7859 
                 0.9042 
                 0.5553 
                 ARFGEF2 
               
               
                 0 
                 0.7864 
                 0.9803 
                 0.4765 
                 L3MBTL2 
               
               
                 0 
                 0.7874 
                 0.9729 
                 0.5098 
                 FAM127A 
               
               
                 0 
                 0.7875 
                 0.9916 
                 0.4092 
                 FOXC1 
               
               
                 0 
                 0.7884 
                 0.9889 
                 0.4946 
                 MYO1F 
               
               
                 0 
                 0.7887 
                 0.9734 
                 0.6167 
                 ENO1 
               
               
                 0 
                 0.7887 
                 0.995 
                 0.4453 
                 IFI27L1 
               
               
                 0 
                 0.7893 
                 0.9625 
                 0.2405 
                 GNPTG 
               
               
                 0 
                 0.7893 
                 0.9634 
                 0.485 
                 HDAC4 
               
               
                 0 
                 0.7907 
                 0.9676 
                 0.5618 
                 C15orf52 
               
               
                 0 
                 0.7908 
                 0.9863 
                 0.3637 
                 SCUBE2 
               
               
                 0 
                 0.791 
                 0.9069 
                 0.414 
                 ERBB2IP 
               
               
                 0 
                 0.7912 
                 0.9935 
                 0.5062 
                 VPS18 
               
               
                 0 
                 0.7924 
                 0.9953 
                 0.4164 
                 TMCO1 
               
               
                 0 
                 0.7927 
                 0.9936 
                 0.5016 
                 UQCR 
               
               
                 0 
                 0.7928 
                 0.9551 
                 0.481 
                 DOT1L 
               
               
                 0 
                 0.794 
                 0.9783 
                 0.511 
                 FAM110A 
               
               
                 0 
                 0.7942 
                 0.9611 
                 0.5636 
                 ADCYAP1 
               
               
                 0 
                 0.7942 
                 0.9928 
                 0.3832 
                 GSR 
               
               
                 0 
                 0.7943 
                 0.9775 
                 0.561 
                 FAM173B 
               
               
                 0 
                 0.7944 
                 0.9635 
                 0.3841 
                 PIAS1 
               
               
                 0 
                 0.7946 
                 0.9932 
                 0.563 
                 NAT10 
               
               
                 0 
                 0.7948 
                 0.9923 
                 0.2763 
                 MAP3K13 
               
               
                 0 
                 0.7949 
                 0.9865 
                 0.2372 
                 KIAA1920 
               
               
                 0 
                 0.7955 
                 0.9871 
                 0.4835 
                 LOC100128062 
               
               
                 0 
                 0.7956 
                 0.9578 
                 0.6056 
                 CIZ1 
               
               
                 0 
                 0.7961 
                 0.992 
                 0.4024 
                 CDC20 
               
               
                 0 
                 0.7966 
                 0.9016 
                 0.5901 
                 POLD4 
               
               
                 0 
                 0.7978 
                 0.9634 
                 0.4638 
                 LARS 
               
               
                 0 
                 0.7978 
                 0.9967 
                 0.5465 
                 ARHGEF11 
               
               
                 0 
                 0.7982 
                 0.9963 
                 0.4248 
                 HEATR3 
               
               
                 0 
                 0.7988 
                 0.9866 
                 0.5443 
                 ATXN2 
               
               
                 0 
                 0.7998 
                 0.9941 
                 0.512 
                 TAF2 
               
               
                 0 
                 0.8001 
                 0.9855 
                 0.4593 
                 HIST1H2AA 
               
               
