Patent Publication Number: US-2009221671-A1

Title: Modulation of lmw-ptpase expression

Description:
FIELD OF THE INVENTION 
     Disclosed herein are compounds, compositions and methods for modulating the expression of LMW-PTPase in a cell, tissue or animal. 
     BACKGROUND OF THE INVENTION 
     Considerable attention has been devoted to the characterization of tyrosine kinases and tyrosine phosphatases and their associations with disease states (Zhang, Crit. Rev. Biochem. Mol. Biol., 1998, 33, 1-52). LMW-PTPase [also known ACP1: Acid phosphatase 1, soluble, Bf isoform; Bs isoform; HAAP; HCPTP; Cytoplasmic Phosphotyrosyl Protein Phosphatase; MGC3499; RCAP; Red sell acid phosphatase 1, isozyme F; Red cell acid phosphatase 1, isozyme S; acid phosphatase of erythrocyte adipocyte acid phosphatase; low molecular weight phosphotyrosine protein phosphatase; red cell acid phosphatase 1] was originally isolated as an acid phosphatase from red blood cells and was subsequently found to be expressed in many additional tissues, including placenta, brain, kidney, liver, and leukocytes (Bryson et al., Genomics, 1995, 30, 133-140; Dissing and Svensmark, Biochim. Biophys. Acta., 1990, 1041, 232-242; Hopkinson et al., Nature, 1963, 199, 969-971; Wo et al., J. Biol. Chem., 1992, 267, 10856-10865). The LMW-PTPase locus was mapped to chromosome 2 at 2p23-p25 (Bryson et al., Genomics, 1995, 30, 133-140; Magenis et al., Birth. Defects Orig. Artic. Ser., 1976, 12, 326-327; Wakita et al., Hunt Genet., 1985, 71, 259-260) and found to contain seven exons spanning 18 kilobases (Emanuel et al., Am. J. Med. Genet., 1979, 4, 167-172: Junien et al., Hum. Genet., 1979, 48, 17-21). 
     LMW-PTPase interacts directly with insulin stimulated insulin receptors, and negatively modulates metabolic and mitogenic insulin signaling (Chiarugi et al., Biochem. Biophys. Res. Commun., 1997, 238, 676-682). A recombinant form of one LMW-PTPase isoforma, HAAPβ, dephosphorylates the adipocyte lipid binding protein (ALBP), which may be a substrate for insulin receptor kinase (Shekels et al., Protein Sci., 1992, 1, 710-721). Together, these findings suggest a role for LMW-PTPase in regulating insulin signaling. LMW-PTPase also modulates flavin mononucleotide (FMN) levels, and dephosphorylates Band 3, the erythrocyte anion transporter. These functions regulate red blood cell metabolism and integrity and account for the association between LMW-PTPase and diseases such as hemolytic favism, a disease characterized by an acute idiosyncratic hemolytic response to molecules derived from fava beans (Bottini et al, Arch. Immuol. Ther. Exp. (Warsz), 2002, 50, 95-104). 
     The most common genetic polymorphisms of LMW-PTPase (also known as ACP1) result in the occurrence of three alleles (ACP1*A, ACP1*B and ACP1*C) and account for six different genotypes, each of which exhibits strong variations in total enzymatic activity (Golden and Sensabaugh, Hum. Genet., 1986, 72, 340-343; Hopkinson et al., Nature, 1963, 199, 969-971). Numerous rare alleles have been reported, including ACP1*D, E, F, G, H, I, K, M, R, TIC1, GUA, and a silent allele, ACP1*Q0 (Miller et al., Hum. Hered, 1987, 37, 371-375). Alternative splicing accounts for two isoforms which have been labeled fast (F) and slow (S), based on their electrophoretic mobility (Dissing, Biochem. Genet., 1987, 25, 901-918). The F and S isoforms exhibit different enzymatic properties, which, coupled with differences in the ratios of these isozymes, results in variations in activity modulation (Bottini et al., Hum. Genet., 1995, 96, 629-637). 
     Protein tyrosine phosphatases are signaling molecules that regulate a variety of cellular processes, including cell growth and differentiation, cell cycle progression and growth factor signaling. For example, a number of protein tyrosine phosphatases have been implicated as negative regulators of insulin signaling (Zhang, Crit. Rev. Biochem. Mol. Biol., 1998, 33, 1-52). LMW-PTPase is a phosphotyrosine phosphatase that is involved in multiple signal transduction pathways. For example, LMW-PTPase interacts directly with insulin stimulated insulin receptors, and negatively modulates metabolic and mitogenic insulin signaling (Chiarugi et al., Biochem. Biophys. Res. Commun., 1997, 238, 676-682). A recombinant form of one LMW-PTPase isoforma, HAAPβ, dephosphorylates the adipocyte lipid binding protein (ALBP), which may be a substrate for insulin receptor kinase (Shekels et al., Protein Sci., 1992, 1, 710-721). Together, these findings suggest a role for LMW-PTPase in regulating insulin signaling. LMW-PTPase also modulates flavin mononucleotide (FMN) levels, and dephosphorylates Band 3, the erythrocyte anion transporter. These functions regulate red blood cell metabolism and integrity and account for the association between LMW-PTPase and diseases such as hemolytic favism, a disease characterized by an acute idiosyncratic hemolytic response to molecules derived from fava beans (Bottini et al., Arch. Immunol. Ther. Exp. (Warsz), 2002, 50, 95-104). 
     The most common genetic polymorphisms of LMW-PTPase (also known as ACP1) result in the occurrence of three alleles (ACP1*A, ACP1*B and ACP1*C) and account for six different genotypes, each of which exhibits strong variations in total enzymatic activity (Golden and Sensabaugh, Hum. Genet., 1986, 72, 340-343; Hopkinson et al., Nature, 1963, 199, 969-971). Numerous rare alleles have been reported, including ACP1*D, E, F, G, H, I, K, M, R, TIC1, GUA, and a silent allele, ACP1*Q0 (Miller et al., Hum. Hered., 1987, 37, 371-375). Alternative splicing accounts for two isoforms which have been labeled fast (F) and slow (S), based on their electrophoretic mobility (Dissing, Biochem. Genet., 1987, 25, 901-918). The F and S isoforms exhibit different enzymatic properties, which, coupled with differences in the ratios of these isozymes, results in variations in activity modulation (Bottini et al., Hum. Genet., 1995, 96, 629-637). LMW-PTPase genotypes, and consequently isoform levels and total enzymatic activity, show correlation to a number of disease states. (See, e.g., Bottini et al., Arch. Immunol. Ther. Exp. (Warsz), 2002, 50, 95-104). These findings, together with the evidence that LMW-PTPase participates in insulin signaling, support a role for LMW-PTPase in metabolic disorders such as diabetes. 
     Given the genetic evidence for the involvement of LMW-PTPase in human disease, pharmacological modulation of LMW-PTPase activity and/or expression is an appropriate point of therapeutic intervention in these and other pathological conditions. Currently, there are no known therapeutic agents which effectively inhibit the synthesis of LMW-PTPase. Consequently, there remains a long felt need for agents capable of effectively inhibiting LMW-PTPase function. 
     Antisense technology is an effective means for reducing the expression of LMW-PTPase and is uniquely useful in a number of therapeutic, diagnostic, and research applications. Generally, the principle behind antisense technology is that an antisense compound hybridizes to a target nucleic acid and effects the modulation of gene expression activity, or function, such as transcription or translation. The modulation of gene expression can be achieved by, for example, target RNA degradation or occupancy-based inhibition. An example of modulation of target RNA function by degradation is RNase H-based degradation of the target RNA upon hybridization with a DNA-like antisense compound. Another example of modulation of gene expression by target degradation is RNA interference (RNAi) using small interfering RNAs (siRNAs). RNAi is a form of antisense-mediated gene silencing involving the introduction of double stranded (ds)RNA-like oligonucleotides leading to the sequence-specific reduction of targeted endogenous mRNA levels. This sequence-specificity makes antisense compounds extremely attractive as tools for target validation and gene functionalization, as well as therapeutics to selectively modulate the expression of genes involved in diseases. 
     SUMMARY OF THE INVENTION 
     Disclosed herein are antisense compounds targeted to and hybridizable with a nucleic acid molecule encoding LMW-PTPase and which modulate the expression of LMW-PTPase. In a preferred embodiment the nucleic acid molecule encoding LMW-PTPase has a nucleotide sequence that is substantially similar to one or more of GenBank Accession Nos.: NM — 004300.2, NM — 007099.2, NM — 177554.1, and NT — 022327.13_ (SEQ ID NOS: 3-6, respectively), presented in table 1, below and incorporated herein by reference. In a further aspect, the antisense compounds are targeted to and hybridizable with a region of a nucleic acid molecule encoding LMW-PTPase. Still further, the antisense compounds are targeted to and hybridizable with a segment of a nucleic acid molecule encoding LMW-PTPase. Still further the antisense compounds are targeted to and hybridizable with a site of a nucleic acid molecule encoding LMW-PTPase. 
     Further disclosed herein are active target segments comprising segments of a nucleic acid molecule encoding LMW-PTPase, the active target segments being accessible to antisense hybridization, and so, suitable for antisense modulation. In one embodiment, the active target segments have been discovered herein using empirical data that is presented below, wherein at least two chimeric oligonucleotides are shown to hybridize within the active target segment and reduce expression of the target nucleic acid (hereinafter, “active antisense compound”). The at least two active antisense compounds are preferably separated by about 60 nucleobases on the nucleic acid molecule encoding LMW-PTPase. In another embodiment, antisense compounds are designed to target the active target segments and modulate expression of the nucleic acid molecule encoding LMW-PTPase. 
     In one aspect there are herein provided antisense compounds comprising sequences 12 to 35 nucleotides in length comprising at least two chemical modifications selected from a modified internucleoside linkage, a modified nucleobase or a modified sugar. Provided herein are chimeric oligonucleotides comprising a deoxynucleotide mid-region flanked on each of the 5′ and 3′ ends by wing regions, each wing region comprising at least one high affinity nucleotide. 
     In one embodiment there is herein provided chimeric oligonucleotides comprising ten deoxynucleotide mid-regions flanked on each of the 5′ and 3′ ends with wing regions comprising five 2′-O-(2-methoxyethyl) nucleotides and wherein each internucleoside linkage of the chimeric oligonucleotide is a phosphorothioate. In another embodiment there is herein provided chimeric oligonucleotides comprising fourteen deoxynucleotide mid-regions flanked on each of the 5′ and 3′ ends with wing regions comprising three locked nucleic acid nucleotides and wherein each internucleoside linkage of the chimeric oligonucleotide is a phosphorothioate. In a further embodiment there are herein provided chimeric oligonucleotides comprising fourteen deoxynucleotide mid-regions flanked on each of the 5′ and 3′ ends by wing regions comprising two 2′-O-(2-methoxyethyl) nucleotides and wherein each internucleoside linkage of the chimeric oligonucleotide is a phosphorothioate. In a further embodiment, the antisense compounds may comprise at least one 5-methylcytosine. 
     Further provided are methods of modulating the expression of LMW-PTPase in cells, tissues or animals comprising contacting said cells, tissues or animals with one or more of the compounds or compositions of the present invention. For example, in one embodiment, the compounds can be used to inhibit the expression of LMW-PTPase in cells, tissues or animals. In this aspect of the invention cells are analyzed for indicators of a decrease in expression of LMW-PTPase mRNA and/or protein by direct measurement of mRNA and/or protein levels, and/or indicators of a disease or condition, such as glucose levels, lipid levels, weight, or a combination thereof. 
     One embodiment provides methods of lowering glucose and triglycerides. Glucose may be blood, plasma or serum glucose. Triglycerides may be blood, plasma, or serum triglycerides. Another embodiment provides methods of improving insulin sensitivity. Another embodiment provides methods of lowering cholesterol. In some embodiments, cholesterol is LDL or VLDL cholesterol. An embodiment provides methods of improving glucose tolerance. 
     Other embodiments are directed to methods of ameliorating or lessening the severity of a condition in an animal comprising contacting said animal with an effective amount of an antisense compound so that expression of LMW-PTPase is inhibited and measurement of one or more physical indicator of said condition indicates a lessening of the severity of said condition. In some embodiments, the conditions include, but are not limited to, diabetes, insulin resistance, insulin deficiency, hypercholesterolemia, hyperglycemia, dyslipidemia, hyperlipidemia, hypertriglyceridemia, and hyperfattyacidemia. In some embodiments, the diabetes is type II diabetes. In another embodiment, the condition is metabolic syndrome. In another embodiment, the condition is prediabetes. In another embodiment the condition is steatosis. In one embodiment, the steatosis is steatohepatitis. In another embodiment, the steatosis is NASH. In another embodiment, the condition is a cardiovascular disease. In another embodiment, the cardiovascular disease is coronary heart disease. In another embodiment, the condition is a cardiovascular risk factor. 
     In another embodiment, there is provided a method of decreasing hepatic glucose output in an animal comprising administering an oligomeric compound of the invention. In one embodiment, the present invention provides a method of decreasing hepatic glucose-6-phosphatase expression comprising administering an oligomeric compound of the invention. Another aspect of the present invention is a method of reducing LMW-PTPase expression in liver, fat, or in both tissues. 
     Also provided are methods of ameliorating or lessening the severity of a condition in an animal comprising contacting said animal with an oligomeric compound of the invention in combination with a glucose-lowering, lipid-lowering, or anti-obesity agent to achieve an additive therapeutic effect. 
     Also provided are methods for the prevention, amelioration, and/or treatment of diabetes, type II diabetes, prediabetes, insulin resistance, insulin deficiency, hypercholesterolemia, hyperglycemia, dyslipidemia, hyperlipidemia, hypertriglyceridemia, metabolic syndrome, hyperfattyacidemia, steatosis, steatohepatitis, NASH, cardiovascular disease, coronary heart disease, a cardiovascular risk factor or combinations thereof comprising administering at least one compound of the instant invention to an individual in need of such intervention. 
     The invention also provides a method of use of the compositions of the instant invention for the preparation of a medicament for the prevention, amelioration, and/or treatment disease, especially a disease associated with and including at least one indicator of diabetes, type II diabetes, prediabetes, insulin resistance, insulin deficiency, hypercholesterolemia, hyperglycemia, dyslipidemia, hyperlipidemia, hypertriglyceridemia, metabolic syndrome, hyperfattyacidemia, steatosis, steatohepatitis, NASH, cardiovascular disease, coronary heart disease, a cardiovascular risk factor or combinations thereof. 
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     LMW-PTPase is shown to effect in vivo glucose levels, triglyceride levels, cholesterol levels, insulin sensitivity and glucose tolerance, therefore, LMW-PTPase is indicated in diseases and conditions related thereto and including, but not limited to, diabetes, type II diabetes, obesity, insulin resistance, insulin deficiency, hypercholesterolemia, hyperglycemia, hyperlipidemia, hypertriglyceridemia, hyperfattyacidemia, liver steatosis, steatohepatitis, non-alcoholic steatohepatitis, metabolic syndrome, cardiovascular disease and coronary heart disease. Provided herein are antisense compounds for the prevention, amelioration, and/or treatment of diseases and conditions relating to LMW-PTPase function. As used herein, the term “prevention” means to delay or forestall onset or development of a condition or disease for a period of time from hours to days, preferably weeks to months. As used herein, the term “amelioration” means a lessening of at least one indicator of the severity of a condition or disease. The severity of indicators may be determined by subjective or objective measures which are known to those skilled in the art. As used herein, “treatment” means to administer a composition of the invention to effect an alteration or improvement of the disease or condition. Prevention, amelioration, and/or treatment may require administration of multiple doses at regular intervals, or prior to exposure to an agent to alter the course of the condition or disease. 
     Disclosed herein are antisense compounds, including antisense oligonucleotides and other antisense compounds for use in modulating the expression of nucleic acid molecules encoding LMW-PTPase. This is accomplished by providing antisense compounds that hybridize with one or more target nucleic acid molecules encoding LMW-PTPase. As used herein, the terms “target nucleic acid” and “nucleic acid molecule encoding LMW-PTPase” have been used for convenience to encompass RNA (including pre-mRNA and mRNA or portions thereof) transcribed from DNA encoding LMW-PTPase, and also cDNA derived from such RNA. In a preferred embodiment, the target nucleic acid is an mRNA encoding LMW-PTPase. 
     Target Nucleic Acids 
     “Targeting” an antisense compound to a particular target nucleic acid molecule can be a multistep process. The process usually begins with the identification of a target nucleic acid whose expression is to be modulated. For example, the target nucleic acid can be a cellular gene (or mRNA transcribed from the gene) whose expression is associated with a particular disorder or disease state, or a nucleic acid molecule from an infectious agent. As disclosed herein, the target nucleic acid encodes LMW-PTPase and has a polynucleotide sequence that is substantially similar to one or more of SEQ ID NOS: 1-4. 
     It is also known in the art that alternative RNA transcripts can be produced from the same genomic region of DNA. These alternative transcripts are generally known as “variants.” More specifically, “pre-mRNA variants” are transcripts produced from the same genomic DNA that differ from other transcripts produced from the same genomic DNA in either their start or stop position and contain both intronic and exonic sequence. Variants can result in mRNA variants including, but not limited to, those with alternate splice junctions, or alternate initiation and termination codons. Variants in genomic and mRNA sequences can result in disease. Antisense compounds targeted to such variants are within the scope of the instant invention. 
     In accordance with the present invention are compositions and methods for modulating the expression of LMW-PTPase. Table 1 lists the GenBank accession numbers of sequences corresponding to nucleic acid molecules encoding LMW-PTPase (nt=nucleotide), the date the version of the sequence was entered in GenBank, and the corresponding SEQ ID NO in the instant application, when assigned, each of which is incorporated herein by reference. 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Gene Targets 
               
            
           
           
               
               
               
               
            
               
                 Species 
                 Genbank # 
                 Genbank Date 
                 SEQ ID NO 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Human 
                 M83653.1 
                 Apr 27 1993 
                 1 
               
               
                 Human 
                 M83654.1 
                 Apr 27 1993 
                 2 
               
               
                 Human 
                 NM_004300.2 
                 Apr 24 2003 
                 3 
               
               
                 Human 
                 NM_007099.2 
                 Apr 24 2003 
                 4 
               
               
                 Human 
                 NM_177554.1 
                 Apr 24 2003* 
                 5 
               
               
                 Human 
                 nucleotides 254496 to 268683 of NT_022327.13 
                 Oct 7 2003 
                 6 
               
               
                 Human 
                 U25847.1 
                 Jan 5 1996 
                 7 
               
               
                 Human 
                 U25848.1 
                 Jan 5 1996 
                 8 
               
               
                 Human 
                 U25849.1 
                 Jan 5 1996 
                 9 
               
               
                 Human 
                 Y16846.1 
                 Jun 16 1998 
                 10 
               
               
                 Mouse 
                 BF167197.1 
                 Oct 27 2000 
                 11 
               
               
                 Mouse 
                 NM_021330.1 
                 Oct 23 2000 
                 12 
               
               
                 Mouse 
                 the complement of nucleotides 6757610 to 
                 Oct 30 2003 
                 13 
               
               
                   
                 6776640 of NT_039548.2 
               
               
                 Mouse 
                 Y17343.1 
                 Jul 8 1998 
                 14 
               
               
                 Mouse 
                 Y17344.1 
                 Jul 8 1998 
                 355 
               
               
                 Mouse 
                 Y17345.1 
                 Jul 8 1998 
                 15 
               
               
                 Rat 
                 NM_021262.2 
                 Jan 1 2004 
                 16 
               
               
                 Rat 
                 the complement of nucleotides 905000 to 921000 
                 Sep 22 2003 
                 17 
               
               
                   
                 of NW_047759.1 
               
               
                 Rat 
                 XM_343044.1 
                 Sep 22 2003 
                 18 
               
               
                   
               
               
                 *NM_177554.1 was permanently suppressed because it is a nonsense-mediated mRNA decay (NMD) candidate. 
               
