Patent Publication Number: US-2011064730-A1

Title: Method of modulating angiogenesis

Description:
RELATED APPLICATIONS 
     This application is a continuation-in-part patent application which claims the benefit of the filing date of U.S. patent application Ser. No. 10/550,533, filed Sep. 22, 2005, which claims the benefit of the filing date of PCT International Patent Application Serial No. PCT/AU2004/000383, filed Mar. 26, 2004, which claims the benefit of the filing date of Australian Patent Application Serial No. 2003901511, filed Mar. 28, 2003, the contents of each of which are herein incorporated by reference in their entirety. 
    
    
     TECHNICAL FIELD 
     The present invention relates to novel nucleic acid sequences (“angiogenic genes”) involved in the process of angiogenesis. Each of the angiogenic genes encode a polypeptide that has a role in angiogenesis. In view of the realisation that these genes play a role in angiogenesis, the invention is also concerned with the therapy of pathologies associated with angiogenesis, the screening of drugs for pro- or anti-angiogenic activity, the diagnosis and prognosis of pathologies associated with angiogenesis, and in some cases the use of the nucleic acid sequences to identify and obtain full-length angiogenesis-related genes. 
     BACKGROUND 
     The formation of new blood vessels from pre-existing vessels, a process termed angiogenesis, is essential for normal growth. Important angiogenic processes include those taking place in embryogenesis, renewal of the endometrium, formation and growth of the corpus luteum of pregnancy, wound healing and in the restoration of tissue structure and function after injury. 
     The formation of new capillaries requires a co-ordinated series of events mediated through the expression of multiple genes which may have either pro- or anti-angiogenic activities. The process begins with an angiogenic stimulus to existing vasculature, usually mediated by growth factors such as vascular endothelial growth factor or basic fibroblast growth factor. This is followed by degradation of the extracellular matrix, cell adhesion changes (and disruption), an increase in cell permeability, proliferation of endothelial cells (ECs) and migration of ECs towards the site of blood vessel formation. Subsequent processes include capillary tube or lumen formation, stabilisation and differentiation by the migrating ECs. 
     In the (normal) healthy adult, angiogenesis is virtually arrested and occurs only when needed. However, a number of pathological situations are characterised by enhanced, uncontrolled angiogenesis. These conditions include cancer, rheumatoid arthritis, diabetic retinopathy, psoriasis and cardiovascular diseases such as atherosclerosis. In other pathologies such as ischaemic limb disease or in coronary artery disease, growing new vessels through the promotion of an expanding vasculature would be of benefit. 
     A number of in vitro assays have been established which are thought to mimic angiogenesis and these have provided important tools to examine the mechanisms by which the angiogenic process takes place and the genes most likely to be involved. 
     Lumen formation is a key step in angiogenesis. The presence of vacuoles within ECs undergoing angiogenesis have been reported and their involvement in lumen formation has been postulated (Folkman and Haudenschild, 1980; Gamble et al., 1993). The general mechanism of lumen formation suggested by Folkman and Haudenschild (1980), has been that vacuoles form within the cytoplasm of a number of aligned ECs which are later converted to a tube. The union of adjacent tubes results in the formation of a continuous unicellular capillary lumen. However, little is known about the changes in cell morphology leading to lumen formation or the signals required for ECs to construct this feature. 
     An in vitro model of angiogenesis has been created from human umbilical vein ECs plated onto a 3 dimensional collagen matrix (Gamble et al., 1993). In the presence of phorbol myristate acetate (PMA) these cells form capillary tubes within 24 hours. With the addition of anti-integrin antibodies, the usually unicellular tubes (thought to reflect an immature, poorly differentiated phenotype) are converted to form a multicellular lumen through the inhibition of cell-matrix interactions and promotion of cell-cell interactions. This model has subsequently allowed the investigation of the morphological events which occur in lumen formation. 
     For the treatment of diseases associated with angiogenesis, understanding the molecular genetic mechanisms of the process is of paramount importance. The use of the in vitro model described above (Gamble et al., 1993), a model that reflects the critical events that occur during angiogenesis in vivo in a time dependant and broadly synchronous manner, has provided a tool for the identification of the key genes involved. 
     SUMMARY 
     This Summary lists several embodiments of the presently disclosed subject matter, and in many cases lists variations and permutations of these embodiments. This Summary is merely exemplary of the numerous and varied embodiments. Mention of one or more representative features of a given embodiment is likewise exemplary. Such an embodiment can typically exist with or without the feature(s) mentioned; likewise, those features can be applied to other embodiments of the presently disclosed subject matter, whether listed in this Summary or not. To avoid excessive repetition, this Summary does not list or suggest all possible combinations of such features. 
     In some embodiments, the presently disclosed subject matter provides a method of modulating angiogenesis comprising modulating the expression or activity of a BNO802 polypeptide in a cell, wherein the BNO802 polypeptide is encoded by a BNO802 nucleic acid molecule set forth in Table 1. In some embodiments, the expression or activity of the BNO802 polypeptide is modulated by introducing into the cell an antisense to the BNO802 nucleic acid molecule. In some embodiments, the expression or activity of the BNO802 polypeptide is modulated by introducing into the cell a nucleic acid which is an siRNA. In some embodiments, the expression or activity of the BNO802 polypeptide is modulated by an antibody capable of binding the BNO802 polypeptide. In some embodiments, the antibody is a fully human antibody. In some embodiments, the antibody is selected from the group consisting of a monoclonal antibody, a humanised antibody, a chimaeric antibody or an antibody fragment including a Fab fragment, (Fab′)2 fragment, Fv fragment, single chain antibodies and single domain antibodies. 
     In some embodiments, the presently disclosed subject matter provides a method for the treatment of an angiogenesis-related disorder, comprising modulating the expression or activity of a BNO802 polypeptide encoded by a BNO802 nucleic acid molecule set forth in Table 1. In some embodiments, the expression or activity of the BNO802 polypeptide is modulated by introducing into the cell an antisense to the BNO802 nucleic acid molecule. In some embodiments, the expression or activity of the BNO802 polypeptide is modulated by introducing into the cell a nucleic acid which is an siRNA. In some embodiments, the expression or activity of the BNO802 polypeptide is modulated by an antibody capable of binding the BNO802 polypeptide. In some embodiments, the antibody is a fully human antibody. In some embodiments, the antibody is selected from the group consisting of a monoclonal antibody, a humanised antibody, a chimaeric antibody or an antibody fragment including a Fab fragment, (Fab′)2 fragment, Fv fragment, single chain antibodies and single domain antibodies. In some embodiments, the disorder is selected from the group consisting of cancer, rheumatoid arthritis, diabetic retinopathy, psoriasis, and cardiovascular diseases such as atherosclerosis, ischaemic limb disease or coronary artery disease. 
     In some embodiments, the presently disclosed subject matter provides a method of screening for a candidate pharmaceutical compound for the treatment of an angiogenesis-related disorder, comprising the steps of: (1) providing a BNO802 polypeptide set forth in Table 1; (2) adding a candidate pharmaceutical compound to said BNO802 polypeptide; and (3) determining the binding of said candidate compound to said BNO802 polypeptide; wherein a compound that binds to the polypeptide is a candidate for the treatment of an angiogenesis-related disorder. 
     In some embodiments, the presently disclosed subject matter provides a method of screening for a candidate pharmaceutical compound useful in the treatment of an angiogenesis-related disorder, comprising the steps of: (1) providing a cell transformed with an expression vector comprising a BNO802 nucleic acid molecule set forth in Table 1; (2) adding a candidate pharmaceutical compound to said cell; and (3) determining the effect of said candidate pharmaceutical compound on the expression or activity of the polypeptide encoded by the BNO802 nucleic acid molecule that is part of the expression vector in said cell; wherein a compound that alters the expression or activity of the polypeptide encoded by the BNO802 nucleic acid molecule that is part of the expression vector in said cell is a candidate for the treatment of an angiogenesis-related disorder. 
     It is an object of the presently disclosed subject matter to provide methods of modulating angiogenesis comprising modulating the expression or activity of a BNO802 polypeptide in a cell. 
     An object of the presently disclosed subject matter having been stated above, other objects and advantages will become apparent to those of ordinary skill in the art after a study of the following description of the presently disclosed subject matter and non-limiting Examples. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIGS. 1A-1D . Example of the expression profile of selected differentially expressed clones during defined time points in the in vitro model of angiogenesis. Time points at the defined stages of 0.5 hours, 3 hours, 6 hours and 24 hours of the in vitro tube formation assay were plotted against the log ratio of cy5 (red) and cy3 (green) dyes used for microarray hybridizations.  FIG. 1A : example of a clone with peak expression at the 0.5 hour time point;  FIG. 1B : example of a clone with peak expression at the 3 hour time point;  FIG. 1C : example of a clone with peak expression at the 6 hour time point; and  FIG. 1D : example of a clone with peak expression at the 24 hour time point. 
         FIGS. 2A and 2B . Expression profile of differentially expressed genes BNO782 and BNO481. Both genes show peak expression at the 6 hour time point of the in vitro tube formation assay.  FIG. 2A : BNO782;  FIG. 2B : BNO481. 
         FIGS. 3A and 3B . Analysis of the level of BNO782 expression knock-down mediated by BNO782 siRNA2 and BNO481 expression knock-down mediated by BNO481 siRNA1, as measured by real-time RT-PCR. The three siRNA oligonucleotides targeted to each gene were able to reduce expression of the gene to varying degrees with BNO781 siRNA2 inhibiting BNO781 expression by 24% ( FIG. 3A ) and BNO481 siRNA1 inhibiting expression of BNO481 by 36% ( FIG. 3B ). 
         FIGS. 4A-4D . Reducing BNO782 or BNO481 mRNA expression inhibits HUVEC tube formation. HUVECs infected with BNO782 siRNA2, BNO481 siRNA1, or a vector control were plated on Matrigel for 24 hrs. Vector infected cells formed extensive networks of tube structures ( FIGS. 4A and 4C ). In contrast, cells infected with BNO782 siRNA2 or BNO481 siRNA1 exhibited tube structure networks of significantly reduced complexity with a high number of incomplete tube extensions ( FIGS. 4B and 4D ). 
         FIGS. 5A-5C . Evaluation of the consequences of siRNA-mediated knockdown of BNO802 on the formation of capillary tubes by endothelial cells on Matrigel. Pictures of endothelial capillary tubes were taken after 22 hours incubation using a Olympus BX51 microscope with 4× objective and CCD Optronics high resolution camera and Olympus CKX41 inverted microscope with DP11 digital camera. 
         FIGS. 6A and 6B . Evaluation of the consequences of siRNA-mediated knockdown of BNO802 on the ability of endothelial cells to proliferate. Endothelial cells are seeded at 1000 cells/well and cultured for 72 hours. Cell growth was evaluated using a colorimetric assay. Error bars are base on standard deviation derived from triplicate wells. 
         FIG. 7 . RealTime-RTPCR analysis evaluating the degree of BNO802 gene knockdown achieved with RNAi. Total RNA was extracted from cells and reverse transcribed into cDNA followed by RealTime PCR amplification using gene specific primers. Expression levels were normalised to the house-keeping gene POLR2K and expressed as a percent of the vector control (n=3). 
         FIG. 8 . Evaluation of BNO802 gene expression in normal human tissues using RealTime RTPCR analysis. Human RNA samples (Ambion) were reverse transcribed into cDNA followed by RealTime PCR using gene specific primers. Gene expression data was normalised to the expression of the house-keeping gene POLR2K. The level of gene expression in each tissue was expressed relative to the gene expression found in a homogeneous endothelial cell population (HUVEC) (n=4). 
     
    
    
     DETAILED DESCRIPTION 
     Total RNA from cells harvested at specific time points from a biological model, in this case the Gamble et al (1993) model for angiogenesis, were used to prepare cDNAs, which were subjected to a novel process incorporating suppression subtractive hybridization (SSH) to identify cDNAs derived from differentially expressed genes. 
     According to one aspect of the present invention there is provided a method for the identification of a gene differentially expressed in an in vitro model of a biological system, comprising the steps of: 
     (1) harvesting cells from the model system at predetermined time points; 
     (2) obtaining total RNA from the cells harvested at each time point; 
     (3) preparing cDNA from the total RNA from each time point to provide a plurality of pools of cDNA; 
     (4) performing a suppression subtractive hybridization (SSH) on the cDNA pools from each time point sequentially so as to progressively amplify cDNAs derived from genes differentially expressed from one time period to the next. 
     Thus, up-regulation of a gene whose expression subsequently remains up-regulated at the same level will be detected (and the cDNA amplified) only in the first time period where the level cDNA is elevated, as the quantity of cDNA in pools is from the subsequent time points will be the same. This reduction in redundancy reduces the possibility that other genes of lower representation in the cell mRNA expression pool will be masked. In a particularly preferred embodiment of the present invention the model system is an in vitro model for angiogenesis (Gamble et al., 1993). 
     Those cDNAs identified to be differentially expressed in the SSH process were cloned and subjected to microarray analysis, which lead to the identification of a number of genes that are up-regulated in their expression during the angiogenesis process. 
     According to a further aspect of the present invention there is provided a method for the identification of a gene up-regulated in an in vitro model of a biological system, comprising the steps of: 
     (1) harvesting cells from the model system at predetermined time points; 
     (2) obtaining total RNA from the cells harvested at each time point; 
     (3) preparing cDNA from the total RNA from each time point to provide a plurality of pools of cDNA; 
     (4) performing a suppression subtractive hybridization (SSH) on the cDNA pools from each time point sequentially so as to progressively amplify cDNAs derived from genes differentially expressed from one time period to the next. 
     (5) cloning the amplified cDNAs; 
     (6) locating DNA from each clone on a microarray; 
     (7) generating antisense RNA by reverse transcription of total RNA from cells harvested from the in vitro model at said predetermined time intervals and labelling the antisense RNA; and 
     (8) probing the microarray with labelled antisense RNA from 0 hours and each of the other time points separately to identify clones containing cDNA derived from genes which are up-regulated at said time points in the in vitro model. 
     Functional analysis of a subset of these up-regulated angiogenic genes and their effect on endothelial cell function and capillary tube formation is described in detail below. 
     Accordingly, the present invention provides isolated nucleic acid molecules, which have been shown to be up-regulated in their expression during angiogenesis (see Tables 1 and 2). The isolation of these angiogenic genes has provided novel targets for the treatment of angiogenesis-related disorders. 
     In a first aspect of the present invention there is provided an isolated nucleic acid molecule as defined by SEQ ID Numbers: 1 to 44. 
     Following the realisation that these molecules, and those listed in Tables 1 and 2, are up-regulated in their expression during angiogenesis, the invention provides isolated nucleic acid molecules as defined by SEQ ID Numbers: 1 to 44, and laid out in Tables 1 and 2, or fragments thereof, that play a role in an angiogenic process. Such a process may include, but is not restricted to, embryogenesis, menstrual cycle, wound repair, tumour angiogenesis and exercise induced muscle hypertrophy. 
     In addition, the present invention provides isolated nucleic acid molecules as defined by SEQ ID Numbers: 1 to 44, and laid out in Tables 1 and 2 (hereinafter referred to as “angiogenic genes”, “angiogenic nucleic acid molecules” or “angiogenic polypeptides” for the sake of convenience), or fragments thereof, that play a role in diseases associated with the angiogenic process. Diseases may include, but are not restricted to, cancer, rheumatoid arthritis, diabetic retinopathy, psoriasis, and cardiovascular diseases such as atherosclerosis, ischaemic limb disease and coronary artery disease. Useful fragments may include those which are unique and which do not overlap any previously identified genes, unique fragments which do overlap with a known sequence, and fragments which span alternative splice junctions etc. 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Novel Angiogenesis Genes 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 Peak 
               
               
                 BNO 
                   
                   
                 UniGene 
                   
                 Expression 
               
               
                 Number 
                 Symbol 
                 Gene Description - Homology 
                 Number 
                 GenBank Number 
                 (h) 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 BNO605 
                 BNO605 
                 EST, UI-HF-BR0p-ajy-c-08-0-UI.s1  Homo sapiens  cDNA 
                 None 
                 AW576601 
                   
               
               
                 BNO612 
                 FLJ20445 
                 hypothetical protein FLJ20445 
                 Hs.343748 
                 NM_017824 
                 6 
               
               
                 BNO616 
                 MGC2747 
                 hypothetical protein MGC2747 
                 Hs.194017 
                 NM_024104 
                 0.5, 6   
               
               
                 BNO617 
                 FLJ20986 
                 hypothetical protein FLJ20986 
                 Hs.324507 
                 NM_024524 
                 6 
               
               
                 BNO618 
                 FLJ14834 
                 hypothetical protein FLJ14834 
                 Hs.62905 
                 NM_032849 
                 3 
               
               
                 BNO620 
                 FLJ22746 
                 hypothetical protein FLJ22746 
                 Hs.147585 
                 NM_024785 
                 0.5 
               
               
                 BNO622 
                 KIAA1376 
                 KIAA1376 protein 
                 Hs.24684 
                 BC015928 
                 3, 24 
               
               
                 BNO627 
                 BNO627 
                 EST, AV756199 BM  Homo sapiens  cDNA clone 
                 None 
                 SEQ ID NO: 1 
                 6 
               
               
                   
                   
                 BMFAUH02 5′ 
               
               
                 BNO628 
                 BNO628 
                 EST, QV1-BT0631-130300-111-e03 BT0631  Homo   
                 None 
                 SEQ ID NO: 2 
                 6 
               
               
                   
                   
                   sapiens  cDNA 
               
               
                 BNO629 
                 BNO629 
                 EST,  Homo sapiens  cDNA clone IMAGE: 2664022 3′ 
                 None 
                 SEQ ID NO: 3 
                 6 
               
               
                 BNO630 
                 BNO630 
                 EST,  Homo sapiens  cDNA clone IMAGE: 2357465 3′ 
                 None 
                 SEQ ID NO: 4, 51 
                 6 
               
               
                 BNO632 
                 BNO632 
                 ESTs 
                 Hs.404198 
                 SEQ ID NO: 5 
                 6 
               
               
                 BNO633 
                 BNO633 
                 ESTs, Weakly similar to hypothetical protein FLJ20378 
                 Hs.310598 
                 SEQ ID NO: 6 
                 24 
               
               
                 BNO634 
                 BNO634 
                 ESTs 
                 Hs.345443 
                 SEQ ID NO: 7 
                 6 
               
               
                 BNO635 
                 BNO635 
                 Hypothetical protein 
                 Hs.54347 
                 BC057847 
                 6 
               
               
                 BNO636 
                 BNO636 
                 ESTs 
                 Hs.105636 
                 SEQ ID NO: 8 
                 3 
               
               
                 BNO637 
                 BNO637 
                 ESTs 
                 Hs.486928 
                 SEQ ID NO: 9, 52 
                 6 
               
               
                 BNO638 
                 BNO638 
                 EST 
                 None 
                 SEQ ID NO: 10 
                 6 
               
               
                 BNO639 
                 BNO639 
                 None 
                 None 
                 SEQ ID NO: 11, 53 
                 6 
               
               
                 BNO640 
                 BNO640 
                 None 
                 None 
                 SEQ ID NO: 12 
                 6 
               
               
                 BNO645 
                 FLJ10498 
                 hypothetical protein FLJ10498 
                 Hs.270107 
                 NM_018115 
                 24 
               
               
                 BNO648 
                 LOC57146 
                 hypothetical protein from clone 24796 
                 Hs.27191 
                 NM_020422 
                 0.5 
               
               
                 BNO652 
                 FLJ31051 
                 hypothetical protein FLJ31051 
                 Hs.406199 
                 NM_153687 
                 6 
               
               
                 BNO655 
                 LOC51122 
                 HSPC042 protei 
                 Hs.432729 
                 NM_016094 
                 3 
               
               
                 BNO659 
                 FLJ32123 
                 FLJ32123 
                 Hs.349397 
                 AK056685 
                 6 
               
               
                 BNO662 
                 BNO662 
                 ESTs 
                 Hs.444495 
                 BX647355 
                 6 
               
               
                 BNO664 
                 FLJ10312 
                 FLJ10312 
                 None 
                 NM_030672 
                 3 
               
               
                 BNO669 
                 BNO669 
                 ESTs 
                 Hs.172998 
                 BC030094 
                 3 
               
               
                 BNO671 
                 KIAA0882 
                 KIAA0882 protein 
                 Hs.411317 
                 AB020689 
                 3 
               
               
                 BNO673 
                 BNO673 
                 hypothetical protein DKFZp434L142 
                 Hs.323583 
                 NM_016613 
                 6 
               
               
                 BNO675 
                 FLJ10700 
                 hypothetical protein FLJ10700 
                 Hs.295909 
                 NM_018182 
                 3 
               
               
                 BNO677 
                 FLJ30135 
                 FLJ30135 
                 Hs.34906 
                 BC020494 
                 3, 24 
               
               
                 BNO685 
                 FLJ10849 
                 hypothetical protein FLJ10849 
                 Hs.386784 
                 NM_018243 
               
               
                 BNO687 
                 MGC45416 
                 hypothetical protein MGC45416 
                 Hs.95835 
                 NM_152398 
                 24 
               
               
                 BNO690 
                 C15orf15 
                 chromosome 15 open reading frame 15 
                 Hs.274772 
                 NM_016304 
                 3 
               
               
                 BNO694 
                 BNO694 
                 cDNA DKFZp566E0124 
                 None 
                 AL050030 
                 6 
               
               
                 BNO697 
                 BNO697 
                 Hypothetical protein MGC45871 
                 Hs.345588 
                 BC014203 
                 24 
               