                 0 
                 0.8001 
                 0.9997 
                 0.5497 
                 PHB2 
               
               
                 0 
                 0.8002 
                 0.9594 
                 0.5511 
                 MARVELD2 
               
               
                 0 
                 0.8006 
                 0.9752 
                 0.5392 
                 FKSG30 
               
               
                 0 
                 0.8011 
                 0.9741 
                 0.527 
                 PIAS1 
               
               
                 0 
                 0.8012 
                 0.9275 
                 0.701 
                 ERBB4 
               
               
                 0 
                 0.8018 
                 0.9315 
                 0.5793 
                 SCUBE2 
               
               
                 0 
                 0.8028 
                 0.9998 
                 0.3789 
                 GUSB 
               
               
                 0 
                 0.8029 
                 0.9209 
                 0.6017 
                 MFSD1 
               
               
                 0 
                 0.8032 
                 0.9991 
                 0.3723 
                 CENPF 
               
               
                 0 
                 0.8033 
                 0.9838 
                 0.5098 
                 ADCYAP1 
               
               
                 0 
                 0.8039 
                 0.9871 
                 0.5352 
                 KRT5 
               
               
                 0 
                 0.8045 
                 0.9884 
                 0.4947 
                 NUDCD3 
               
               
                 0 
                 0.8045 
                 0.98 
                 0.6506 
                 SLC25A28 
               
               
                 0 
                 0.8045 
                 0.9543 
                 0.6513 
                 SMS 
               
               
                 0 
                 0.8051 
                 0.9972 
                 0.4344 
                 TBXAS1 
               
               
                 0 
                 0.8055 
                 0.9735 
                 0.5294 
                 MGC4093 
               
               
                 0 
                 0.8056 
                 0.9717 
                 0.6619 
                 PLEKHF2 
               
               
                 0 
                 0.8057 
                 0.9784 
                 0.4543 
                 IRGC 
               
               
                 0 
                 0.8066 
                 0.9421 
                 0.5289 
                 FKSG30 
               
               
                 0 
                 0.8074 
                 0.9622 
                 0.6389 
                 NAT10 
               
               
                 0 
                 0.8077 
                 0.9586 
                 0.4516 
                 PHB2 
               
               
                 0 
                 0.8079 
                 0.9646 
                 0.5062 
                 SFRS1 
               
               
                 0 
                 0.8081 
                 0.9591 
                 0.4882 
                 PHGDH 
               
               
                 0 
                 0.8085 
                 0.9918 
                 0.5035 
                 CNFN 
               
               
                 0 
                 0.8087 
                 0.9482 
                 0.5376 
                 POLD4 
               
               
                 0 
                 0.8087 
                 0.975 
                 0.5217 
                 ADAMTSL5 
               
               
                 0 
                 0.809 
                 0.9991 
                 0.3924 
                 IGJ 
               
               
                 0 
                 0.8092 
                 0.9619 
                 0.5416 
                 BIRC5 
               
               
                 0 
                 0.8094 
                 0.9991 
                 0.5216 
                 LOC643159 
               
               
                 0 
                 0.8102 
                 0.9987 
                 0.5383 
                 CSNK1A1 
               
               
                 0 
                 0.8105 
                 0.9991 
                 0.645 
                 PLD4 
               
               
                 0 
                 0.8114 
                 0.9584 
                 0.6001 
                 FLJ22659 
               
               
                 0 
                 0.8118 
                 0.9781 
                 0.5185 
                 MYC 
               
               
                 0 
                 0.8125 
                 0.9773 
                 0.4933 
                 Kua-UEV 
               