            
           
         
       
     
     Modulation of Target Expression 
     Modulation of expression of a target nucleic acid can be achieved through alteration of any number of nucleic acid (DNA or RNA) functions. “Modulation” means a perturbation of function, for example, either an increase (stimulation or induction) or a decrease (inhibition or reduction) in expression. As another example, modulation of expression can include perturbing splice site selection of pre-mRNA processing. “Expression” includes all the functions by which a gene&#39;s coded information is converted into structures present and operating in a cell. These structures include the products of transcription and translation. “Modulation of expression” means the perturbation of such functions. The functions of RNA to be modulated can include translocation functions, which include, but are not limited to, translocation of the RNA to a site of protein translation, translocation of the RNA to sites within the cell which are distant from the site of RNA synthesis, and translation of protein from the RNA. RNA processing functions that can be modulated include, but are not limited to, splicing of the RNA to yield one or more RNA species, capping of the RNA, 3′ maturation of the RNA and catalytic activity or complex formation involving the RNA which may be engaged in or facilitated by the RNA. Modulation of expression can result in the increased level of one or more nucleic acid species or the decreased level of one or more nucleic acid species, either temporally or by net steady state level. One result of such interference with target nucleic acid function is modulation of the expression of LMW-PTPase. Thus, in one embodiment modulation of expression can mean increase or decrease in target RNA or protein levels. In another embodiment modulation of expression can mean an increase or decrease of one or more RNA splice products, or a change in the ratio of two or more splice products. 
     The effect of antisense compounds of the present invention on target nucleic acid expression can be tested in any of a variety of cell types provided that the target nucleic acid is present at measurable levels. The effect of antisense compounds of the present invention on target nucleic acid expression can be routinely determined using, for example, PCR or Northern blot analysis. Cell lines are derived from both normal tissues and cell types and from cells associated with various disorders (e.g. hyperproliferative disorders). Cell lines derived from multiple tissues and species can be obtained from American Type Culture Collection (ATCC, Manassas, Va.) and other public sources, and are well known to those skilled in the art. Primary cells, or those cells which are isolated from an animal and not subjected to continuous culture, can be prepared according to methods known in the art, or obtained from various commercial suppliers. Additionally, primary cells include those obtained from donor human subjects in a clinical setting (i.e. blood donors, surgical patients). Primary cells prepared by methods known in the art. 
     Assaying Modulation of Expression 
     Modulation of LMW-PTPase expression can be assayed in a variety of ways known in the art. LMW-PTPase mRNA levels can be quantitated by, e.g., Northern blot analysis, competitive polymerase chain reaction (PCR), or real-time PCR. RNA analysis can be performed on total cellular RNA or poly(A)+mRNA by methods known in the art. Methods of RNA isolation are taught in, for example, Ausubel, F. M. et al.,  Current Protocols in Molecular Biology , Volume 1, pp. 4.1.1-4.2.9 and 4.5.1-4.5.3, John Wiley &amp; Sons, Inc., 1993. 
     Northern blot analysis is routine in the art and is taught in, for example, Ausubel, F. M. et al.,  Current Protocols in Molecular Biology , Volume 1, pp. 4.2.1-4.2.9, John Wiley &amp; Sons, Inc., 1996. Real-time quantitative (PCR) can be conveniently accomplished using the commercially available ABI PRISM™ 7700 Sequence Detection System, available from PE-Applied Biosystems, Foster City, Calif. and used according to manufacturer&#39;s instructions. The method of analysis of modulation of RNA levels is not a limitation of the instant invention. 
     Levels of a protein encoded by LMW-PTPase can be quantitated in a variety of ways well known in the art, such as immunoprecipitation, Western blot analysis (immunoblotting), ELISA or fluorescence-activated cell sorting (FACS). Antibodies directed to a protein encoded by LMW-PTPase can be identified and obtained from a variety of sources, such as the MSRS catalog of antibodies (Aerie Corporation, Birmingham, Mich.), or can be prepared via conventional antibody generation methods. Methods for preparation of polyclonal antisera are taught in, for example, Ausubel, F. M. et al.,  Current Protocols in Molecular Biology , Volume 2, pp. 11.12.1-11.12.9, John Wiley &amp; Sons, Inc., 1997. Preparation of Monoclonal Antibodies is Taught in, for Example, Ausubel, F. M. et al.,  Current Protocols in Molecular Biology , Volume 2, pp. 11.4.1-11.11.5, John Wiley &amp; Sons, Inc., 1997. 
     Immunoprecipitation methods are standard in the art and can be found at, for example, Ausubel, F. M. et al.,  Current Protocols in Molecular Biology , Volume 2, pp. 10.16.1-10.16.11, John Wiley &amp; Sons, Inc., 1998. Western blot (immunoblot) analysis is standard in the art and can be found at, for example, Ausubel, F. M. et al.,  Current Protocols in Molecular Biology , Volume 2, pp. 10.8.1-10.8.21, John Wiley &amp; Sons, inc., 1997. 
     Active Target Segments 
     The locations on the target nucleic acid defined by having at least two active antisense compounds targeted thereto are referred to as “active target segments.” An active target segment is defined by one of the at least two active antisense compounds hybridizing at the 5′ end of the active target segment and the other hybridizing at the 3′ end of the active target segment. Additional active antisense compounds may hybridize within this defined active target segment. The compounds are preferably separated by no more than about 60 nucleotides on the target sequence, more preferably no more than about 30 nucleotides on the target sequence, even more preferably the compounds are contiguous, most preferably the compounds are overlapping. There may be substantial variation in activity (e.g., as defined by percent inhibition) of the antisense compounds within an active target segment. Active antisense compounds are those that modulate the expression of their target RNA. In one of the assays provided herein, active antisense compounds inhibit expression of their target RNA at least 10%, preferably 20%. In a preferred embodiment, at least about 50%, preferably about 70% of the oligonucleotides targeted to the active target segment modulate expression of their target RNA at least 40%. In a more preferred embodiment, the level of inhibition required to define an active antisense compound is defined based on the results from the screen used to define the active target segments. One ordinarily skilled in the art will readily understand that values received from any single assay will vary in comparison to other similar assays due to assay-to-assay conditions. 
     Hybridization 
     As used herein, “hybridization” means the pairing of complementary strands of antisense compounds to their target sequence. While not limited to a particular mechanism, the most common mechanism of pairing involves hydrogen bonding, which may be Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding, between complementary nucleoside or nucleotide bases (nucleobases). For example, the natural base adenine is complementary to the natural nucleobases thymidine and uracil which pair through the formation of hydrogen bonds. The natural base guanine is complementary to the natural base 5-methyl cytosine and the artificial base known as a G-clamp. Hybridization can occur under varying circumstances. 
     An antisense compound is specifically hybridizable when there is a sufficient degree of complementarity to avoid non-specific binding of the antisense compound to non-target nucleic acid sequences under conditions in which specific binding is desired, i.e., under physiological conditions in the case of in vivo assays or therapeutic treatment, and under conditions in which assays are performed in the case of in vitro assays. 
     As used herein, “stringent hybridization conditions” or “stringent conditions” refers to conditions under which an antisense compound will hybridize to its target sequence, but to a minimal number of other sequences. Stringent conditions are sequence-dependent and will be different in different circumstances, and “stringent conditions” under which antisense compounds hybridize to a target sequence are determined by the nature and composition of the antisense compounds and the assays in which they are being investigated. 
     Complementarity 
     “Complementarity,” as used herein, refers to the capacity for precise pairing between two nucleobases on either two oligomeric compound strands or an antisense compound with its target nucleic acid. For example, if a nucleobase at a certain position of an antisense compound is capable of hydrogen bonding with a nucleobase at a certain position of a target nucleic acid, then the position of hydrogen bonding between the oligonucleotide and the target nucleic acid is considered to be a complementary position. The antisense compound and the further DNA or RNA are complementary to each other when a sufficient number of complementary positions in each molecule are occupied by nucleobases which can hydrogen bond with each other. Thus, “specifically hybridizable” and “complementary” are terms which are used to indicate a sufficient degree of precise pairing or complementarity over a sufficient number of nucleobases such that stable and specific binding occurs between the antisense compound and a target nucleic acid. 
     Those in the art understand that for an antisense compound to be active it need not be 100% complementary to the target nucleic acid site wherein it hybridizes. Often, once an antisense compound has been identified as an active antisense compound, the compounds are routinely modified to include mismatched nucleobases compared to the sequence of the target nucleic acid site. The art teaches methods for introducing mismatches into an antisense compound without substantially altering its activity. Antisense compounds may be able to tolerate up to about 20% mismatches without significant alteration of activity, particularly so when a high affinity modification accompanies the mismatches. 
     Identity 
     Antisense compounds, or a portion thereof, may have a defined percent identity to a SEQ ID NO, or a compound having a specific compound number. As used herein, a sequence is identical to the sequence disclosed herein if it has the same nucleobase pairing ability. For example, a RNA which contains uracil in place of thymidine in the disclosed sequences of the instant invention would be considered identical as they both pair with adenine. Similarly, a G-clamp modified heterocyclic base would be considered identical to a cytosine or a 5-Me cytosine in the sequences of the instant application as it pairs with a guanine. This identity may be over the entire length of the oligomeric compound, or in a portion of the antisense compound (e.g., nucleobases 1-20 of a 27-mer may be compared to a 20-mer to determine percent identity of the oligomeric compound to the SEQ ID NO.) It is understood by those skilled in the art that an antisense compound need not have an identical sequence to those described herein to function similarly to the antisense compound described herein. Shortened versions of antisense compound taught herein, or non-identical versions of the antisense compound taught herein fall within the scope of the invention. Non-identical versions are those wherein each base does not have the same pairing activity as the antisense compounds disclosed herein. Bases do not have the same pairing activity by being shorter or having at least one a basic site. Alternatively, a non-identical version can include at least one base replaced with a different base with different pairing activity (e.g., G can be replaced by C, A, or T). Percent identity is calculated according to the number of bases that have identical base pairing corresponding to the SEQ ID NO or antisense compound to which it is being compared. The non-identical bases may be adjacent to each other, dispersed through out the oligonucleotide, or both. 
     For example, a 16-mer having the same sequence as nucleobases 2-17 of a 20-mer is 80% identical to the 20-mer. Alternatively, a 20-mer containing four nucleobases not identical to the 20-mer is also 80% identical to the 20-mer. A 14-mer having the same sequence as nucleobases 1-14 of an 18-mer is 78% identical to the 18-mer. Such calculations are well within the ability of those skilled in the art. 
     The percent identity is based on the percent of nucleobases in the original sequence present in a portion of the modified sequence. Therefore, a 30 nucleobase antisense compound comprising the full sequence of the complement of a 20 nucleobase active target segment would have a portion of 100% identity with the complement of the 20 nucleobase active target segment, while further comprising an additional 10 nucleobase portion. In the context of the invention, the complement of an active target segment may constitute a single portion. In a preferred embodiment, the oligonucleotides of the instant invention are at least about 80%, more preferably at least about 85%, even more preferably at least about 90%, most preferably at least 95% identical to at least a portion of the complement of the active target segments presented herein. 
     It is well known by those skilled in the art that it is possible to increase or decrease the length of an antisense compound and/or introduce mismatch bases without eliminating activity. For example, in Woolf et al. (Proc. Natl. Acad. Sci. USA 89:7305-7309, 1992, incorporated herein by reference), a series of ASOs 13-25 nucleobases in length were tested for their ability to induce cleavage of a target RNA in an oocyte injection model. ASOs 25 nucleobases in length with 8 or 11 mismatch bases near the ends of the ASOs were able to direct specific cleavage of the target mRNA, albeit to a lesser extent than the ASOs that contained no mismatches. Similarly, target specific cleavage was achieved using a 13 nucleobase ASOs, including those with 1 or 3 mismatches. Maher and Dolnick (Nuc. Acid. Res. 16:3341-3358, 1988, incorporated herein by reference) tested a series of tandem 14 nucleobase ASOs, and a 28 and 42 nucleobase ASOs comprised of the sequence of two or three of the tandem ASOs, respectively, for their ability to arrest translation of human DBFR in a rabbit reticulocyte assay. Each of the three 14 nucleobase ASOs alone were able to inhibit translation, albeit at a more modest level than the 28 or 42 nucleobase ASOs. 
     Therapeutics 
     Antisense compounds of the invention can be used to modulate the expression of LMW-PTPase in an animal, such as a human. In one non-limiting embodiment, the methods comprise the step of administering to said animal in need of therapy for a disease or condition associated with LMW-PTPase an effective amount of an antisense compound that inhibits expression of LMW-PTPase. A disease or condition associated with LMW-PTPase includes, but is not limited to, diabetes, type II diabetes, obesity, insulin resistance, insulin deficiency, hypercholesterolemia, hyperglycemia, hyperlipidemia, hypertriglyceridemia, hyperfattyacidemia, liver steatosis, steatohepatitis, non-alcoholic steatohepatitis, metabolic syndrome, cardiovascular disease and coronary heart disease. The diseases or conditions are associated with clinical indicators that include, but are not limited to blood glucose levels, blood lipid levels, hepatic lipid levels, insulin levels, cholesterol levels, transaminase levels, electrocardiogram, glucose uptake, gluconeogenesis, insulin sensitivity, body weight and combinations thereof. In one embodiment, the antisense compounds of the present invention effectively inhibit the levels or function of LMW-PTPase RNA. Because reduction in LMW-PTPase mRNA levels can lead to alteration in LMW-PTPase protein products of expression as well, such resultant alterations can also be measured. Antisense compounds of the present invention that effectively inhibit the level or function of LMW-PTPase RNA or protein products of expression are considered an active antisense compounds. In one embodiment, the antisense compounds of the invention inhibit the expression of LMW-PTPase causing a reduction of RNA by at least 30%, by at least 40%, by at least 50%, by at least 60%, by at least 70%, by at least 75%, by at least 80%, by at least 85%, by at least 90%, by at least 95%, by at least 98%, by at least 99%, or by 100%. 
     For example, the reduction of the expression of LMW-PTPase can be measured in a bodily fluid, tissue or organ of the animal. Methods of obtaining samples for analysis, such as body fluids (e.g., blood), tissues (e.g., biopsy), or organs, and methods of preparation of the samples to allow for analysis are well known to those skilled in the art. Methods for analysis of RNA and protein levels are discussed above and are well known to those skilled in the art. The effects of treatment can be assessed by measuring biomarkers associated with the LMW-PTPase expression in the aforementioned fluids, tissues or organs, collected from an animal contacted with one or more compounds of the invention, by routine clinical methods known in the art. These biomarkers include but are not limited to: liver transaminases, bilirubin, albumin, blood urea nitrogen, creatine and other markers of kidney and liver function; glucose levels, triglyceride levels, insulin levels, fatty acid levels, cholesterol levels, electrocardiogram, glucose uptake, gloconeogenesis, insulin sensitivity and body weight, and other markers of diabetes, type II diabetes, obesity, insulin resistance, insulin deficiency, hypercholesterolemia, hyperglycemia, hyperlipidemia, hypertriglyceridemia, hyperfattyacidemia, liver steatosis, steatohepatitis, non-alcoholic steatohepatitis, metabolic syndrome, cardiovascular disease and coronary heart disease. Additionally, the effects of treatment can be assessed using non-invasive indicators of improved disease state or condition, such as electrocardiogram, body weight, and the like. 
     The antisense compounds of the present invention can be utilized in pharmaceutical compositions by adding an effective amount of a compound to a suitable pharmaceutically acceptable diluent or carrier. Acceptable carriers and diluents are well known to those skilled in the art. Selection of a dilutent or carrier is based on a number of factors, including, but not limited to, the solubility of the compound and the route of administration. Such considerations are well understood by those skilled in the art. In one aspect, the compounds of the present invention inhibit the expression of LMW-PTPase. The compounds of the invention can also be used in the manufacture of a medicament for the treatment of diseases and disorders related to LMW-PTPase expression by restoring glucose levels, triglyceride levels, insulin levels, fatty acid levels, cholesterol levels, glucose uptake, gloconeogenesis and insulin sensitivity to non-disease state profiles. 
     Methods whereby bodily fluids, organs or tissues are contacted with an effective amount of one or more of the antisense compounds or compositions of the invention are also contemplated. Bodily fluids, organs or tissues can be contacted with one or more of the compounds of the invention resulting in modulation of LMW-PTPase expression in the cells of bodily fluids, organs or tissues. 
     Kits, Research Reagents, and Diagnostics 
     The antisense compounds of the present invention can be utilized for diagnostics, and as research reagents and kits. Furthermore, antisense compounds, which are able to inhibit gene expression with specificity, are often used by those of ordinary skill to elucidate the function of particular genes or to distinguish between functions of various members of a biological pathway. 
     For use in kits and diagnostics, the antisense compounds of the present invention, either alone or in combination with other compounds or therapeutics, can be used as tools in differential and/or combinatorial analyses to elucidate expression patterns of a portion or the entire complement of genes expressed within cells and tissues. Methods of gene expression analysis are well known to those skilled in the art. 
     Antisense Compounds 
     The term “antisense compound” refers to a polymeric structure capable of hybridizing to a region of a nucleic acid molecule. As is used herein, the term “active antisense compound” is an antisense compound that has been shown to hybridize with the target nucleic acid and modulate it expression. Generally, antisense compounds comprise a plurality of monomeric subunits linked together by internucleoside linking groups and/or internucleoside linkage mimetics. Each of the monomeric subunits comprises a sugar, abasic sugar, modified sugar, or a sugar mimetic, and except for the abasic sugar includes a nucleobase, modified nucleobase or a nucleobase mimetic. Preferred monomeric subunits comprise nucleosides and modified nucleosides. An antisense compound is at least partially complementary to the region of a target nucleic acid molecule to which it hybridizes and which modulates (increases or decreases) its expression. This term includes oligonucleotides, oligonucleosides, oligonucleotide analogs, oligonucleotide mimetics, antisense compounds, antisense oligomeric compounds, and chimeric combinations of these. An “antisense oligonucleotide” is an antisense compound that is a nucleic acid-based oligomer. An antisense oligonucleotide can, in some cases, include one or more chemical modifications to the sugar, base, and/or internucleoside linkages. Nonlimiting examples of antisense compounds include antisense compounds, antisense oligonucleotides, external guide sequence (EGS) oligonucleotides, alternate splicers, and siRNAs. As such, these compounds can be introduced in the form of single-stranded, double-stranded, circular, branched or hairpins and can contain structural elements such as internal or terminal bulges or loops. In some embodiments it is desirous to take advantage of alternate antisense mechanisms (such as RNAi). Antisense compounds that use these alternate mechanisms may optionally comprise a second compound which is complementary to the antisense compound. In other words, antisense double-stranded compounds can be two strands hybridized to form double-stranded compounds or a single strand with sufficient self complementarity to allow for hybridization and formation of a fully or partially double-stranded compound. The compounds of the instant invention are not auto-catalytic. As used herein, “auto-catalytic” means a compound has the ability to promote cleavage of the target RNA in the absence of accessory factors, e.g. proteins. 
     In one embodiment of the invention, double-stranded antisense compounds encompass short interfering RNAs (siRNAs). As used herein, the term “siRNA” is defined as a double-stranded compound having a first and second strand, each strand having a central portion and two independent terminal portions. The central portion of the first strand is complementary to the central portion of the second strand, allowing hybridization of the strands. The terminal portions are independently, optionally complementary to the corresponding terminal portion of the complementary strand. The ends of the strands may be modified by the addition of one or more natural or modified nucleobases to form an overhang 
     Each strand of the siRNA duplex may be from about 12 to about 35 nucleobases. In a preferred embodiment, each strand of the siRNA duplex is about 17 to about 25 nucleobases. The two strands may be fully complementary (i.e., form a blunt ended compound), or include a 5′ or 3′ overhang on one or both strands. Double-stranded compounds can be made to include chemical modifications as discussed herein. 
     In one embodiment of the invention, the antisense compound comprises a single stranded oligonucleotide. In some embodiments of the invention the antisense compound contains chemical modifications. In a preferred embodiment, the antisense compound is a single stranded, chimeric oligonucleotide wherein the modifications of sugars, bases, and internucleoside linkages are independently selected. 
     The antisense compounds may comprise a length from about 12 to about 35 nucleobases (i.e. from about 12 to about 35 linked nucleosides). In other words, a single-stranded compound of the invention comprises from about 12 to about 35 nucleobases, and a double-stranded antisense compound of the invention (such as a siRNA, for example) comprises two strands, each of which is independently from about 12 to about 35 nucleobases. This includes oligonucleotides 15 to 35 and 16 to 35 nucleobases in length. Contained within the antisense compounds of the invention (whether single or double stranded and on at least one strand) are antisense portions. The “antisense portion” is that part of the antisense compound that is designed to work by one of the aforementioned antisense mechanisms. One of ordinary skill in the art will appreciate that about 12 to about 35 nucleobases includes 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 nucleobases. For convenience we describe antisense compounds, but one ordinarily skilled in the art will understand that analogues and mimetics can have a length within this same range. 