               
                 BNO700 
                 C7orf30 
                 chromosome 7 open reading frame 30 
                 Hs.87385 
                 NM_138446 
                 24 
               
               
                 BNO704 
                 KIAA1102 
                 KIAA1102 protein 
                 Hs.156761 
                 AB029025 
               
               
                 BNO705 
                 BNO705 
                 ESTs 
                 Hs.30280 
                 SEQ ID NO: 13 
                 3 
               
               
                 BNO706 
                 LOC116441 
                 hypothetical protein BC014339 
                 Hs.22026 
                 NM_138786 
                 24 
               
               
                 BNO708 
                 BNO708 
                 ESTs 
                 Hs.12876 
                 SEQ ID NO: 14 
                 6 
               
               
                 BNO710 
                 BNO710 
                 FLJ23228 
                 Hs.170623 
                 AK026881 
                 6 
               
               
                 BNO712 
                 BNO712 
                 FLJ21592 
                 Hs.5921 
                 AK025245 
                 3 
               
               
                 BNO713 
                 KIAA0970 
                 KIAA0970 protein 
                 Hs.103329 
                 NM_014923 
                 6 
               
               
                 BNO714 
                 KIAA0121 
                 KIAA0121 gene product 
                 Hs.155584 
                 D50911 
                 6 
               
               
                 BNO723 
                 C14orf123 
                 chromosome 14 open reading frame 123 
                 Hs.279761 
                 NM_014169 
                 6 
               
               
                 BNO725 
                 KIAA0582 
                 KIAA0582 protein 
                 Hs.146007 
                 NM_015147 
                 24 
               
               
                 BNO730 
                 BNO730 
                 ESTs 
                 Hs.158753 
                 SEQ ID NO: 15 
                 6 
               
               
                 BNO731 
                 C6orf166 
                 chromosome 6 open reading frame 166 
                 Hs.201864 
                 NM_018064 
                 3 
               
               
                 BNO735 
                 FLJ32029 
                 Unnamed protein product 
                 Hs.26612 
                 NM_173582 
                 6 
               
               
                 BNO737 
                 BNO737 
                 hypothetical protein DKFZp434F0318 
                 Hs.23388 
                 NM_030817 
               
               
                 BNO740 
                 KIAA1728 
                 KIAA1728 protein 
                 Hs.437362 
                 AB051515 
                 24 
               
               
                 BNO742 
                 BNO742 
                 hypothetical protein FLJ11795 
                 Hs.84560 
                 NM_024669 
                 24 
               
               
                 BNO745 
                 BNO745 
                 hypothetical protein DKFZp547A023 
                 Hs.374649 
                 NM_018704 
                 6 
               
               
                 BNO747 
                 MGC23937 
                 hypothetical protein MGC23937 similar to CG4798 
                 Hs.91612 
                 NM_145052 
                 6 
               
               
                 BNO753 
                 BNO753 
                 cDNA DKFZp667P1024 
                 Hs.127811 
                 AL832835 
                 3 
               
               
                 BNO754 
                 KIAA0303 
                 KIAA0303 protein 
                 Hs.212787 
                 AB002301 
                 3 
               
               
                 BNO756 
                 BNO756 
                 ESTs 
                 Hs.443155 
                 SEQ ID NO: 16, 54 
               
               
                 BNO759 
                 KIAA1416 
                 KIAA1416 protein 
                 Hs.397426 
                 AB037837 
                 6 
               
               
                 BNO761 
                 C7orf24 
                 chromosome 7 open reading frame 24 
                 Hs.444840 
                 NM_024051 
                 6 
               
               
                 BNO762 
                 FLJ11223 
                 cDNA FLJ11223 
                 Hs.92308 
                 AL832083 
                 3 
               
               
                 BNO768 
                 FLJ30478 
                 cDNA FLJ30478 
                 Hs.298258 
                 AK092048 
                 6 
               
               
                 BNO772 
                 FLJ10525 
                 Hypothetical protein FLJ10525 
                 Hs.31082 
                 NM_018126 
                 6 
               
               
                 BNO780 
                 LOC58489 
                 hypothetical protein from EUROIMAGE 588495 
                 Hs.26765 
                 AL390079 
                 3 
               
               
                 BNO782 
                 MGC26717 
                 Hypothetical protein 
                 Hs.406060 
                 BC024188 
                 6 
               
               
                 BNO791 
                 KIAA1053 
                 KIAA1053 protein 
                 Hs.98259 
                 NM_015589 
                 6 
               
               
                 BNO793 
                 KIAA0766 
                 KIAA0766 gene product 
                 Hs.28020 
                 NM_014805 
                 24 
               
               
                 BNO795 
                 BNO795 
                 ESTs moderately similar to MDC-3.13 isoform 2 mRNA 
                 Hs.306343 
                 AK123281 
                 6 
               
               
                 BNO800 
                 KIAA1577 
                 KIAA1577 protein 
                 Hs.449290 
                 AB046797 
                 6 
               
               
                 BNO802 
                 KIAA0877 
                 KIAA0877 protein 
                 Hs.408623 
                 SEQ ID NO: 59, 60 
                 24 
               
               
                 BNO812 
                 KIAA0372 
                 KIAA0372 gene product 
                 Hs.435330 
                 NM_014639 
                 6 
               
               
                 BNO816 
                 BNO816 
                 cDNA clone 4052238 
                 Hs.348514 
                 BC014384 
                 6 
               
               
                 BNO818 
                 MGC10067 
                 hypothetical protein MGC10067 
                 Hs.42251 
                 NM_145049 
                 3 
               
               
                 BNO819 
                 KIAA1191 
                 KIAA1191 protein 
                 Hs.8594 
                 NM_020444 
                 24 
               
               
                 BNO821 
                 BNO821 
                 ESTs 
                 Hs.87606 
                 SEQ ID NO: 17 
                 24 
               
               
                 BNO825 
                 FBXO30 
                 F-box protein 30 
                 Hs.421095 
                 NM_032145 
                 3 
               
               
                 BNO831 
                 C8orf1 
                 chromosome 8 open reading frame 1 
                 Hs.436445 
                 NM_004337 
                 24 
               
               
                 BNO833 
                 C6orf115 
                 Chromosome 6 open reading frame 115 
                 Hs.238205 
                 BC014953 
                 24 
               
               
                 BNO838 
                 BNO838 
                 ESTs 
                 Hs.319095 
                 SEQ ID NO: 18 
                 3 
               
               
                 BNO845 
                 FLJ23728 
                 cDNA FLJ23728 
                 Hs.191094 
                 AK074308 
                 6 
               
               
                 BNO848 
                 C10orf45 
                 Chromosome 10 open reading frame 45 
                 Hs.103378 
                 NM_031453 
                 24 
               
               
                 BNO849 
                 BNO849 
                 cDNA DKFZp434G0972 
                 Hs.106148 
                 AL133577 
                 24 
               
               
                 BNO852 
                 CGI-111 
                 CGI-111 protein 
                 Hs.11085 
                 NM_016048 
                 6 
               
               
                 BNO856 
                 LOC116068 
                 hypothetical protein LOC116068 
                 Hs.136235 
                 AL832721 
                 24 
               
               
                 BNO857 
                 C12orf2 
                 chromosome 12 open reading frame 2 
                 Hs.140821 
                 NM_007211 
                 6 
               
               
                 BNO862 
                 BNO862 
                 DKFZP434C212 protein 
                 Hs.287266 
                 AK023841 
               
               
                 BNO868 
                 BNO868 
                 DKFZP566C134 protein 
                 Hs.20237 
                 AB040922 
                 3 
               
               
                 BNO870 
                 LOC57228 
                 hypothetical protein from clone 643 
                 Hs.206501 
                 NM_020467 
                 24 
               
               
                 BNO871 
                 KIAA1463 
                 KIAA1463 protein 
                 Hs.21104 
                 AB040896 
                 6 
               
               
                 BNO873 
                 KIAA1376 
                 KIAA1376 protein 
                 Hs.24684 
                 NM_020801 
                 0.5, 24   
               
               
                 BNO876 
                 FLJ10326 
                 hypothetical protein FLJ10326 
                 Hs.262823 
                 NM_018060 
                 24 
               
               
                 BNO878 
                 BNO878 
                 hypothetical protein DKFZp761L1417 
                 Hs.270753 
                 NM_152913 
                 6 
               
               
                 BNO881 
                 MGC11349 
                 hypothetical protein MGC11349 
                 Hs.288697 
                 NM_025112 
                 6 
               
               
                 BNO883 
                 FLJ39541 
                 similar to RIKEN cDNA 9130404H11 gene 
                 Hs.21388 
                 NM_178566 
                 6 
               
               
                 BNO886 
                 BNO886 
                 cDNA DKFZp686D04119 
                 Hs.30258 
                 BX537597 
                 6 
               
               
                 BNO887 
                 KIAA0648 
                 KIAA0648 protein 
                 Hs.31921 
                 NM_015200 
                 24 
               
               
                 BNO890 
                 KIAA1160 
                 KIAA1160 protein 
                 Hs.512661 
                 NM_020701 
                 3 
               
               
                 BNO892 
                 C20orf108 
                 chromosome 20 open reading frame 108 
                 Hs.143736 
                 NM_080821 
                 3 
               
               
                 BNO894 
                 KIAA0205 
                 KIAA0205 gene product 
                 Hs.528724 
                 NM_014873 
                 6 
               
               
                 BNO895 
                 C20orf112 
                 chromosome 20 open reading frame 112 
                 Hs.335142 
                 NM_080616 
                 0.5 
               
               
                 BNO898 
                 BNO898 
                 clone IMAGE: 5243590 
                 Hs.454832 
                 BC036880 
                 6 
               
               
                 BNO905 
                 KIAA1462 
                 KIAA1462 protein 
                 Hs.192726 
                 AB040895 
                 3 
               
               
                 BNO906 
                 KIAA1199 
                 KIAA1199 protein 
                 Hs.212584 
                 AB033025 
                 6 
               
               
                 BNO908 
                 C15orf12 
                 chromosome 15 open reading frame 12 
                 Hs.513041 
                 NM_018285 
               
               
                 BNO910 
                 BNO910 
                 cDNA DKFZp564F053 
                 Hs.529772 
                 AL049265 
                 6 
               
               
                 BNO917 
                 BNO917 
                 hypothetical protein dJ465N24.2.1 
                 Hs.259412 
                 NM_020317 
                 24 
               
               
                 BNO926 
                 KIAA1238 
                 KIAA1238 protein 
                 Hs.372288 
                 AB033064 
               
               
                 BNO928 
                 BNO928 
                 EST 
                 None 
                 SEQ ID NO: 19 
                 3 
               
               
                 BNO929 
                 BNO929 
                 EST 
                 None 
                 SEQ ID NO: 20 
                 6 
               
               
                 BNO930 
                 BNO930 
                 EST 
                 Hs.478376 
                 SEQ ID NO: 21 
                 6 
               
               
                 BNO932 
                 BNO932 
                 EST 
                 Hs.492501 
                 SEQ ID NO: 22, 55 
                 3 
               
               
                 BNO933 
                 BNO933 
                 EST 
                 None 
                 SEQ ID NO: 23 
                 6 
               
               
                 BNO934 
                 BNO934 
                 EST 
                 None 
                 SEQ ID NO: 24 
                 6 
               
               
                 BNO935 
                 BNO935 
                 EST 
                 None 
                 SEQ ID NO: 25 
                 6 
               
               
                 BNO936 
                 BNO936 
                 EST 
                 None 
                 SEQ ID NO: 26, 56 
                 6 
               
               
                 BNO937 
                 BNO937 
                 alpha gene sequence 
                 None 
                 AF203815 
                 6 
               
               
                 BNO938 
                 BNO938 
                 EST 
                 None 
                 SEQ ID NO: 27 
                 0.5 
               
               
                 BNO939 
                 BNO939 
                 EST 
                 None 
                 SEQ ID NO: 28 
                 6 
               
               
                 BNO940 
                 BNO940 
                 EST 
                 None 
                 SEQ ID NO: 29 
                 6 
               
               
                 BNO941 
                 BNO941 
                 EST 
                 None 
                 SEQ ID NO: 30 
                 3 
               
               
                 BNO942 
                 BNO942 
                 EST 
                 None 
                 SEQ ID NO: 31 
                 6 
               
               
                 BNO943 
                 BNO943 
                 EST 
                 None 
                 SEQ ID NO: 32 
                 6 
               
               
                 BNO944 
                 BNO944 
                 EST 
                 None 
                 SEQ ID NO: 33 
                 6 
               
               
                 BNO945 
                 BNO945 
                 EST 
                 None 
                 SEQ ID NO: 34 
                 6 
               
               
                 BNO946 
                 BNO946 
                 EST 
                 None 
                 SEQ ID NO: 35, 57 
                 6 
               
               
                 BNO948 
                 BNO948 
                 EST 
                 None 
                 SEQ ID NO: 36 
                 6 
               
               
                 BNO949 
                 BNO949 
                 EST 
                 None 
                 SEQ ID NO: 37, 58 
                 3 
               
               
                 BNO950 
                 BNO950 
                 EST 
                 None 
                 SEQ ID NO: 38 
                 24 
               
               
                 BNO951 
                 BNO951 
                 EST 
                 None 
                 SEQ ID NO: 39 
                 24 
               
               
                 BNO953 
                 BNO953 
                 EST 
                 None 
                 SEQ ID NO: 40 
                 24 
               
               
                 BNO961 
                 BNO961 
                 FLJ00138 protein 
                 Hs.199749 
                 AK074067 
                 3, 24 
               
               
                 BNO1018 
                 BNO1018 
                 EST 
                 Hs.485935 
                 SEQ ID NO: 41 
                 3 
               
               
                 BNO1019 
                 BNO1019 
                 EST 
                 None 
                 SEQ ID NO: 42 
                 24 
               
               
                 BNO1020 
                 BNO1020 
                 EST 
                 None 
                 SEQ ID NO: 43 
                 3 
               
               
                 BNO1021 
                 BNO1021 
                 EST 
                 None 
                 SEQ ID NO: 44 
                 3 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 Genes with a Previously Unknown Role in Angiogenesis 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 Peak 
               
               
                 BNO 
                   
                   
                 UniGene 
                   
                 Expression 
               
               
                 Number 
                 Symbol 
                 Gene Description - Homology 
                 Number 
                 GenBank Number 
                 (h) 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 BNO436 
                 NP 
                 nucleoside phosphorylase 
                 Hs.75514 
                 NM_000270 
                 6 
               
               
                 BNO438 
                 CD59 
                 CD59 antigen p18-20 
                 Hs.278573 
                 NM_000611 
                 24 
               
               
                 BNO441 
                 BIRC3 
                 baculoviral IAP repeat-containing 3 
                 Hs.127799 
                 NM_001165 
                 3 
               
               
                 BNO442 
                 FABP5 
                 fatty acid binding protein 5 (psoriasis-associated) 
                 Hs.408061 
                 NM_001444 
                 24 
               
               
                 BNO443 
                 CBFB 
                 core-binding factor, beta subunit 
                 Hs.179881 
                 NM_001755 
                 6 
               
               
                 BNO446 
                 INHBA 
                 inhibin, beta A (activin A, activin AB alpha polypeptide) 
                 Hs.727 
                 NM_002192 
                 6 
               
               
                 BNO447 
                 MGST2 
                 microsomal glutathione S-transferase 2 
                 Hs.81874 
                 NM_002413 
                 24 
               
               
                 BNO448 
                 RAB6A 
                 RAB6A, member RAS oncogene family 
                 Hs.5636 
                 NM_002869 
                 6 
               
               
                 BNO449 
                 SAT 
                 spermidine/spermine N1-acetyltransferase 
                 Hs.28491 
                 NM_002970 
                 6 
               
               
                 BNO451 
                 TXNRD1 
                 thioredoxin reductase 1 
                 Hs.13046 
                 NM_003330 
                 6 
               
               
                 BNO452 
                 SLC4A7 
                 solute carrier family 4, sodium bicarbonate cotransporter, 
                 Hs.132904 
                 NM_003615 
                 6 
               
               
                   
                   
                 member 7 
               
               
                 BNO453 
                 PPAP2B 
                 phosphatidic acid phosphatase type 2B 
                 Hs.432840 
                 NM_003713 
                 3 
               
               
                 BNO454 
                 BCL10 
                 B-cell CLL/lymphoma 10 
                 Hs.193516 
                 NM_003921 
                 3 
               
               
                 BNO455 
                 DUSP1 
                 dual specificity phosphatase 1 
                 Hs.171695 
                 NM_004417 
                 0.5 
               
               
                 BNO456 
                 KIF5B 
                 kinesin family member 5B 
                 Hs.149436 
                 NM_004521 
                 6 
               
               
                 BNO457 
                 WTAP 
                 Wilms&#39; tumour 1-associating protein 
                 Hs.119 
                 NM_004906 
                 0.5 
               
               
                 BNO459 
                 FOS 
                 v-fos FBJ murine osteosarcoma viral oncogene homolog 
                 Hs.25647 
                 NM_005252 
                 0.5 
               
               
                 BNO460 
                 GATA6 
                 GATA binding protein 6 
                 Hs.50924 
                 NM_005257 
                 3 
               
               
                 BNO461 
                 HRY 
                 hairy and enhancer of split 1, ( Drosophila ) 
                 Hs.250666 
                 NM_005524 
                 0.5 
               
               
                 BNO462 
                 SGK 
                 serum/glucocorticoid regulated kinase 
                 Hs.296323 
                 NM_005627 
                 3 
               
               
                 BNO463 
                 TIEG 
                 TGFB inducible early growth response 
                 Hs.82173 
                 NM_005655 
                 0.5 
               
               
                 BNO464 
                 BCAP31 
                 B-cell receptor-associated protein 31 
                 Hs.381232 
                 NM_005745 
               
               
                 BNO465 
                 CALCRL 
                 calcitonin receptor-like 
                 Hs.152175 
                 NM_005795 
                 24 
               
               
                 BNO466 
                 SUI1 
                 putative translation initiation factor 
                 Hs.150580 
                 NM_005801 
                 3 
               
               
                 BNO467 
                 TSC22 
                 transforming growth factor beta-stimulated protein TSC-22 
                 Hs.114360 
                 NM_006022 
                 6 
               
               
                 BNO468 
                 RAN 
                 RAN, member RAS oncogene family 
                 Hs.426035 
                 NM_006325 
               
               
                 BNO469 
                 LYPLA1 
                 lysophospholipase I 
                 Hs.12540 
                 NM_006330 
                 6 
               
               
                 BNO470 
                 SSFA2 
                 sperm specific antigen 2 
                 Hs.351355 
                 NM_006751 
                 6 
               
               
                 BNO472 
                 CLIC4 
                 chloride intracellular channel 4 
                 Hs.25035 
                 NM_013943 
                 24 
               
               
                 BNO473 
                 SLC7A11 
                 solute carrier family 7, member 11 
                 Hs.6682 
                 NM_014331 
                 3 
               
               
                 BNO474 
                 RAI14 
                 retinoic acid induced 14 
                 Hs.15165 
                 NM_015577 
                 6 
               
               
                 BNO475 
                 HSPC014 
                 chromosome 13 open reading frame 12 
                 Hs.279813 
                 NM_015932 
                 24 
               
               
                 BNO476 
                 UMP- 
                 UMP-CMP kinase 
                 Hs.11463 
                 NM_016308 
                 3 
               
               
                   
                 CMPK 
               
               
                 BNO477 
                 SLC38A2 
                 solute carrier family 38, member 2 
                 Hs.298275 
                 NM_018976 
                 3 
               
               
                 BNO478 
                 ZNF317 
                 zinc finger protein 317 
                 Hs.18587 
                 NM_020933 
                 24 
               
               
                 BNO479 
                 RAB6C 
                 RAB6C, member RAS oncogene family 
                 Hs.333139 
                 NM_032144 
                 24 
               
               
                 BNO480 
                 MKI67IP 
                 MKI67 (FHA domain) interacting nucleolar phosphoprotein 
                 Hs.142838 
                 NM_032390 
                 3 
               
               
                 BNO481 
                 KPNA4 
                 karyopherin alpha 4 (importin alpha 3) 
                 Hs.288193 
                 NM_002268 
                 3 
               
               
                 BNO483 
                 C14orf32 
                 chromosome 14 open reading frame 32 
                 Hs.406401 
                 NM_144578 
                 3 
               
               
                 BNO484 
                 SMARCA2 
                 SWI/SNF related, matrix associated, regulator of 
                 Hs.198296 
                 NM_003070 
                 0.5 
               
               
                   
                   
                 chromatin, A2 
               
               
                 BNO485 
                 SOX4 
                   Homo sapiens  SRY (sex determining region Y)-box 4 
                 Hs.83484 
                 NM_003107 
                 3 
               
               
                   
                   
                 (SOX4), mRNA 
               
               
                 BNO487 
                 NR4A3 
                 nuclear receptor subfamily 4, group A, member 3 
                 Hs.80561 
                 NM_006981 
                 0.5 
               
               
                 BNO488 
                 NTN4 
                 netrin 4 
                 Hs.102541 
                 NM_021229 
               
               
                 BNO489 
                 DNCI2 
                 dynein, cytoplasmic, intermediate polypeptide 2 (DNCI2), 
                 Hs.66881 
                 XM_027780 
                 0.5 
               
               
                   
                   
                 mRNA 
               
               
                 BNO490 
                 UGCG 
                 UDP-glucose ceramide glucosyltransferase 
                 Hs.432605 
                 NM_003358 
                 0.5, 24 
               