               
                 0 
                 0.8126 
                 0.9912 
                 0.4778 
                 LOC643159 
               
               
                 0 
                 0.8126 
                 0.9523 
                 0.6253 
                 PDZK1IP1 
               
               
                 0 
                 0.8132 
                 0.9899 
                 0.5081 
                 MAD2L2 
               
               
                 0 
                 0.8145 
                 0.9697 
                 0.54 
                 MSN 
               
               
                 0 
                 0.8146 
                 0.995 
                 0.5387 
                 FABP5 
               
               
                 0 
                 0.816 
                 0.9743 
                 0.4729 
                 NME4 
               
               
                 0 
                 0.8166 
                 0.9423 
                 0.4208 
                 ANAPC1 
               
               
                 0 
                 0.8173 
                 0.9896 
                 0.5609 
                 METTL3 
               
               
                 0 
                 0.8177 
                 0.9082 
                 0.582 
                 FAM173B 
               
               
                 0 
                 0.8191 
                 0.9887 
                 0.5152 
                 MYC 
               
               
                 0 
                 0.8191 
                 0.9742 
                 0.5227 
                 CCDC74A 
               
               
                 0 
                 0.8192 
                 0.9726 
                 0.5983 
                 GHR 
               
               
                 0 
                 0.8198 
                 0.95 
                 0.5253 
                 OGFR 
               
               
                 0 
                 0.82 
                 0.985 
                 0.6715 
                 CD68 
               
               
                 0 
                 0.8202 
                 0.973 
                 0.657 
                 HNRPA3 
               
               
                 0 
                 0.8203 
                 0.9702 
                 0.4825 
                 PPP2R2D 
               
               
                 0 
                 0.8205 
                 0.9733 
                 0.5151 
                 GSR 
               
               
                 0 
                 0.8206 
                 0.9986 
                 0.5146 
                 ADCY7 
               
               
                 0 
                 0.8212 
                 0.9688 
                 0.5202 
                 SMS 
               
               
                 0 
                 0.8224 
                 0.9819 
                 0.6383 
                 FARP2 
               
               
                 0 
                 0.823 
                 0.9939 
                 0.5167 
                 MVP 
               
               
                 0 
                 0.8237 
                 0.9959 
                 0.6136 
                 ARFGEF2 
               
               
                 0 
                 0.8257 
                 0.9757 
                 0.6619 
                 GUSBL2 
               
               
                 0 
                 0.8259 
                 0.9843 
                 0.6225 
                 CLIP3 
               
               
                 0 
                 0.8264 
                 0.9883 
                 0.6808 
                 HRH2 
               
               
                 0 
                 0.8268 
                 0.9875 
                 0.6082 
                 CD68 
               
               
                 0 
                 0.8275 
                 0.9936 
                 0.6562 
                 LOC400590 
               
               
                 0 
                 0.8275 
                 0.9781 
                 0.599 
                 RRM2 
               
               
                 0 
                 0.8278 
                 0.9674 
                 0.5771 
                 NEBL 
               
               
                 0 
                 0.8282 
                 0.9683 
                 0.5229 
                 TMEM19 
               
               
                 0 
                 0.8287 
                 0.9992 
                 0.5074 
                 SULT1A2 
               
               
                 0 
                 0.8295 
                 0.9874 
                 0.4886 
                 PCSK6 
               
               
                 0 
                 0.8301 
                 0.9704 
                 0.611 
                 C14orf1 
               
               
                 0 
                 0.8306 
                 0.9989 
                 0.5225 
                 AGPS 
               
               
                 0 
                 0.8308 
                 0.9686 
                 0.4749 
                 FLJ22795 
               
               
                 0 
                 0.8317 
                 0.959 
                 0.6623 
                 LOC442019 
               
               
                 0 
                 0.8322 
                 0.9763 
                 0.6096 
                 TMCO1 
               
               
                 0 
                 0.8331 
                 0.9889 
                 0.6527 
                 FLJ35390 
               
               
                 0 
                 0.8334 
                 0.995 
                 0.5498 
                 KRT17 
               
               
                 0 
                 0.8341 
                 0.9754 
                 0.5523 
                 SNX5 
               
               
                 0 
                 0.8343 
                 0.9954 
                 0.4872 
                 C20orf20 
               