     Antisense compounds about 12 to 35 nucleobases in length, preferably about 15 to 35 nucleobases in length, comprising a stretch of at least eight (8), preferably at least 12, more preferably at least 15 consecutive nucleobases selected from within the active target regions are considered to be suitable antisense compounds as well. 
     Modifications can be made to the antisense compounds of the instant invention and may include conjugate groups attached to one of the termini, selected nucleobase positions, sugar positions or to one of the internucleoside linkages. Possible modifications include, but are not limited to, 2′-fluoro (2′-F), 2′-OMethyl (2′-OMe), 2′-Methoxy ethoxy (2′-MOE) sugar modifications, inverted abasic caps, deoxynucleobases, and bicyclice nucleobase analogs such as locked nucleic acids (LNA.sup™) and ENA. 
     Chemical Modifications 
     As is known in the art, a nucleoside is a base-sugar combination. The base portion of the nucleoside is normally a heterocyclic base (sometimes referred to as a “nucleobase” or simply a “base”). The two most common classes of such heterocyclic bases are the purines and the pyrimidines. Nucleotides are nucleosides that further include a phosphate group covalently linked to the sugar portion of the nucleoside. For those nucleosides that include a pentofuranosyl sugar, the phosphate group can be linked to the 2′, 3′ or 5′ hydroxyl moiety of the sugar. In forming oligonucleotides, the phosphate groups covalently link adjacent nucleosides to one another to form a linear polymeric compound. Within oligonucleotides, the phosphate groups are commonly referred to as forming the internucleoside backbone of the oligonucleotide. The normal linkage or backbone of RNA and DNA is a 3′ to 5′ phosphodiester linkage. It is often preferable to include chemical modifications in oligonucleotides to alter their activity. Chemical modifications can alter oligonucleotide activity by, for example: increasing affinity of an antisense oligonucleotide for its target RNA, increasing nuclease resistance, and/or altering the pharmacokinetics of the oligonucleotide. The use of chemistries that increase the affinity of an oligonucleotide for its target can allow for the use of shorter oligonucleotide compounds. 
     The term “nucleobase” or “heterocyclic base moiety” as used herein, refers to the heterocyclic base portion of a nucleoside. In general, a nucleobase is any group that contains one or more atom or groups of atoms capable of hydrogen bonding to a base of another nucleic acid. In addition to “unmodified” or “natural” nucleobases such as the purine nucleobases adenine (A) and guanine (G), and the pyrimidine nucleobases thymine (T), cytosine (C) and uracil (U), many modified nucleobases or nucleobase mimetics known to those skilled in the art are amenable to the present invention. The terms modified nucleobase and nucleobase mimetic can overlap but generally a modified nucleobase refers to a nucleobase that is fairly similar in structure to the parent nucleobase, such as for example a 7-deaza purine or a 5-methyl cytosine, whereas a nucleobase mimetic would include more complicated structures, such as for example a tricyclic phenoxazine nucleobase mimetic. Methods for preparation of the above noted modified nucleobases are well known to those skilled in the art. 
     Antisense compounds may also contain one or more nucleosides having modified sugar moieties. The furanosyl sugar ring of a nucleoside can be modified in a number of ways including, but not limited to, addition of a substituent group, bridging of two non-germinal ring atoms to form a bicyclic nucleic acid (BNA) and substitution of an atom or group such as —S—, —N(R)— or —C(R 1 )(R 2 ) for the ring oxygen at the 4′-position. Modified sugar moieties are well known and can be used to alter, typically increase, the affinity of the antisense compound for its target and/or increase nuclease resistance. A representative list of preferred modified sugars includes but is not limited to bicyclic modified sugars (BNA&#39;s), including LNA and ENA (4′-(CH 2 ) 2 —O-2′ bridge); and substituted sugars, especially 2′-substituted sugars having a 2′-F, 2′-OCH 2  or a 2′-O(CH 2 ) 2 —OCH 3  substituent group. Sugars can also be replaced with sugar mimetic groups among others. Methods for the preparations of modified sugars are well known to those skilled in the art. 
     Internucleoside linking groups link the nucleosides or otherwise modified monomer units together thereby forming an antisense compound. The two main classes of internucleoside linking groups are defined by the presence or absence of a phosphorus atom. Representative phosphorus containing internucleoside linkages include, but are not limited to, phosphodiesters, phosphotriesters, methylphosphonates, phosphoramidate, and phosphorothioates. Representative non-phosphorus containing internucleoside linking groups include, but are not limited to, methylenemethylimino (—CH.sub.2-N(CH.sub.3)-O—CH.sub.2-), thiodiester (—O—C(O)—S—), thionocarbamate (—O—C(O)(NH)—S—); siloxane (—O—Si(H).sub.2-O—); and N,N′-dimethylhydrazine (—CH.sub.2-N(CH.sub.3)-N(CH.sub.3)-). Antisense compounds having non-phosphorus internucleoside linking groups are referred to as oligonucleosides. Modified internucleoside linkages, compared to natural phosphodiester linkages, can be used to alter, typically increase, nuclease resistance of the antisense compound. Internucleoside linkages having a chiral atom can be prepared racemic, chiral, or as a mixture. Representative chiral internucleoside linkages include, but are not limited to, alkylphosphonates and phosphorothioates. Methods of preparation of phosphorous-containing and non-phosphorous-containing linkages are well known to those skilled in the art. 
     As used herein the term “mimetic” refers to groups that are substituted for a sugar, a nucleobase, and/or internucleoside linkage. Generally, a mimetic is used in place of the sugar or sugar-internucleoside linkage combination, and the nucleobase is maintained for hybridization to a selected target. Representative examples of a sugar mimetic include, but are not limited to, cyclohexenyl or morpholino. Representative examples of a mimetic for a sugar-internucleoside linkage combination include, but are not limited to, peptide nucleic acids (PNA) and morpholino groups linked by uncharged achiral linkages. In some instances a mimetic is used in place of the nucleobase. Representative nucleobase mimetics are well known in the art and include, but are not limited to, tricyclic phenoxazine analogs and universal bases (Berger et al., Nuc Acid Res. 2000, 28:2911-14, incorporated herein by reference). Methods of synthesis of sugar, nucleoside and nucleobase mimetics are well known to those skilled in the art. 
     As used herein the term “nucleoside” includes, nucleosides, abasic nucleosides, modified nucleosides, and nucleosides having mimetic bases and/or sugar groups. 
     In the context of this disclosure, the term “oligonucleotide” refers to an oligomeric compound which is an oligomer or polymer of ribonucleic acid (RNA) or deoxyribonucleic acid (DNA). This term includes oligonucleotides composed of naturally- and non-naturally-occurring nucleobases, sugars and covalent internucleoside linkages, possibly further including non-nucleic acid conjugates. 
     Provided are compounds having reactive phosphorus groups useful for forming internucleoside linkages including for example phosphodiester and phosphorothioate internucleoside linkages. Methods of preparation and/or purification of precursors or antisense compounds of the instant invention are not a limitation of the compositions or methods of the invention. Methods for synthesis and purification of DNA, RNA, and the antisense compounds are well known to those skilled in the art. 
     As used herein the term “chimeric antisense compound” refers to an antisense compound, having at least one sugar, nucleobase and/or internucleoside linkage that is differentially modified as compared to the other sugars, nucleobases and internucleoside linkages within the same oligomeric compound. The remainder of the sugars, nucleobases and internucleoside linkages can be independently modified or unmodified. In general a chimeric oligomeric compound will have modified nucleosides that can be in isolated positions or grouped together in regions that will define a particular motif. Any combination of modifications and or mimetic groups can comprise a chimeric oligomeric compound. 
     Chimeric antisense compounds typically contain at least one region modified so as to confer increased resistance to nuclease degradation, increased cellular uptake, and/or increased binding affinity for the target nucleic acid. An additional region of the oligomeric compound may serve as a substrate for enzymes capable of cleaving RNA:DNA or RNA:RNA hybrids. By way of example, RNase H is a cellular endonuclease that cleaves the RNA strand of an RNA:DNA duplex. Activation of RNase H, therefore, results in cleavage of the RNA target, thereby greatly enhancing the efficiency of inhibition of gene expression. Consequently, comparable results can often be obtained with shorter antisense compounds when chimeras are used, compared to for example phosphorothioate deoxyoligonucleotides hybridizing to the same target region. Cleavage of the RNA target can be routinely detected by gel electrophoresis and, if necessary, associated nucleic acid hybridization techniques known in the art. 
     Certain chimeric as well as non-chimeric antisense compounds can be further described as having a particular motif. As used herein, the term “motif” refers to the orientation of modified sugar moieties and/or sugar mimetic groups in an aitisense compound relative to like or differentially modified or unmodified nucleosides. As used herein, the terms “sugars”, “sugar moieties” and “sugar mimetic groups&#39; are used interchangeably. Such motifs include, but are not limited to, gapped motifs, alternating motifs, fully modified motifs, hemimer motifs, blockmer motifs, and positionally modified motifs. The sequence and the structure of the nucleobases and type of internucleoside linkage is not a factor in determining the motif of an antisense compound. 
     As used herein, the term “gapped motif” refers to an antisense compound comprising a contiguous sequence of nucleosides that is divided into 3 regions, an internal region (gap) flanked by two external regions (wings). The regions are differentiated from each other at least by having differentially modified sugar groups that comprise the nucleosides. In some embodiments, each modified region is uniformly modified (e.g. the modified sugar groups in a given region are identical); however, other motifs can be applied to regions. For example, the wings in a gapmer could have an alternating motif. The nucleosides located in the gap of a gapped antisense compound have sugar moieties that are different than the modified sugar moieties in each of the wings. 
     As used herein, the term “alternating motif” refers to an antisense compound comprising a contiguous sequence of nucleosides comprising two differentially sugar modified nucleosides that alternate for essentially the entire sequence of the antisense compound, or for essentially the entire sequence of a region of an antisense compound. 
     As used herein, the term “fully modified motif” refers to an antisense compound comprising a contiguous sequence of nucleosides wherein essentially each nucleoside is a sugar modified nucleoside having uniform modification. 
     As used herein, the term “hemimer motif” refers to a sequence of nucleosides that have uniform sugar moieties (identical sugars, modified or unmodified) and wherein one of the 5′-end or the 3′-end has a sequence of from 2 to 12 nucleosides that are sugar modified nucleosides that are different from the other nucleosides in the hemimer modified antisense compound. 
     As used herein, the term “blockmer motif” refers to a sequence of nucleosides that have uniform sugars (identical sugars, modified or unmodified) that is internally interrupted by a block of sugar modified nucleosides that are uniformly modified and wherein the modification is different from the other nucleosides. Methods of preparation of chimeric oligonucleotide compounds are well known to those skilled in the art. 
     As used herein, the term “positionally modified motif” comprises all other motifs. Methods of preparation of positionally modified oligonucleotide compounds are well known to those skilled in the art. 
     The compounds described herein contain one or more asymmetric centers and thus give rise to enantiomers, diastereomers, and other stereoisomeric configurations that may be defined, in terms of absolute stereochemistry, as (R) or (S), alpha. or beta., or as (D) or (L) such as for amino acids et al. This is meant to include all such possible isomers, as well as their racemic and optically pure forms. 
     In one aspect, antisense compounds are modified by covalent attachment of one or more conjugate groups. Conjugate groups may be attached by reversible or irreversible attachments. Conjugate groups may be attached directly to antisense compounds or by use of a linker. Linkers may be mono- or bifunctional linkers. Such attachment methods and linkers are well known to those skilled in the art. In general, conjugate groups are attached to antisense compounds to modify one or more properties. Such considerations are well known to those skilled in the art. 
     Oligomer Synthesis 
     Oligomerization of modified and unmodified nucleosides can be routinely performed according to literature procedures for DNA (Protocols for Oligonucleotides and Analogs, Ed. Agrawal (1993), Humana Press) and/or RNA (Scaringe, Methods (2001), 23, 206-217. Gait et al., Applications of Chemically synthesized RNA in RNA: Protein Interactions, Ed. Smith (1998), 1-36. Gallo et al., Tetrahedron (2001), 57, 5707-5713). 
     Antisense compounds can be conveniently and routinely made through the well-known technique of solid phase synthesis. Equipment for such synthesis is sold by several vendors including, for example, Applied Biosystems (Foster City, Calif.). Any other means for such synthesis known in the art may additionally or alternatively be employed. It is well known to use similar techniques to prepare oligonucleotides such as the phosphorothioates and alkylated derivatives. The invention is not limited by the method of antisense compound synthesis. 
     Oligomer Purification and Analysis 
     Methods of oligonucleotide purification and analysis are known to those skilled in the art. Analysis methods include capillary electrophoresis (CE) and electrospray-mass spectroscopy. Such synthesis and analysis methods can be performed in multi-well plates. The compositions and methods disclosed herein not limited by the method of oligomer purification. 
     Salts, Prodrugs and Bioequivalents 
     The antisense compounds may comprise any pharmaceutically acceptable salts, esters, or salts of such esters, or any other functional chemical equivalent which, upon administration to an animal including a human, is capable of providing (directly or indirectly) the biologically active metabolite or residue thereof. Accordingly, for example, the disclosure is also drawn to prodrugs and pharmaceutically acceptable salts of the antisense compounds, pharmaceutically acceptable salts of such prodrugs, and other bioequivalents. 
     The term “prodrug” indicates a therapeutic agent that is prepared in an inactive or less active form that is converted to an active form (i.e., drug) within the body or cells thereof by the action of endogenous enzymes, chemicals, and/or conditions. In particular, prodrug versions of the oligonucleotides of the invention are prepared as SATE ((S-acetyl-2-thioethyl) phosphate) derivatives according to the methods disclosed in WO 93/24510 or WO 94/26764. Prodrugs can also include antisense compounds wherein one or both ends comprise nucleobases that are cleaved (e.g., phosphodiester backbone linkages) to produce the smaller active compound. 
     The term “pharmaceutically acceptable salts” refers to physiologically and pharmaceutically acceptable salts of the antisense compounds: i.e., salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects thereto. Sodium salts of antisense oligonucleotides are useful and are well accepted for therapeutic administration to humans. In another embodiment, sodium salts of dsRNA compounds are also provided. 
     Formulations 
     The antisense compounds may also be admixed, encapsulated, conjugated or otherwise associated with other molecules, molecule structures or mixtures of compounds. 
     The antisense compounds may also include pharmaceutical compositions and formulations. The pharmaceutical compositions of the present invention may be administered in a number of ways depending upon whether local or systemic treatment is desired and upon the area to be treated. 
     The pharmaceutical formulations, which may conveniently be presented in unit dosage form, may be prepared according to conventional techniques well known in the pharmaceutical industry. Such techniques include the step of bringing into association the active ingredients with the pharmaceutical carrier(s) or excipient(s). In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredients with liquid carriers, finely divided solid carriers, or both, and then, if necessary, shaping the product (e.g., into a specific particle size for delivery). 
     A “pharmaceutical carrier” or “excipient” can be a pharmaceutically acceptable solvent, suspending agent or any other pharmacologically inert vehicle for delivering one or more nucleic acids to an animal and are known in the art. The excipient may be liquid or solid and is selected, with the planned manner of administration in mind, so as to provide for the desired bulk, consistency, etc., when combined with a nucleic acid and the other components of a given pharmaceutical composition. 
     Combinations 
     Compositions provided herein can contain two or more antisense compounds. In another related embodiment, compositions can contain one or more antisense compounds, particularly oligonucleotides, targeted to a first nucleic acid and one or more additional antisense compounds targeted to a second nucleic acid target. Alternatively, compositions can contain two or more antisense compounds targeted to different regions of the same nucleic acid target. Two or more combined compounds may be used together or sequentially. Compositions of the instant invention can also be combined with other non-antisense compound therapeutic agents. 
     Nonlimiting Disclosure and Incorporation by Reference 
     While certain compounds, compositions and methods have been described with specificity in accordance with certain embodiments, the following examples serve only as illustrations of the compounds and methods and are not intended to limit the claims of the invention. Each of the references, GenBank accession numbers, and the like recited in the present application is incorporated herein by reference in its entirety. 
     EXAMPLE 1 
     Cell Types and Transfection Methods 
     Cell types—The effect of oligomeric compounds on target nucleic acid expression was tested in one or more of the following cell types. 
     A549: The human lung carcinoma cell line A549 was obtained from the American Type Culture Collection (Manassas, Va.). A549 cells were routinely cultured in DMEM, high glucose (Invitrogen Life Technologies, Carlsbad, Calif.) supplemented with 10% fetal bovine serum, 100 units per ml penicillin, and 100 micrograms per ml streptomycin (Invitrogen Life Technologies, Carlsbad, Calif.). Cells were routinely passaged by trypsinization and dilution when they reached approximately 90% confluence. Cells were seeded into 96-well plates (Falcon-Primaria #3872) at a density of approximately 5000 cells/well for use in oligomeric compound transfection experiments. 
     b.END. The mouse brain endothelial cell line b.END was obtained from Dr. Werner Risau at the Max Plank Institute (Bad Nauheim, Germany). b.END cells were routinely cultured in DMEM, high glucose (Invitrogen Life Technologies, Carlsbad, Calif.) supplemented with 10% fetal bovine serum (Invitrogen Life Technologies, Carlsbad, Calif.). Cells were routinely passaged by trypsinization and dilution when they reached approximately 90% confluence. Cells were seeded into 96-well plates (Falcon-Primaria #353872, BD Biosciences, Bedford, Mass.) at a density of approximately 3000 cells/well for use in oligomeric compound transfection experiments. 
     A10: The rat aortic smooth muscle cell line A10 was obtained from the American Type Culture Collection (Manassas, Va.). A10 cells were routinely cultured in DMEM, high glucose (American Type Culture Collection, Manassas, Va.) supplemented with 10% fetal bovine serum (Invitrogen Life Technologies, Carlsbad, Calif.). Cells were routinely passaged by trypsinization and dilution when they reached approximately 80% confluence. Cells were seeded into 96-well plates (Falcon-Primaria #3872) at a density of approximately 2500 cells/well for use in oligomeric compound transfection experiments. 
     Primary Mouse Hepatocytes: Primary mouse hepatocytes were prepared from CD-1 mice purchased from Charles River Labs. Primary mouse hepatocytes were routinely cultured in Hepatocyte Attachment Media supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin, 1% antibiotic-antimitotic (Invitrogen Life Technologies, Carlsbad, Calif.) and 10 nM bovine insulin (Sigma-Aldrich, St. Louis, Mo.). Cells were seeded into 96-well plates (Falcon-Primaria #353872, BD Biosciences, Bedford, Mass.) coated with 0.1 mg/ml collagen at a density of approximately 10,000 cells/well for use in oligomeric compound transfection experiments. 
     Primary Rat Hepatocytes: Primary rat hepatocytes are prepared from Sprague-Dawley rats purchased from Charles River Labs (Wilmington, Mass.) and are routinely cultured in DMEM, high glucose (Invitrogen Life Technologies, Carlsbad, Calif.) supplemented with 10% fetal bovine serum (Invitrogen Life Technologies, Carlsbad, Calif.), 100 units per mL penicillin, and 100.micro.g/mL streptomycin (Invitrogen Life Technologies, Carlsbad, Calif.). Cells are seeded into 96-well plates (Falcon-Primaria #353872, BD Biosciences, Bedford, Mass.) at a density of 4000-6000 cells/well treatment with the oligomeric compounds of the invention. 
     For Northern blotting or other analysis, cells may be seeded onto 100 mm or other standard tissue culture plates and treated similarly, using appropriate volumes of medium and oligonucleotide. 
     Treatment with oligomeric compounds: When cells reach appropriate confluency, they are treated with oligonucleotide using a transfection method as described. 
     Lipofectin™ When cells reached 65-75% confluency, they were treated with oligonucleotide. Oligonucleotide was mixed with LIPOFECTIN™ Invitrogen Life Technologies, Carlsbad, Calif.) in Opti-MEM™-1 reduced serum medium (Invitrogen Life Technologies, Carlsbad, Calif.) to achieve the desired concentration of oligonucleotide and a LIPOFECTIN™ concentration of 2.5 or 3 μg/mL per 100 nM oligonucleotide. This transfection mixture was incubated at room temperature for approximately 0.5 hours. For cells grown in 96-well plates, wells were washed once with 100 μL OPTI-MEM™-1 and then treated with 130 μL of the transfection mixture. Cells grown in 24-well plates or other standard tissue culture plates are treated similarly, using appropriate volumes of medium and oligonucleotide. Cells are treated and data are obtained in duplicate or triplicate. After approximately 4-7 hours of treatment at 37° C., the medium containing the transfection mixture was replaced with fresh culture medium. Cells were harvested 16-24 hours after oligonucleotide treatment. 
     CYTOFECTIN™: When cells reached 65-75% confluency, they were treated with oligonucleotide. Oligonucleotide was mixed with CYTOFECTIN™ (Gene Therapy Systems, San Diego, Calif.) in OPTI-MEM-1.sup™ reduced serum medium (Invitrogen Life Technologies, Carlsbad, Calif.) to achieve the desired concentration of oligonucleotide and a CYTOFECTIN™ concentration of 2 or 4.micro.g/mL per 100 nM oligonucleotide. This transfection mixture was incubated at room temperature for approximately 0.5 hours. For cells grown in 96-well plates, wells were washed once with 100.micro.L OPTI-MEM-1.sup™ and then treated with 130.micro.L of the transfection mixture. Cells grown in 24-well plates or other standard tissue culture plates are treated similarly, using appropriate volumes of medium and oligonucleotide. Cells are treated and data are obtained in duplicate or triplicate. After approximately 4-7 hours of treatment at 37° C., the medium containing the transfection mixture was replaced with fresh culture medium. Cells were harvested 16-24 hours after oligonucleotide treatment. 
     Control Oligonucleotides 
     Control oligonucleotides are used to determine the optimal oligomeric compound concentration for a particular cell line. Furthermore, when oligomeric compounds of the invention are tested in oligomeric compound screening experiments or phenotypic assays, control oligonucleotides are tested in parallel with compounds of the invention. 
     