               
                 BNO491 
                 P125 
                 Sec23-interacting protein p125 
                 Hs.300208 
                 NM_007190 
                 3 
               
               
                 BNO492 
                 NUDT4 
                 nudix (nucleoside diphosphate linked moiety X)-type motif 4 
                 Hs.355399 
                 NM_019094 
                 6 
               
               
                 BNO495 
                 SATB1 
                 special AT-rich sequence binding protein 1 
                 Hs.74592 
                 NM_002971 
                 6 
               
               
                 BNO496 
                 BZW1 
                 basic leucine zipper and W2 domains 1 
                 Hs.155291 
                 NM_014670 
                 3 
               
               
                 BNO497 
                 TDG 
                 thymine-DNA glycosylase 
                 Hs.173824 
                 NM_003211 
                 6 
               
               
                 BNO498 
                 ACTR3 
                 ARP3 actin-related protein 3 homolog (yeast) 
                 Hs.380096 
                 NM_005721 
                 24 
               
               
                 BNO499 
                 LAMP2 
                 lysosomal-associated membrane protein 2 
                 Hs.8262 
                 NM_013995 
                 6 
               
               
                 BNO500 
                 ERBB2IP 
                 erbb2 interacting protein 
                 Hs.8117 
                 NM_018695 
                 6 
               
               
                 BNO501 
                 DNAJB6 
                 DnaJ (Hsp40) homolog, subfamily B, member 6 
                 Hs.181195 
                 NM_005494 
                 3 
               
               
                 BNO502 
                 EMP1 
                 epithelial membrane protein 1 
                 Hs.79368 
                 NM_001423 
                 6 
               
               
                 BNO503 
                 MAPK1 
                 mitogen-activated protein kinase 1 
                 Hs.324473 
                 NM_002745 
                 24 
               
               
                 BNO504 
                 CYP1A1 
                 cytochrome P450, subfamily 1, polypeptide 1 
                 Hs.72912 
                 NM_000499 
                 6 
               
               
                 BNO505 
                 ACVR1 
                 activin A receptor, type I 
                 Hs.150402 
                 NM_001105 
                 3 
               
               
                 BNO506 
                 TPT1 
                 tumor protein, translationally-controlled 1 
                 Hs.401448 
                 NM_003295 
                 0.5, 24 
               
               
                 BNO507 
                 VAV3 
                 vav 3 oncogene 
                 Hs.267659 
                 NM_006113 
                 3 
               
               
                 BNO508 
                 CAP 
                 adenylyl cyclase-associated protein 
                 Hs.104125 
                 NM_006367 
                 24 
               
               
                 BNO509 
                 HSPA5 
                 Heat shock 70 kDa protein 5 (glucose-regulated protein, 
                 Hs.75410 
                 NM_005347 
                 6 
               
               
                   
                   
                 78 kDa) 
               
               
                 BNO510 
                 TIA1 
                 TIA1 cytotoxic granule-associated RNA binding protein 
                 Hs.239489 
                 NM_022173 
                 6 
               
               
                 BNO511 
                 CCNT2 
                 cyclin T2 
                 Hs.155478 
                 NM_001241 
                 6 
               
               
                 BNO512 
                 CHC1L 
                 chromosome condensation 1-like 
                 Hs.27007 
                 NM_001268 
                 0.5 
               
               
                 BNO513 
                 SFPQ 
                 splicing factor proline/glutamine rich 
                 Hs.180610 
                 NM_005066 
                 3 
               
               
                 BNO514 
                 PRKAR1A 
                 protein kinase, cAMP-dependent, regulatory, type I, alpha 
                 Hs.183037 
                 NM_002734 
                 24 
               
               
                 BNO515 
                 RALA 
                 v-ral simian leukemia viral oncogene homolog A (ras 
                 Hs.6906 
                 NM_005402 
                 6 
               
               
                   
                   
                 related) 
               
               
                 BNO516 
                 ANXA2 
                 annexin A2 
                 Hs.217493 
                 NM_004039 
                 0.5 
               
               
                 BNO517 
                 NUP153 
                 nucleoporin 153 kDa 
                 Hs.211608 
                 NM_005124 
                 3 
               
               
                 BNO518 
                 RANBP9 
                 RAN binding protein 9 
                 Hs.279886 
                 NM_005493 
                 24 
               
               
                 BNO519 
                 PRPF4B 
                 PRP4 pre-mRNA processing factor 4 homolog B (yeast) 
                 Hs.198891 
                 NM_003913 
                 6 
               
               
                 BNO520 
                 TSN 
                 translin 
                 Hs.75066 
                 NM_004622 
                 6 
               
               
                 BNO521 
                 H3F3A 
                 H3 histone, family 3A 
                 Hs.181307 
                 NM_002107 
                 24 
               
               
                 BNO523 
                 PROS1 
                 protein S (alpha) 
                 Hs.64016 
                 NM_000313 
                 6 
               
               
                 BNO524 
                 DDX3 
                 DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide 3 
                 Hs.380774 
                 NM_001356 
                 3 
               
               
                 BNO525 
                 TCF4 
                 transcription factor 4 
                 Hs.359289 
                 NM_003199 
                 6 
               
               
                 BNO526 
                 PTP4A1 
                 Protein tyrosine phosphatase type IVA, member 1 
                 Hs.227777 
                 NM_003463 
                 6 
               
               
                 BNO527 
                 BMPR2 
                 bone morphogenetic protein receptor, type II 
                 Hs.53250 
                 NM_001204 
                 3 
               
               
                   
                   
                 (serine/threonine kinase) 
               
               
                 BNO528 
                 NFE2L2 
                 nuclear factor (erythroid-derived 2)-like 2 
                 Hs.155396 
                 NM_006164 
                 3 
               
               
                 BNO531 
                 AHR 
                 aryl hydrocarbon receptor 
                 Hs.170087 
                 NM_001621 
                 3 
               
               
                 BNO532 
                 RANBP7 
                 RAN binding protein 7 
                 Hs.5151 
                 NM_006391 
                 3 
               
               
                 BNO533 
                 ARF6 
                 ADP-ribosylation factor 6 
                 Hs.89474 
                 NM_001663 
                 3 
               
               
                 BNO534 
                 SCARF1 
                 SCARF1 Scavenger receptor class F, member 1 
                 Hs.57735 
                 NM_003693E 
                 24 
               
               
                 BNO535 
                 PLU-1 
                 putative DNA/chromatin binding motif 
                 Hs.143323 
                 NM_006618 
                 24 
               
               
                 BNO536 
                 TOMM20 
                 translocase of outer mitochondrial membrane 20 (yeast) 
                 Hs.75187 
                 NM_014765 
                 6 
               
               
                   
                   
                 homolog 
               
               
                 BNO537 
                 B2M 
                 beta-2-microglobulin 
                 Hs.48516 
                 NM_004048 
                 24 
               
               
                 BNO538 
                 zizimin1 
                 zizimin1 
                 Hs.8021 
                 NM_015296 
                 6 
               
               
                 BNO539 
                 ARPP-19 
                 cyclic AMP phosphoprotein, 19 kD 
                 Hs.7351 
                 NM_006628 
                 3 
               
               
                 BNO540 
                 RAP1B 
                 RAP1B, member of RAS oncogene family 
                 Hs.156764 
                 NM_015646 
                 3 
               
               
                 BNO541 
                 MCP 
                 membrane cofactor protein 
                 Hs.83532 
                 NM_153826 
                 6 
               
               
                 BNO542 
                 IFI16 
                 interferon, gamma-inducible protein 16 
                 Hs.155530 
                 NM_005531 
                 0.5 
               
               
                 BNO543 
                 PRG1 
                 proteoglycan 1, secretory granule 
                 Hs.1908 
                 NM_002727 
               
               
                 BNO544 
                 KIT 
                 v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene 
                 Hs.81665 
                 NM_000222 
                 0.5, 24 
               
               
                   
                   
                 homolog 
               
               
                 BNO545 
                 SYBL1 
                 synaptobrevin-like 1 
                 Hs.24167 
                 NM_005638 
                 6 
               
               
                 BNO546 
                 TCF8 
                 transcription factor 8 (represses interleukin 2 expression) 
                 Hs.232068 
                 NM_030751E 
                 6 
               
               
                 BNO548 
                 NXF1 
                 nuclear RNA export factor 1 
                 Hs.323502 
                 NM_006362 
                   3, 24 
               
               
                 BNO549 
                 RAP2B 
                 RAP2B, member of RAS oncogene family 
                 Hs.239527 
                 NM_002886 
                 3 
               
               
                 BNO551 
                 IL6ST 
                 interleukin 6 signal transducer (gp130, oncostatin M 
                 Hs.82065 
                 NM_002184 
                 6 
               
               
                   
                   
                 receptor) 
               
               
                 BNO552 
                 REST 
                 RE1-silencing transcription factor 
                 Hs.401145 
                 NM_005612 
                 6 
               
               
                 BNO553 
                 SLC19A2 
                 solute carrier family 19 (thiamine transporter), member 2 
                 Hs.30246 
                 NM_006996 
                 3 
               
               
                 BNO554 
                 EIF4G2 
                 eukaryotic translation initiation factor 4 gamma, 2 
                 Hs.183684 
                 NM_001418 
                 3 
               
               
                 BNO555 
                 PTPRE 
                 protein tyrosine phosphatase, receptor type, E 
                 Hs.31137 
                 NM_006504 
                 3 
               
               
                 BNO556 
                 PDE3A 
                 phosphodiesterase 3A, cGMP-inhibited 
                 Hs.777 
                 NM_000921 
                 3 
               
               
                 BNO557 
                 C1QR1 
                 complement component 1, q subcomponent, receptor 1 
                 Hs.97199 
                 NM_012072 
                 24 
               
               
                 BNO558 
                 RANBP2 
                 RAN binding protein 2 
                 Hs.199179 
                 NM_006267 
               
               
                 BNO559 
                 KIS 
                 kinase interacting with leukemia-associated gene (stathmin) 
                 Hs.127310 
                 NM_144624 
                 24 
               
               
                 BNO560 
                 HMGCR 
                 3-hydroxy-3-methylglutaryl-Coenzyme A reductase 
                 Hs.11899 
                 NM_000859 
                 6 
               
               
                 BNO561 
                 PDCD4 
                 programmed cell death 4 (neoplastic transformation 
                 Hs.326248 
                 NM_145341 
                 3 
               
               
                   
                   
                 inhibitor) 
               
               
                 BNO562 
                 TACC1 
                 transforming, acidic coiled-coil containing protein 1 
                 Hs.173159 
                 NM_006283 
                 0.5 
               
               
                 BNO564 
                 DIS3 
                 mitotic control protein dis3 homolog 
                 Hs.323346 
                 NM_014953 
                 6 
               
               
                 BNO565 
                 TOP2A 
                 topoisomerase (DNA) II alpha 170 kDa 
                 Hs.156346 
                 NM_001067 
                 6 
               
               
                 BNO566 
                 SLC7A2 
                 solute carrier family 7, member 2 
                 Hs.153985 
                 NM_003046 
                 6 
               
               
                 BNO567 
                 FH 
                 fumarate hydratase 
                 Hs.75653 
                 NM_000143 
                 6 
               
               
                 BNO568 
                 IL1RL1 
                 interleukin 1 receptor-like 1 
                 Hs.66 
                 NM_003856 
                 6 
               
               
                 BNO569 
                 HPRP3P 
                 U4/U6-associated RNA splicing factor 
                 Hs.11776 
                 NM_004698 
                 6 
               
               
                 BNO570 
                 DDX5 
                 DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide 5 
                 Hs.76053 
                 NM_004396 
               
               
                 BNO571 
                 MAD2L1 
                 MAD2 mitotic arrest deficient-like 1 (yeast) 
                 Hs.79078 
                 NM_002358 
                 0.5, 24 
               
               
                 BNO572 
                 MADH7 
                 MAD, mothers against decapentaplegic homolog 7 
                 Hs.100602 
                 NM_005904 
                 3 
               
               
                   
                   
                 ( Drosophila ) 
               
               
                 BNO573 
                 E2F3 
                 E2F transcription factor 3 
                 Hs.1189 
                 NM_001949 
                 3 
               
               
                 BNO574 
                 CSNK2A2 
                 CSNK2A2 Casein kinase 2, alpha prime polypeptide 
                 Hs.82201 
                 NM_001896 
                 6 
               
               
                 BNO575 
                 MAX 
                 MAX protein 
                 Hs.42712 
                 NM_002382 
                 6 
               
               
                 BNO576 
                 ERAP140 
                 140 kDa estrogen receptor associated protein 
                 Hs.339283 
                 AF493978 
                 3 
               
               
                 BNO577 
                 CD9 
                 CD9 antigen (p24) 
                 Hs.1244 
                 NM_001769 
                 24 
               
               
                 BNO578 
                 ATRX 
                 alpha thalassemia/mental retardation syndrome X-linked 
                 Hs.96264 
                 NM_000489 
                 6 
               
               
                 BNO579 
                 YWHAZ 
                 tyrosine/tryptophan activation protein, zeta polypeptide 
                 Hs.75103 
                 NM_003406 
                 3 
               
               
                 BNO580 
                 IDS 
                 iduronate 2-sulfatase (Hunter syndrome) 
                 Hs.172458 
                 NM_000202 
                 24 
               
               
                 BNO581 
                 SERPINE2 
                 serine (or cysteine) proteinase inhibitor, clade E, member 2 
                 Hs.21858 
                 NM_006216 
                 6 
               
               
                 BNO582 
                 DDEF1 
                 development and differentiation enhancing factor 1 
                 Hs.10669 
                 NM_018482 
                 6 
               
               
                 BNO583 
                 GLRX 
                 glutaredoxin (thioltransferase) 
                 Hs.28988 
                 NM_002064 
                 24 
               
               
                 BNO584 
                 MAP3K1 
                 MAP3K1 Mitogen-activated protein kinase kinase kinase 1 
                 Hs.170610 
                 XM_042066 
                 3 
               
               
                 BNO585 
                 ANKH 
                 ankylosis, progressive homolog (mouse) 
                 Hs.168640 
                 NM_054027 
                 3 
               
               
                 BNO586 
                 RBX1 
                 ring-box 1 
                 Hs.279919 
                 NM_014248 
                 24 
               
               
                 BNO587 
                 NAB1 
                 NGFI-A binding protein 1 (EGR1 binding protein 1) 
                 Hs.107474 
                 NM_005966 
                 3 
               
               
                 BNO588 
                 TNFSF10 
                 tumor necrosis factor (ligand) superfamily, member 10 
                 Hs.83429 
                 NM_003810 
                 3 
               
               
                 BNO589 
                 PRDX3 
                 peroxiredoxin 3 
                 Hs.75454 
                 NM_006793 
                 6 
               
               
                 BNO590 
                 MAP2K1 
                 mitogen-activated protein kinase kinase 1 
                 Hs.3446 
                 NM_002755 
                 3 
               
               
                 BNO591 
                 NFATC1 
                 nuclear factor of activated T-cells, calcineurin-dependent 1 
                 Hs.96149 
                 NM_006162 
                 24 
               
               
                 BNO594 
                 USP7 
                 ubiquitin specific protease 7 (herpes virus-associated) 
                 Hs.78683 
                 NM_003470 
               
               
                 BNO595 
                 ARHB 
                 ras homolog gene family, member B 
                 Hs.406064 
                 NM_004040 
                 3 
               
               
                 BNO596 
                 PTEN 
                 phosphatase and tensin homolog 
                 Hs.10712 
                 NM_000314 
               
               
                 BNO597 
                 UBL1 
                 ubiquitin-like 1 (sentrin) 
                 Hs.81424 
                 NM_003352 
                 24 
               
               
                 BNO598 
                 RAB5A 
                 RAB5A, member RAS oncogene family 
                 Hs.73957 
                 NM_004162 
                 3 
               
               
                 BNO599 
                 ITGB1 
                 integrin, beta 1 
                 Hs.287797 
                 NM_002211 
                 24 
               
               
                 BNO600 
                 PRDM2 
                 PR domain containing 2, with ZNF domain 
                 Hs.26719 
                 NM_012231 
                 6 
               
               
                 BNO602 
                 ITGA2 
                 integrin, alpha 2 (CD49B, alpha 2 subunit of VLA-2 
                 Hs.271986 
                 NM_002203 
                 6 
               
               
                   
                   
                 receptor) 
               
               
                 BNO603 
                 ETV5 
                 ets variant gene 5 (ets-related molecule) 
                 Hs.43697 
                 NM_004454 
                 3 
               
               
                 BNO604 
                 ZFHX1B 
                 zinc finger homeobox 1b 
                 Hs.34871 
                 NM_014795 
                 3 
               
               
                 BNO606 
                 LOC157713 
                 lysophospholipase I-like pseudogene on chromosome 6 
                 None 
                 NG_001063 
               
               
                 BNO607 
                 RBM3 
                 RNA binding motif protein 3 
                 Hs.301404 
                 NM_006743 
                 0.5 
               
               
                 BNO609 
                 NET-6 
                 transmembrane 4 superfamily member tetraspan NET-6 
                 Hs.364544 
                 NM_014399 
                 6 
               
               
                 BNO610 
                 EHD3 
                 EH-domain containing 3 
                 Hs.87125 
                 NM_014600 
                 24 
               
               
                 BNO611 
                 KIAA0992 
                 palladin 
                 Hs.194431 
                 NM_016081 
                 6 
               
               
                 BNO613 
                 METL 
                 methyltransferase like 2 
                 Hs.433213 
                 NM_018396 
                 3 
               
               
                 BNO614 
                 HT010 
                 uncharacterized hypothalamus protein HT010 
                 Hs.6375 
                 NM_018471 
                 0.5 
               
               
                 BNO615 
                 C3orf4 
                 chromosome 3 open reading frame 4 
                 Hs.107393 
                 NM_019895 
                 6 
               
               
                 BNO619 
                 RPL27A 
                 ribosomal protein L27a 
                 Hs.76064 
                 NM_000990 
                 6 
               
               
                 BNO621 
                 MIB 
                 Ubiquitin ligase mind bomb 
                 Hs.34892 
                 AY149908 
                 0.5 
               
               
                 BNO623 
                 KIAA0261 
                 KIAA0261 protein 
                 Hs.154978 
                 XM_042946 
                 24 
               
               
                 BNO624 
                 KIAA1199 
                 KIAA1199 protein 
                 Hs.50081 
                 XM_051860 
                 6 
               
               
                 BNO625 
                 HIF1 
                 huntingtin interacting protein B 
                 Hs.6947 
                 NM_014159 
               
               
                 BNO642 
                 ETL 
                 EGF-TM7-latrophilin-related protein 
                 Hs.57958 
                 NM_022159 
                 24 
               
               
                 BNO643 
                 VMP1 
                 likely ortholog of rat vacuole membrane protein 1 
                 Hs.166254 
                 NM_030938 
                 3 
               
               
                 BNO644 
                 TAF9 
                 TATA box binding protein (TBP)-associated factor, 32 kDa 
                 Hs.60679 
                 NM_016283 
                 24 
               
               
                 BNO646 
                 MAN1A1 
                 mannosidase, alpha, class 1A, member 1 
                 Hs.432931 
                 NM_005907 
                 6 
               
               
                 BNO647 
                 DOCK4 
                 Dedicator of cytokinesis 4 
                 Hs.118140 
                 NM_014705 
                 24 
               
               
                 BNO649 
                 ADAMTS9 
                 a disintegrin-like and metalloprotease (thrombospondin type 
                 Hs.126855 
                 NM_020249 
                 24 
               
               
                   
                   
                 1 motif, 9) 
               
               
                 BNO650 
                 CSNK2A2 
                 Casein kinase 2, alpha prime polypeptide 
                 Hs.82201 
                 NM_001896 
                 6 
               
               
                 BNO651 
                 RPLP0 
                 ribosomal protein, large, P0 
                 Hs.406511 
                 NM_001002 
                 6 
               
               
                 BNO653 
                 GALNT4 
                 N-acetylgalactosaminyltransferase 4 
                 Hs.271923 
                 NM_003774 
                 3 
               
               
                 BNO654 
                 GNG2 
                 guanine nucleotide binding protein (G protein), gamma 2 
                 Hs.289026 
                 BC020774 
                 6 
               
               
                 BNO656 
                 MBNL 
                 muscleblind-like ( Drosophila ) 
                 Hs.28578 
                 NM_021038 
               
               
                 BNO657 
                 ARL8 
                 ADP-ribosylation factor-like 8 
                 Hs.25362 
                 BC024163 
                 3 
               
               
                 BNO658 
                 ASB3 
                 ankyrin repeat and SOCS box-containing 3 
                 Hs.9893 
                 NM_016115 
                 6 
               
               
                 BNO660 
                 GG2-1 
                 TNF-induced protein 
                 Hs.17839 
                 NM_014350 
                 3 
               
               
                 BNO661 
                 ELL2 
                 ELL-related RNA polymerase II, elongation factor 
                 Hs.98124 
                 NM_012081 
                 3 
               
               
                 BNO663 
                 ATP5J2 
                 ATP synthase, H+ transporting, mitochondrial F0 complex, 
                 Hs.235557 
                 NM_004889 
                 24 
               
               
                   
                   
                 subunit f 2 
               
               
                 BNO665 
                 SDCBP 
                 syndecan binding protein (syntenin) 
                 Hs.8180 
                 NM_005625 
                 3 
               
               
                 BNO666 
                 KIAA1959 
                 Nm23-phosphorylated unknown substrate 
                 Hs.55067 
                 NM_032873 
                 3 
               
               
                 BNO667 
                 GNPNAT1 
                 glucosamine-phosphate N-acetyltransferase 1 
                 Hs.478025 
                 NM_198066 
                 6 
               