               
                 0 
                 0.8347 
                 0.9954 
                 0.4962 
                 FOXC1 
               
               
                 0 
                 0.8347 
                 0.9672 
                 0.7422 
                 GSR 
               
               
                 0 
                 0.8355 
                 0.9891 
                 0.6917 
                 EVL 
               
               
                 0 
                 0.837 
                 0.9727 
                 0.5342 
                 LOC200810 
               
               
                 0 
                 0.8384 
                 0.9918 
                 0.6627 
                 BMP2K 
               
               
                 0 
                 0.8387 
                 0.9783 
                 0.6471 
                 DPY19L4 
               
               
                 0 
                 0.8415 
                 0.9997 
                 0.6035 
                 SULT1A2 
               
               
                 0 
                 0.8423 
                 0.9859 
                 0.6327 
                 TMBIM1 
               
               
                 0 
                 0.843 
                 0.999 
                 0.6451 
                 ANAPC1 
               
               
                 0 
                 0.845 
                 0.9743 
                 0.6192 
                 LOC653391 
               
               
                 0 
                 0.8455 
                 0.9924 
                 0.709 
                 PRELID1 
               
               
                 0 
                 0.846 
                 0.9775 
                 0.6775 
                 MVP 
               
               
                 0 
                 0.8465 
                 0.9855 
                 0.5448 
                 ETS2 
               
               
                 0 
                 0.8468 
                 0.9994 
                 0.7006 
                 HNRPA3 
               
               
                 0 
                 0.847 
                 0.9761 
                 0.5791 
                 CLPB 
               
               
                 0 
                 0.8482 
                 0.9834 
                 0.4922 
                 MGAT4B 
               
               
                 0 
                 0.8497 
                 0.9957 
                 0.5635 
                 IMPAD1 
               
               
                 0 
                 0.8501 
                 0.9438 
                 0.6114 
                 PHRF1 
               
               
                 0 
                 0.851 
                 0.9683 
                 0.66 
                 VPS18 
               
               
                 0 
                 0.8523 
                 0.9642 
                 0.6895 
                 SELO 
               
               
                 0 
                 0.8528 
                 0.9957 
                 0.6256 
                 GHR 
               
               
                 0 
                 0.8538 
                 0.9849 
                 0.7121 
                 PHGDH 
               
               
                 0 
                 0.8547 
                 0.9979 
                 0.6107 
                 PLD4 
               
               
                 0 
                 0.8585 
                 0.958 
                 0.7646 
                 ZNF592 
               
               
                 0 
                 0.8601 
                 0.9967 
                 0.6851 
                 MYO1F 
               
               
                 0 
                 0.8602 
                 0.9796 
                 0.5052 
                 OGFR 
               
               
                 0 
                 0.8603 
                 0.9876 
                 0.712 
                 LOC200810 
               
               
                 0 
                 0.8608 
                 0.9784 
                 0.5134 
                 RASD2 
               
               
                 0 
                 0.8671 
                 0.9935 
                 0.6818 
                 PLEKHF2 
               
               
                 0 
                 0.8677 
                 0.9848 
                 0.6755 
                 FLJ22795 
               
               
                 0 
                 0.8685 
                 0.9998 
                 0.5926 
                 PTPRA 
               
               
                 0 
                 0.8697 
                 0.988 
                 0.7056 
                 BAG1 
               
               
                 0 
                 0.8708 
                 0.997 
                 0.6107 
                 ATXN2 
               
               
                 0 
                 0.871 
                 0.9679 
                 0.4552 
                 ANAPC1 
               
               
                 0 
                 0.8727 
                 0.9988 
                 0.5969 
                 RELB 
               
               
                 0 
                 0.8732 
                 0.9981 
                 0.7045 
                 CD68 
               
               
                 0 
                 0.8755 
                 0.9856 
                 0.7283 
                 TRIB3 
               
               
                 0 
                 0.8776 
                 0.9917 
                 0.6808 
                 SLC25A29 
               
               
                 0 
                 0.8807 
                 0.9743 
                 0.6968 
                 GRB7 
               
               
                 0 
                 0.8855 
                 0.9921 
                 0.6874 
                 STK11IP 
               
               
                 0 
                 0.8868 
                 0.9881 
                 0.6278 
                 ALG13 
               
               
                 0 
                 0.8956 
                 0.9952 
                 0.6646 
                 NEDD8 
               
               
                 0 
                 0.9076 
                 0.9843 
                 0.7082 
                 RELB 
               
               
                   