       
         
           
               
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 Control oligonucleotides for cell line testing, oligomeric 
                   
               
               
                 compound screening and phenotypic assays 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                 Species 
                   
                   
                 SEQ 
                   
               
               
                 Compound 
                 Target 
                 of 
                   
                   
                 ID 
               
               
                 No. 
                 Name 
                 Target 
                 Sequence (5′ to 3′) 
                 Motif 
                 NO 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 113131 
                 CD86 
                 Human 
                 CGTGTGTCTGTGCTAGTCCC 
                 5-10-5 
                 19 
                   
               
               
                 289865 
                 forkhead 
                 Human 
                 GGCAACGTGAACAGGTCCAA 
                 5-10-5 
                 20 
               
               
                   
                 box O1A 
               
               
                   
                 (rhabdo- 
               
               
                   
                 myosarcoma) 
               
               
                   
               
               
                 25237 
                 integrin 
                 Human 
                 GCCCATTGCTGGACATGC 
                 4-10-4 
                 21 
               
               
                   
                 beta 3 
               
               
                   
               
               
                 196103 
                 integrin 
                 Human 
                 AGCCCATTGCTGGACATGCA 
                 5-10-5 
                 22 
               
               
                   
                 beta 3 
               
               
                   
               
               
                 148715 
                 Jagged 2 
                 Human; 
                 TTGTCCCAGTCCCAGGCCTC 
                 5-105 
                 23 
               
               
                   
                   
                 Mouse; 
               
               
                   
                   
                 Rat 
               
               
                   
               
               
                 18076 
                 Jun N- 
                 Human 
                 CTTTC u CGTTGGA u C u CCCTGGG 
                 5-9-6 
                 24 
               
               
                   
                 Terminal 
               
               
                   
                 Kinase - 1 
               
               
                   
               
               
                   
               
               
                 18078 
                 Jun N- 
                 Human 
                 GTGCG u CG u CGAC u C u C u CGAAATC 
                 5-9-6 
                 25 
               
               
                   
                 Terminal 
               
               
                   
                 Kinase - 2 
               
               
                   
               
               
                 183881 
                 kinesin- 
                 Human 
                 ATCCAAGTGCTACTGTAGTA 
                 5105 
                 26 
               
               
                   
                 like 1 
               
               
                   
               
               
                 29848 
                 none 
                 none 
                 NNNNNNNNNNNNNNNNNNNN 
                 5-10-5 
                 27 
               
               
                   
               
               
                 226844 
                 Notch 
                 Human; 
                 GCCCTCCATGCTGGCACAGG 
                 5-10-5 
                 28 
               
               
                   
                 (Drosophila) 
                 Mouse 
               
               
                   
                 homolog 1 
               
               
                   
               
               
                 105990 
                 Peroxisome 
                 Human 
                 AGCAAAAGATCAATCCGTTA 
                 5-10-5 
                 29 
               
               
                   
                 proliferator- 
               
               
                   
                 activated 
               
               
                   
                 receptor 
               
               
                   
                 gamma 
               
               
                   
               
               
                 336806 
                 Raf kinase 
                 Human 
                 TACAGAAGGCTGGGCCTTGA 
                 5-10-5 
                 30 
               
               
                   
                 C 
               
               
                   
               
               
                 15770 
                 Raf kinase 
                 Mouse; 
                 ATGCATT u CTG u C u C u C u C u CAAGGA 
                 5-10-5 
                 31 
               
               
                   
                 C 
                 Murine 
               
               
                   
                   
                 sarcoma 
               
               
                   
                   
                 virus; 
               
               
                   
                   
                 Rat 
               
               
                   
               
            
           
         
       
     
     The concentration of oligonucleotide used varies from cell line to cell line. To determine the optimal oligonucleotide concentration for a particular cell line, the cells are treated with a positive control oligonucleotide at a range of concentrations. Positive controls are shown in Table 2. For human and non-human primate cells, the positive control oligonucleotide is selected from Compound No. 13650, Compound No. 336806, or Compound No. 18078. For mouse or rat cells the positive control oligonucleotide is Compound No. 15770 or Compound No. 15346. The concentration of positive control oligonucleotide that results in 80% inhibition of the target mRNA, for example, human Raf kinase C for Compound No. 13650, is then utilized as the screening concentration for new oligonucleotides in subsequent experiments for that cell line. If 80% inhibition is not achieved, the lowest concentration of positive control oligonucleotide that results in 60% inhibition of the target mRNA is then utilized as the oligonucleotide screening concentration in subsequent experiments for that cell line. If 60% inhibition is not achieved, that particular cell line is deemed as unsuitable for oligonucleotide transfection experiments. The concentrations of antisense oligonucleotides used herein are from 50 nM to 300 nM when the antisense oligonucleotide is transfected using a liposome reagent and 1.micro.M to 40.micro.M when the antisense oligonucleotide is transfected by electroporation. 
     EXAMPLE 2 
     Real-time Quantitative PCR Analysis of LMW-PTPase mRNA Levels 
     Quantitation of LMW-PTPase mRNA levels was accomplished by real-time quantitative PCR using the ABI PRISM™ 7600, 7700, or 7900 Sequence Detection System (PE-Applied Biosystems, Foster City, Calif.) according to manufacturer&#39;s instructions. 
     Prior to quantitative PCR analysis, primer-probe sets specific to the LMW-PTPase being measured were evaluated for their ability to be “multiplexed” with a GAPDH amplification reaction. After isolation the RNA is subjected to sequential reverse transcriptase (RT) reaction and real-time PCR, both of which are performed in the same well. RT and PCR reagents were obtained from Invitrogen Life Technologies (Carlsbad, Calif.). RT, real-time PCR was carried out in the same by adding 20.micro.L PCR cocktail (2.5×PCR buffer minus MgCl.sub.2, 6.6 mM MgCl.sub.2, 375.micro.M each of dATP, dCTP, dCTP and dGTP, 375 nM each of forward primer and reverse primer, 125 nM of probe, 4 Units RNAse inhibitor, 1.25 Units PLATINUM® Taq, 5 Units MuLV reverse transcriptase, and 2.5×ROX dye) to 96-well plates containing 30.micro.L total RNA solution (20-200 ng). The RT reaction was carried out by incubation for 30 minutes at 48.deg.C. Following a 10 minute incubation at 95.deg.C. to activate the PLATINUM® Taq, 40 cycles of a two-step PCR protocol were carried out: 95.deg.C. for 15 seconds (denaturation) followed by 60.deg.C. for 1.5 minutes (annealing/extension). 
     Gene target quantities obtained by RT, real-time PCR were normalized using either the expression level of GAPDH, a gene whose expression is constant, or by quantifying total RNA using RiboGreen™ (Molecular Probes, Inc. Eugene, Oreg.). GAPDH expression was quantified by RT, real-time PCR, by being run simultaneously with the target, multiplexing, or separately. Total RNA was quantified using RiboGreen™ RNA quantification reagent (Molecular Probes, Inc. Eugene, Oreg.). 
     170.micro.L of RiboGreen™ working reagent (RiboGreen™ reagent diluted 1:350 in 10 mM Tris-HCl, 1 mM EDTA, pH 7.5) was pipetted into a 96-well plate containing 30.micro.L purified cellular RNA. The plate was read in a CytoFluor 4000 (PE Applied Biosystems) with excitation at 485 nm and emission at 530 nm. 
     The GAPDH PCR probes have JOE covalently linked to the 5′ end and TAMRA or MGB covalently linked to the 3′ end, where JOE is the fluorescent reporter dye and TAMRA or MGB is the quencher dye. In some cell types, primers and probe designed to a GAPDH sequence from a different species are used to measure GAPDH expression. For example, a human GAPDH primer and probe set is used to measure GAPDH expression in monkey-derived cells and cell lines. 
     Probes and primers for use in real-time PCR were designed to hybridize to target-specific sequences. The primers and probes and the target nucleic acid sequences to which they hybridize are presented in Table 3. The target-specific PCR probes have FAM covalently linked to the 5′ end and TAMRA or MGB covalently linked to the 3′ end, where FAM is the fluorescent dye and TAMRA or MGB is the quencher dye. 
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 LMW-PTPase-specific primers and probes for use in real-time PCR 
               
            
           
           
               
               
               
               
               
               
            
               
                 Target 
                   
                 Sequence 
                   
                 SEQ ID 
                   
               
               
                 Name 
                 Species 
                 Description 
                 Sequence (5′ to 3′) 
                 NO 
               
               
                   
               
               
                 GAPDH 
                 Human 
                 Forward Primer 
                 CAACGGAT1TGGTCGTATTGG 
                 32 
                   
               
               
                   
               
               
                 GAPDH 
                 Human 
                 Reverse Primer 
                 GGCAACAATATCCACTTTACCAGAGT 
                 33 
               
               
                   
               
               
                 GAPDH 
                 Human 
                 Probe 
                 CGCCTGGTCACCAGGGCTGCT 
                 34 
               
               
                   
               
               
                 GAPDH 
                 Human 
                 Forward Primer 
                 GAAGGTGAAGGTCGGAGTC 
                 35 
               
               
                   
               
               
                 GAPDH 
                 Human 
                 Reverse Primer 
                 GAAGATGGTGATGGGATTTC 
                 36 
               
               
                   
               
               
                 GAPDH 
                 Human 
                 Probe 
                 CAAGCTTCCCGTTGTCAGCC 
                 37 
               
               
                   
               
               
                 GAPDH 
                 Human 
                 Forward Primer 
                 GAAGGTGAAGGTCGGAGTC 
                 35 
               
               
                   
               
               
                 GAPDH 
                 Human 
                 Reverse Primer 
                 GAAGATGGTGATGGGATTTC 
                 36 
               
               
                   
               
               
                 GAPDH 
                 Human 
                 Probe 
                 TGGAATCATATTGGAACATG 
                 38 
               
               
                   
               
               
                 GAPDH 
                 Mouse 
                 Forward Primer 
                 GGCAAATTCAACGGCACAGT 
                 39 
               
               
                   
               
               
                 GAPDH 
                 Mouse 
                 Reverse Primer 
                 GGGTCTCGCTCCTGGAAGAT 
                 40 
               
               
                   
               
               
                 GAPDH 
                 Mouse 
                 Probe 
                 AAGGCCGAGAATGGGAAGCTTGTCATC 
                 41 
               
               
                   
               
               
                 GAPDH 
                 Rat 
                 Forward Primer 
                 TGTFPCTAGAGACAGCCGCATCTT 
                 42 
               
               
                   
               
               
                 GAPDH 
                 Rat 
                 Reverse Primer 
                 CACCGACCTTCACCATCTTGT 
                 43 
               
               
                   
               
               
                 GAPDH 
                 Rat 
                 Probe 
                 TTGTGCAGTGCCAGCCTCGTCTCA 
                 44 
               
               
                   
               
            
           
         
       
     
     EXAMPLE 3 
     Antisense Inhibition of Human LMW-PTPase Expression by Oligomeric Compounds 
     A series of antisense compounds was designed to target different regions of human LMW-PTPase RNA, using published sequences or portions of published sequences as cited in Table 1. The designed antisense compounds are complementary to one or more of the target nucleic acids in Table 1. The start and stop sites on the target nucleic acids for each antisense compound are presented in Tables 4a, b, c and d. 
     