               
                 BNO668 
                 SPRED1 
                 Sprouty-related, EVH1 domain containing 1 
                 Hs.132804 
                 NM_152594 
                   3, 24 
               
               
                 BNO670 
                 Nbak2 
                 homeodomain interacting protein kinase 1-like protein 
                 Hs.12259 
                 NM_152696 
                 6 
               
               
                 BNO672 
                 GABPA 
                 GA binding protein transcription factor, alpha subunit 60 kDa 
                 Hs.78 
                 NM_002040 
                 3 
               
               
                 BNO674 
                 V-1 
                 likely ortholog of rat V-1 protein 
                 Hs.21321 
                 NM_145808 
                 24 
               
               
                 BNO676 
                 C8FW 
                 phosphoprotein regulated by mitogenic pathways 
                 Hs.7837 
                 NM_025195 
                 3 
               
               
                 BNO678 
                 TBC1D4 
                 TBC1 domain family, member 4 
                 Hs.173802 
                 NM_014832 
                 6 
               
               
                 BNO679 
                 ACATE2 
                 likely ortholog of mouse acyl-Coenzyme A thioesterase 2 
                 Hs.18625 
                 NM_012332 
                 24 
               
               
                 BNO680 
                 CRYZ 
                 crystallin, zeta (quinone reductase) 
                 Hs.83114 
                 NM_001889 
                 6 
               
               
                 BNO681 
                 KPNB1 
                 karyopherin (importin) beta 1 
                 Hs.180446 
                 NM_002265 
                 24 
               
               
                 BNO682 
                 RPL23A 
                 ribosomal protein L23a 
                 Hs.350046 
                 NM_000984 
                 0.5 
               
               
                 BNO683 
                 LIMS1 
                 LIM and senescent cell antigen-like domains 1 
                 Hs.112378 
                 NM_004987 
                 6 
               
               
                 BNO684 
                 WW45 
                 WW45 protein 
                 Hs.288906 
                 NM_021818 
                 3 
               
               
                 BNO686 
                 ST3GALVI 
                 alpha2,3-sialyltransferase 
                 Hs.34578 
                 NM_006100 
                 6 
               
               
                 BNO688 
                 CPR8 
                 cell cycle progression 8 protein 
                 Hs.283753 
                 NM_004748 
                 24 
               
               
                 BNO689 
                 HDCL 
                 hHDC for homolog of  Drosophila  headcase 
                 Hs.6679 
                 NM_016217 
                 3 
               
               
                 BNO691 
                 UBC 
                 ubiquitin C 
                 Hs.183704 
                 NM_021009 
                 3 
               
               
                 BNO692 
                 RDX 
                 radixin 
                 Hs.263671 
                 NM_002906 
                 24 
               
               
                 BNO693 
                 PELI1 
                 pellino homolog 1 ( Drosophila ) 
                 Hs.7886 
                 NM_020651 
                 3 
               
               
                 BNO695 
                 MCC 
                 mutated in colorectal cancers 
                 Hs.1345 
                 NM_002387 
                 6 
               
               
                 BNO696 
                 RetSDR2 
                 RetSDR2 Retinal short-chain dehydrogenase/reductase 2 
                 Hs.282984 
                 NM_016245 
                 3 
               
               
                 BNO698 
                 CSS3 
                 Chondroitin sulfate synthase 3 
                 Hs.165050 
                 AB086062 
                 3 
               
               
                 BNO699 
                 BRE 
                 brain and reproductive organ-expressed (TNFRSF1A 
                 Hs.80426 
                 NM_004899 
                 6 
               
               
                   
                   
                 modulator) 
               
               
                 BNO701 
                 BAZ1A 
                 bromodomain adjacent to zinc finger domain, 1A 
                 Hs.8858 
                 NM_013448 
                 3 
               
               
                 BNO702 
                 HNRPDL 
                 heterogeneous nuclear ribonucleoprotein D-like 
                 Hs.372673 
                 NM_005463 
                 3 
               
               
                 BNO703 
                 PREI3 
                 preimplantation protein 3 
                 Hs.107942 
                 NM_015387 
                 6 
               
               
                 BNO707 
                 BNO707 
                 Human XIST, coding sequence “a” 
                 Hs.83623 
                 X56199 
                 3 
               
               
                 BNO709 
                 ROD1 
                 ROD1 regulator of differentiation 1 ( S. pombe ) 
                 Hs.374634 
                 NM_005156 
                 6 
               
               
                 BNO711 
                 SMAP-5 
                 golgi membrane protein SB140 
                 Hs.5672 
                 NM_030799 
                 6 
               
               
                 BNO715 
                 M-RIP 
                 Myosin phosphatase-Rho interacting protein 
                 Hs.430725 
                 AB020671 
                 0.5, 24 
               
               
                 BNO716 
                 HIVEP2 
                 human immunodeficiency virus type I enhancer binding 
                 Hs.75063 
                 NM_006734 
                 3 
               
               
                   
                   
                 protein 2 
               
               
                 BNO717 
                 DC42 
                 hypothetical protein DC42 
                 None 
                 NM_030921 
                 3 
               
               
                 BNO718 
                 GRPEL2 
                 GrpE-like 2, mitochondrial 
                 Hs.17121 
                 NM_152407 
                 6 
               
               
                 BNO719 
                 PCMF 
                 potassium channel modulatory factor 
                 Hs.5392 
                 NM_020122 
                 3 
               
               
                 BNO720 
                 UBE2E1 
                 ubiquitin-conjugating enzyme E2E 1 (UBC4/5 homolog, 
                 Hs.163546 
                 NM_003341 
                 24 
               
               
                   
                   
                 yeast) 
               
               
                 BNO721 
                 KLHL4 
                 kelch-like 4 ( Drosophila ) 
                 Hs.49075 
                 NM_019117 
               
               
                 BNO722 
                 MANEA 
                 Mannosidase, endo-alpha 
                 Hs.46903 
                 NM_024641 
                 3 
               
               
                 BNO724 
                 TCF12 
                 transcription factor 12 (HTF4, helix-loop-helix transcription 
                 Hs.21704 
                 NM_003205 
                 6 
               
               
                   
                   
                 factors 4) 
               
               
                 BNO726 
                 STAF42 
                 SPT3-associated factor 42 
                 Hs.435967 
                 NM_053053 
                 6 
               
               
                 BNO727 
                 CYFIP1 
                 cytoplasmic FMR1 interacting protein 1 
                 Hs.77257 
                 NM_014608 
                 6 
               
               
                 BNO728 
                 NOL5A 
                 nucleolar protein 5A (56 kDa with KKE/D repeat) 
                 Hs.376064 
                 NM_006392 
                 6 
               
               
                 BNO729 
                 GSA7 
                 ubiquitin activating enzyme E1-like protein 
                 Hs.278607 
                 NM_006395 
                 6 
               
               
                 BNO732 
                 P66 Alpha 
                 P66 Alpha 
                 Hs.118964 
                 NM_017660 
               
               
                 BNO733 
                 STAG1 
                 stromal antigen 1 
                 Hs.286148 
                 NM_005862 
               
               
                 BNO734 
                 MYCT1 
                 Myc target 1 
                 Hs.18160 
                 NM_025107 
                 6 
               
               
                 BNO736 
                 SCAMP1 
                 secretory carrier membrane protein 1 
                 Hs.31218 
                 NM_004866 
                 3 
               
               
                 BNO738 
                 ACTG1 
                 actin, gamma 1 
                 Hs.14376 
                 NM_001614 
                 0.5 
               
               
                 BNO739 
                 HRB2 
                 HIV-1 rev binding protein 2 
                 Hs.154762 
                 NM_007043 
                 6 
               
               
                 BNO741 
                 VMP1 
                 Likely orthologue of rat vacuole membrane protein 1 
                 Hs.166254 
                 NM_030938 
                 6 
               
               
                 BNO743 
                 BCAT1 
                 branched chain aminotransferase 1, cytosolic 
                 Hs.438993 
                 NM_005504 
                 0.5, 24 
               
               
                 BNO744 
                 PJA2 
                 Praja 2, RING-H2 motif containing 
                 Hs.224262 
                 NM_014819 
               
               
                 BNO746 
                 FKSG14 
                 leucine zipper protein FKSG14 
                 Hs.192843 
                 NM_022145 
                 6 
               
               
                 BNO748 
                 KLHL6 
                 kelch-like 6 ( Drosophila ) 
                 Hs.43616 
                 NM_130446 
                 6 
               
               
                 BNO749 
                 TTL 
                 Tubulin tyrosine ligase 
                 Hs.358997 
                 NM_153712 
                 6 
               
               
                 BNO750 
                 CDC23 
                 CDC23 (cell division cycle 23, yeast, homolog) 
                 Hs.153546 
                 NM_004661 
                 24 
               
               
                 BNO751 
                 ULK2 
                 unc-51-like kinase 2 ( C. elegans ) 
                 Hs.151406 
                 NM_014683 
                 3 
               
               
                 BNO752 
                 SCARB2 
                 SCARB2 Scavenger receptor class B, member 2 
                 Hs.323567 
                 NM_005506E 
                 3 
               
               
                 BNO755 
                 ZMPSTE24 
                 zinc metalloproteinase (STE24 homolog, yeast) 
                 Hs.25846 
                 NM_005857 
               
               
                 BNO757 
                 U5-100K 
                 prp28, U5 snRNP 100 kd protein 
                 Hs.184771 
                 NM_004818 
               
               
                 BNO758 
                 CHD4 
                 chromodomain helicase DNA binding protein 4 
                 Hs.74441 
                 NM_001273 
                 6 
               
               
                 BNO760 
                 CGI-127 
                 yippee protein 
                 Hs.184542 
                 NM_016061 
                   3, 24 
               
               
                 BNO763 
                 BET1 
                 BET1 homolog ( S. cerevisiae ) 
                 Hs.23103 
                 NM_005868 
                 24 
               
               
                 BNO764 
                 ARHGAP5 
                 Rho GTPase activating protein 5 
                 Hs.267831 
                 NM_001173 
               
               
                 BNO765 
                 TUBA 
                 Scaffold protein TUBA 
                 Hs.429994 
                 NM_015221 
                 3 
               
               
                 BNO766 
                 NUMB 
                 numb homolog ( Drosophila ) 
                 Hs.78890 
                 NM_003744 
                 6 
               
               
                 BNO767 
                 P5 
                 protein disulfide isomerase-related protein 
                 Hs.182429 
                 NM_005742 
                 0.5 
               
               
                 BNO769 
                 SFRS2IP 
                 splicing factor, arginine/serine-rich 2, interacting protein 
                 Hs.51957 
                 NM_004719 
                 6 
               
               
                 BNO770 
                 OXA1L 
                 oxidase (cytochrome c) assembly 1-like 
                 Hs.151134 
                 NM_005015 
                 0.5, 24 
               
               
                 BNO771 
                 POH1 
                 26S proteasome-associated pad1 homolog 
                 Hs.178761 
                 NM_005805 
                 6 
               
               
                 BNO773 
                 AHCYL1 
                 S-adenosylhomocysteine hydrolase-like 1 
                 Hs.4113 
                 NM_006621 
                 3 
               
               
                 BNO774 
                 UAP1 
                 UDP-N-acteylglucosamine pyrophosphorylase 1 
                 Hs.21293 
                 NM_003115 
                 3 
               
               
                 BNO775 
                 PLS3 
                 plastin 3 (T isoform) 
                 Hs.4114 
                 NM_005032 
                 6 
               
               
                 BNO776 
                 TSNAX 
                 translin-associated factor X 
                 Hs.96247 
                 NM_005999 
                 0.5 
               
               
                 BNO777 
                 HELO1 
                 homolog of yeast long chain polyunsaturated fatty acid 
                 Hs.250175 
                 NM_021814 
                 6 
               
               
                   
                   
                 elong. enz. 2 
               
               
                 BNO778 
                 MAN2A1 
                 mannosidase, alpha, class 2A, member 1 
                 Hs.377915 
                 NM_002372 
                 3 
               
               
                 BNO779 
                 RAB21 
                 RAB21, member RAS oncogene family 
                 Hs.184627 
                 NM_014999 
                 6 
               
               
                 BNO781 
                 WAC 
                 WW domain-containing adapter with a coiled-coil region 
                 Hs.70333 
                 NM_016628 
                 3 
               
               
                 BNO783 
                 POSH 
                 likely ortholog of mouse plenty of SH3 domains 
                 Hs.301804 
                 AB040927 
                 6 
               
               
                 BNO784 
                 RBM9 
                 RNA binding motif protein 9 
                 Hs.433574 
                 NM_014309 
               
               
                 BNO785 
                 CSRP2 
                 cysteine and glycine-rich protein 2 
                 Hs.10526 
                 NM_001321 
                 3 
               
               
                 BNO786 
                 COPA 
                 coatomer protein complex, subunit alpha 
                 Hs.75887 
                 NM_004371 
                 6 
               
               
                 BNO787 
                 TIMM17A 
                 translocase of inner mitochondrial membrane 17 homolog A 
                 Hs.20716 
                 NM_006335 
                 6 
               
               
                   
                   
                 (yeast) 
               
               
                 BNO788 
                 RIN2 
                 Ras and Rab interactor 2 
                 Hs.62349 
                 NM_018993 
                 24 
               
               
                 BNO789 
                 KLHL5 
                 kelch-like 5 ( Drosophila ) 
                 Hs.272239 
                 NM_015990 
                 24 
               
               
                 BNO790 
                 IPLA2(γ) 
                 intracellular memb.-assoc. calcium-independent 
                 Hs.44198 
                 AF263613 
                 6 
               
               
                   
                   
                 phospholipase A2γ 
               
               
                 BNO794 
                 SMARCA5 
                 SWI/SNF related regulator of chromatin, a5 
                 Hs.9456 
                 NM_003601 
               
               
                 BNO796 
                 FBXL3A 
                 F-box and leucine-rich repeat protein 3A 
                 Hs.7540 
                 NM_012158 
                 24 
               
               
                 BNO797 
                 SART2 
                 squamous cell carcinoma antigen recognized by T cell 
                 Hs.58636 
                 NM_013352E 
                 6 
               
               
                 BNO798 
                 YWHAZ 
                 14-3-3zeta 
                 Hs.386834 
                 NM_145690 
               
               
                 BNO799 
                 SH3BGRL2 
                 SH3 domain binding glutamic acid-rich protein like 2 
                 Hs.9167 
                 NM_031469 
                   3, 24 
               
               
                 BNO801 
                 PUM1 
                 pumilio homolog 1 ( Drosophila ) 
                 Hs.153834 
                 NM_014676 
                 3 
               
               
                 BNO803 
                 CCT2 
                 chaperonin containing TCP1, subunit 2 (beta) 
                 Hs.432970 
                 NM_006431 
                 6 
               
               
                 BNO804 
                 PTPRK 
                 protein tyrosine phosphatase, receptor type, K 
                 Hs.79005 
                 NM_002844 
                 6 
               
               
                 BNO806 
                 TM4SF1 
                 transmembrane 4 superfamily member 1 
                 Hs.351316 
                 NM_014220 
                 6 
               
               
                 BNO807 
                 CHSY1 
                 carbohydrate (chondroitin) synthase 1 
                 Hs.110488 
                 NM_014918 
                 24 
               
               
                 BNO808 
                 TERF2IP 
                 telomeric repeat binding factor 2, interacting protein 
                 Hs.274428 
                 NM_018975 
                 6 
               
               
                 BNO809 
                 RDC1 
                 G protein-coupled receptor 
                 Hs.23016 
                 BC036661 
                 3 
               
               
                 BNO810 
                 CD59 
                 CD59 antigen p18-20 
                 Hs.278573 
                 AK095453 
                 0.5, 6  
               
               
                 BNO811 
                 UBE2D1 
                 ubiquitin-conjugating enzyme E2D 1 (UBC4/5 homolog, 
                 Hs.129683 
                 NM_003338 
                 6 
               
               
                   
                   
                 yeast) 
               
               
                 BNO813 
                 CUL4B 
                 cullin 4B 
                 Hs.155976 
                 NM_003588 
                 24 
               
               
                 BNO814 
                 LCHN 
                 LCHN protein 
                 Hs.233044 
                 AB032973 
                 3 
               
               
                 BNO815 
                 PELO 
                 pelota homolog ( Drosophila ) 
                 Hs.5798 
                 NM_015946 
                 3 
               
               
                 BNO817 
                 MRPS10 
                 mitochondrial ribosomal protein S10 
                 Hs.380887 
                 NM_018141 
                 6 
               
               
                 BNO820 
                 EIF3S2 
                 eukaryotic translation initiation factor 3, subunit 2 beta, 
                 Hs.192023 
                 NM_003757 
                 3 
               
               
                   
                   
                 36 kDa 
               
               
                 BNO822 
                 UBQLN1 
                 ubiquilin 1 
                 Hs.9589 
                 NM_013438 
                 3 
               
               
                 BNO823 
                 PSMB3 
                 proteasome (prosome, macropain) subunit, beta type, 3 
                 Hs.82793 
                 NM_002795 
                 0.5, 24 
               
               
                 BNO826 
                 UBE2J1 
                 ubiquitin-conjugating enzyme E2, J1 (UBC6 homolog, 
                 Hs.184325 
                 NM_016336 
                 24 
               
               
                   
                   
                 yeast) 
               
               
                 BNO827 
                 CDK2AP1 
                 CDK2-associated protein 1 
                 Hs.433201 
                 NM_004642 
                 24 
               
               
                 BNO828 
                 CRY1 
                 cryptochrome 1 (photolyase-like) 
                 Hs.151573 
                 NM_004075 
                 3 
               
               
                 BNO830 
                 HSPC051 
                 ubiquinol-cytochrome c reductase complex (7.2 kD) 
                 Hs.284292 
                 NM_013387 
                 6 
               
               
                 BNO832 
                 GNG11 
                 guanine nucleotide binding protein (G protein), gamma 11 
                 Hs.83381 
                 NM_004126 
                 0.5, 24 
               
               
                 BNO834 
                 ZNF198 
                 zinc finger protein 198 
                 Hs.109526 
                 NM_003453 
                 6 
               
               
                 BNO835 
                 RAB11A 
                 RAB11A, member RAS oncogene family 
                 Hs.75618 
                 NM_004663 
                 6 
               
               
                 BNO836 
                 SMAP1 
                 stromal membrane-associated protein 
                 Hs.373517 
                 NM_021940 
                 6 
               
               
                 BNO837 
                 COPG 
                 Coatomer protein complex, subunit gamma 
                 Hs.368056 
                 NM_016128 
                 3 
               
               
                 BNO839 
                 MTHFD2 
                 methylene tetrahydrofolate dehydrogenase (NAD+ 
                 Hs.154672 
                 NM_006636 
                 3 
               
               
                   
                   
                 dependent) 
               
               
                 BNO840 
                 PODXL 
                 podocalyxin-like 
                 Hs.16426 
                 NM_005397 
                 6 
               
               
                 BNO841 
                 SLC30A7 
                 Solute carrier family 30 (zinc transporter), member 7 
                 Hs.38856 
                 NM_133496 
                 3 
               
               
                 BNO842 
                 API5 
                 apoptosis inhibitor 5 
                 Hs.227913 
                 NM_006595 
                 3 
               
               
                 BNO843 
                 ERdj5 
                 ER-resident protein ERdj5 
                 Hs.1098 
                 NM_018981 
                 3 
               
               
                 BNO844 
                 HDGFRP3 
                 Hepatoma-derived growth factor, related protein 3 
                 Hs.127842 
                 NM_016073 
                 6 
               
               
                 BNO847 
                 TUCAN 
                 tumor up-regulated CARD-containing antagonist of caspase 
                 Hs.10031 
                 NM_014959 
                 6 
               
               
                   
                   
                 nine 
               
               
                 BNO850 
                 PCDH17 
                 protocadherin 17 
                 Hs.106511 
                 NM_014459 
                 24 
               
               
                 BNO851 
                 GALNT10 
                 N-acetylgalactosaminyltransferase 10 
                 Hs.107260 
                 NM_017540 
                 24 
               
               
                 BNO853 
                 UQCRC1 
                 ubiquinol-cytochrome c reductase core protein I 
                 Hs.119251 
                 NM_003365 
                 6 
               
               
                 BNO854 
                 RPL3 
                 ribosomal protein L3 
                 Hs.119598 
                 NM_000967 
                 24 
               
               
                 BNO855 
                 CMT2 
                 gene predicted from cDNA with a complete coding 
                 Hs.124 
                 NM_014628 
                 24 
               
               
                   
                   
                 sequence 
               
               
                 BNO858 
                 PSMD7 
                 proteasome 26S subunit, non-ATPase, 7 
                 Hs.155543 
                 NM_002811 
                 6 
               
               
                 BNO859 
                 CCT5 
                 chaperonin containing TCP1, subunit 5 (epsilon) 
                 Hs.1600 
                 NM_012073 
                 3 
               
               
                 BNO860 
                 SEC5 
                 homolog of yeast Sec5 
                 Hs.16580 
                 NM_018303 
                 6 
               
               
                 BNO861 
                 SKP1A 
                 S-phase kinase-associated protein 1A (p19A) 
                 Hs.171626 
                 NM_006930 
                 24 
               
               
                 BNO863 
                 CAPZA1 
                 capping protein (actin filament) muscle Z-line, alpha 1 
                 Hs.184270 
                 NM_006135 
                 24 
               
               
                 BNO864 
                 YES1 
                 v-yes-1 Yamaguchi sarcoma viral oncogene homolog 1 
                 Hs.194148 
                 NM_005433 
                 24 
               