               
            
           
         
       
     
     Since each gene was treated as categorical variable based on quartiles with lowest quartile as reference, there are three categories for each gene. Mean, minimum, and maximum interaction p-values from 10-fold jack knifing process are shown. Fifteen genes were significant 100% of the time (FLOT2, CA12, TUBB2C, UNC119, GATA3, SUPT6H, RPL23A, SLC39A14, ABHD2, FTH1, FAM84B, ACVR1B, ZACN). Clustering of these or any other combination of genes selected purely based on statistical significance did not allow for robust identification of subsets with differential benefit from trastuzumab. In light of this, it was decided to attempt an additional approach to identify subsets with differential benefit from trastuzumab. 
     From among all of the results of gene assessment performed, it was noticed that the top predictive genes included several estrogen receptor associated genes, CA12 (mean interaction p=0.0059), GATA3 (p=0.007), PIK3A (p=0.0388) as well as genes from HER2 amplicon: ERBB2 (p=0.0485) and C17orf37 (p=0.0442). Using this information and the facts that ER status has been associated with lower rates of complete pathological response in several published studies (Untch, et al., supra; Bhargava, et al.,  Mod. Pathol.  24:367-374, 2011) and that HER2 (ERBB2) is the target for trastuzumab, it was decided to select, as the basis to develop a predictive algorithm, genes whose expression levels were correlated with ESR1 mRNA or with ERBB2 mRNA having Spearman&#39;s correlation coefficient over 0.7 and also a minimum interaction P value below 0.1. The top genes correlated with ESR1 and ERBB2 are shown in Table 5. From this pool, 8 genes met the criteria of a correlation coefficient over 0.7 and a minimum interaction P value below 0.1. These genes included ESR1, NAT1, GATA3, CA12, IGFR1, ERBB2, c17orf37 and GRB7. 
     
       
         
           
               
               
               
               
             
               
                   
                 TABLE 5 
               
               
                   
                   
               
               
                   
                   
                   
                 Minimum 
               
               
                   
                 Gene 
                   
                 Interaction 
               
               
                   
                 Symbol 
                   
                 P Value 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
            
               
                   
                   
                 Correlation with 
                   
               
               
                   
                   
                 ERBB2 
               
               
                   
                 ERBB2 
                 1 
                 0.025 
               
               
                   
                 GRB7 
                 0.912 
                 0.06 
               
               
                   
                 C17orf37 
                 0.833 
                 0.0003 
               
               
                   
                 KRT7 
                 0.498 
                 0.047 
               
               
                   
                 TMEM45B 
                 0.453 
                 0.29 
               
               
                   
                 ORMDL3 
                 0.448 
                 0.076 
               
               
                   
                 C1orf93 
                 0.427 
                 0.1 
               
               
                   
                 SPDEF 
                 0.4 
                 0.013 
               
               
                   
                 VEGFA 
                 0.395 
                 0.24 
               
               
                   
                 FGFR4 
                 0.347 
                 0.35 
               
               
                   
                   
                 Correlation with 
               
               
                   
                   
                 ESR1 
               
               
                   
                 ESR1 
                 1 
                 0.064 
               
               
                   
                 TBC1D9 
                 0.757 
                 0.49 
               
               
                   
                 CA12 
                 0.733 
                 0.0024 
               
               
                   
                 IGF1R 
                 0.731 
                 0.042 
               
               
                   
                 GATA3 
                 0.727 
                 0.0036 
               
               
                   