       
         
           
               
             
               
                 TABLE 4a 
               
             
            
               
                   
               
               
                 SEQ ID NO: 3 
               
            
           
           
               
               
               
            
               
                 Compound # 
                 Start Site 
                 Stop Site 
               
               
                   
               
            
           
           
               
               
               
            
               
                 356739 
                 65 
                 84 
               
               
                 356740 
                 73 
                 92 
               
               
                 356741 
                 78 
                 97 
               
               
                 356742 
                 87 
                 106 
               
               
                 356743 
                 103 
                 122 
               
               
                 356744 
                 117 
                 136 
               
               
                 288247 
                 127 
                 146 
               
               
                 356745 
                 132 
                 151 
               
               
                 356746 
                 148 
                 167 
               
               
                 356747 
                 170 
                 189 
               
               
                 356748 
                 190 
                 209 
               
               
                 356801 
                 291 
                 310 
               
               
                 356755 
                 312 
                 331 
               
               
                 288270 
                 328 
                 347 
               
               
                 288271 
                 333 
                 352 
               
               
                 288273 
                 338 
                 357 
               
               
                 288274 
                 340 
                 359 
               
               
                 288275 
                 343 
                 362 
               
               
                 288276 
                 345 
                 364 
               
               
                 356756 
                 353 
                 372 
               
               
                 356757 
                 381 
                 400 
               
               
                 356758 
                 415 
                 434 
               
               
                 356759 
                 441 
                 460 
               
               
                 356760 
                 451 
                 470 
               
               
                 356761 
                 459 
                 478 
               
               
                 356762 
                 464 
                 483 
               
               
                 356763 
                 473 
                 492 
               
               
                 356764 
                 489 
                 508 
               
               
                 356765 
                 524 
                 543 
               
               
                 356766 
                 536 
                 555 
               
               
                 356767 
                 547 
                 566 
               
               
                 356768 
                 567 
                 586 
               
               
                 356769 
                 591 
                 610 
               
               
                 356770 
                 601 
                 620 
               
               
                 356771 
                 619 
                 638 
               
               
                 356772 
                 637 
                 656 
               
               
                 356773 
                 668 
                 687 
               
               
                 356774 
                 727 
                 746 
               
               
                 356775 
                 746 
                 765 
               
               
                 356776 
                 751 
                 770 
               
               
                 356777 
                 757 
                 776 
               
               
                 356778 
                 840 
                 859 
               
               
                 356779 
                 860 
                 879 
               
               
                 356780 
                 873 
                 892 
               
               
                 356781 
                 888 
                 907 
               
               
                 356782 
                 905 
                 924 
               
               
                 356783 
                 961 
                 980 
               
               
                 356784 
                 1022 
                 1041 
               
               
                 356785 
                 1030 
                 1049 
               
               
                 356786 
                 1050 
                 1069 
               
               
                 356787 
                 1058 
                 1077 
               
               
                 356788 
                 1093 
                 1112 
               
               
                 356789 
                 1111 
                 1130 
               
               
                 356790 
                 1118 
                 1137 
               
               
                 356791 
                 1125 
                 1144 
               
               
                 356792 
                 1170 
                 1189 
               
               
                 356793 
                 1184 
                 1203 
               
               
                 356794 
                 1227 
                 1246 
               
               
                 356795 
                 1259 
                 1278 
               
               
                 356796 
                 1288 
                 1307 
               
               
                 356797 
                 1387 
                 1406 
               
               
                 356798 
                 1414 
                 1433 
               
               
                 356799 
                 1478 
                 1497 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 4b 
               
             
            
               
                   
               
               
                 SEQ ID NO: 4 
               
            
           
           
               
               
               
            
               
                 Compound # 
                 Start Site 
                 Stop Site 
               
               
                   
               
            
           
           
               
               
               
            
               
                 356739 
                 65 
                 84 
               
               
                 356740 
                 73 
                 92 
               
               
                 356741 
                 78 
                 97 
               
               
                 356742 
                 87 
                 106 
               
               
                 356743 
                 103 
                 122 
               
               
                 356744 
                 117 
                 136 
               
               
                 288247 
                 127 
                 146 
               
               
                 356745 
                 132 
                 151 
               
               
                 356746 
                 148 
                 167 
               
               
                 356747 
                 170 
                 189 
               
               
                 356800 
                 177 
                 196 
               
               
                 356750 
                 201 
                 220 
               
               
                 356751 
                 242 
                 261 
               
               
                 356752 
                 253 
                 272 
               
               
                 356753 
                 272 
                 291 
               
               
                 356754 
                 277 
                 296 
               
               
                 356755 
                 312 
                 331 
               
               
                 288270 
                 328 
                 347 
               
               
                 288271 
                 333 
                 352 
               
               
                 288273 
                 338 
                 357 
               
               
                 288274 
                 340 
                 359 
               
               
                 288275 
                 343 
                 362 
               
               
                 288276 
                 345 
                 364 
               
               
                 356756 
                 353 
                 372 
               
               
                 356757 
                 381 
                 400 
               
               
                 356758 
                 415 
                 434 
               
               
                 356759 
                 441 
                 460 
               
               
                 356760 
                 451 
                 470 
               
               
                 356761 
                 459 
                 478 
               
               
                 356762 
                 464 
                 483 
               
               
                 356763 
                 473 
                 492 
               
               
                 356764 
                 489 
                 508 
               
               
                 356765 
                 524 
                 543 
               
               
                 356766 
                 536 
                 555 
               
               
                 356767 
                 547 
                 566 
               
               
                 356768 
                 567 
                 586 
               
               
                 356769 
                 591 
                 610 
               
               
                 356770 
                 601 
                 620 
               
               
                 356771 
                 619 
                 638 
               
               
                 356772 
                 637 
                 656 
               
               
                 356773 
                 668 
                 687 
               
               
                 356774 
                 727 
                 746 
               
               
                 356775 
                 746 
                 765 
               
               
                 356776 
                 751 
                 770 
               
               
                 356777 
                 757 
                 776 
               
               
                 356778 
                 840 
                 859 
               
               
                 356779 
                 860 
                 879 
               
               
                 356780 
                 873 
                 892 
               
               
                 356781 
                 888 
                 907 
               
               
                 356782 
                 905 
                 924 
               
               
                 356783 
                 961 
                 980 
               
               
                 356784 
                 1022 
                 1041 
               
               
                 356785 
                 1030 
                 1049 
               
               
                 356786 
                 1050 
                 1069 
               
               
                 356787 
                 1058 
                 1077 
               
               
                 356788 
                 1093 
                 1112 
               
               
                 356789 
                 1111 
                 1130 
               
               
                 356790 
                 1118 
                 1137 
               
               
                 356791 
                 1125 
                 1144 
               
               
                 356792 
                 1170 
                 1189 
               
               
                 356793 
                 1184 
                 1203 
               
               
                 356794 
                 1227 
                 1246 
               
               
                 356795 
                 1259 
                 1278 
               
               
                 356796 
                 1288 
                 1307 
               
               
                 356797 
                 1387 
                 1406 
               
               
                 356798 
                 1414 
                 1433 
               
               
                 356799 
                 1478 
                 1497 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 4c 
               
             
            
               
                   
               
               
                 SEQ ID NO: 5 
               
            
           
           
               
               
               
            
               
                 Compound # 
                 Start Site 
                 Stop Site 
               
               
                   
               
            
           
           
               
               
               
            
               
                 356739 
                 65 
                 84 
               
               
                 356740 
                 73 
                 92 
               
               
                 356741 
                 78 
                 97 
               
               
                 356742 
                 87 
                 106 
               
               
                 356743 
                 103 
                 122 
               
               
                 356744 
                 117 
                 136 
               
               
                 288247 
                 127 
                 146 
               
               
                 356745 
                 132 
                 151 
               
               
                 356746 
                 148 
                 167 
               
               
                 356747 
                 170 
                 189 
               
               
                 356748 
                 190 
                 209 
               
               
                 356749 
                 204 
                 223 
               
               
                 356750 
                 230 
                 249 
               
               
                 356751 
                 271 
                 290 
               
               
                 356752 
                 282 
                 301 
               
               
                 356753 
                 301 
                 320 
               
               
                 356754 
                 306 
                 325 
               
               
                 356755 
                 341 
                 360 
               
               
                 288270 
                 357 
                 376 
               
               
                 288271 
                 362 
                 381 
               
               
                 288273 
                 367 
                 386 
               
               
                 288274 
                 369 
                 388 
               
               
                 288275 
                 372 
                 391 
               
               
                 288276 
                 374 
                 393 
               
               
                 356756 
                 382 
                 401 
               
               
                 356757 
                 410 
                 429 
               
               
                 356758 
                 444 
                 463 
               
               
                 356759 
                 470 
                 489 
               
               
                 356760 
                 480 
                 499 
               
               
                 356761 
                 488 
                 507 
               
               
                 356762 
                 493 
                 512 
               
               
                 356763 
                 502 
                 521 
               
               
                 356764 
                 518 
                 537 
               
               
                 356765 
                 553 
                 572 
               
               
                 356766 
                 565 
                 584 
               
               
                 356767 
                 576 
                 595 
               
               
                 356768 
                 596 
                 615 
               
               
                 356769 
                 620 
                 639 
               
               
                 356770 
                 630 
                 649 
               
               
                 356771 
                 648 
                 667 
               
               
                 356772 
                 666 
                 685 
               
               
                 356773 
                 697 
                 716 
               
               
                 356774 
                 756 
                 775 
               
               
                 356775 
                 775 
                 794 
               
               
                 356776 
                 780 
                 799 
               
               
                 356777 
                 786 
                 805 
               
               
                 356778 
                 869 
                 888 
               
               
                 356779 
                 889 
                 908 
               
               
                 356780 
                 902 
                 921 
               
               
                 356781 
                 917 
                 936 
               
               
                 356782 
                 934 
                 953 
               
               
                 356783 
                 990 
                 1009 
               
               
                 356784 
                 1051 
                 1070 
               
               
                 356785 
                 1059 
                 1078 
               
               
                 356786 
                 1079 
                 1098 
               
               
                 356787 
                 1087 
                 1106 
               
               
                 356788 
                 1122 
                 1141 
               
               
                 356789 
                 1140 
                 1159 
               
               
                 356790 
                 1147 
                 1166 
               
               
                 356791 
                 1154 
                 1173 
               
               
                 356792 
                 1199 
                 1218 
               
               
                 356793 
                 1213 
                 1232 
               
               
                 356794 
                 1256 
                 1275 
               
               
                 356795 
                 1288 
                 1307 
               
               
                 356796 
                 1317 
                 1336 
               
               
                 356797 
                 1416 
                 1435 
               
               
                 356798 
                 1443 
                 1482 
               
               
                 356799 
                 1507 
                 1526 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 4d 
               
             
            
               
                   
               
               
                 SEQ ID NO: 6 
               
            
           
           
               
               
               
            
               
                 Compound # 
                 Start Site 
                 Stop Site 
               
               
                   
               
            
           
           
               
               
               
            
               
                 356739 
                 465 
                 484 
               
               
                 356740 
                 473 
                 492 
               
               
                 356741 
                 478 
                 497 
               
               
                 356742 
                 487 
                 506 
               
               
                 356731 
                 1499 
                 1518 
               
               
                 356732 
                 2753 
                 2772 
               
               
                 356733 
                 7057 
                 7076 
               
               
                 356744 
                 7375 
                 7394 
               
               
                 288247 
                 7385 
                 7404 
               
               
                 356745 
                 7390 
                 7409 
               
               
                 356746 
                 7406 
                 7425 
               
               
                 356734 
                 7435 
                 7454 
               
               
                 356748 
                 7545 
                 7564 
               
               
                 356750 
                 7711 
                 7730 
               
               
                 356751 
                 7752 
                 7771 
               
               
                 356752 
                 7763 
                 7782 
               
               
                 356753 
                 7782 
                 7801 
               
               
                 356754 
                 7787 
                 7806 
               
               
                 356735 
                 10635 
                 10654 
               
               
                 356755 
                 10656 
                 10675 
               
               
                 288270 
                 10672 
                 10691 
               
               
                 288271 
                 10677 
                 10696 
               
               
                 288273 
                 10682 
                 10701 
               
               
                 288274 
                 10684 
                 10703 
               
               
                 288275 
                 10687 
                 10706 
               
               
                 356736 
                 10697 
                 10716 
               
               
                 356737 
                 10721 
                 10740 
               
               
                 356738 
                 12475 
                 12494 
               
               
                 356757 
                 12503 
                 12522 
               
               
                 356758 
                 12537 
                 12556 
               
               
                 356759 
                 12563 
                 12582 
               
               
                 356763 
                 12736 
                 12755 
               
               
                 356764 
                 12752 
                 12771 
               
               
                 356765 
                 12787 
                 12806 
               
               
                 356766 
                 12799 
                 12818 
               
               
                 356767 
                 12810 
                 12829 
               
               
                 356768 
                 12830 
                 12849 
               
               
                 356769 
                 12854 
                 12873 
               
               
                 356770 
                 12864 
                 12883 
               
               
                 356771 
                 12882 
                 12901 
               
               
                 356772 
                 12900 
                 12919 
               
               
                 356773 
                 12931 
                 12950 
               
               
                 356774 
                 12990 
                 13009 
               
               
                 356775 
                 13009 
                 13028 
               
               
                 356776 
                 13014 
                 13033 
               
               
                 356777 
                 13020 
                 13039 
               
               
                 356778 
                 13103 
                 13122 
               
               
                 356779 
                 13123 
                 13142 
               
               
                 356780 
                 13136 
                 13155 
               
               
                 356781 
                 13151 
                 13170 
               
               
                 356782 
                 13168 
                 13187 
               
               
                 356783 
                 13224 
                 13243 
               
               
                 356784 
                 13285 
                 13304 
               
               
                 356785 
                 13293 
                 13312 
               
               
                 356786 
                 13313 
                 13332 
               
               
                 356787 
                 13321 
                 13340 
               
               
                 356788 
                 13356 
                 13375 
               
               
                 356789 
                 13374 
                 13393 
               
               
                 356790 
                 13381 
                 13400 
               
               
                 356791 
                 13388 
                 13407 
               
               
                 356792 
                 13433 
                 13452 
               
               
                 356793 
                 13447 
                 13466 
               
               
                 356794 
                 13490 
                 13509 
               
               
                 356795 
                 13522 
                 13541 
               
               
                 356796 
                 13551 
                 13570 
               
               
                 356797 
                 13650 
                 13669 
               
               
                 356798 
                 13677 
                 13696 
               
               
                 356799 
                 13741 
                 13760 
               
               
                   
               
            
           
         
       
     
     As stated above, antisense oligonucleotides directed to a target or more preferably to an active target segment can be from about 13 to about 80 linked nucleobases. The following Table 4e provides a non-limiting example of such antisense oligonucleotides targeting SEQ ID NO 1. 
     
       
         
           
               
             
               
                 TABLE 4e 
               
             
            
               
                   
               
               
                 Antisense Oligonucleotides from about 13 to about 35 Nucleobases 
               
            
           
           
               
               
            
               
                 Sequence 
                 Length 
               
               
                   
               
            
           
           
               
               
               
            
               
                         CCATGATTTCTTAGGCAGCT 
                 20 nucleobases (SEQ D NO: 76) 
                   
               
               
                   
               
               
                     AATGCCATGATTTCT 
                 15 nucleobases (SEQ ID NO: 356) 
               
               
                   
               
               
                        GCCATGATTTCTTAC 
                 15 nucleobases (SEQ ID NO: 357) 
               
               
                   
               
               
                      ATGCCATGATTTC 
                 13 nucleobases (SEQ ID NO: 358) 
               
               
                   
               
               
                     AATGCCATGATTTCTTAGGCAGCTC 
                 24 nucleobases (SEQ ID NO: 359) 
               
               
                   
               
               
                               TTTCTTAGGCAGCT 
                 14 nucleobases (SEQ ID NO: 360) 
               
               
                   
               
               
                 TGTGAATGCCATGATTTCTTAGGCAGCTCACAGCT 
                 35 nucleobases (SEQ ID NO: 361) 
               
               
                   
               
               
                 CCATGATTTCTTAGGCAGCTCACAGCT 
                 27 nucleobases (SEQ ID NO: 362) 
               
               
                   
               
               
                 GATTTCTTAGGCAGCTCACAGCT 
                 22 nucleobases (SEQ ID NO: 363) 
               
               
                   
               
            
           
         
       
     
     Antisense oligonucleotides directed to a target or more preferably to an active target segment can also contain mismatched nucleobases when compared to the target sequence. The following Table 4f provides a non-limiting example of such antisense oligonucleotides targeting nucleobases 282 to 301 of SEQ ID NO 5. Mismatched nucleobases are underlined. One ordinarily skilled in the art understands that antisense compounds can tolerate mismatches yet still retain their ability to hybridize with a target site and modulate the target nucleic acid through antisense mechanisms. 
     
       
         
           
               
               
             
               
                 TABLE 4f 
               
             
            
               
                   
               
               
                 Antisense Oligonucleotides from about 1-3 
                   
               
               
                 Nucleobases Mismatched to the Target Sequence 
               
            
           
           
               
               
               
            
               
                   
                 Number of mismatches to 
                   
               
               
                 Sequence 
                 SEQ ID NO:5 
               
               
                   
               
               
                 CCATGATTTCTTAGGCAGCT 
                 None 
                   
               
               
                 (SEQ ID NO: 76) 
               
               
                   
               
               
                 CCATGATTTCTTAGGCATCT 
                 One mismatch 
               
               
                 (SEQ ID NO: 364) 
               
               
                   
               
               
                 CCATGATCTCTTAGGCAGCT 
                 One mismatch 
               
               
                 (SEQ ID NO: 365) 
               
               
                   
               
               
                 CCATGATTTCTTAGGCACAT 
                 Two mismatches 
               
               
                 (SEQ ID NO: 366) 
               
               
                   
               
               
                 GCATGATTTCTTAGGCCGCT 
                 Two mismatches 
               
               
                 (SEQ ID NO: 367) 
               
               
                   
               
               
                 CGTTGATACCTTAGGCAGCT 
                 Three mismatches 
               
               
                 (SEQ ID NO: 368) 
               
               
                   
               
            
           
         
       
     
     Antisense compounds were designed against one or more of the human LMW-PTPase target nucleic acids sequences published in table 1 and were screened in vitro to determine the compound&#39;s ability to modulate expression of a target nucleic acid that encodes LMW-PTPase. The compounds shown in Table 5 are all chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of 10 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′) by five-nucleotide “wings”. The wings are composed of 2′-O-(2-methoxyethyl) nucleotides, also known as 2′-MOE nucleotides. The internucleoside (backbone) linkages are phosphorothioate throughout the oligonucleotide. All cytidine residues are 5-methylcytidines. The compounds were analyzed for their effect on gene target mRNA levels by quantitative real-time PCR as described in other examples herein, using the primer-probe set designed to hybridize to human LMW-PTPase (Table 2). Data are averages from two experiments in which A549 cells were treated with 65 nM of the disclosed oligomeric compounds using LIPOFECTIN™. A reduction in expression is expressed as percent inhibition in Table 5. If present, “N.D.” indicates “not determined”. The control oligomeric compound used was SEQ ID NO: 25. 
     