               
                 BNO865 
                 DAAM1 
                 dishevelled associated activator of morphogenesis 1 
                 Hs.197751 
                 NM_014992 
                 6 
               
               
                 BNO866 
                 BCL6B 
                 B-cell CLL/lymphoma 6, member B (zinc finger protein) 
                 Hs.22575 
                 NM_181844 
                 6 
               
               
                 BNO872 
                 AF5Q31 
                 ALL1 fused gene from 5q31 
                 Hs.231967 
                 NM_014423 
                 6 
               
               
                 BNO874 
                 ALDH9A1 
                 aldehyde dehydrogenase 9 family, member A1 
                 Hs.2533 
                 NM_000696 
                 24 
               
               
                 BNO875 
                 CDC42EP3 
                 CDC42 effector protein (Rho GTPase binding) 3 
                 Hs.260024 
                 NM_006449 
                 0.5, 24 
               
               
                 BNO877 
                 MIS12 
                 homolog of yeast Mis12 
                 Hs.267194 
                 NM_024039 
                 6 
               
               
                 BNO879 
                 ATP6V1D 
                 ATPase, H+ transporting, lysosomal 34 kDa, V1 subunit D 
                 Hs.272630 
                 NM_015994 
                 6 
               
               
                 BNO880 
                 VCIP135 
                 valosin-containing protein (p97)/p47 complex-interacting 
                 Hs.287727 
                 NM_025054 
                 6 
               
               
                   
                   
                 protein p135 
               
               
                 BNO882 
                 D10S170 
                 DNA segment on chromosome 10 (unique) 170 
                 Hs.288862 
                 NM_005436 
                 6 
               
               
                 BNO884 
                 ARPC3 
                 actin related protein 2/3 complex, subunit 3, 21 kDa 
                 Hs.293750 
                 NM_005719 
                 24 
               
               
                 BNO885 
                 RPS19 
                 ribosomal protein S19 
                 Hs.298262 
                 NM_001022 
                 6 
               
               
                 BNO888 
                 NEUGRIN 
                 mesenchymal stem cell protein DSC92 
                 Hs.323467 
                 NM_016645 
                 6 
               
               
                 BNO889 
                 CALD1 
                 caldesmon 1 
                 Hs.325474 
                 NM_033138 
                 0.5 
               
               
                 BNO891 
                 NFIB 
                 nuclear factor I/B 
                 Hs.33287 
                 NM_005596 
                 0.5 
               
               
                 BNO893 
                 HSPCA 
                 heat shock 90 kDa protein 1, alpha 
                 Hs.356531 
                 NM_005348 
                 6 
               
               
                 BNO896 
                 NSAP1 
                 NS1-associated protein 1 
                 Hs.373499 
                 NM_006372 
                 6 
               
               
                 BNO897 
                 SYT11 
                 synaptotagmin XI 
                 Hs.380439 
                 NM_152280 
                 6 
               
               
                 BNO899 
                 HNRPC 
                 heterogeneous nuclear ribonucleoprotein C (C1/C2) 
                 Hs.406125 
                 NM_006321 
                 24 
               
               
                 BNO900 
                 STMN1 
                 stathmin 1/oncoprotein 18 
                 Hs.406269 
                 NM_005563 
                 6 
               
               
                 BNO901 
                 ATP5B 
                 ATP synthase, H+ transporting, mitochondrial F1 complex, 
                 Hs.406510 
                 NM_001686 
                 0.5, 24 
               
               
                   
                   
                 beta 
               
               
                 BNO902 
                 PSMB1 
                 proteasome (prosome, macropain) subunit, beta type, 1 
                 Hs.407981 
                 NM_002793 
                 0.5, 24 
               
               
                 BNO903 
                 DDX10 
                 DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide 10 (RNA 
                 Hs.41706 
                 NM_004398 
                 6 
               
               
                   
                   
                 helicase) 
               
               
                 BNO904 
                 RPL36AL 
                 ribosomal protein L36a-like 
                 Hs.419465 
                 NM_001001 
                 24 
               
               
                 BNO907 
                 NDUFV2 
                 NADH dehydrogenase (ubiquinone) flavoprotein 2, 24 kDa 
                 Hs.51299 
                 NM_021074 
                 0.5, 24 
               
               
                 BNO909 
                 DCK 
                 deoxycytidine kinase 
                 Hs.709 
                 NM_000788 
                 24 
               
               
                 BNO911 
                 MDH1 
                 malate dehydrogenase 1, NAD (soluble) 
                 Hs.75375 
                 NM_005917 
                 24 
               
               
                 BNO912 
                 SERP1 
                 stress-associated endoplasmic reticulum protein 1 
                 Hs.76698 
                 NM_014445 
                 0.5 
               
               
                 BNO913 
                 RPS3A 
                 ribosomal protein S3A 
                 Hs.77039 
                 NM_001006 
                 0.5 
               
               
                 BNO914 
                 ARHA 
                 ras homolog gene family, member A 
                 Hs.77273 
                 NM_001664 
                 0.5 
               
               
                 BNO915 
                 LAMA4 
                 laminin, alpha 4 
                 Hs.78672 
                 NM_002290 
                 6 
               
               
                 BNO916 
                 SNX9 
                 sorting nexin 9 
                 Hs.7905 
                 NM_016224 
                 6 
               
               
                 BNO918 
                 RAD21 
                 RAD21 homolog ( S. pombe ) 
                 Hs.81848 
                 NM_006265 
                 0.5, 24 
               
               
                 BNO920 
                 PHLDA1 
                 pleckstrin homology-like domain, family A, member 1 
                 Hs.82101 
                 NM_007350 
                 6 
               
               
                 BNO921 
                 ARHGDIB 
                 Rho GDP dissociation inhibitor (GDI) beta 
                 Hs.83656 
                 NM_001175 
                 24 
               
               
                 BNO922 
                 ELP2 
                 elongator protein 2 
                 Hs.8739 
                 NM_018255 
                 6 
               
               
                 BNO924 
                 ATP6V1G1 
                 ATPase, H+ transporting, lysosomal 13 kDa, V1 subunit G 
                 Hs.90336 
                 NM_004888 
                 24 
               
               
                   
                   
                 isoform 1 
               
               
                 BNO925 
                 DNAJA1 
                 DnaJ (Hsp40) homolog, subfamily A, member 1 
                 Hs.94 
                 NM_001539 
                 3 
               
               
                 BNO927 
                 CYB561 
                 cytochrome b-561 
                 None 
                 NM_001915 
                 24 
               
               
                 BNO947 
                 HNRPDL 
                 Heterogeneous nuclear ribonucleoprotein D-like 
                 Hs.372673 
                 NM_005463 
                 3 
               
               
                 BNO952 
                 ARHB 
                 Ras homolog gene family, member B 
                 Hs.406064 
                 NM_004040 
                 3 
               
               
                 BNO955 
                 CYB561 
                 Cytochrome b-561 
                 Hs.355264 
                 AK095244 
                 24 
               
               
                 BNO958 
                 ATP6 
                 ATP synthase F0 subunit 6 - mitochondrial gene 
                 None 
                 NC_001807 
                 24 
               
               
                 BNO969 
                 ND4L 
                 NADH dehydrogenase subunit 4L - mitochondrial gene 
                 None 
                 NC_001807 
                 6 
               
               
                 BNO960 
                 COX2 
                 cytochrome C oxidase subunit II - mitochondrial gene 
                 None 
                 NC_001807 
                 0.5, 24 
               
               
                 BNO1014 
                 SET 
                 SET translocation (myeloid leukemia-associated) 
                 Hs.145279 
                 NM_003011 
                 6 
               
               
                 BNO1015 
                 JUNB 
                 jun B proto-oncogene 
                 Hs.400124 
                 NM_002229 
                 0.5 
               
               
                 BNO1016 
                 HMGB1 
                 high-mobility group box 1 
                 Hs.6727 
                 NM_002128 
                 6 
               
               
                 BNO1017 
                 PAFAH1B2 
                 Platelet-activating factor acetylhydrolase, isoform Ib, beta 
                 Hs.93354 
                 NM_002572 
                 24 
               
               
                   
                   
                 subunit 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 Genes Previously Associated with Angiogenesis 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 Peak 
               
               
                 BNO 
                   
                   
                 UniGene 
                   
                 Expression 
               
               
                 Number 
                 Symbol 
                 Gene Description - Homology 
                 Number 
                 GenBank Number 
                 (h) 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 BNO435 
                 ICAM1 
                 intercellular adhesion molecule 1 (CD54), human rhinovirus 
                 Hs.168383 
                 NM_000201 
                 3 
               
               
                   
                   
                 receptor 
               
               
                 BNO437 
                 IL8 
                 interleukin 8 
                 Hs.624 
                 NM_000584 
                 3 
               
               
                 BNO439 
                 VCAM1 
                 vascular cell adhesion molecule 1 
                 Hs.109225 
                 NM_001078 
                 3 
               
               
                 BNO440 
                 ANGPT2 
                 angiopoietin 2 
                 Hs.115181 
                 NM_001147 
                 6 
               
               
                 BNO444 
                 CTNNB1 
                 catenin (cadherin-associated protein), beta 1, 88 kDa 
                 Hs.171271 
                 NM_001904 
                 3 
               
               
                 BNO445 
                 F3 
                 coagulation factor III (thromboplastin, tissue factor) 
                 Hs.62192 
                 NM_001993 
                 3 
               
               
                 BNO450 
                 STC1 
                 stanniocalcin 1 
                 Hs.25590 
                 NM_003155 
                 24 
               
               
                 BNO458 
                 ADAMTS4 
                 a disintegrin-like and metalloprotease (thrombospondin type 
                 Hs.211604 
                 NM_005099 
                 6 
               
               
                   
                   
                 1 motif, 4) 
               
               
                 BNO471 
                 ESM1 
                 endothelial cell-specific molecule 1 
                 Hs.41716 
                 NM_007036 
                 3, 24 
               
               
                 BNO482 
                 CMG2 
                 capillary morphogenesis protein 2 
                 Hs.5897 
                 NM_058172 
                 6 
               
               
                 BNO486 
                 EFNB2 
                 ephrin-B2 
                 Hs.30942 
                 NM_004093 
                 3 
               
               
                 BNO493 
                 PTGS1 
                 prostaglandin-endoperoxide synthase 1 
                 Hs.88474 
                 NM_000962 
                 6 
               
               
                 BNO494 
                 KDR 
                 kinase insert domain receptor (a type III receptor tyrosine 
                 Hs.12337 
                 NM_002253 
               
               
                   
                   
                 kinase) 
               
               
                 BNO522 
                 F2R 
                 coagulation factor II (thrombin) receptor 
                 Hs.128087 
                 NM_001992 
                 3 
               
               
                 BNO529 
                 CTSB 
                 cathepsin B 
                 Hs.297939 
                 NM_001908 
                 24 
               
               
                 BNO530 
                 LIF 
                 leukemia inhibitory factor (cholinergic differentiation factor) 
                 Hs.2250 
                 NM_002309 
                 3 
               
               
                 BNO547 
                 EDN1 
                 endothelin 1 
                 Hs.2271 
                 NM_001955E 
                 0.5 
               
               
                 BNO550 
                 JAK1 
                 Janus kinase 1 (a protein tyrosine kinase) 
                 Hs.50651 
                 NM_002227 
                 24 
               
               
                 BNO563 
                 THBD 
                 thrombomodulin 
                 Hs.2030 
                 NM_000361 
                 24 
               
               
                 BNO592 
                 PSEN1 
                 presenilin 1 (Alzheimer disease 3) 
                 Hs.3260 
                 NM_000021 
                 0.5 
               
               
                 BNO593 
                 STAT3 
                 signal transducer and activator of transcription 3 
                 Hs.321677 
                 NM_139276 
                 6 
               
               
                 BNO601 
                 GJA1 
                 gap junction protein, alpha 1, 43 kDa (connexin 43) 
                 Hs.74471 
                 NM_000165 
                 3 
               
               
                 BNO608 
                 HEY1 
                 hairy/enhancer-of-split related with YRPW motif 1 
                 Hs.234434 
                 NM_012258 
                 0.5 
               
               
                 BNO846 
                 CXCR4 
                 chemokine (C—X—C motif) receptor 4 
                 Hs.89414 
                 NM_003467 
                 24 
               
               
                 BNO869 
                 ENTPD1 
                 ectonucleoside triphosphate diphosphohydrolase 1 
                 Hs.205353 
                 NM_001776 
                 0.5 
               
               
                 BNO919 
                 SERPINE1 
                 serine (or cysteine) proteinase inhibitor, clade E, member 1 
                 Hs.82085 
                 NM_000602 
                 3 
               
               
                 BNO923 
                 THBS1 
                 thrombospondin 1 
                 Hs.87409 
                 NM_003246 
                 0.5 
               
               
                   
               
            
           
         
       
     