                 THSD4 
                 0.727 
                 0.12 
               
               
                   
                 NAT1 
                 0.701 
                 0.075 
               
               
                   
                 SLC39A6 
                 0.685 
                 0.21 
               
               
                   
                 SCUBE2 
                 0.637 
                 0.47 
               
               
                   
                 SIAH2 
                 0.632 
                 0.19 
               
               
                   
                   
               
            
           
         
       
     
     In order to identify subsets with different degree of benefit from trastuzumab while accommodating the non-linearity of interaction between genes and trastuzumab, the first two principal components (PC1 and PC2) obtained from the 8 selected predictive genes were used to create a three dimensional subset treatment effect pattern plot with spline interpolation to smooth the plot with hazard ratio for trastuzumab on Z-axis. Hazard ratios were color coded as green if less than 0.5 (large benefit from trastuzumab), brown for 0.5-1.0 (moderate benefit), or red for over 1.0 (no benefit). This plot readily identified subsets with differential benefit from trastuzumab. Cut-points were derived for two principal components (PC1 and PC2) that defined three subsets based on TDSTEPP and the event rate in each subgroup. 
     The cut-points for two principal components (PC1 and PC2) that defined these three subsets were determined as follows: No benefit group if PC1&gt;0.6 and PC2&gt;0.1; Large benefit group if −0.12&lt;PC1&lt;=0.6 and 0.1&lt;PC2&lt;=0.6 and PC2&gt;PC1+0.22, if −0.6&lt;PC1&lt;=0.6 and PC2&gt;=0.6, or if PC1&lt;=−0.12 and −0.55&lt;PC2&lt;0.6. Remaining patients were classified as the moderate benefit group. 
     Kaplan-Meier plots were created for three subsets identified using these cut-points for PC1 and PC2. The no benefit group (Group 1, N=81) had a hazard ratio of 1.56. The moderate benefit group (Group 2, N=255) had a HR of 0.56, and the large benefit group (Group 3, N=252) had a HR of 0.27. It should be noted that p-values and confidence intervals for these data are not appropriate, because these plots are for the discovery cohort that was used to develop the algorithm. The plots were used to illustrate the degree of differentiation in trastuzumab effect that is achieved with the algorithm. 
     Assessment of the Pre-Defined Cut-Points for the Prediction Model in the Confirmation Cohort 
     The pre-defined cut-points from the 8-gene prediction model described above were assessed in the remaining 991 B-31 patients not included in the discovery phase for whom specimens were available. Since the algorithm has not yet been developed into a formal clinical test, a formal NCI registered date stamped protocol was not developed before proceeding to the cut-points assessment. Kaplan-Meier plots were created based on the pre-defined cut-off values for the two principal components created by applying the eigen-vector coefficients from the candidate discovery set to the confirmation dataset. Applying the pre-defined cut-points for the 8-gene prediction model readily identified: a subset with no benefit from trastuzumab (Group 1) with a hazard ratio of 1.58 (95% CI: 0.67-3.69, p=0.29, N=100), a subset with moderate benefit (Group 2) with a hazard ratio of 0.60 (95% CI: 0.41-0.89, p=0.011, N=449), and a subset with large benefit (Group 3) with a hazard ratio of 0.28 (95% CI: 0.20-0.41, p&lt;0.0001, N=442). The p-value for the interaction between predictive algorithm and trastuzumab was 0.0002. 
     Distribution of Central HER2Assay Negative Cases Among Categories Defined by the Prediction Model 
     Because HER2 is the target for trastuzumab, it is expected that Group 1 with no benefit should express the lowest levels of ERBB2 mRNA. A correlation analysis was performed between ERBB2 and ESR1 mRNA levels in which each subgroup defined by the 8-gene prediction model. Surprisingly, the subset with no benefit expressed high levels of ESR1 mRNA and intermediate levels of ERBB2 mRNA rather than the lowest levels in both candidate discovery and confirmation cohorts. 