       
         
           
               
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 Inhibition of human LMW-PTPase inRNA levels by chimeric 
                   
               
               
                 oligonucleotides having 2′-MOE wings and deoxy gap 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 Target 
                   
                   
                   
                   
                   
               
               
                 Compound 
                 SEQ ID 
                 Target 
                   
                   
                 SEQ ID 
               
               
                 No. 
                 NO 
                 Site 
                 Sequence (5′ to 3′) 
                 % Inhib 
                 NO 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 356301 
                 3 
                 291 
                 CTTTGGTAATCTGCCGGGCA 
                 36 
                 54 
                   
               
               
                   
               
               
                 288247 
                 4 
                 127 
                 ACTGCTTCTGCAATGGGTGA 
                 67 
                 55 
               
               
                   
               
               
                 356800 
                 4 
                 177 
                 CAATGACCCAATTCTCTGAG 
                 16 
                 56 
               
               
                   
               
               
                 288270 
                 4 
                 328 
                 TCCATACATAGTATATAATC 
                 39 
                 57 
               
               
                   
               
               
                 288271 
                 4 
                 333 
                 TTTCATCCATACATAGTATA 
                 29 
                 58 
               
               
                   
               
               
                 288273 
                 4 
                 338 
                 ATTGCTTTCATCCATACATA 
                 54 
                 59 
               
               
                   
               
               
                 288274 
                 4 
                 340 
                 AGATTGCTTTGATCCATACA 
                 51 
                 60 
               
               
                   
               
               
                 288275 
                 4 
                 343 
                 CTCAGATTGCTTTCATCCAT 
                 63 
                 61 
               
               
                   
               
               
                 288276 
                 4 
                 345 
                 CTCTCAGATTGCTTTCATCC 
                 56 
                 62 
               
               
                   
               
               
                 356739 
                 5 
                 65 
                 AGCCTGTTCCGCCATCTTCC 
                 74 
                 63 
               
               
                   
               
               
                 356740 
                 5 
                 73 
                 GACTTGGTAGCCTGTTCCGC 
                 67 
                 64 
               
               
                   
               
               
                 356741 
                 5 
                 78 
                 GCACGGACTTGGTAGGCTGT 
                 68 
                 65 
               
               
                   
               
               
                 356742 
                 5 
                 87 
                 ACACAAACAGCACGGACTTG 
                 35 
                 66 
               
               
                   
               
               
                 356743 
                 5 
                 103 
                 CAAATGTTACCCAGACACAC 
                 33 
                 67 
               
               
                   
               
               
                 356744 
                 5 
                 117 
                 CAATGGGTGATCGACAAATG 
                 55 
                 68 
               
               
                   
               
               
                 356745 
                 5 
                 132 
                 TGAAAACTGCTTCTGCAATG 
                 54 
                 69 
               
               
                   
               
               
                 356746 
                 5 
                 148 
                 TCGGTTACAAGTTTCCTGAA 
                 64 
                 70 
               
               
                   
               
               
                 356747 
                 5 
                 170 
                 CCAATTCTCTGAGATGTTTT 
                 68 
                 71 
               
               
                   
               
               
                 356748 
                 5 
                 190 
                 GTTGCCGCGCTGTCTACCCT 
                 70 
                 72 
               
               
                   
               
               
                 356749 
                 5 
                 204 
                 ATGACCCACCGGAAGTTGCC 
                 22 
                 73 
               
               
                   
               
               
                 356750 
                 5 
                 230 
                 TCCAGTCAGAAACAGCACCG 
                 50 
                 74 
               
               
                   
               
               
                 356751 
                 5 
                 271 
                 TAGGCAGCTCACAGCTCTTG 
                 59 
                 75 
               
               
                   
               
               
                 356752 
                 5 
                 282 
                 CCATGTTTCTTAGGCAGCT 
                 76 
                 76 
               
               
                   
               
               
                 356753 
                 5 
                 301 
                 TTTATGGGCTGTGTGAATGC 
                 56 
                 77 
               
               
                   
               
               
                 356754 
                 5 
                 306 
                 CTTGCTTTATGGGCTGTGTG 
                 70 
                 78 
               
               
                   
               
               
                 356755 
                 5 
                 341 
                 AATCAAATGTGGCAAAATCT 
                 51 
                 79 
               
               
                   
               
               
                 356756 
                 5 
                 382 
                 ATTCAAATCTCTCAGATTGC 
                 52 
                 80 
               
               
                   
               
               
                 356757 
                 5 
                 410 
                 TGCAGGTTTTAACTTGATTA 
                 57 
                 81 
               
               
                   
               
               
                 356758 
                 5 
                 444 
                 GGATCATAGCTCCCAAGTAG 
                 57 
                 82 
               
               
                   
               
               
                 356759 
                 5 
                 470 
                 GATCTTCAATAATAAGTTGT 
                 55 
                 83 
               
               
                   
               
               
                 356760 
                 5 
                 480 
                 CCATAATAGGGATCTTCAAT 
                 26 
                 84 
               
               
                   
               
               
                 356761 
                 5 
                 488 
                 AGTCATTCCCATAATAGGGA 
                 52 
                 85 
               
               
                   
               
               
                 356762 
                 5 
                 493 
                 GTCAGAGTCATTCCCATAAT 
                 60 
                 86 
               
               
                   
               
               
                 356763 
                 5 
                 502 
                 CGTCTCAAAGTCAGAGTCAT 
                 62 
                 87 
               
               
                   
               
               
                 356764 
                 5 
                 518 
                 CACACTGCTGGTACACCGTC 
                 74 
                 88 
               
               
                   
               
               
                 356765 
                 5 
                 553 
                 GTGGGCCTTCTCCAAGAACG 
                 43 
                 89 
               
               
                   
               
               
                 356766 
                 5 
                 565 
                 GAACCTGCCTCAGTGGGCCT 
                 74 
                 90 
               
               
                   
               
               
                 356767 
                 5 
                 576 
                 CAGCAGGGCACGAACCTGCC 
                 38 
                 91 
               
               
                   
               
               
                 356768 
                 5 
                 596 
                 GGGTCTAGTCAGGCTGGCCG 
                 67 
                 92 
               
               
                   
               
               
                 356769 
                 5 
                 620 
                 TGAGAAATGCAGGACCTCAG 
                 80 
                 93 
               
               
                   
               
               
                 356770 
                 5 
                 630 
                 ACACACCGACTGAGAAATGC 
                 64 
                 94 
               
               
                   
               
               
                 356771 
                 5 
                 648 
                 GGGCCCTGGAACGTGATTAC 
                 55 
                 95 
               
               
                   
               
               
                 356772 
                 5 
                 666 
                 AACAAAGAGCTGGGCTTTGG 
                 71 
                 96 
               
               
                   
               
               
                 356773 
                 5 
                 697 
                 CTTTTTAAGGTAAGAAACAG 
                 25 
                 97 
               
               
                   
               
               
                 356774 
                 5 
                 756 
                 TGAATCAAAGATTTTTAYPG 
                 15 
                 98 
               
               
                   
               
               
                 356775 
                 5 
                 775 
                 AAATACCCCATAAGCTGTCT 
                 45 
                 99 
               
               
                   
               
               
                 356776 
                 5 
                 780 
                 GCTTAAAATACCCCATAAGC 
                 61 
                 100 
               
               
                   
               
               
                 356777 
                 5 
                 786 
                 AAGAATGCTTAAAATACCCC 
                 27 
                 101 
               
               
                   
               
               
                 356778 
                 5 
                 869 
                 CAAGTGAGGTTTTCCTTCAT 
                 67 
                 102 
               
               
                   
               
               
                 356779 
                 5 
                 889 
                 TAGATGTTGACCTGGGCCTT 
                 61 
                 103 
               
               
                   
               
               
                 356780 
                 5 
                 902 
                 GTCTCAACAGGGTTAGATGT 
                 53 
                 104 
               
               
                   
               
               
                 356781 
                 5 
                 917 
                 GACTCGATTATCTAAGTCTC 
                 57 
                 105 
               
               
                   
               
               
                 356782 
                 5 
                 934 
                 AACCTACTGAAGAGGTAGAC 
                 76 
                 106 
               
               
                   
               
               
                 356783 
                 5 
                 990 
                 AAGAGAGAGGTAGCACTGGG 
                 45 
                 107 
               
               
                   
               
               
                 356784 
                 5 
                 1051 
                 CTAATCTAGACTGTGAGCTC 
                 79 
                 108 
               
               
                   
               
               
                 356785 
                 5 
                 1059 
                 AAACACTTCTAATCTAGACT 
                 21 
                 109 
               
               
                   
               
               
                 356786 
                 5 
                 1079 
                 CTATGGGTGTGTAGAAATTA 
                 67 
                 110 
               
               
                   
               
               
                 356787 
                 5 
                 1087 
                 AGTGTGCACTATGGGTGTGT 
                 51 
                 111 
               
               
                   
               
               
                 356788 
                 5 
                 1122 
                 AAATGTTTCTCTCTTCCCTA 
                 31 
                 112 
               
               
                   
               
               
                 356789 
                 5 
                 1140 
                 GCCAAGGACTGATTCCATAA 
                 87 
                 113 
               
               
                   
               
               
                 356790 
                 5 
                 1147 
                 TGAAGGTGCCAACGACTGAT 
                 79 
                 114 
               
               
                   
               
               
                 356791 
                 5 
                 1154 
                 GAAGTATTGAAGGTGCGAAC 
                 80 
                 115 
               
               
                   
               
               
                 356792 
                 5 
                 1199 
                 GCCAATGGGCTGACCTCCTC 
                 77 
                 116 
               
               
                   
               
               
                 356793 
                 5 
                 1213 
                 TGGTTCAGATGGGAGCCAAT 
                 66 
                 117 
               
               
                   
               
               
                 356794 
                 5 
                 1256 
                 AAGTGTCCTTCTTTCTGGAT 
                 67 
                 118 
               
               
                   
               
               
                 356795 
                 5 
                 1288 
                 CATATTCCTCAACTGACCAT 
                 31 
                 119 
               
               
                   
               
               
                 356796 
                 5 
                 1317 
                 TTTGGGTTACATGTGCATAT 
                 73 
                 120 
               
               
                   
               
               
                 356797 
                 5 
                 1416 
                 TGATGAAGAATACTTATTCA 
                 49 
                 121 
               
               
                   
               
               
                 356798 
                 5 
                 1443 
                 ACATCTGCCTATACATTTAT 
                 24 
                 122 
               
               
                   
               
               
                 356799 
                 5 
                 1507 
                 TCCCCAGTTTATTTTGAAAT 
                 37 
                 123 
               
               
                   
               
               
                 356731 
                 6 
                 1499 
                 GGAAGCAACTCATGATCTGG 
                 63 
                 124 
               
               
                   
               
               
                 356732 
                 6 
                 2753 
                 AATGCATGCCATATAGTAGA 
                 43 
                 125 
               
               
                   
               
               
                 356733 
                 6 
                 7057 
                 CTAATGATCCAGGAGTGAAT 
                 39 
                 126 
               
               
                   
               
               
                 356734 
                 6 
                 7435 
                 TGGTACTTACATTCTCTGAG 
                 23 
                 127 
               
               
                   
               
               
                 356735 
                 6 
                 10635 
                 CTTTGGTAATCTAAAATTGA 
                 15 
                 128 
               
               
                   
               
               
                 356736 
                 6 
                 10697 
                 ACAGGATTACCTCAGATTGC 
                 53 
                 129 
               
               
                   
               
               
                 356737 
                 6 
                 10721 
                 GTTGAACAGAAATATTCTTC 
                 13 
                 130 
               
               
                   
               
               
                 356738 
                 6 
                 12475 
                 ATTCAAATCTCTGTAAAATT 
                 14 
                 131 
               
               
                   
               
            
           
         
       
     
     The screen identified active target segments within the human LMW-PTPase mRNA sequence, specifically SEQ ID NOS: 3, 4 and 5. Each active target segment was targeted by at least one active antisense oligonucleotide. These active target regions identified for SEQ ID NO: 3 include nucleotides 1111 to 1189 (Region A) with an average inhibition of 80.5%, nucleotides 489 to 656 (Region B) with an average inhibition of 62.8%, nucleotides 536 to 656 (Region C) with an average inhibition of 64.1%, nucleotides 489 to 610 (Region D) with an average inhibition of 62.6%, nucleotides 1111 to 1203 (Region E) with an average inhibition of 77.6%, nucleotides 840 to 924 (Region F) with an average inhibition of 62.8%, nucleotides 1022 to 1069 (Region G) with an average inhibition of 55.6%, nucleotides 65 to 209 (Region H) with an average inhibition of 59.5%, nucleotides 65 to 136 (Region I) with an average inhibition of 55.2%, nucleotides 117 to 209 (Region J) with an average inhibition of 63.0% and nucleotides 338 to 460 (Region K) with an average inhibition of 55.6%. Over half of the oligonucleotides tested in this region inhibited expression by greater than 63%. Identification of these regions allows for the design of antisense oligonucleotides that modulate the expression of LMW-PTPase. 
     The active target regions identified for SEQ ID NO: 4 include nucleotides 65 to 189 (Region AA) with an average inhibition of 58.4%, nucleotides 65 to 146 (Region AB) with an average inhibition of 56.8%, nucleotides 127 to 189 (Region AC) with an average inhibition of 63.2%, nucleotides 338 to 460 (Region AD) with an average inhibition of 55.6%, nucleotides 489 to 610 Region AE) with an average inhibition of 62.6%, nucleotides 536 to 656 (Region AF) with an average inhibition of 64.1%, nucleotides 489 to 656 (Region AG) with an average inhibition of 62.8%, nucleotides 840 to 924 (Region AH) with an average inhibition of 62.8%, nucleotides 1022 to 1069 (Region AI) with an average inhibition of 55.6%, nucleotides 1111 to 1189 (Region AJ) with an average inhibition of 80.5% and nucleotides 1111 to 1203 (Region AK) with an average inhibition of 77.6%. 
     Active target regions have also been identified for SEQ ID NO: 5. These active target regions include nucleotides 65 to 136 (Region BA) with an average inhibition of 55.2%, nucleotides 117 to 209 (Region BB) with an average inhibition of 63.0%, nucleotides 65 to 209 (Region BC) with an average inhibition of 59.5%, nucleotides 367 to 489 (Region BD) with an average inhibition of 55.6%, nucleotides 518 to 639 (Region BE) with an average inhibition of 62.6%, nucleotides 565 to 685 (Region BF) with an average inhibition of 64.1%, nucleotides 518 to 685 (Region BG) with an average inhibition of 62.8%, nucleotides 689 to 953 (Region BH) with an average inhibition of 62.8%, nucleotides 1051 to 1098 (Region BI) with an average inhibition of 55.6%, nucleotides 1140 to 1218 (Region BJ) with an average inhibition of 80.53% and nucleotides 1140 to 1232 (Region BK) with an average inhibition of 77.6%. 
     EXAMPLE 4 
     Antisense Inhibition of Mouse LMW-PTPase Expression by Oligomeric Compounds 
     Antisense compounds were designed against one or more of the mouse LMW-PTPase target nucleic acid sequences cited in Table 1 and were screened in vitro to determine the compound&#39;s ability to modulate expression of a target nucleic acid that encodes LMW-PTPase. The compounds shown in Table 6 are all chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of 10 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′) by five-nucleotide “wings”. The wings are composed of 2′-O-(2-methoxyethyl) nucleotides, also known as 2′-MOE nucleotides. The internucleoside (backbone) linkages are phosphorothioate throughout the oligonucleotide. All cytidine residues are 5-methylcytidines. The compounds were analyzed for their effect on gene target mRNA levels by quantitative real-time PCR as described in other examples herein, using the primer-probe set designed to hybridize to mouse LMW-PTPase (Table 2). Data are averages from two experiments in which b.END cells were treated with 75 nM of the disclosed oligomeric compounds using LIPOFECTIN™. A reduction in expression is expressed as percent inhibition in Table 6. The control oligomeric compound used was SEQ ID NO: 25. If present, “N.D.” indicates “not determined”. 
     
       
         
           
               
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                 Inhibition of mouse LMW-PTPase mRNA levels by chimeric 
                   
               
               
                 oligonucleotides having 2′-MOE wings and deoxy gap 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 Target 
                   
                   
                   
                   
                   
               
               
                 Compound 
                 SEQ ID 
                 Target 
                   
                   
                 SEQ ID 
               
               
                 No. 
                 NO 
                 Site 
                 Sequence (5′ to 3′) 
                 % Inhib 
                 NO 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 288290 
                 11 
                 207 
                 CTGTCTGACTCAAATGCTTT 
                 69 
                 132 
                   
               
               
                   
               
               
                 288291 
                 11 
                 252 
                 GGTCAGAGGTTTAGTTAGTC 
                 80 
                 133 
               
               
                   
               
               
                 288292 
                 11 
                 493 
                 TCCGTCTGCGGTTTTATGTA 
                 4 
                 134 
               
               
                   
               
               
                 288293 
                 11 
                 982 
                 GTGGTGCTCTGTTGAGGTGT 
                 0 
                 135 
               
               
                   
               
               
                 288216 
                 12 
                 3 
                 TTGCTTAGTCTATAACTGAC 
                 0 
                 136 
               
               
                   
               
               
                 288217 
                 12 
                 13 
                 ATGATGGAGATTGCTTAGTC 
                 0 
                 137 
               
               
                   
               
               
                 288218 
                 12 
                 24 
                 AAATATGCTAAATGATGGAG 
                 0 
                 138 
               
               
                   
               
               
                 288219 
                 12 
                 40 
                 GCTTCCTGTGCACCAGAAAT 
                 64 
                 139 
               
               
                   
               
               
                 288220 
                 12 
                 48 
                 CTCACGTTGCTTCCTGTGCA 
                 0 
                 140 
               
               
                   
               
               
                 288221 
                 12 
                 79 
                 ACTTTGTAATGGGAGTAGAT 
                 3 
                 141 
               
               
                   
               
               
                 288222 
                 12 
                 90 
                 TATAATGGTAGACTTPGTAA 
                 0 
                 142 
               
               
                   
               
               
                 288223 
                 12 
                 114 
                 TAGAGAATGCAAGCATATCA 
                 0 
                 143 
               
               
                   
               
               
                 288224 
                 12 
                 123 
                 TTCAATTAATAGAGAATGCA 
                 0 
                 144 
               
               
                   
               
               
                 288225 
                 12 
                 149 
                 ACATATACACATGAGTTGTA 
                 0 
                 145 
               
               
                   
               
               
                 288226 
                 12 
                 159 
                 CTTTGTAATGACATATACAC 
                 0 
                 146 
               
               
                   
               
               
                 288227 
                 12 
                 164 
                 AAACTCTTTGTAATGACATA 
                 0 
                 147 
               
               
                   
               
               
                 288228 
                 12 
                 179 
                 TGCTTCCATGAAGCAAAACT 
                 0 
                 148 
               
               
                   
               
               
                 288229 
                 12 
                 189 
                 GATACTTTCATGCTTCCATG 
                 0 
                 149 
               
               
                   
               
               
                 288230 
                 12 
                 196 
                 AATATGTGATACTTTCATGC 
                 0 
                 150 
               
               
                   
               
               
                 288231 
                 12 
                 202 
                 GCCATAAATATGTGATACTT 
                 0 
                 151 
               
               
                   
               
               
                 288232 
                 12 
                 232 
                 AGACCCTCAATTTCTCTAAT 
                 14 
                 152 
               
               
                   
               
               
                 288233 
                 12 
                 248 
                 TGCCATGTTTCGGTGCAGAC 
                 75 
                 153 
               
               
                   
               
               
                 288234 
                 12 
                 253 
                 ACCTCTGCCATGTIICGGTG 
                 64 
                 154 
               
               
                   
               
               
                 288235 
                 12 
                 266 
                 TGACTTGGACCCAACCTCTG 
                 59 
                 155 
               
               
                   
               
               
                 288236 
                 12 
                 273 
                 ACAGGACTGACTTGGACCCA 
                 75 
                 156 
               
               
                   
               
               