     The invention also encompasses an isolated nucleic acid molecule that is at least 70% identical to any one of the angiogenic genes of the invention and which plays a role in the angiogenic process. 
     Such variants will have preferably at least about 85%, and most preferably at least about 95% sequence identity to the angiogenic genes. Any one of the polynucleotide variants described above can encode an amino acid sequence, which contains at least one functional or structural characteristic of the relevant angiogenic gene of the invention. 
     Sequence identity is typically calculated using the BLAST algorithm, described in Altschul et al (1997) with the BLOSUM62 default matrix. 
     The invention also encompasses an isolated nucleic acid molecule which hybridizes under stringent conditions with any one of the angiogenic genes of the invention and which plays a role in an angiogenic process. 
     Hybridization with PCR probes which are capable of detecting polynucleotide sequences, including genomic sequences, may be used to identify nucleic acid sequences which encode the relevant angiogenic gene. The specificity of the probe, whether it is made from a highly specific region, e.g., the 5′ regulatory region, or from a less specific region, e.g., a conserved motif, and the stringency of the hybridization or amplification will determine whether the probe identifies only naturally occurring sequences encoding the angiogenic gene, allelic variants, or related sequences. 
     Probes may also be used for the detection of related sequences, and should preferably have at least 50% sequence identity to any of the angiogenic gene-encoding sequences of the invention. The hybridization probes of the present invention may be DNA or RNA and may be derived from any one of the angiogenic gene sequences or from genomic sequences including promoters, enhancers, and introns of the angiogenic genes. 
     Means for producing specific hybridization probes for DNAs encoding any one of the angiogenic genes include the cloning of polynucleotide sequences encoding the relevant angiogenic gene or its derivatives into vectors for the production of mRNA probes. Such vectors are known in the art, and are commercially available. Hybridization probes may be labelled by radionuclides such as  32 P or  35 S, or by enzymatic labels, such as alkaline phosphatase coupled to the probe via avidin/biotin coupling systems, or other methods known in the art. 
     Under stringent conditions, hybridization with  32 P labelled probes will most preferably occur at 42° C. in 750 mM NaCl, 75 mM trisodium citrate, 2% SDS, 50% formamide, 1×Denhart&#39;s, 10% (w/v) dextran sulphate and 100 μg/ml denatured salmon sperm DNA. Useful variations on these conditions will be readily apparent to those skilled in the art. The washing steps which follow hybridization most preferably occur at 65° C. in 15 mM NaCl, 1.5 mM trisodium citrate, and 1% SDS. Additional variations on these conditions will be readily apparent to those skilled in the art. 
     The nucleic acid molecules, or fragments thereof, of the present invention have a nucleotide sequence obtainable from a natural source. They therefore include naturally occurring normal, naturally occurring mutant, naturally occurring polymorphic alleles, differentially spliced transcripts, splice variants etc. Natural sources include animal cells and tissues, body fluids, tissue culture cells etc. 
     The nucleic acid molecules of the present invention can also be engineered using methods accepted in the art so as to alter the angiogenic gene-encoding sequences for a variety of purposes. These include, but are not limited to, modification of the cloning, processing, and/or expression of the gene product. PCR reassembly of gene fragments and the use of synthetic oligonucleotides allow the engineering of angiogenic gene nucleotide sequences. For example, oligonucleotide-mediated site-directed mutagenesis can introduce mutations that create new restriction sites, alter glycosylation patterns and produce splice variants etc. 
     As a result of the degeneracy of the genetic code, a number of nucleic acid sequences encoding the angiogenic genes of the invention, some that may have minimal similarity to the nucleic acid sequences of any known and naturally occurring gene, may be produced. Thus, the invention includes each and every possible variation of polynucleotide sequence that could be made by selecting combinations based on possible codon choices. These combinations are made in accordance with the standard triplet genetic code as applied to the polynucleotide sequence of the naturally occurring angiogenic gene, and all such variations are to be considered as being specifically disclosed. 
     The nucleic acid molecules of this invention are typically DNA molecules, and include cDNA, genomic DNA, synthetic forms, and mixed polymers, both sense and antisense strands, and may be chemically or biochemically modified, or may contain non-natural or derivatised nucleotide bases as will be appreciated by those skilled in the art. Such modifications include labels, methylation, intercalators, alkylators and modified linkages. In some instances it may be advantageous to produce nucleotide sequences encoding an angiogenic gene or its derivatives possessing a substantially different codon usage than that of the naturally occurring gene. For example, codons may be selected to increase the rate of expression of the peptide in a particular prokaryotic or eukaryotic host corresponding with the frequency that the host utilizes particular codons. Other reasons to alter the nucleotide sequence encoding an angiogenic gene or its derivatives without altering the encoded amino acid sequence include the production of RNA transcripts having more desirable properties, such as a greater half-life, than transcripts produced from the naturally occurring sequence. 
     The invention also encompasses production of the nucleic acid molecules of the invention, entirely by synthetic chemistry. Synthetic sequences may be inserted into expression vectors and cell systems that contain the necessary elements for transcriptional and translational control of the inserted coding sequence in a suitable host. These elements may include regulatory sequences, promoters, 5′ and 3′ untranslated regions and specific initiation signals (such as an ATG initiation codon and Kozak consensus sequence) which allow more efficient translation of sequences encoding the angiogenic genes. In cases where the complete coding sequence including its initiation codon and upstream regulatory sequences are inserted into the appropriate expression vector, additional control signals may not be needed. However, in cases where only coding sequence, or a fragment thereof, is inserted, exogenous translational control signals as described above should be provided by the vector. Such signals may be of various origins, both natural and synthetic. The efficiency of expression may be enhanced by the inclusion of enhancers appropriate for the particular host cell system used (Scharf et al., 1994). 
     The invention also includes nucleic acid molecules that are the complements of the sequences described herein. 
     The present invention allows for the preparation of purified polypeptides or proteins. In order to do this, host cells may be transfected with a nucleic acid molecule as described above. Typically, said host cells are transfected with an expression vector comprising a nucleic acid molecule according to the invention. A variety of expression vector/host systems may be utilized to contain and express the sequences. These include, but are not limited to, microorganisms such as bacteria transformed with plasmid or cosmid DNA expression vectors; yeast transformed with yeast expression vectors; insect cell systems infected with viral expression vectors (e.g., baculovirus); or mouse or other animal or human tissue cell systems. Mammalian cells can also be used to express a protein that is encoded by a specific angiogenic gene of the invention using various expression vectors including plasmid, cosmid and viral systems such as a vaccinia virus expression system. The invention is not limited by the host cell or vector employed. 
     The nucleic acid molecules, or variants thereof, of the present invention can be stably expressed in cell lines to allow long term production of recombinant proteins in mammalian systems. Sequences encoding any one of the angiogenic genes of the invention can be transformed into cell lines using expression vectors which may contain viral origins of replication and/or endogenous expression elements and a selectable marker gene on the same or on a separate vector. The selectable marker confers resistance to a selective agent, and its presence allows growth and recovery of cells which successfully express the introduced sequences. Resistant clones of stably transformed cells may be propagated using tissue culture techniques appropriate to the cell type. 
     The protein produced by a transformed cell may be secreted or retained intracellularly depending on the sequence and/or the vector used. As will be understood by those of skill in the art, expression vectors containing polynucleotides which encode a protein may be designed to contain signal sequences which direct secretion of the protein through a prokaryotic or eukaryotic cell membrane. 
     In addition, a host cell strain may be chosen for its ability to modulate expression of the inserted sequences or to process the expressed protein in the desired fashion. Such modifications of the polypeptide include, but are not limited to, acetylation, glycosylation, phosphorylation, and acylation. Post-translational cleavage of a “prepro” form of the protein may also be used to specify protein targeting, folding, and/or activity. Different host cells having specific cellular machinery and characteristic mechanisms for post-translational activities (e.g., CHO or HeLa cells), are available from the American Type Culture Collection (ATCC) and may be chosen to ensure the correct modification and processing of the foreign protein. 
     According to still another aspect of the present invention there is provided an expression vector comprising a nucleic acid molecule of the invention as described above. 
     According to still another aspect of the present invention there is provided a cell comprising a nucleic acid molecule of the invention as described above. 
     When large quantities of protein are needed such as for antibody production, vectors which direct high levels of expression may be used such as those containing the T5 or T7 inducible bacteriophage promoter. The present invention also includes the use of the expression systems described above in generating and isolating fusion proteins which contain important functional domains of the protein. These fusion proteins are used for binding, structural and functional studies as well as for the generation of appropriate antibodies. 
     In order to express and purify the protein as a fusion protein, the appropriate polynucleotide sequences of the present invention are inserted into a vector which contains a nucleotide sequence encoding another peptide (for example, glutathionine succinyl transferase). The fusion protein is expressed and recovered from prokaryotic or eukaryotic cells. The fusion protein can then be purified by affinity chromatography based upon the fusion vector sequence and the relevant protein can subsequently be obtained by enzymatic cleavage of the fusion protein. 
     Fragments of polypeptides of the present invention may also be produced by direct peptide synthesis using solid-phase techniques. Automated synthesis may be achieved by using the ABI 431A Peptide Synthesizer (Perkin-Elmer). Various fragments of polypeptide may be synthesized separately and then combined to produce the full length molecule. 
     In instances where the isolated nucleic acid molecules of the invention represent only partial gene sequence, these partial sequences can be used to obtain the corresponding sequence of the full-length angiogenic gene. Therefore, the present invention further provides the use of a partial nucleic acid molecule of the invention comprising a nucleotide sequence defined by any one of SEQ ID Numbers: 1 to 15, 17 to 37, and 39 to 44 to identify and/or obtain full-length human genes involved in the angiogenic process. Full-length angiogenic genes may be cloned using the partial nucleotide sequences of the invention by methods known per se to those skilled in the art. For example, in silico analysis of sequence databases such as those hosted at the National Centre for Biotechnology Information can be searched in order to obtain overlapping nucleotide sequence. This provides a “walking” strategy towards obtaining the full-length gene sequence. Appropriate databases to search at this site include the expressed sequence tag (EST) database (database of GenBank, EMBL and DDBJ sequences from their EST divisions) or the non redundant (nr) database (contains all GenBank, EMBL, DDBJ and PDB sequences but does not include EST, STS, OSS, or phase 0, 1 or 2 HTGS sequences). Typically searches are performed using the BLAST algorithm described in Altschul et al (1997) with the BLOSUM62 default matrix. In instances where in silico “walking” approaches fail to retrieve the complete gene sequence, additional strategies may be employed. These include the use of “restriction-site PCR” will allows the retrieval of unknown sequence adjacent to a portion of DNA whose sequence is known. In this technique universal primers are used to retrieve unknown sequence. Inverse PCR may also be used, in which primers based on the known sequence are designed to amplify adjacent unknown sequences. These upstream sequences may include promoters and regulatory elements. In addition, various other PCR-based techniques may be used, for example a kit available from Clontech (Palo Alto, Calif.) allows for a walking PCR technique, the 5′RACE kit (Gibco-BRL) allows isolation of additional 5′ gene sequence, while additional 3′ sequence can be obtained using practised techniques (for example see Gecz et al., 1997). 
     In a further aspect of the present invention there is provided an isolated polypeptide as defined by SEQ ID Numbers: 51 to 58 and laid out in Table 1. 
     The present invention also provides isolated polypeptides, which have been shown to be up-regulated in their expression during angiogenesis (see Tables 1 and 2). 
     More specifically, following the realisation that these polypeptides are up-regulated in their expression during angiogenesis, the invention provides isolated polypeptides as defined by SEQ ID Numbers: 51 to 58, and as laid out in Tables 1 and 2, or fragments thereof, that play a role in an angiogenic process. Such a process may include, but is not restricted to, embryogenesis, menstrual cycle, wound repair, tumour angiogenesis and exercise induced muscle hypertrophy. 
     In addition, the present invention provides isolated polypeptides as defined by SEQ ID Numbers: 51 to 58, and as laid out in Tables 1 and 2, or fragments thereof, that play a role in diseases associated with the angiogenic process. Diseases may include, but are not restricted to, cancer, rheumatoid arthritis, diabetic retinopathy, psoriasis, and cardiovascular diseases such as atherosclerosis, ischaemic limb disease and coronary artery disease. 
     The invention also encompasses an isolated polypeptide having at least 70%, preferably 85%, and more preferably 95%, identity to any one of SEQ ID Numbers: 51 to 58, and which plays a role in an angiogenic process. 
     Sequence identity is typically calculated using the BLAST algorithm, described in Altschul et al (1997) with the BLOSUM62 default matrix. 
     In a further aspect of the invention there is provided a method of preparing a polypeptide as described above, comprising the steps of: 
     (1) culturing cells as described above under conditions effective for production of the polypeptide; and 
     (2) harvesting the polypeptide. 
     According to still another aspect of the invention there is provided a polypeptide which is the product of the process described above. 
     Substantially purified protein or fragments thereof can then be used in further biochemical analyses to establish secondary and tertiary structure. Such methodology is known in the art and includes, but is not restricted to, X-ray crystallography of crystals of the proteins or by nuclear magnetic resonance (NMR). Determination of structure allows for the rational design of pharmaceuticals to interact with the protein, alter protein charge configuration or charge interaction with other proteins, or to alter its function in the cell. 
     The invention has provided a number of genes likely to be involved in angiogenesis and therefore enables methods for the modulation of angiogenesis. As angiogenesis is critical in a number of pathological processes, the invention therefore also enables therapeutic methods for the treatment of all angiogenesis-related disorders, and may enable the diagnosis or prognosis of all angiogenesis-related disorders associated with abnormalities in expression and/or function of any one of the angiogenic genes. 
     Examples of such disorders include, but are not limited to, cancer, rheumatoid arthritis, diabetic retinopathy, psoriasis, and cardiovascular diseases such as atherosclerosis, ischaemic limb disease and coronary artery disease. 
     Therapeutic Applications 
     According to another aspect of the present invention there is provided a method of treating an angiogenesis-related disorder as described above, comprising administering a selective antagonist or agonist of an angiogenic gene or protein of the invention to a subject in need of such treatment. 
     In still another aspect of the invention there is provided the use of a selective antagonist or agonist of an angiogenic gene or protein of the invention in is the manufacture of a medicament for the treatment of an angiogenesis-related disorder as described above. 
     For the treatment of angiogenesis-related disorders which result in uncontrolled or enhanced angiogenesis, including but not limited to, cancer, rheumatoid arthritis, diabetic retinopathy, psoriasis and cardiovascular diseases such as atherosclerosis, therapies which inhibit the expanding vasculature are desirable. This would involve inhibition of any one of the angiogenic genes or proteins that are able to promote angiogenesis, or enhancement, stimulation or re-activation of any one of the angiogenic genes or proteins that are able to inhibit angiogenesis. 
     For the treatment of angiogenesis-related disorders which are characterised by inhibited or decreased angiogenesis, including but not limited to, ischaemic limb disease and coronary artery disease, therapies which enhance or promote vascular expansion are desirable. This would involve inhibition of any one of the angiogenic genes or proteins that are able to restrict angiogenesis or enhancement, stimulation or re-activation of any one of the angiogenic genes or proteins that are able to promote angiogenesis. 
     For instance, decreasing the expression of BNO782 and BNO481 has been shown to disrupt endothelial cell activity leading to an inhibition of capillary tube formation and angiogenesis. Therefore, in the treatment of disorders where angiogenesis needs to be restricted, it would be desirable to inhibit the function of these genes. Alternatively, in the treatment of disorders where angiogenesis needs to be stimulated it may be desirable to enhance the function of these genes. 
     In some embodiments, a method of modulating angiogenesis comprising modulating the expression or activity of a BNO802 polypeptide in a cell, wherein the BNO802 polypeptide is encoded by a BNO802 nucleic acid molecule set forth in Table 1, is provided. 
     For each of these cases, the relevant therapy will be useful in treating angiogenesis-related disorders regardless of whether there is a lesion in the angiogenic gene. 
     Inhibiting Gene or Protein Function 
     Inhibiting the function of a gene or protein can be achieved in a variety of ways. Antisense nucleic acid methodologies represent one approach to inactivate genes that are causative of a disorder. Antisense or gene-targeted silencing strategies may include, but are not limited to, the use of antisense oligonucleotides, injection of antisense RNA, transfection of antisense RNA expression vectors, and the use of RNA interference (RNAi) or short interfering RNAs (siRNA). RNAi can be used in vitro and in vivo to silence a gene when its expression contributes to angiogenesis (Sharp and Zamore, 2000; Grishok et al., 2001). Still further, catalytic nucleic acid molecules such as DNAzymes and ribozymes may be used for gene silencing (Breaker and Joyce, 1994; Haseloff and Gerlach, 1988). These molecules function by cleaving their target mRNA molecule rather than merely binding to it as in traditional antisense approaches. 
     In one aspect of the invention an isolated nucleic acid molecule, which is the complement of any one of the relevant angiogenic nucleic acid molecules described above may be administered to a subject in need of such treatment. Typically, a complement to any relevant one of the angiogenic genes is administered to a subject to treat or prevent an angiogenesis-related disorder. In a further aspect the complement may encode an RNA molecule that hybridizes with the mRNA encoded by the relevant angiogenic gene of the invention or may be a short interfering oligonucleotide (siRNA) that hybridizes with the mRNA encoded by the relevant angiogenic gene of the invention. 
     In a further aspect of the invention there is provided the use of an isolated nucleic acid molecule which is the complement of any one of the relevant nucleic acid molecules of the invention and which encodes an RNA molecule or a short interfering oligonucleotide (siRNA) that hybridizes with the mRNA encoded by the relevant angiogenic gene of the invention, in the manufacture of a medicament for the treatment of an angiogenesis-related disorder. 
     Typically, a vector expressing the complement of a polynucleotide encoding any one of the relevant angiogenic genes may be administered to a subject to treat or prevent an angiogenesis-related disorder including, but not limited to, those described above. Many methods for introducing vectors into cells or tissues are available and equally suitable for use in vivo, in vitro, and ex vivo. For ex vivo therapy, vectors may be introduced into stem cells taken from the patient and clonally propagated for autologous transplant back into that same patient. Delivery by transfection, by liposome injections, or by polycationic amino polymers may be achieved using methods which are well known in the art. (For example, see Goldman et al., 1997). 
     In a further aspect purified protein according to the invention may be used to produce antibodies which specifically bind any relevant angiogenic protein of the invention. These antibodies may be used directly as an antagonist or indirectly as a targeting or delivery mechanism for bringing a pharmaceutical agent (such as a cytotoxic agent) to cells or tissues that express the relevant angiogenic protein. Such antibodies may include, but are not limited to, polyclonal, monoclonal, chimeric and single chain antibodies as would be understood by the person skilled in the art. 
     For the production of antibodies, various hosts including rabbits, rats, goats, mice, humans, and others may be immunized by injection with a protein of the invention or with any fragment or oligopeptide thereof, which has immunogenic properties. Various adjuvants may be used to increase immunological response and include, but are not limited to, Freund&#39;s, mineral gels such as aluminum hydroxide, and surface-active substances such as lysolecithin. Adjuvants used in humans include BCG (bacilli Calmette-Guerin) and  Corynebacterium parvum.    
     It is preferred that the oligopeptides, peptides, or fragments used to induce antibodies to the relevant angiogenic protein have an amino acid sequence consisting of at least about 5 amino acids, and, more preferably, of at least about 10 amino acids. It is also preferable that these oligopeptides, peptides, or fragments are identical to a portion of the amino acid sequence of the natural protein and contain the entire amino acid sequence of a small, naturally occurring molecule. Short stretches of amino acids from these proteins may be fused with those of another protein, such as KLH, and antibodies to the chimeric molecule may be produced. 
     Monoclonal antibodies to any relevant angiogenic protein may be prepared using any technique which provides for the production of antibody molecules by continuous cell lines in culture. These include, but are not limited to, the hybridoma technique, the human B-cell hybridoma technique, and the EBV-hybridoma technique. (For example, see Kohler and Milstein, 1975; Kozbor et al., 1985; Cote et al., 1983; Cole et al., 1984). 
     Monoclonal antibodies produced may include, but are not limited to, mouse-derived antibodies, humanised antibodies and fully-human antibodies. For example, antibodies are obtained from transgenic mice that have been engineered to produce specific human antibodies in response to antigenic challenge. In one example of this technique, elements of the human heavy and light chain loci are introduced into strains of mice derived from embryonic stem cell lines that contain targeted disruptions of the endogenous heavy and light chain loci. These transgenic mice can synthesise human antibodies specific for human antigens and can be used to produce human antibody-secreting hybridomas. Methods for obtaining human antibodies from transgenic mice are described for example in Lonberg et al., 1994; Green et al., 1994; Taylor et al., 1994. 
     Antibodies may also be produced by inducing in vivo production in the lymphocyte population or by screening immunoglobulin libraries or panels of highly specific binding reagents as disclosed in the literature. (For example, see Orlandi et al., 1989; Winter et al., 1991). 
     Antibody fragments which contain specific binding sites for any relevant angiogenic protein may also be generated. For example, such fragments include, F(ab′)2 fragments produced by pepsin digestion of the antibody molecule and Fab fragments generated by reducing the disulfide bridges of the F(ab′)2 fragments. Alternatively, Fab expression libraries may be constructed to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity. (For example, see Huse et al., 1989). 
     Various immunoassays may be used for screening to identify antibodies having the desired specificity. Numerous protocols for competitive binding or immunoradiometric assays using either polyclonal or monoclonal antibodies with established specificities are well known in the art. Such immunoassays typically involve the measurement of complex formation between a protein and its specific antibody. A two-site, monoclonal-based immunoassay utilizing monoclonal antibodies reactive to two non-interfering epitopes is preferred, but a competitive binding assay may also be employed. 
     In a further aspect, antagonists may include peptides, phosphopeptides or small organic or inorganic compounds. These antagonists should disrupt the function of any relevant angiogenic gene of the invention so as to provide the necessary therapeutic effect. 
     Peptides, phosphopeptides or small organic or inorganic compounds suitable for therapeutic applications may be identified using nucleic acids and polypeptides of the invention in drug screening applications as described below. 
     Enhancing Gene or Protein Function 
     Enhancing, stimulating or re-activating a gene&#39;s or protein&#39;s function can be achieved in a variety of ways. In one aspect of the invention administration of an isolated nucleic acid molecule, as described above, to a subject in need of such treatment may be initiated. Typically, any relevant angiogenic gene of the invention can be administered to a subject to treat or prevent an angiogenesis-related disorder. 
     In a further aspect, there is provided the use of an isolated nucleic acid molecule, as described above, in the manufacture of a medicament for the treatment of an angiogenesis-related disorder. 
     Typically, a vector capable of expressing any relevant angiogenic gene, or a fragment or derivative thereof, may be administered to a subject to treat or prevent a disorder including, but not limited to, those described above. Transducing retroviral vectors are often used for somatic cell gene therapy because of their high efficiency of infection and stable integration and expression. Any relevant full-length gene, or portions thereof, can be cloned into a retroviral vector and expression may be driven from its endogenous promoter or from the retroviral long terminal repeat or from a promoter specific for the target cell type of interest. Other viral vectors can be used and include, as is known in the art, adenoviruses, adeno-associated viruses, vaccinia viruses, papovaviruses, lentiviruses and retroviruses of avian, murine and human origin. 
     Gene therapy would be carried out according to established methods (Friedman, 1991; Culver, 1996). A vector containing a copy of any relevant angiogenic gene linked to expression control elements and capable of replicating inside the cells is prepared. Alternatively the vector may be replication deficient and may require helper cells for replication and use in gene therapy. 