     An unexpected finding from the B-31 trial was that central HER2 assay negative patients also derived benefit from trastuzumab. Because the 8-gene prediction model was developed independent of the knowledge of centrally performed HER2 testing results, it was tested whether central HER2 assay negative cases belong to the Group 1 defined by the predictive model with no expected benefit. When central HER2 negative results were overlaid on these subsets, only a few HER2 negative patients belonged to the subgroup with no benefit, while a majority belonged to the moderate-benefit subgroup. 
     These results support the hypothesis that HER2 negative patients may derive benefit from trastuzumab. 
     Discussion 
     Using multiplexed gene expression profiling with RNA extracted from archived formalin fixed paraffin embedded tumor blocks from NSABP trial B-31, a predictive algorithm for the degree of benefit from trastuzumab added to adjuvant chemo-endocrine therapy of HER2 positive breast cancer was developed. In the internal confirmation set of 991 patients, this algorithm and pre-defined cut-points were validated with interaction p-value of 0.0002. 
     The data demonstrate a complex relationship between HER2 and ER as determinants of clinical benefit from trastuzumab added to adjuvant chemo-endocrine therapy. ERBB2 mRNA-by-trastuzumab interaction was not linear and was also modulated by other genes, especially those from estrogen receptor pathway. Most surprisingly, the identified subgroup with no clinical benefit from adjuvant trastuzumab actually expressed intermediate—not the lowest—levels of ERBB2 mRNA, together with the highest levels of ESR1-associated genes. This subgroup also had an excellent baseline prognosis, which was similar to the prognosis of others treated with trastuzumab. 
     While not bound to any particular theory, there could be at least two explanations for the lack of benefit in this subgroup. In NSABP trial B-14, it was observed that ESR1 mRNA level is a linear predictor of the degree of benefit from tamoxifen (Kim, et al.,  J. Clin. Oncol.  29:4160-4167, 2011). Therefore, one explanation may be that patients with tumors that express highest levels of ESR1 and its associated mRNAs may have already derived maximum clinical benefit from antiestrogen therapy. An alternative explanation is that such tumors are biologically resistant to trastuzumab. Lower rate of complete pathological response to neoadjuvant trastuzumab in ER-positive tumors compared to ER-negative tumors supports the second interpretation. It is possible that estrogen receptor is directly responsible by inducing anti-apoptotic proteins such as Bcl-2 or IGF1R. Overexpressed IGF1R can hetero-trimerize with HER2 and EGFR, and cause resistance to trastuzumab in vitro and in vivo (Huang, et al., Cancer Res. 70:1204-1214, 2010; Lu, et al.,  J. Natl. Cancer Inst.  93:1852-1857, 2001). In reality, due to a close association of expression levels among these genes, it is impossible to separate them. 
     Regardless of the mechanisms responsible for no clinical benefit, therapeutic strategies to improve the outcome of this subgroup need to be developed because, although their prognosis is favorable, patients still suffer from over 10% recurrences in 5 years, which is not improved by the addition of trastuzumab. A combination of HER2, ER, and IGF1R targeting, HER2 targeting combined with complete blockage of ER pathway using fulvestrant (because IGF1R is induced by ER; Osborne, et al.,  Br. J. Cancer  90 Suppl. 1:S2-S6, 2004), or a SRC inhibitor (Zhang, et al.,  Nat. Med.  17:461-469, 2011) may be a potential strategy. 
     The data also support the hypothesis based on central HER2 testing results from B-31 that HER2 negative patients may benefit from adjuvant trastuzumab. Because HER2 negative patients belong to Group 2, approximately 40 percent reduction in recurrences is expected from the addition of trastuzumab to adjuvant chemotherapy with minor side effects. This hypothesis is currently being tested through a randomized clinical trial (NSABP protocol B-47: NCT01275677). 
     All of the compositions and/or methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and/or methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.