                 288237 
                 12 
                 277 
                 ACGAACAGCACTGACTTGGA 
                 61 
                 157 
               
               
                   
               
               
                 288238 
                 12 
                 281 
                 ACACACGAACAGCACTGACT 
                 62 
                 158 
               
               
                   
               
               
                 288239 
                 12 
                 289 
                 TTACCGAGACACAGGAACAG 
                 52 
                 159 
               
               
                   
               
               
                 288240 
                 12 
                 293 
                 AATGTTACCGAGACACACGA 
                 53 
                 160 
               
               
                   
               
               
                 288241 
                 12 
                 296 
                 GCAAATGTTACCGAGACACA 
                 56 
                 161 
               
               
                   
               
               
                 288242 
                 12 
                 304 
                 GGTGACCGGCAAATGTTACC 
                 68 
                 162 
               
               
                   
               
               
                 288243 
                 12 
                 306 
                 TGGGTGACCGGCAAATGTTA 
                 61 
                 163 
               
               
                   
               
               
                 288244 
                 12 
                 309 
                 CAATGGGTGACCGGCAAATG 
                 62 
                 164 
               
               
                   
               
               
                 288245 
                 12 
                 310 
                 GCAATGGGTGACCGGCAAAT 
                 81 
                 165 
               
               
                   
               
               
                 288246 
                 12 
                 317 
                 TGCTTCTGCAATGGGTGACC 
                 77 
                 166 
               
               
                   
               
               
                 288247 
                 12 
                 319 
                 ACTGCTTCTGCAATGGGTGA 
                 83 
                 55 
               
               
                   
               
               
                 288248 
                 12 
                 321 
                 ATACTGCTTCTGCAATGGGT 
                 73 
                 167 
               
               
                   
               
               
                 288249 
                 12 
                 323 
                 GAATACTGCTTCTGCAATGG 
                 76 
                 168 
               
               
                   
               
               
                 288250 
                 12 
                 325 
                 CTGAATACTGCTTCTGCAAT 
                 59 
                 169 
               
               
                   
               
               
                 288251 
                 12 
                 328 
                 TTCCTGAATACTGCTTCTGC 
                 73 
                 170 
               
               
                   
               
               
                 288252 
                 12 
                 334 
                 ACCAGTTTCGTGAATACTGC 
                 79 
                 171 
               
               
                   
               
               
                 288253 
                 12 
                 338 
                 AGTTACCAGTTTCCTGAATA 
                 63 
                 172 
               
               
                   
               
               
                 288254 
                 12 
                 343 
                 TCATCAGTTACCAGTTTCCT 
                 57 
                 173 
               
               
                   
               
               
                 288255 
                 12 
                 347 
                 CTTTTCATCAGTTACCAGTT 
                 63 
                 174 
               
               
                   
               
               
                 288256 
                 12 
                 350 
                 AACCTTTTCATCAGTTACCA 
                 67 
                 175 
               
               
                   
               
               
                 288257 
                 12 
                 358 
                 TTATCTGAAACCTTTTCATG 
                 48 
                 176 
               
               
                   
               
               
                 288258 
                 12 
                 360 
                 AATTATCTGAAACCTTTTCA 
                 49 
                 177 
               
               
                   
               
               
                 288259 
                 12 
                 365 
                 GGCCCAATTATCTGAAACCT 
                 57 
                 178 
               
               
                   
               
               
                 288260 
                 12 
                 367 
                 ATGGCCCAATTATCTGAAAC 
                 43 
                 179 
               
               
                   
               
               
                 288261 
                 12 
                 375 
                 TGCTGTCAATGGCCCAATTA 
                 62 
                 180 
               
               
                   
               
               
                 288262 
                 12 
                 379 
                 GCGCTGCTGTCAATGGCCCA 
                 61 
                 181 
               
               
                   
               
               
                 288263 
                 12 
                 406 
                 GGCCGGCCCACGTTCCAGTG 
                 65 
                 182 
               
               
                   
               
               
                 288264 
                 12 
                 493 
                 GCAAAGTCTTCTTTTGTAAT 
                 57 
                 183 
               
               
                   
               
               
                 288265 
                 12 
                 499 
                 AATGTGGCAAAGTCTTCTTT 
                 54 
                 184 
               
               
                   
               
               
                 288266 
                 12 
                 505 
                 TAATCGAATGTGGCAAAGTC 
                 59 
                 185 
               
               
                   
               
               
                 288267 
                 12 
                 510 
                 GTATATAATCGAATGTGGCA 
                 80 
                 186 
               
               
                   
               
               
                 288268 
                 12 
                 511 
                 AGTATATAATCGAATGTGGC 
                 76 
                 187 
               
               
                   
               
               
                 288269 
                 12 
                 518 
                 CATACATAGTATATAATCGA 
                 52 
                 188 
               
               
                   
               
               
                 288270 
                 12 
                 520 
                 TCCATACATAGTATATAATC 
                 60 
                 57 
               
               
                   
               
               
                 288271 
                 12 
                 525 
                 TTTCATCCATACATAGTATA 
                 57 
                 58 
               
               
                   
               
               
                 288272 
                 12 
                 526 
                 CTTTCATCCATACATAGTAT 
                 57 
                 189 
               
               
                   
               
               
                 288273 
                 12 
                 530 
                 ATTGCTTTCATCCATACATA 
                 71 
                 59 
               
               
                   
               
               
                 288274 
                 12 
                 532 
                 AGATTGCTTTCATCCATACA 
                 70 
                 60 
               
               
                   
               
               
                 288275 
                 12 
                 535 
                 CTCAGATTGCTTTCATCCAT 
                 73 
                 61 
               
               
                   
               
               
                 288276 
                 12 
                 537 
                 CTCTCAGATTGCTTTCATCC 
                 78 
                 62 
               
               
                   
               
               
                 288277 
                 12 
                 538 
                 TCTCTCAGATTGCTTTCATC 
                 66 
                 190 
               
               
                   
               
               
                 288278 
                 12 
                 540 
                 GATCTCTCAGATTGCTTTCA 
                 70 
                 191 
               
               
                   
               
               
                 288279 
                 12 
                 545 
                 ATTGAGATCTCTCAGATTGC 
                 61 
                 192 
               
               
                   
               
               
                 288280 
                 12 
                 549 
                 TTCTATTGAGATCTCTCAGA 
                 65 
                 193 
               
               
                   
               
               
                 288281 
                 12 
                 572 
                 GCAGTTTTTAACTTGATTAC 
                 61 
                 194 
               
               
                   
               
               
                 288282 
                 12 
                 626 
                 AATGATGAGCTGTTTCTGTG 
                 53 
                 195 
               
               
                   
               
               
                 288283 
                 12 
                 636 
                 AGGGATCTTCAATGATGAGC 
                 59 
                 196 
               
               
                   
               
               
                 288284 
                 12 
                 639 
                 AATAGGGATCTTCAATGATG 
                 48 
                 197 
               
               
                   
               
               
                 288285 
                 12 
                 663 
                 CCTGGAAGTCAGAGTCATTG 
                 82 
                 198 
               
               
                   
               
               
                 288286 
                 12 
                 668 
                 CACCACCTCGAAGTCAGAGT 
                 81 
                 199 
               
               
                   
               
               
                 288287 
                 12 
                 673 
                 TGGTACACCACCTCGAAGTC 
                 74 
                 200 
               
               
                   
               
               
                 288288 
                 12 
                 678 
                 ATTGCTGGTACACCACCTCG 
                 75 
                 201 
               
               
                   
               
            
           
         
       
     
     EXAMPLE 5 
     Antisense Inhibition of Rat LMW-PTPase Expression by Oligomeric Compounds 
     Antisense compounds were designed against one or more of the rat LMW-PTPase target nucleic acid sequences cited in Table 1 and were screened in vitro to determine the compound&#39;s ability to modulate expression of a target nucleic acid that encodes LMW-PTPase. The compounds shown in Table 7 are all chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of 10 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′) by five-nucleotide “wings”. The wings are composed of 2′-O-(2-methoxyethyl) nucleotides, also known as 2′-MOE nucleotides. The internucleoside (backbone) linkages are phosphorothioate throughout the oligonucleotide. All cytidine residues are 5-methylcytidines. The compounds were analyzed for their effect on gene target mRNA levels by quantitative real-time PCR as described in other examples herein, using the primer-probe set designed to hybridize to rat LMW-PTPase (Table 2). Data are averages from two experiments in which A10 cells were treated with 50 nM of the disclosed oligomeric compounds using LIPOFECTIN™. A reduction in expression is expressed as percent inhibition in Table 7. The control oligomeric compound used was SEQ ID NO: 25. If present, “N.D.” indicates “not determined”. 
     
       
         
           
               
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                 Inhibition of rat LMW-PTPase mRNA levels by chimeric 
                   
               
               
                 oligonucleotides having 2′-MOE wings and deoxy gap 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 Target 
                   
                   
                   
                   
                   
               
               
                 Compound 
                 SEQ ID 
                 Target 
                   
                   
                 SEQ ID 
               
               
                 No. 
                 NO 
                 Site 
                 Sequence (5′ to 3′) 
                 % Inhib 
                 NO 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 288289 
                 15 
                 403 
                 ACCTCGAAGTCAGAGTCATT 
                 70 
                 202 
                   
               
               
                   
               
               
                 288233 
                 16 
                 24 
                 TGCCATGTITCGGTGCAGAC 
                 74 
                 153 
               
               
                   
               
               
                 288234 
                 16 
                 29 
                 ACCTCTGCCATGTTTCGGTG 
                 83 
                 154 
               
               
                   
               
               
                 355621 
                 16 
                 36 
                 GGACCCAACCTCTGCCATGT 
                 89 
                 203 
               
               
                   
               
               
                 288235 
                 16 
                 42 
                 TGACTTGGACCCAACCTCTG 
                 73 
                 155 
               
               
                   
               
               
                 288238 
                 16 
                 57 
                 ACACACGAACAGCACTGACT 
                 29 
                 158 
               
               
                   
               
               
                 288239 
                 16 
                 65 
                 TTACCGAGACACACGAACAG 
                 34 
                 159 
               
               
                   
               
               
                 288241 
                 16 
                 72 
                 GCAAATGTTACCGAGACACA 
                 50 
                 161 
               
               
                   
               
               
                 288274 
                 16 
                 308 
                 AGATTGCTTTGATCCATACA 
                 54 
                 60 
               
               
                   
               
               
                 288286 
                 16 
                 444 
                 CACCACCTCGAAGTCAGAGT 
                 69 
                 199 
               
               
                   
               
               
                 288287 
                 16 
                 449 
                 TGGTACACCACCTCGAAGTC 
                 81 
                 200 
               
               
                   
               
               
                 288288 
                 16 
                 454 
                 ATTGCTGGTACAGCACCTCG 
                 69 
                 201 
               
               
                   
               
               
                 355622 
                 16 
                 461 
                 CTAAGGCATTGCTGGTACAC 
                 72 
                 204 
               
               
                   
               
               
                 355623 
                 16 
                 466 
                 AGCACCTAAGGCATTGCTGG 
                 74 
                 205 
               
               
                   
               
               
                 355624 
                 16 
                 471 
                 CTTGGAGCACCTAAGGCATT 
                 74 
                 206 
               
               
                   
               
               
                 355625 
                 16 
                 476 
                 AAGGCCTTGCAGCACCTAAG 
                 83 
                 207 
               
               
                   
               
               
                 355626 
                 16 
                 481 
                 CCAGGAAGGCCTTGCAGCAC 
                 86 
                 208 
               
               
                   
               
               
                 355627 
                 16 
                 486 
                 CTTCTCCAGGAAGGCCTTGC 
                 79 
                 209 
               
               
                   
               
               
                 355628 
                 16 
                 492 
                 GTGAGTCTTCTCCAGGAAGG 
                 82 
                 210 
               
               
                   
               
               
                 355629 
                 16 
                 504 
                 TAGGACCAGGTAGTGAGTCT 
                 74 
                 211 
               
               
                   
               
               
                 355630 
                 16 
                 519 
                 CTCAGTGGTGGTGGTTAGGA 
                 55 
                 212 
               
               
                   
               
               
                 355631 
                 16 
                 556 
                 GCCACCACCCTTGGGCACAG 
                 78 
                 213 
               
               
                   
               
               
                 355632 
                 16 
                 567 
                 GGCTAAGGACTGCCACCACC 
                 78 
                 214 
               
               
                   
               
               
                 355633 
                 16 
                 607 
                 GATATACAGTAAGTCAGCTG 
                 80 
                 215 
               
               
                   
               
               
                 355634 
                 16 
                 625 
                 ACCTACAATTATTTTAAAGA 
                 15 
                 216 
               
               
                   
               
               
                 355635 
                 16 
                 633 
                 TGATTTCCACCTACAATTAT 
                 52 
                 217 
               
               
                   
               
               
                 355636 
                 16 
                 638 
                 ATGCCTGATTTCCACCTACA 
                 91 
                 218 
               
               
                   
               
               
                 355637 
                 16 
                 647 
                 TCTGAACAAATGCCTGATTT 
                 82 
                 219 
               
               
                   
               
               
                 355638 
                 16 
                 674 
                 AATGTCTGCCTCAAATGTTT 
                 73 
                 220 
               
               
                   
               
               
                 355639 
                 16 
                 680 
                 ACCTCAAATGTCTGCCTCAA 
                 78 
                 221 
               
               
                   
               
               
                 355640 
                 16 
                 687 
                 GAGCCACACCTCAAATGTCT 
                 89 
                 222 
               
               
                   
               
               
                 355641 
                 16 
                 700 
                 GTCTAAGAATACTGAGCCAC 
                 87 
                 223 
               
               
                   
               
               
                 355642 
                 16 
                 706 
                 TTGTTAGTCTAAGAATACTG 
                 52 
                 224 
               
               
                   
               
               
                 355643 
                 16 
                 725 
                 TATGGCGAGGCCAGAGCTTT 
                 77 
                 225 
               
               
                   
               
               
                 355644 
                 16 
                 735 
                 ATTTTGTAATTATGGCGAGG 
                 60 
                 226 
               
               
                   
               
               
                 355645 
                 16 
                 753 
                 ACAGTTGGTCGTTCCACTAT 
                 92 
                 227 
               
               
                   
               
               
                 355646 
                 16 
                 759 
                 TGTTCCACAGTTGCTCGTTC 
                 95 
                 228 
               
               
                   
               
               
                 355647 
                 16 
                 795 
                 CCTTGTGGGTCATTCTTACT 
                 71 
                 229 
               
               
                   
               
               
                 355648 
                 16 
                 819 
                 GCTGGGCTCAAAGGCTGATC 
                 67 
                 230 
               
               
                   
               
               
                 355649 
                 16 
                 841 
                 TTAGACCAGACTACCCAGGC 
                 80 
                 231 
               
               
                   
               
               
                 355650 
                 16 
                 853 
                 CTCACACTCCAGTTAGACCA 
                 63 
                 232 
               
               
                   
               
               
                 355651 
                 16 
                 871 
                 CACTGGGTGCTGGCCATGCT 
                 70 
                 233 
               
               
                   
               
               
                 355652 
                 16 
                 890 
                 GTAAGGCAAGCAAACAGGAC 
                 40 
                 234 
               
               
                   
               
               
                 355653 
                 16 
                 924 
                 TTGTCACAATAAGAGACAAT 
                 55 
                 235 
               
               
                   
               
               
                 355654 
                 16 
                 931 
                 GGAGATATTGTCACAATAAG 
                 85 
                 236 
               
               
                   
               
               
                 355655 
                 16 
                 939 
                 CCATGGATGGAGATATTGTC 
                 82 
                 237 
               
               
                   
               
               
                 355656 
                 16 
                 944 
                 GGCTGCCATGGATGGAGATA 
                 79 
                 238 
               
               
                   
               
               
                 355657 
                 16 
                 952 
                 AAATGGAAGGCTGCCATGGA 
                 80 
                 239 
               
               
                   
               
               
                 355658 
                 16 
                 958 
                 AGTGTTAAATGGAAGGCTGC 
                 59 
                 240 
               
               
                   
               
               
                 355659 
                 16 
                 970 
                 TTAAACTCTCCCAGTGTTAA 
                 76 
                 241 
               
               
                   
               
               
                 355660 
                 16 
                 976 
                 CTGGGTTTAAACTCTCCCAG 
                 80 
                 242 
               
               
                   
               
               
                 355661 
                 16 
                 1013 
                 GGTTCTCCTCTCTCAAATAT 
                 82 
                 243 
               
               
                   
               
               
                 355662 
                 16 
                 1038 
                 AGTTCCAGGCCCATCATCAC 
                 61 
                 244 
               
               
                   
               
               
                 355663 
                 16 
                 1050 
                 ATGGCCTGCTGGAGTTCCAG 
                 67 
                 245 
               
               
                   
               
               
                 355664 
                 16 
                 1089 
                 TTTTTATCTTTCAGACAGGG 
                 19 
                 246 
               
               
                   
               
               
                 355665 
                 16 
                 1103 
                 CCCATCTGTTAGCATTTTTA 
                 73 
                 247 
               
               
                   
               
               
                 355666 
                 16 
                 1111 
                 CTGTTGCTCCCATCTGTTAG 
                 80 
                 248 
               
               
                   
               
               
                 355667 
                 16 
                 1125 
                 TTAACTTCACCAACCTGTTG 
                 85 
                 249 
               
               
                   
               
               
                 355668 
                 16 
                 1169 
                 CCAAGCTCAAGAAACTACAC 
                 69 
                 250 
               
               
                   
               
               
                 355669 
                 16 
                 1187 
                 AAGTGGCTCAAATAGGAACC 
                 31 
                 251 
               
               
                   
               
               
                 355670 
                 16 
                 1206 
                 CTTTCTCTTTAAAGAAGCAA 
                 24 
                 252 
               
               
                   
               
               
                 355671 
                 16 
                 1215 
                 GCACACTTAGTTTCTCTTTA 
                 84 
                 253 
               
               
                   
               
               
                 355672 
                 16 
                 1228 
                 CACCACTATTTAAGCACACT 
                 28 
                 254 
               
               
                   
               
               
                 355673 
                 16 
                 1237 
                 ACAAACGCACACCACTATTT 
                 62 
                 255 
               
               
                   
               
               
                 355674 
                 16 
                 1255 
                 GTTGATGAGAGAACACTTAC 
                 79 
                 256 
               
               
                   
               
               
                 355675 
                 16 
                 1270 
                 GTAACTTTGTAAAATGTTGA 
                 46 
                 257 
               
               
                   
               
               
                 355676 
                 16 
                 1286 
                 TTACTCATGGTTGCCTGTAA 
                 90 
                 258 
               
               
                   
               
               
                 355677 
                 16 
                 1312 
                 GTCCGTTTTCTGAAAATACA 
                 78 
                 259 
               
               
                   
               
               
                 355678 
                 16 
                 1323 
                 ATAAATTTGAGGTCCCTTTT 
                 66 
                 260 
               