     Gene transfer using non-viral methods of infection in vitro can also be used. These methods include direct injection of DNA, uptake of naked DNA in the presence of calcium phosphate, electroporation, protoplast fusion or liposome delivery. Gene transfer can also be achieved by delivery as a part of a human artificial chromosome or receptor-mediated gene transfer. This involves linking the DNA to a targeting molecule that will bind to specific cell-surface receptors to induce endocytosis and transfer of the DNA into mammalian cells. One such technique uses poly-L-lysine to link asialoglycoprotein to DNA. An adenovirus is also added to the complex to disrupt the lysosomes and thus allow the DNA to avoid degradation and move to the nucleus. Infusion of these particles intravenously has resulted in gene transfer into hepatocytes. 
     Although not identified to date, it is possible that certain individuals with angiogenesis-related disorders contain an abnormality in any one of the angiogenic genes of the invention. In affected subjects that express a mutated form of any one of the angiogenic genes of the invention it may be possible to prevent the disorder by introducing into the affected cells a wild-type copy of the gene such that it recombines with the mutant gene. This requires a double recombination event for the correction of the gene mutation. Vectors for the introduction of genes in these ways are known in the art, and any suitable vector may be used. Alternatively, introducing another copy of the gene bearing a second mutation in that gene may be employed so as to negate the original gene mutation and block any negative effect. 
     In a still further aspect, there is provided a method of treating an angiogenesis-related disorder comprising administering a polypeptide, as described above, or an agonist thereof, to a subject in need of such treatment. 
     In another aspect the invention provides the use of a polypeptide as described above, or an agonist thereof, in the manufacture of a medicament for the treatment of an angiogenesis-related disorder. Examples of such disorders are described above. 
     In a further aspect, a suitable agonist may also include peptides, phosphopeptides or small organic or inorganic compounds that can mimic the function of any relevant angiogenic gene, or may include an antibody to any relevant angiogenic gene that is able to restore function to a normal level. 
     Peptides, phosphopeptides or small organic or inorganic compounds suitable for therapeutic applications may be identified using nucleic acids and polypeptides of the invention in drug screening applications as described below. 
     In further embodiments, any of the agonists, antagonists, complementary sequences, nucleic acid molecules, proteins, antibodies, or vectors of the invention may be administered in combination with other appropriate therapeutic agents. Selection of the appropriate agents may be made by those skilled in the art, according to conventional pharmaceutical principles. The combination of therapeutic agents may act synergistically to effect the treatment or prevention of the various disorders described above. Using this approach, therapeutic efficacy with lower dosages of each agent may be possible, thus reducing the potential for adverse side effects. 
     Any of the therapeutic methods described above may be applied to any subject in need of such therapy, including, for example, mammals such as dogs, cats, cows, horses, rabbits, monkeys, and most preferably, humans. 
     Modulation of Angiogenesis 
     As the invention has provided a number of genes likely to be involved in angiogenesis it therefore enables methods for the modulation of angiogenesis. In a further aspect of the present invention, any of the methods described above used for the treatment of an angiogenesis-related disorder may be used for the modulation of angiogenesis in any system comprising cells. These systems may include but are not limited to, in vitro assay systems (e.g. Matrigel assays, proliferation assays, migration assays, collagen assays, bovine capillary endothelial cell assay etc), in vivo assay systems (e.g. in vivo Matrigel-type assays, chicken chorioallantoic membrane assay, isolated organs, tissues or cells etc), animal models (e.g. in vivo neovascularisation assays, tumour angiogenesis models etc) or hosts in need of treatment (e.g. hosts suffering from angiogenesis-related disorders as previously described. 
     Drug Screening 
     According to still another aspect of the invention, nucleic acid molecules of the invention as well as peptides of the invention, particularly any relevant purified angiogenic polypeptides or fragments thereof, and cells expressing these are useful for screening of candidate pharmaceutical compounds in a variety of techniques for the treatment of angiogenesis-related disorders. 
     Still further, it provides the use wherein high throughput screening techniques are employed. 
     Compounds that can be screened in accordance with the invention include, but are not limited to peptides (such as soluble peptides), phosphopeptides and small organic or inorganic molecules (such as natural product or synthetic chemical libraries and peptidomimetics). 
     In one embodiment, a screening assay may include a cell-based assay utilising eukaryotic or prokaryotic host cells that are stably transformed with recombinant nucleic acid molecules expressing the relevant angiogenic polypeptide or fragment, in competitive binding assays. Binding assays will measure for the formation of complexes between the relevant polypeptide or fragments thereof and the compound being tested, or will measure the degree to which a compound being tested will interfere with the formation of a complex between the relevant polypeptide or fragment thereof, and its interactor or ligand. 
     Non cell-based assays may also be used for identifying compounds that interrupt binding between the polypeptides of the invention and their interactors. Such assays are known in the art and include for example AlphaScreen technology (PerkinElmer Life Sciences, MA, USA). This application relies on the use of beads such that each interaction partner is bound to a separate bead via an antibody. Interaction of each partner will bring the beads into proximity, such that laser excitation initiates a number of chemical reactions ultimately leading to fluorophores emitting a light signal. Candidate compounds that disrupt the binding of the relevant angiogenic polypeptide with its interactor will result in loss of light emission enabling identification and isolation of the responsible compound. 
     High-throughput drug screening techniques may also employ methods as described in WO84/03564. Small peptide test compounds synthesised on a solid substrate can be assayed through relevant angiogenic polypeptide binding and washing. The relevant bound angiogenic polypeptide is then detected by methods well known in the art. In a variation of this technique, purified angiogenic polypeptides can be coated directly onto plates to identify interacting test compounds. 
     An additional method for drug screening involves the use of host eukaryotic cell lines that carry mutations in any relevant angiogenic gene of the invention. The host cell lines are also defective at the polypeptide level. Other cell lines may be used where the expression of the relevant angiogenic gene can be regulated (i.e. over-expressed, under-expressed, or switched off). The host cell lines or cells are grown in the presence of various drug compounds and the rate of growth of the host cells is measured to determine if the compound is capable of regulating the growth of defective cells. 
     The angiogenic polypeptides of the present invention may also be used for screening compounds developed as a result of combinatorial library technology. This provides a way to test a large number of different substances for their ability to modulate activity of a polypeptide. A substance identified as a modulator of polypeptide function may be peptide or non-peptide in nature. Non-peptide “small molecules” are often preferred for many in vivo pharmaceutical applications. In addition, a mimic or mimetic of the substance may be designed for pharmaceutical use. The design of mimetics based on a known pharmaceutically active compound (“lead” compound) is a common approach to the development of novel pharmaceuticals. This is often desirable where the original active compound is difficult or expensive to synthesise or where it provides an unsuitable method of administration. In the design of a mimetic, particular parts of the original active compound that are important in determining the target property are identified. These parts or residues constituting the active region of the compound are known as its pharmacophore. Once found, the pharmacophore structure is modelled according to its physical properties using data from a range of sources including x-ray diffraction data and NMR. A template molecule is then selected onto which chemical groups that mimic the pharmacophore can be added. The selection can be made such that the mimetic is easy to synthesise, is likely to be pharmacologically acceptable, does not degrade in vivo and retains the biological activity of the lead compound. Further optimisation or modification can be carried out to select one or more final mimetics useful for in vivo or clinical testing. 
     It is also possible to isolate a target-specific antibody and then solve its crystal structure. In principle, this approach yields a pharmacophore upon which subsequent drug design can be based as described above. It may be possible to avoid protein crystallography altogether by generating anti-idiotypic antibodies (anti-ids) to a functional, pharmacologically active antibody. As a mirror image of a mirror image, the binding site of the anti-ids would be expected to be an analogue of the original binding site. The anti-id could then be used to isolate peptides from chemically or biologically produced peptide banks. 
     Another alternative method for drug screening relies on structure-based rational drug design. Determination of the three dimensional structure of the polypeptides of the invention, or the three dimensional structure of the protein complexes which may incorporate these polypeptides allows for structure-based drug design to identify biologically active lead compounds. 
     Three dimensional structural models can be generated by a number of applications, some of which include experimental models such as x-ray crystallography and NMR and/or from in silico studies using information from structural databases such as the Protein Databank (PDB). In addition, three dimensional structural models can be determined using a number of known protein structure prediction techniques based on the primary sequences of the polypeptides (e.g. SYBYL—Tripos Associated, St. Louis, Mo.), de novo protein structure design programs (e.g. MODELER—MSI Inc., San Diego, Calif., or MOE—Chemical Computing Group, Montreal, Canada) or ab initio methods (e.g. see U.S. Pat. Nos. 5,331,573 and 5,579,250). 
     Once the three dimensional structure of a polypeptide or polypeptide complex has been determined, structure-based drug discovery techniques can be employed to design biologically active compounds based on these three dimensional structures. Such techniques are known in the art and include examples such as DOCK (University of California, San Francisco) or AUTODOCK (Scripps Research Institute, La Jolla, Calif.). A computational docking protocol will identify the active site or sites that are deemed important for protein activity based on a predicted protein model. Molecular databases, such as the Available Chemicals Directory (ACD) are then screened for molecules that complement the protein model. 
     Using methods such as these, potential clinical drug candidates can be identified and computationally ranked in order to reduce the time and expense associated with typical ‘wet lab’ drug screening methodologies. 
     Compounds identified from the screening methods described above form a part of the present invention, as do pharmaceutical compositions containing these and a pharmaceutically acceptable carrier. 
     Pharmaceutical Preparations 
     Compounds identified from screening assays as indicated above can be administered to a patient at a therapeutically effective dose to treat or ameliorate a disorder associated with angiogenesis. A therapeutically effective dose refers to that amount of the compound sufficient to result in amelioration of symptoms of the disorder. 
     Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals. The data obtained from these studies can then be used in the formulation of a range of dosages for use in humans. 
     Pharmaceutical compositions for use in accordance with the present invention can be formulated in a conventional manner using one or more physiological acceptable carriers, excipients or stabilisers which are well known. Acceptable carriers, excipients or stabilizers are non-toxic at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including absorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; binding agents including hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and/or non-ionic surfactants such as Tween, Pluronics or polyethylene glycol (PEG). 
     The formulation of pharmaceutical compositions for use in accordance with the present invention will be based on the proposed route of administration. Routes of administration may include, but are not limited to, inhalation, insufflation (either through the mouth or nose), oral, buccal, rectal or parental administration. 
     Diagnostic and Prognostic Applications 
     Should abnormalities in any one of the angiogenic genes of the invention exist, which alter activity and/or expression of the gene to give rise to angiogenesis-related disorders, the polynucleotides and polypeptides of the invention may be used for the diagnosis or prognosis of these disorders, or a predisposition to such disorders. Examples of such disorders include, but are not limited to, cancer, rheumatoid arthritis, diabetic retinopathy, psoriasis, cardiovascular diseases such as atherosclerosis, ischaemic limb disease and coronary artery disease. Diagnosis or prognosis may be used to determine the severity, type or stage of the disease state in order to initiate an appropriate therapeutic intervention. 
     In another embodiment of the invention, the polynucleotides that may be used for diagnostic or prognostic purposes include oligonucleotide sequences, genomic DNA and complementary RNA and DNA molecules. The polynucleotides may be used to detect and quantitate gene expression in biopsied tissues in which abnormal expression or mutations in any one of the angiogenic genes may be correlated with disease. Genomic DNA used for the diagnosis or prognosis may be obtained from body cells, such as those present in the blood, tissue biopsy, surgical specimen, or autopsy material. The DNA may be isolated and used directly for detection of a specific sequence or may be amplified by the polymerase chain reaction (PCR) prior to analysis. Similarly, RNA or cDNA may also be used, with or without PCR amplification. To detect a specific nucleic acid sequence, direct nucleotide sequencing, reverse transcriptase PCR (RT-PCR), hybridization using specific oligonucleotides, restriction enzyme digest and mapping, PCR mapping, RNAse protection, and various other methods may be employed. Oligonucleotides specific to particular sequences can be chemically synthesized and labelled radioactively or nonradioactively and hybridized to individual samples immobilized on membranes or other solid-supports or in solution. The presence, absence or excess expression of any one of the angiogenic genes may then be visualized using methods such as autoradiography, fluorometry, or colorimetry. 
     In a particular aspect, the nucleotide sequences of the invention may be useful in assays that detect the presence of associated disorders, particularly those mentioned previously. The nucleotide sequences may be labelled by standard methods and added to a fluid or tissue sample from a patient under conditions suitable for the formation of hybridization complexes. After a suitable incubation period, the sample is washed and the signal is quantitated and compared with a standard value. If the amount of signal in the patient sample is significantly altered in comparison to a control sample then the presence of altered levels of nucleotide sequences in the sample indicates the presence of the associated disorder. Such assays may also be used to evaluate the efficacy of a particular therapeutic treatment regimen in animal studies, in clinical trials, or to monitor the treatment of an individual patient. 
     In order to provide a basis for the diagnosis or prognosis of an angiogenesis-related disorder associated with a mutation in any one of the angiogenic genes of the invention, the nucleotide sequence of the relevant gene can be compared between normal tissue and diseased tissue in order to establish whether the patient expresses a mutant gene. 
     In order to provide a basis for the diagnosis or prognosis of a disorder associated with abnormal expression of any one of the angiogenic genes of the invention, a normal or standard profile for expression is established. This may be accomplished by combining body fluids or cell extracts taken from normal subjects, either animal or human, with a sequence, or a fragment thereof, encoding the relevant angiogenic gene, under conditions suitable for hybridization or amplification. Standard hybridization may be quantified by comparing the values obtained from normal subjects with values from an experiment in which a known amount of a substantially purified polynucleotide is used. Another method to identify a normal or standard profile for expression of any one of the angiogenic genes is through quantitative RT-PCR studies. RNA isolated from body cells of a normal individual, particularly RNA isolated from endothelial cells, is reverse transcribed and real-time PCR using oligonucleotides specific for the relevant gene is conducted to establish a normal level of expression of the gene. Standard values obtained in both these examples may be compared with values obtained from samples from patients who are symptomatic for a disorder. Deviation from standard values is used to establish the presence of a disorder. 
     Once the presence of a disorder is established and a treatment protocol is initiated, hybridization assays or quantitative RT-PCR studies may be repeated on a regular basis to determine if the level of expression in the patient begins to approximate that which is observed in the normal subject. The results obtained from successive assays may be used to show the efficacy of treatment over a period ranging from several days to months. 
     According to a further aspect of the invention there is provided the use of an angiogenic polypeptide as described above in the diagnosis or prognosis of an angiogenesis-related disorder associated with any one of angiogenic genes of the invention, or a predisposition to such disorders. 
     When a diagnostic or prognostic assay is to be based upon any relevant angiogenic polypeptide, a variety of approaches are possible. For example, diagnosis or prognosis can be achieved by monitoring differences in the electrophoretic mobility of normal and mutant proteins. Such an approach will be particularly useful in identifying mutants in which charge substitutions are present, or in which insertions, deletions or substitutions have resulted in a significant change in the electrophoretic migration of the resultant protein. Alternatively, diagnosis or prognosis may be based upon differences in the proteolytic cleavage patterns of normal and mutant proteins, differences in molar ratios of the various amino acid residues, or by functional assays demonstrating altered function of the gene products. 
     In another aspect, antibodies that specifically bind the relevant angiogenic gene product may be used for the diagnosis or prognosis of disorders characterized by abnormal expression of the gene, or in assays to monitor patients being treated with the relevant angiogenic gene or protein or agonists, antagonists, or inhibitors thereof. Antibodies useful for diagnostic or prognostic purposes may be prepared in the same manner as described above for therapeutics. Diagnostic or prognostic assays may include methods that utilize the antibody and a label to detect the relevant protein in human body fluids or in extracts of cells or tissues. The antibodies may be used with or without modification, and may be labelled by covalent or non-covalent attachment of a reporter molecule. 
     A variety of assays for measuring the relevant angiogenic polypeptide based on the use of antibodies specific for the polypeptide are known in the art and provide a basis for diagnosing altered or abnormal levels of expression. Normal or standard values for expression are established by combining body fluids or cell extracts taken from normal mammalian subjects, preferably human, with antibody to the relevant protein under conditions suitable for complex formation. The amount of standard complex formation may be quantitated by various methods which are known in the art. Examples include, but are not limited to, enzyme-linked immunosorbent assays (ELISAs), radioimmunoassays (RIAs), immunofluorescence, flow cytometry, histology, electron microscopy, in situ assays, immunoprecipitation, Western blot etc. For example, using the ELISA technique an enzyme which is bound to the antibody will react with an appropriate substrate, preferably a chromogenic substrate, in such a manner as to produce a chemical moiety that can be detected for example by spectrophotomeric, fluorimetric or by visual means. Detection may also be accomplished by using other assays such as RIAs where the antibodies or antibody fragments are radioactively labelled. It is also possible to label the antibody with a fluorescent compound. When the fluorescently labelled antibody is exposed to light of a certain wavelength, its presence can then be detected due to fluorescence. The antibody can also be detectably labelled by coupling it to a chemiluminescent compound. The presence of the chemiluminescent-tagged antibody is then determined by detecting the presence of luminescence that arises during the course of a chemical reaction. 
     Quantities of protein expressed in subject, control, and disease samples from biopsied tissues are compared with the standard values. Deviation between standard and subject values establishes the parameters for diagnosing or prognosing disease. 
     Once an individual has been diagnosed or prognosed with a disorder, effective treatments can be initiated, as described above. In the treatment of angiogenesis-related diseases which are characterised by uncontrolled or enhanced angiogenesis, the expanding vasculature needs to be inhibited. This would involve inhibiting the relevant angiogenic genes or proteins of the invention that promote angiogenesis. In addition, treatment may also need to stimulate expression or function of the relevant angiogenic genes or proteins of the invention whose normal role is to inhibit angiogenesis but whose activity is reduced or absent in the affected individual. 
     In the treatment of angiogenesis-related diseases which are characterised by inhibited or decreased angiogenesis, approaches which enhance or promote vascular expansion are desirable. This may be achieved using methods essentially as described above but will involve stimulating the expression or function of the relevant angiogenic gene or protein whose normal role is to promote angiogenesis but whose activity is reduced or absent in the affected individual. Alternatively, inhibiting genes or proteins that restrict angiogenesis may also be an approach to treatment. 
     Microarray 
     In further embodiments, complete cDNAs, oligonucleotides or longer fragments derived from any of the polynucleotide sequences described herein may be used as probes in a microarray. The microarray can be used to monitor the expression level of large numbers of genes simultaneously and to identify genetic variants, mutations, and polymorphisms. This information may be used to determine gene function, to understand the genetic basis of angiogenesis-related disorders, to diagnose or prognose angiogenesis-related disorders, and to develop and monitor the activities of therapeutic agents. Microarrays may be prepared, used, and analysed using methods known in the art. (For example, see Schena et al., 1996; Heller et al., 1997). 
     Transformed Hosts 
     The present invention also provides for the production of genetically modified (knock-out, knock-in and transgenic), non-human animal models comprising the nucleic acid molecules of the invention. These animals are useful for the study of the function of the relevant angiogenic gene, to study the process of angiogenesis, to study the mechanisms of angiogenic disease as related to these genes, for the screening of candidate pharmaceutical compounds for the treatment of angiogenesis-related disorders for the creation of explanted mammalian cell cultures which express the protein or mutant protein, and for the evaluation of potential therapeutic interventions. 
     Animal species which are suitable for use in the animal models of the present invention include, but are not limited to, rats, mice, hamsters, guinea pigs, rabbits, dogs, cats, goats, sheep, pigs, and non-human primates such as monkeys and chimpanzees. For initial studies, genetically modified mice and rats are highly desirable due to the relative ease in generating knock-in, knock-out or transgenics of these animals, their ease of maintenance and their shorter life spans. For certain studies, transgenic yeast or invertebrates may be suitable and preferred because they allow for rapid screening and provide for much easier handling. For longer term studies, non-human primates may be desired due to their similarity with humans. 
     To create an animal model based on any one of the angiogenic genes of the invention, several methods can be employed. These include, but are not limited to, generation of a specific mutation in a homologous animal gene, insertion of a wild type human gene and/or a humanized animal gene by homologous recombination, insertion of a mutant (single or multiple) human gene as genomic or minigene cDNA constructs using wild type, mutant or artificial promoter elements, or insertion of artificially modified fragments of the endogenous gene by homologous recombination. The modifications include insertion of mutant stop codons, the deletion of DNA sequences, or the inclusion of recombination elements (lox p sites) recognized by enzymes such as Cre recombinase. 
     To create transgenic mice in order to study gain of gene function in vivo, any relevant angiogenic gene can be inserted into a mouse germ line using standard techniques such as oocyte microinjection. Gain of gene function can mean the overexpression of a gene and its protein product, or the genetic complementation of a mutation of the gene under investigation. For oocyte injection, one or more copies of the wild type or mutant gene can be inserted into the pronucleus of a just-fertilized mouse oocyte. This oocyte is then reimplanted into a pseudo-pregnant foster mother. The liveborn mice can then be screened for integrants using analysis of tail DNA for the presence of the relevant human angiogenic gene sequence. The transgene can be either a complete genomic sequence injected as a YAC, BAC, PAC or other chromosome DNA fragment, a cDNA with either the natural promoter or a heterologous promoter, or a minigene containing all of the coding region and other elements found to be necessary for optimum expression. 
     To generate knock-out mice or knock-in mice, gene targeting through homologous recombination in mouse embryonic stem (ES) cells may be applied. Knock-out mice are generated to study loss of gene function in vivo while knock-in mice allow the study of gain of function or to study the effect of specific gene mutations. Knock-in mice are similar to transgenic mice however the integration site and copy number are defined in the former. 
     For knock-out mouse generation, gene targeting vectors can be designed such that they disrupt (knock-out) the protein coding sequence of the relevant angiogenic gene in the mouse genome. Knock-out animals of the invention will comprise a functional disruption of a relevant angiogenesis gene of the invention such that the gene does not express a biologically active product. It can be substantially deficient in at least one functional activity coded for by the gene. Expression of the polypeptide encoded by the gene can be substantially absent (i.e. essentially undetectable amounts are made) or may be deficient in activity such as where only a portion of the gene product is produced. In contrast, knock-in mice can be produced whereby a gene targeting vector containing the relevant angiogenic gene can integrate into a defined genetic locus in the mouse genome. For both applications, homologous recombination is catalysed by specific DNA repair enzymes that recognise homologous DNA sequences and exchange them via double crossover. 
     Gene targeting vectors are usually introduced into ES cells using electroporation. ES cell integrants are then isolated via an antibiotic resistance gene present on the targeting vector and are subsequently genotyped to identify those ES cell clones in which the gene under investigation has integrated into the locus of interest. The appropriate ES cells are then transmitted through the germline to produce a novel mouse strain. 
     In instances where gene ablation results in early embryonic lethality, conditional gene targeting may be employed. This allows genes to be deleted in a temporally and spatially controlled fashion. As above, appropriate ES cells are transmitted through the germline to produce a novel mouse strain, however the actual deletion of the gene is performed in the adult mouse in a tissue specific or time controlled manner. Conditional gene targeting is most commonly achieved by use of the cre/lox system. The enzyme cre is able to recognise the 34 base pair loxP sequence such that loxP flanked (or floxed) DNA is recognised and excised by cre. Tissue specific cre expression in transgenic mice enables the generation of tissue specific knock-out mice by mating gene targeted floxed mice with cre transgenic mice. Knock-out can be conducted in every tissue (Schwenk et al., 1995) using the ‘deleter’ mouse or using transgenic mice with an inducible cre gene (such as those with tetracycline inducible cre genes), or knock-out can be tissue specific for example through the use of the CD19-cre mouse (Rickert et al., 1997). 
     According to still another aspect of the invention there is provided the use of genetically modified non-human animals for the screening of candidate pharmaceutical compounds. 
     It will be clearly understood that, although a number of prior art publications are referred to herein, this reference does not constitute an admission that any of these documents forms part of the common general knowledge in the art, in Australia or in any other country. Throughout this specification and the claims, the words “comprise”, “comprises” and “comprising” are used in a non-exclusive sense, except where the context requires otherwise. 
     EXAMPLES 
     Example 1 
     In Vitro Capillary Tube Formation 
     The in vitro model of angiogenesis is essentially as described in Gamble et al (1993). The assay was performed in collagen under the stimulation of phorbol myristate acetate (PMA) and the anti-integrin (α 2 β 1 ) antibody, RMACII. Human umbilical vein endothelial cells (HUVECs) were used in all experiments between passages 2 to 4. 
     Cells were harvested from bulk cultures (t=0), replated onto the collagen gels with stimulation and then harvested from the collagen gels at 0.5, 3.0, 6.0 and 24 hours after commencement of the assay. These time points were chosen since major morphological changes occur at these stages. Briefly, by 0.5 hours, cells have attached to the collagen matrix and have commenced migration into the gel. By 3.0 hours, small intracellular vesicles are visible. By 6.0 hours, these vesicles are coalescing together to form membrane bound vacuoles and the cells in the form of short sprouts have invaded the gel. After this time, these vacuoles fuse with the plasma membrane, thus expanding the intercellular space to generate the lumen (Meyer et al., 1997). The formation of these larger vacuoles is an essential requirement of lumen formation (Gamble et al., 1999). By 24 hours, the overall anastomosing network of capillary tubes has formed and has commenced degeneration. 
     Example 2 
     RNA Isolation, cDNA Synthesis and Amplification 
     Cells harvested at the specified time points were used for the isolation of total RNA using the Trizol reagent (Gibco BRL) according to manufacturers conditions. SMART (Switching mechanism at 5′ end of RNA transcript) technology was used to convert small amounts of total RNA into enough cDNA to enable cDNA subtraction to be performed (see below). This was achieved using the SMART-PCR cDNA synthesis kit (Clontech-user manual PT3041-1) according to manufacturers recommendations. The SMART-PCR cDNA synthesis protocol generated a majority of full length cDNAs which were subsequently PCR amplified for cDNA subtraction. 
     Example 3 
     Suppression Subtractive Hybridization (SSH) 
     SSH was performed on SMART amplified cDNA in order to enrich for cDNAs that were either up-regulated or down-regulated between the cDNA populations defined by the selected time-points. This technique also allowed “normalisation” of the regulated cDNAs, thereby making low abundance cDNAs (i.e. poorly expressed, but important, genes) more easily detectable. To do this, the PCR-Select cDNA synthesis kit (Clontech-user manual PT3041-1) and PCR-Select cDNA subtraction kit (Clontech-user manual PT1117-1) were used based on manufacturers conditions. These procedures relied on subtractive hybridization and suppression PCR amplification. SSH was performed between the following populations: 0-0.5 hours; 0.5-3.0 hours; 3.0-6.0 hours; 6.0-24 hours. 
     Example 4 
     Differential Screening of cDNA Clones 
     Following SSH, the cDNA fragments were digested with EagI and cloned into the compatible unique NotI site in pBluescript KS +  using standard techniques (Sambrook et al., 1989). This generated forward and reverse subtracted libraries for each time period. Initially, the forward subtracted libraries were used in subsequent studies to identify those clones representing genes that were up-regulated in their expression during the in vitro model of angiogenesis. To do this, a microarray analysis procedure was adopted. 
     Microarray Slide Preparation 
     A total of 10,000 clones from the 4 forward subtracted libraries (3,200 clones from 0-0.5 hr; 3,000 clones from 0.5-3 hr; 2,800 clones from 3-6 hr; 1,000 clones from 6-24 hr) were chosen to construct microarray slides. Inserts from these clones were amplified using standard PCR techniques with flanking T3 and T7 pBluescript KS +  vector primers. DNA from each clone was spotted in duplicate onto a single microarray slide. Appropriate positive and negative controls were also incorporated onto the plate. 
     Probe Labelling 
     Human umbilical vein endothelial cells harvested at the specified time points (0, 0.5, 3, 6, and 24 hr) were used for the isolation of total RNA using the Trizol reagent (Gibco BRL) according to manufacturers conditions. From each time point, 0.5 ug of total RNA was used as a template for the amplification of antisense RNA (aRNA) using the Ambion MessageAmp™ aRNA Kit. Briefly, total RNA was reversed transcribed with a T7 oligo(dT) primer in order to synthesize cDNA containing a T7 promoter sequence extending from the poly(A) tails of messages generated by reverse transcription. The cDNA was converted to a double-stranded DNA template and used for in vitro transcription of aRNA, incorporating 5-(3-aminoallyl)-UTP so as to allow coupling of fluorescent CyDyes. A typical amplification reaction would yield approximately 10 ug of mRNA (&gt;400× amplification, assuming the initial total RNA contained &lt;5% mRNA). 
     Microarray Hybridization 
     After coupling of CyDyes, the synthesized aRNA was used as a probe (3.0-3.5 ug) for hybridization to a microarray slide. The hybrizations performed were as follows: 
     1. 0 vs 0.5 h (6 slides, 3 Dye swaps) 
     2. 0 vs 3 h (4 slides, 2 Dye swaps) 
     3. 0 vs 6 h (4 slides, 2 Dye swaps) 
     4. 0 vs 24 h (4 slides, 2 Dye swaps) 
     Multiple slides were hybridized for each time point in order to verify the result from any one hybridization. Slides were hybridized in chambers for 16 hours, washed, and then scanned using the GenePix 2000 scanner. Those clones that were shown to be highly up-regulated were chosen for further analysis. 
     In summary, SSH was used in combination with microarray analyses to identify genes that are up-regulated and may be involved in biological processes underlying endothelial cell activation and blood vessel formation. This approach is novel in that it involves nucleotide hybridization steps that aim to reduce gene detection redundancy and enhance the chances of detecting genes that are of low overall representation in the endothelial cell transcriptome. The nucleotide-based sequential time-points aims to detect the timepoint at which the up-regulation of a particular gene takes place in a way that reduces redundancy of detection. For example, a gene that is up-regulated at 3 hrs, and its expression remains up-regulated in subsequent time-points, will only be detected in the 0.5-3 hr subtraction step. In contrast, if subtractions were done with the 0 hr timepoint for all subsequent timepoints then this example gene would be detected at all subtraction steps following the 3 hrs timepoint subtraction. This would introduce redundancy that could result in masking the possible detection of other genes of lower representation in the endothelial cell mRNA expression pool. The subsequent use of microarray analysis is based on the comparison subtraction hybridization in the SSH step involving each timepoint with the 0 hrs timepoint. This enables the expression profiling of each gene across all timepoints in relation to 0 hrs, irrespective of the timepoint at which it is up-regulated. 
     Example 5 
     Clone Selection 
     From analysis of the microarray hybridizations, a total of 1,963 clones were identified to be up-regulated in their expression at specified time points during the in vitro model of angiogenesis.  FIG. 1  provides an example of the expression profiles observed during defined time points in the in vitro model for a selection of clones. Each of the 1,963 clones were sequenced and subsequent in silico database analysis was used to remove clones containing vector sequences only and clones for which poor sequence was obtained. Following this, redundancy screens were used to group clones according to individual genes that they represented. This left a total of 523 genes that were found to be up-regulated in their expression during the process of angiogenesis. 
     Tables 1, 2 and 3 provide information on the up-regulated clones that were sequenced. Table 1 includes those clones which represent previously uncharacterised or novel genes, while Table 2 includes clones that correspond to previously identified genes which have not before been associated with angiogenesis. Also identified were a number of genes that have previously been shown to be involved in the process of angiogenesis (Table 3). The identification of these clones provides a validation or proof of principle of the effectiveness of the angiogenic gene identification strategy employed and suggests that the clones listed in Tables 1 and 2 are additional angiogenic gene candidates. 
     Example 6 
     Analysis of the Angiogenic Genes 
     Further evidence for the involvement of the genes in Tables 1 and 2 in angiogenesis can be obtained through the functional analysis of each gene, for example by examining the effect that knock-down of their expression has on endothelial cell (EC) function and capillary tube formation. 
     A number of knock-down technologies and assays may be used. For example full-length coding sequences of the genes can be cloned into suitable expression vectors such as retroviruses or adenoviruses in both sense and anti-sense orientations and used for infection into ECs. Retrovirus infection gives long-term EC lines expressing the gene of interest whereas adenovirus infection gives transient gene expression. Infected cells can then be subjected to a number of EC assays including proliferation and capillary tube formation to confirm the role of each gene in angiogenesis. 
     In this study RNA interference (RNAi) gene knock-down technology was used for the analysis of gene function (see detailed description below). In this technique, short gene-specific RNA oligonucleotides are delivered to ECs in culture mediated by retroviral infection. These oligonucleotides bind to the gene transcript under study and induce its degradation resulting in silencing or reduction of gene expression. The consequences of this alteration to gene expression can be subsequently studied using assays that examine the ability of ECs to proliferate, migrate and form capillaries in vitro. The RNAi procedure adopted in this study is described below in detail and documents the analysis of two of the identified up-regulated angiogenesis genes. One of these genes is BNO782 shown in Table 1, a novel gene whose expression peaks at the 6 hour time point of the in vitro angiogenesis model ( FIG. 2A ), while the other gene is BNO481 (KPNA4) as shown in Table 2, which is a previously identified gene that has not before been shown to have a role in angiogenesis. The expression of BNO481 also peaks at the 6 hour time point of the in vitro angiogenesis model ( FIG. 2B ). 
     RNAi Oligonucleotide Design 
     Short interfering RNA (siRNA) oligonucleotides for RNAi-mediated knock-down of BNO782 and BNO481 were identified through application of in-house computer software. This software incorporates a series of parameters for selecting appropriate siRNA oligonucleotides. These parameters ensure that the siRNA sequence starts after an AA dinucleotide, the siRNA is in the open reading frame of the gene and 100 bp downstream the ATG start codon, the GC content of the siRNA is between 35% and 60%, and the siRNA does not have stretches of more than three T, A, C or G nucleotides. siRNA sequences that harbour low complexity regions were not used. In addition, BLAST analysis was used to select against probes that cross-hybridize with a number of genes (Blastn_refseq at “expect 500” and “word size 7” and alignment scores accepted at 19&gt;score&gt;15 where: alignment score=length_match−(gap+mismatch). siRNAs were synthesised in hair-pin format for cloning into retroviral vectors. For each gene, three siRNA oligonucleotides were selected with each one being examined individually for their effects on gene-knock-down and EC function. 
     Retroviral Infection of HUVE Cells 
     Each siRNA oligonucleotide was cloned into a retroviral vector for the delivery of the oligonucleotide to human umbilical vein endothelial cells (HUVECs). The siRNA vector was constructed through a modification of pMSCVpuro (BD Biosciences). Briefly, the 3′LTR of pMSCVpuro was inactivated by removal of the XbaI/NheI fragment. A H1-RNA Polymerase III promoter cassette was then inserted into the MCS of the vector. Annealed siRNA primers were ligated into the modified vector (pMSCVpuro(H1)) digested with BglII and HindIII restriction enzymes. 
     For virus production prior to infection of HUVECs, 293T cells were plated at a density of 1×10 6  cells per well of a 6 well plate 18-24 hours before transfection in RPMI media (Invitrogen) supplemented with 10% FCS (Invitrogen) and 1.0 M Hepes (Invitrogen) without antibiotics. Cells were co-transfected with 2 μg retroviral DNA and 1.5 μg pVPack-VSV-G (Stratagene), 1.5 μg pVPack-GP (Stratagene) using Lipofectamine 2000 reagent (Invitrogen). Transfected cells were incubated overnight in 5% CO 2  at 37° C. The following day, media containing the DNA/LF2000 complexes was removed and replaced with RPMI supplemented with 10% FCS, 1.0 M Hepes and 1% PSG (Invitrogen). Virus containing supernatants were collected 48-72 hours post transfection and filtered using a 0.45 μM filter. Virus was aliquoted and stored at −80° C. 
     For the retroviral infection of HUVECs (Clonetics), cells were plated 24 hours before infection in EGM-2 media (Clonetics) at a density of 1.3×10 5  cells per well of a 6 well plate. The following day, 500 μl of virus supernatant was combined with 500 μl of EGM-2 complete media. Polybrene (Sigma) was added to a final concentration of 8.0 μg/ml. Media was aspirated from the cells and replaced with the viral mix. Cells were incubated with the viral mix in 5% CO 2  at 37° C. After 3 hours incubation, an additional 1.0 ml of EGM-2 media was added and cells were incubated for a further 24 hours. After this time HUVE cells were split 1:2 and replated into a 6 well plate. Cells were incubated for 24 hours following splitting to allow them to recover and adhere. To select for infected cells, medium was replaced with EGM-2 complete medium containing puromycin (Sigma) at a 0.4 μg/ml final concentration. Cells were incubated until uninfected cells treated with puromycin had died and infected resistant cells had grown to confluence. Media containing puromycin was replaced every 48 hours to replenish puromycin and remove cell debris. Once resistant cells were grown to confluence (approximately 4-5 days after starting selection), cells were washed in PBS, trypsinised and their properties analysed using the Matrigel capillary tube formation assay. 
     Capillary Tube Formation Assay 
     96 well tissue culture plates were coated with 50 μl of cold Matrigel (BD Biosciences) at 4° C. in a two layer process. Matrigel was allowed to polymerize at 37° C. for a minimum of 30 minutes before being used. Trypsinised cells were collected in 500 μl of EGM-2 media then centrifuged at 400 rcf for 3 minutes to pellet cells. This allows for the removal of trypsin that may interfere with the assay. Cell pellets were resuspended in 500 μl EGM-2 media then counted using a heamocytometer. Cells were diluted to 2.5×10 5  cells/ml in EGM-2 media. 100 μl of the diluted cell suspension was added to duplicate Matrigel coated wells. The final cell density was 25,000 cells/well. Plates were incubated for 22 hours in a humidified incubator at 37° C. with 5% CO 2 . Images were obtained using an Olympus BX-51 microscope with a 4× objective and Optronics MagnaFire software. Remaining cells were pelleted at 400 rcf for 3 minutes, then media was removed and pellets stored at −80° C. for extraction of RNA for real-time RT-PCR analysis (see below). For all assays performed, a vector control was included. This consisted of HUVECs undergoing the infection and selection process with virus made for the vector containing no siRNA insert. This allows for comparison of capillary tube formation ability between a control (vector) and the individual siRNA under analysis. 
     Real-Time RT-PCR Analysis 
     To determine the level of gene knock-down (mediated by the siRNAs) occurring in the HUVECs, real-time RT-PCR was employed. This involved isolation of RNA from infected cells using the RNeasy Mini or Midi kits (Qiagen) as per manufacturer&#39;s instructions (including the on-column DNase treatment). Total RNA was visualised on a 1.2% TBE agarose gel containing ethidium bromide to check for quality and purity. Total RNA concentration was determined by A 260  on a spectrophotometer. 
     For the synthesis of cDNA, total RNA (at least 1 ug and preferably at a concentration &gt;1.0 ug/ul) was reverse transcribed using M-MLV (Promega) as per manufacturer&#39;s directions. Briefly, the RNA sample to be analysed was made up to 13 ul with water and 1.0 ul of oligo-dT primer (500 ng/ul) was added. After incubating at 70° C. for 5 minutes, the tubes were placed on ice for 5 minutes and 11 ul of a pre-made master mix containing 5.0 ul M-MLV RT 5× Reaction Buffer, 1.25 ul 10 mM dNTP mix, 1.0 ul of M-MLV RT (H″ point mutant) enzyme, and 3.75 ul water was added. This mix was incubated at 40° C. for one hour, and the reaction terminated by incubating at 70° C. for 15 minutes. 
     Real-Time PCRs were run on the RotorGene™ 2000 system (Corbett Research). Reactions used AmpliTaq Gold enzyme (Applied Biosystems) and followed the manufacturers instructions. Real-Time PCR reactions were typically performed in a volume of 25 ul and consisted of 1× AmpliTaq Gold Buffer, 200 nM dNTP mix, 2.0 mM MgCl 2  (may vary for primer combination used), 0.3 uM of each primer, 1×SYBR Green mix (Cambrex BioScience Rockland Inc), 1.2 ul of AmpliTaq Gold Enzyme, and 10 ul of a 1 in 5 dilution of the cDNA template. 
     Cycling conditions were typically performed at 94° C. for 12 minutes, followed by 35 cycles of 94° C. for 15 seconds, 60° C. for 15 seconds, and 72° C. for 20 seconds. The annealing temperature of the primers may vary depending on the properties of the primers used. 
     The PCR cycling was followed by the generation of a melt curve using the RotorGene™ 2000 software where the amount of annealed product was determined by holding at each degree between 50° C. and 99° C. and measuring the absorbance. All products were run on a 1.2% agarose gel containing ethidium bromide to check specificity in addition to observing the melt curve. 
     The level of knock-down of a particular gene was then measured by a comparison of its expression level in HUVECs infected with the relevant siRNA under investigation as opposed to HUVECs infected with the retroviral vector alone. 
     In Vitro Regulation of HUVEC Function—BNO782 and BNO481 
     The siRNA oligonucleotides designed to knock-down BNO782 and BNO481 expression are represented by SEQ ID Numbers: 45-47 and SEQ ID Numbers: 48-50 respectively. Real-time RT-PCR analysis of HUVECs retrovirally infected with these siRNAs revealed that each siRNA was able to knock-down the expression of BNO782 or BNO481 to varying degrees. The level of BNO782 expression knock-down mediated by BNO782 siRNA2 (SEQ ID NO: 46) was 24% ( FIG. 3A ), while expression of BNO481 was reduced by 36% ( FIG. 3B ) using BNO481 siRNA1 (SEQ ID NO: 48). Both of these siRNAs were subsequently used separately in Matrigel assays to examine the effects that this level of knock-down for each gene had on the ability of HUVECs to participate in capillary tube formation. As can be seen in  FIG. 4 , reducing BNO782 or BNO481 mRNA levels inhibits HUVEC tube formation. Vector infected cells formed extensive networks of tube structures ( FIGS. 4A and 4C ) while cells infected with BNO782 siRNA2 or BNO481 siRNA1 exhibited tube structure networks of significantly reduced complexity with a high number of incomplete tube extensions ( FIGS. 4B and 4D ). This result confirms a role for both BNO782 and BNO481 in the process of angiogenesis. 
     Protein Interaction Studies 
     The ability of any one of the angiogenic proteins of the invention, including BNO782 and BNO481, to bind known and unknown proteins can be examined. Procedures such as the yeast two-hybrid system are used to discover and identify any functional partners. The principle behind the yeast two-hybrid procedure is that many eukaryotic transcriptional activators, including those in yeast, consist of two discrete modular domains. The first is a DNA-binding domain that binds to a specific promoter sequence and the second is an activation domain that directs the RNA polymerase II complex to transcribe the gene downstream of the DNA binding site. Both domains are required for transcriptional activation as neither domain can activate transcription on its own. In the yeast two-hybrid procedure, the gene of interest or parts thereof (BAIT), is cloned in such a way that it is expressed as a fusion to a peptide that has a DNA binding domain. A second gene, or number of genes, such as those from a cDNA library (TARGET), is cloned so that it is expressed as a fusion to an activation domain. Interaction of the protein of interest with its binding partner brings the DNA-binding peptide together with the activation domain and initiates transcription of the reporter genes. The first reporter gene will select for yeast cells that contain interacting proteins (this reporter is usually a nutritional gene required for growth on selective media). The second reporter is used for confirmation and while being expressed in response to interacting proteins it is usually not required for growth. 
     The nature of the interacting genes and proteins can also be studied such that these partners can also be targets for drug discovery. 
     Structural Studies 
     Recombinant angiogenic proteins of the invention can be produced in bacterial, yeast, insect and/or mammalian cells and used in crystallographical and NMR studies. Together with molecular modeling of the protein, structure-driven drug design can be facilitated. 
     Example 7 
     BNO802 
     In order to select for such genes an assay that involves the formation of blood capillary counterparts in an extracellular matrix medium derived from tumour cells was used. In this assay endothelial cells were incubated on an extracellular matrix derived from mouse sarcoma cells (Matrigel). Endothelial cells cultured on Matrigel matrices undergo sequential morphological changes culminating in the formation of tube structures that are thought to represent a good in vitro model of blood capillaries. This in vitro behaviour constitutes a phenotypic manifestation of a number of temporally and well-orchestrated cellular events that involve endothelial cell activation, migration and cellular remodelling. Endothelial cell proliferation is a key process underlying the early stages of blood capillary formation. Any compounds interfering with this process are likely to inhibit angiogenesis. The differentiation of endothelial cells into networks of tubes in vitro is preceded by a stop in proliferation. Laminin is one of the components of the extracellular matrix in the natural mileu of the endothelial cell as well as in the Matrigel matrix responsible for imparting anti-proliferative signals and enhancing differentiation into tube structures. Consequently, the tube formation assay does not encompass the proliferative events that take place during parts of the angiogenic process. Thus the proliferation assay was used to complement the Matrigel assay for the validation of potential angiogenesis targets. 
     Using this strategy, a gene is considered to be involved in angiogenesis, and potentially a good drug target, if the RNAi-mediated silencing of the gene resulted in the inhibition of endothelial cell capillary formation on Matrigel and/or is the inhibition of endothelial cell proliferation in culture. 
       FIG. 5  gives an evaluation of the consequences of siRNA-mediated knockdown of BNO802 on the formation of capillary tubes by endothelial cells on Matrigel. The pictures demonstrate a significant reduction in capillary formation. The observation of  FIG. 5  is quantified in the graph of  FIG. 6 , which gives an evaluation of the consequences of siRNA-mediated knockdown of BNO802 on the ability of endothelial cells to proliferate. Again, a significant knockdown is demonstrated in siRNA. 
       FIG. 7  is a realTime-RTPCR analysis evaluating the degree of BNO802 gene knockdown achieved with RNAi. Total RNA was extracted from cells and reverse transcribed into cDNA followed by RealTime PCR amplification using gene specific primers. Expression levels were normalised to the house-keeping gene POLR2K and expressed as a percent of the vector control (n=3). The analysis is as performed in example 5 and the level of knockdown of a particular gene is measured by comparison of its expression level in cells infected with the relevant siRNA as opposed to those infected with retroviral vector alone.  FIG. 7  demonstrates a reduction in expression to a level of less than 20 percent. 
       FIG. 8  provides an evaluation of BNO802 gene expression in normal human tissues using RealTime RTPCR analysis. Human RNA samples (Ambion) were reverse transcribed into cDNA followed by RealTime PCR using gene specific primers. Gene expression data was normalised to the expression of the house-keeping gene POLR2K. The level of gene expression in each tissue was expressed relative to the gene expression found in a homogeneous endothelial cell population (HUVEC) (n=4). 
     REFERENCES 
     References cited herein are listed on the following pages, and are incorporated herein by this reference.
     Altschul, S F. et al. (1997).  Nucleic Acids Res.  25: 3389-3402.   Breaker, R R. and Joyce, G F. (1995).  Chem. Biol.  2: 655-600.   Cole, S R et al. (1984).  Mol. Cell Biol.  62: 109-120.   Cote, R J. et al. (1983).  Proc. Natl. Acad. Sci. USA  80: 2026-2030.   Culver, K. (1996).  Gene Therapy: A Primer for Physicians . Second Edition. (Mary Ann Liebert).   Folkman, J. and Haudenschild, C. (1980).  Nature  ( Lond .) 288: 551-556.   Friedman, T. (1991). In  Therapy for Genetic Diseases . (T Friedman (Ed) Oxford University Press. pp 105-121.   Gamble, J R. et al. (1993).  J. Cell Biol.  121: 931-943.   Gamble, J R. et al. (1999).  Endothelium  7: 23-34.   Gecz, J. et al. (1997).  Genomics  44: 201-213.   Goldman, C K. et al. (1997).  Nature Biotechnology  15: 462-466.   Green, L L. et al. (1994).  Nature Genet.  7: 13-21.   Grishok, A. et al. (2001).  Science  287: 2494-2497.   Haseloff, J. and Gerlach, W L. (1988).  Nature  334: 585-591.   Heller, R A. et al. (1997).  Proc. Natl. Acad. Sci. USA  94: 2150-2155.   Huse, W D. et al. (1989).  Science  246: 1275-1281.   Kohler, G. and Milstein, C. (1975).  Nature  256: 495-497.   Kozbor, D. et al. (1985).  J. Immunol. Methods  81:31-42.   Lonberg, N. et al. (1994).  Nature  368: 856-859.   Meyer, G T. et al. (1997).  The Anatomical Record  249: 327-340.   Orlandi, R. et al. (1989).  Proc. Natl. Acad. Sci. USA  86: 3833-3837.   Rickert, R C. et al. (1997).  Nucleic Acids Res.  25: 1317-1318.   Sambrook, J. et al. (1989).  Molecular cloning: a laboratory manual . Second Edition. (Cold Spring Harbour Laboratory Press, New York).   Scharf, D. et al. (1994).  Results Probl. Cell Differ.  20: 125-162.   Schena, M. et al. (1996).  Proc. Natl. Acad. Sci. USA  93: 10614-10619.   Schwenk, F. et al. (1995).  Nucleic Acids Res.  23: 5080-5081.   Sharp, P A. and Zamore, P D. (2000).  Science  287: 2431-2432.   Taylor, L D. et al. (1994).  Int. Immunol.  6: 579-591.   Winter, G. et al. (1991).  Nature  349: 293-299.   

     All references listed herein including but not limited to all patents, patent applications and publications thereof, scientific journal articles, and database entries are incorporated herein by reference in their entireties to the extent that they supplement, explain, provide a background for, or teach methodology, techniques, and/or compositions employed herein. 
     It will be understood that various details of the presently disclosed subject matter may be changed without departing from the scope of the presently disclosed subject matter. Furthermore, the foregoing description is for the purpose of illustration only, and not for the purpose of limitation.