               
                   
               
               
                 355679 
                 16 
                 1331 
                 ATATCCACATAAATTTGAGG 
                 68 
                 261 
               
               
                   
               
               
                 355680 
                 16 
                 1345 
                 ATCTTTTCTGACATATATCC 
                 64 
                 262 
               
               
                   
               
               
                 355613 
                 17 
                 3444 
                 TCTCCAGTGGCAAAGACAAA 
                 30 
                 263 
               
               
                   
               
               
                 355614 
                 17 
                 5834 
                 AAGCAAGAAACTATGCGGGA 
                 45 
                 264 
               
               
                   
               
               
                 355615 
                 17 
                 8275 
                 CATACGGTACCTGCCGTGCA 
                 53 
                 265 
               
               
                   
               
               
                 355616 
                 17 
                 8312 
                 CAATGGCCCACTGTAACACA 
                 70 
                 266 
               
               
                   
               
               
                 355617 
                 17 
                 13156 
                 GAGTACAT1TGAAGTTAAAA 
                 45 
                 267 
               
               
                   
               
               
                 355618 
                 17 
                 13309 
                 CTCTTGTAATCTACAATTAA 
                 3 
                 268 
               
               
                   
               
               
                 355619 
                 17 
                 13459 
                 TATACCTGAGTTCAAGGTCA 
                 57 
                 269 
               
               
                   
               
               
                 355620 
                 18 
                 204 
                 CTCTTGTAATCTGTCTTGGC 
                 27 
                 270 
               
               
                   
               
            
           
         
       
     
     EXAMPLE 6 
     Inhibition of Mouse LMW-PTPase mRNA Levels by Chimeric Phosphorothioate Oligonucleotides Having 2′-MOE Wings and a Deoxy Gap:Dose Response Studies 
     In a further embodiment, six oligonucleotides were selected for dose-response studies: Compound No. 288285, Compound No. 288276, Compound No. 288268, Compound No. 288286, Compound No. 288267 and Compound No. 288291. Compound No. 129689 (GAGGTCTCGACTTACCCGCT, incorporated herein as SEQ ID NO: 271) and Compound No. 129695 (TTCTACCTCGCGCGATTTAC, incorporated herein as SEQ ID NO: 272, which are not targeted to LMW-PTPase, served as negative controls. Compound No. 129689 and Compound No. 129695 are chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by five-nucleotide “wings”. The wings are composed of 2′-O-(2-methoxyethyl) (2′-MOE) nucleotides. The internucleoside (backbone) linkages are phosphorothioate (PUS) throughout the oligonucleotide. All cytidine residues are 5-methylcytidines. 
     Oligonucleotides were transfected into cells using the CYTOFECTIN™ Reagent (Gene Therapy Systems, San Diego, Calif.). b.END cells were treated with 12.5, 25, 50 or 100 nM of oligonucleotide. Untreated control cells served as the control to which data were normalized. Quantitative real-time PCR to measure LMW-PTPase levels was performed as described herein. 
     Data were averaged from 3 experiments and the results are shown in Table 8 as percent inhibition relative to untreated control. Neither control oligonucleotide (Compound No. 129689 or Compound No. 129695) inhibited LMW-PTPase mRNA expression in this experiment. 
     
       
         
           
               
             
               
                 TABLE 8 
               
             
            
               
                   
               
               
                 Inhibition of mouse LMW-PTPase mRNA expression in mouse 
               
               
                 b.END cells: dose response 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 Compound 
                 SEQ ID 
                 Dose of oligonucleotide (nM) 
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 No. 
                 NO 
                 12.5 
                 25 
                 50 
                 100 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 288267 
                 186 
                 60 
                 74 
                 88 
                 93 
               
               
                   
                 288268 
                 187 
                 54 
                 72 
                 80 
                 93 
               
               
                   
                 288276 
                 62 
                 23 
                 47 
                 68 
                 85 
               
               
                   
                 288285 
                 198 
                 11 
                 41 
                 67 
                 86 
               
               
                   
                 288286 
                 199 
                 50 
                 69 
                 86 
                 96 
               
               
                   
                 288291 
                 133 
                 72 
                 82 
                 90 
                 92 
               
               
                   
                   
               
            
           
         
       
     
     As demonstrated in Table 8, Compound No. 288267, Compound No. 288268, Compound No. 288276, Compound No. 288285, Compound No. 288286 and Compound No. 288291 inhibited mouse LMW-PTPase mRNA expression in a dose-dependent manner. 
     EXAMPLE 7 
     Inhibition of Rat LMW-PTPase mRNA Levels by Chimeric Phosphorothioate Oligonucleotides Having 2′-MOE Wings and a Deoxy Gap:Dose Response Studies 
     In a further embodiment, seven oligonucleotides were selected for dose-response studies: Compound No. 355621, Compound No. 355636, Compound No. 355676, Compound No. 355626, Compound No. 355654, Compound No. 355640, and Compound No. 355641. Compound No. 15770, Compound No. 129690 (TTAGAATACGTCGCGTTATG, incorporated herein as SEQ ID NO: 273), and Compound No. 141923 (CCTTCCCTGAAGGTTCCTCC, incorporated herein as SEQ ID NO: 274), which are not targeted to LMW-PTPase, served as controls. Compound No. 129690 and Compound No. 141923 are chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by five-nucleotide “wings”. The wings are composed of 2′-O-methoxyethyl (2′-MOE) nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. All cytidine residues are 5-methylcytidines. 
     Rat primary hepatocytes were treated with 12.5, 25, 50, 100 or 200 nM of oligonucleotide, using LIPOFECTIN™ as described herein. Untreated control cells served as the control to which data were normalized. Treatment with the transfection mixture and quantitative real-time PCR to measure rat LMW-PTPase levels were both performed as described herein. 
     Results of these studies are shown in Table 9. Data are averaged from four experiments and are expressed as percent inhibition relative untreated control. None of the control oligonucleotides tested (Compound No. 141923, Compound No. 15770 or Compound No. 129690) resulted greater than 4% inhibition of rat LMW-PTPase; data from cells treated with Compound No. 15770 is shown in Table 8 and is representative of the control oligonucleotide treatments. 
     
       
         
           
               
             
               
                 TABLE 9 
               
             
            
               
                   
               
               
                 Inhibition of rat LMW-PTPase mRNA expression in rat 
               
               
                 primary hepatocyte cells: dose response 
               
            
           
           
               
               
            
               
                   
                 % Inhibition 
               
            
           
           
               
               
               
            
               
                 Compound 
                 SEQ ID 
                 Dose of oligonucleotide (nM) 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 No. 
                 NO 
                 12.5 
                 25 
                 50 
                 100 
                 200 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 355621 
                 203 
                 17 
                 29 
                 52 
                 70 
                 82 
               
               
                 355626 
                 208 
                 6 
                 17 
                 33 
                 48 
                 70 
               
               
                 355636 
                 218 
                 17 
                 27 
                 43 
                 64 
                 80 
               
               
                 355640 
                 222 
                 19 
                 31 
                 55 
                 73 
                 84 
               
               
                 355641 
                 223 
                 14 
                 31 
                 46 
                 65 
                 80 
               
               
                 355654 
                 236 
                 18 
                 25 
                 42 
                 66 
                 81 
               
               
                 355676 
                 258 
                 15 
                 24 
                 49 
                 62 
                 77 
               
               
                 15770 
                 31 
                 0 
                 1 
                 0 
                 4 
                 0 
               
               
                   
               
            
           
         
       
     
     As demonstrated in Table 9, Compound No. 355621, Compound No. 355626 Compound No. 355636, Compound No. 355640, Compound No. 355641, Compound No. 355654 and Compound No. 355676 inhibited LMW-PTPase mRNA expression in a dose-dependent manner. 
     EXAMPLE 8 
     Antisense Inhibition of LMW-PTPase Expression In Vivo: ob/ob Mice 
     Leptin is a hormone produced by fat that regulates appetite. Deficiencies in this hormone in both humans and non-human animals leads to obesity. ob/ob mice have a mutation in the leptin gene which results in obesity and hyperglycemia. As such, these mice are a useful model for the investigation of obesity and diabetes and treatments designed to treat these conditions. ob/ob mice have higher circulating levels of insulin and are less hyperglycemic than db/db mice, which harbor a mutation in the leptin receptor. In accordance with the present invention, the oligomeric compounds of the invention are tested in the ob/ob model of obesity and diabetes. 
     C57Bl/6J-Lep ob/ob mice (Jackson Laboratory, Bar Harbor, Me.) are subcutaneously injected with Compound No. 288267 (SEQ ID NO: 186) at a dose of 25 mg/kg two times per week for 4 weeks (n—5). Saline-injected animals serve as controls (n—4). After the treatment period, mice are sacrificed and target levels were evaluated in liver and in fat. RNA isolation and target mRNA expression level quantitation were performed as described by other examples herein. Animals treated with Compound No. 288267 on average showed 90% reduction in liver LMW-PTPase levels as compared to saline treated control animals. LMW-PTPase mRNA levels in epididymal fat were reduced 70% on average in animals treated with Compound No. 288267. 
     To assess the physiological effects resulting from inhibition of target mRNA, the ob/ob mice were evaluated at the end of the treatment period (day 28) for serum triglycerides and serum glucose levels. These parameters were measured by routine clinical analyzer instruments (e.g. Olympus Clinical Analyzer, Melville, N.Y.). At day 28, the average triglyceride levels measured for saline-treated control animals was 168 mg/dL, while the average for animals treated with Compound No. 288267 was 75 mg/dL. At day 28, glucose was 491 mg/dL for animals treated with saline alone and 258 mg/dL for animals treated with Compound No. 288267. Therefore, treatment with Compound No. 288267 caused substantial decreases in glucose and in triglyceride levels. Therefore, one embodiment of the present invention is a method of lowering glucose by administering an oligomeric compound of the invention, and another embodiment of the present invention is a method of lowering triglycerides by administering an oligomeric compound of the invention. In one embodiment, the triglycerides are blood, plasma, or serum triglycerides. Another embodiment of the current invention is a method of ameliorating or lessening the severity of a condition in an animal. In some embodiments, the condition is diabetes. In some embodiments, the diabetes is type II diabetes. In other embodiments, the condition is metabolic syndrome. 
     To further assess the effects of inhibition of target mRNA on glucose metabolism, fasted serum glucose was measured via routine clinical analysis. The average fasted serum glucose level for animals treated with Compound No. 288267 was 194 mg/dL, while the average fasted level measured for animals treated with saline alone was 330 mg/dL. Therefore, another embodiment of the present invention is a method of lowering serum glucose. 
     Insulin levels were also measured after four weeks of treatment using a commercially available kit (e.g. Alpco insulin-specific ELISA kit, Windham, N.H.). Treatment with Compound No. 288267 caused about a 45% reduction in circulating plasma insulin levels. Decreased insulin levels can indicate improvement in insulin sensitivity. In one embodiment, the present invention provides methods of improving insulin sensitivity. In a further embodiment, decreased insulin levels are indicative of improved insulin sensitivity. 
     At the end of the study, mice were sacrificed and tissues were weighed. The average liver and spleen weights were not substantially altered by treatment with Compound No. 288267 as compared to saline-treated controls. Epididymal white adipose tissue weight was reduced by about 10% in animals treated with Compound No. 288267. Therefore, another embodiment of the present invention is a method of reducing adiposity in an animal by administering an oligomeric compound of the invention. 
     EXAMPLE 9 
     Effect of Antisense Inhibition of LMW-PTPase on Insulin Receptor Phosphorylation in ob/ob Mice 
     To assess the effects of inhibition of LMW-PTPase on receptor phosphorylation, a bolus of insulin (2 U/kg) was administered about 8 to 9 minutes prior to sacrifice to a group of the ob/ob mice treated with Compound No. 288267 (SEQ ID NO: 186) or saline as described in Example 8. Liver samples were pooled and subjected to standard immunoprecipitation and Western blot procedures and analyses. Briefly, tissues were lysed in lysis buffer (150 mM NaCl, 50 mM Tris, pH 7.5, 1% Triton X-100, 0.5% NP-40, 0.25% Nadeoxycholate, 1 mM EDTA, 1 mM EGTA, 1 mM NaOV, 1 mM NaF, and protease inhibitor cocktail I (Calbiochem). The lysates were clarified by centrifugation for 15 min. at 12000 g. The clarified lysates were first incubated with protein agarose A/G beads (1:1 ratio) for 3-4 h at 4.deg.C. followed by incubation with anti-phosphotyrosine antibody for another 3-4 h at 4.deg.C. The immune complex was then washed with lysis buffer, boiled in Laemmli&#39;s sample buffer, and analyzed by Western blot. The membrane was blotted with a commercially available anti-insulin receptor beta (IR-.beta. subunit antibody (Santa Cruz, Calif.). The signal was detected by using a commercially available HRP-conjugated goat anti-rabbit IgG antibody and ECL. 
     Quantitation of the resulting bands showed approximately a 4 to 6-fold increase in phosphorylation of the insulin receptor upon insulin stimulation in the samples from animals treated with Compound No. 288267 as compared to that measured for saline-treated controls. These data suggest improved insulin sensitivity upon treatment with antisense oligonucleotides targeted to LMW-PTPase. 
     EXAMPLE 10 
     Effect of Antisense Inhibition of LMW-PTPase on PI3-K Activity in ob/ob Mice 
     To further assess the role of LMW-PTPase in the insulin receptor signaling pathway and to assess the effects of antisense inhibition of LMW-PTPase on insulin action, clarified lysates prepared as described in Example 11 from livers or fat tissue of ob/ob mice treated as described in Examples 8 and 9, and were subjected to further analyses. 
     Phosphatidyl inositol 3-kinase (PI3-K) is an enzyme activated downstream of insulin-receptor stimulation. PI3-kinase activity was measured using methods known in the art (Pandey et al., Biochemistry, 1998, 37, 7006-7014). Briefly, the clarified lysates were subjected to immunoprecipitation with IRS-1/2 antibody (1 ug) for 2 h at 4.deg.C., followed by incubation with protein A/G sepharose for an additional 2 h. The immune complexes were washed and subjected to in vitro PI3-Kinase assay using L-.alpha.-phosphatidylinositol (PI) as an exogenous substrate. The phosphorylated lipid was isolated and separated by thin-layer chromatography (TLC). The TLC plate was exposed to Kodak film, and the radioactive spots associated with the product of PI3-K activity (PIP2) was scratched off of the TLC plates and counted in a scintillation counter. Average results from the scintillation counts are shown in Table 10 in arbitrary units for the livers from each treatment group with or without insulin. The antibody used for the immunoprecipitation is shown in the column designated “IP” (IRS-1 or IRS-2). 
     
       
         
           
               
             
               
                 TABLE 10 
               
             
            
               
                   
               
               
                 Effects of antisense inhibition of LMW-PTPase 
               
               
                 on PI3-K activity in ob/ob mouse liver 
               
            
           
           
               
               
               
            
               
                   
                 PI3-K activity 
                   
               
               
                   
                 (arbitrary units) 
               
            
           
           
               
               
               
               
               
            
               
                   
                 Treatment group 
                 IP 
                 −Insulin 
                 +Insulin 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 Saline 
                 IRS-1 
                 260853 
                 257907 
               
               
                   
                 Compound No. 288267 
                 IRS-1 
                 291982 
                 674202 
               
               
                   
                 Saline 
                 IRS-2 
                 596 
                 677 
               
               
                   
                 Compound No. 288267 
                 IRS-2 
                 679 
                 1092 
               
               
                   
                   
               
            
           
         
       
     
     As shown in Table 10, Compound No. 288267 (SEQ ID NO: 186) caused increases in insulin-stimulated PI3-K activity above the increases observed for animals treated with saline. 
     Approximate results from the scintillation counts are shown in Table 11 as averages in arbitrary units for adipose tissue from each treatment group with or without insulin. The antibody used for the immunoprecipitation was IRS-1. 
     
       
         
           
               
             
               
                 TABLE 11 
               
             
            
               
                   
               
               
                 Effects of antisense inhibition of LMW-PTPase 
               
               
                 on PI3-K activity in ob/ob mouse fat tissue 
               
            
           
           
               
               
               
            
               
                   
                 PI3-K activity 
                   
               
               
                   
                 (arbitrary units) 
               
            
           
           
               
               
               
               
            
               
                   
                 Treatment group 
                 −Insulin 
                 +Insulin 
               
               
                   
                   
               
               
                   
                 Saline 
                 319 
                 365 
               
               
                   
                 Compound No. 288267 
                 438 
                 725 
               
               
                   
                   
               
            
           
         
       
     
     As shown in Table 11, Compound No. 288267 caused increases in insulin-stimulated PI3-K activity above the increases observed for animals treated with saline. Taken together, these results demonstrate a novel role for LMW-PTPase in insulin action and show that antisense inhibition of LMW-PTPase improves insulin signaling in ob/ob mice. Therefore another embodiment of the present invention is a method of improving insulin signaling in an animal by administering an oligomeric compound of the invention. 
     EXAMPLE 11 
     Effect of Antisense Inhibition of LMW-PTPase on Insulin Signaling: In Vitro Studies 
     In accord with the present invention, the effects of antisense oligonucleotides targeted to LMW-PTPase on insulin-signaling were investigated in primary hepatocytes cultured from ob/ob mice using methods described herein. The ob/ob primary hepatocytes were treated with Compound No. 288267 (SEQ ID NO: 186) or the control oligonucleotide Compound No. 141923 (SEQ ID NO: 274) by transfection methods described herein. Treated cells were incubated for 10 minutes in the absence or presence of 100 nM insulin. Cells treated with transfection reagent alone served as controls. 
     Cell lysates were prepared and subjected to immunoprecipitation with anti-phosphotyrosine antibody followed by Western blot analysis using an anti-IR-.beta. antibody (Santa Cruz, Calif.) via standard methods. Cells treated with Compound No. 288267 showed a larger increase in insulin-stimulated IR-.beta. phosphorylation than was observed for control cells or cells treated with Compound No. 141923. 
     In a similar experiment using a commercially available antibody which recognizes phosphorylated Akt (Cell Signaling, Boston, Mass.), cells treated with Compound No. 288267 showed a larger increase in insulin-stimulated Akt phosphorylation than was observed for control cells or cells treated with Compound No. 141923. These data demonstrate that LMW-PTPase plays a role in the insulin signaling pathway. 
     Antisense compound modulation of LMW-PTPase expression levels was determined using primary mouse hepatocytes by treating the cells with 100 ng of Compound No. 288267 or with vehicle as described above. Following incubation, the cells were lysed in RTL buffer and total RNA was isolated using QIAGEN RNA easy kits (Qiagen, Valencia, Calif.). RT-PCR was performed as described above. These data showed an approximate 90% reduction in LMW-PTPase mRNA levels compared to control. A corresponding reduction in LMW-PTPase protein levels was seen by western blot analysis of the LMW-PTPase protein. Interestingly, there was no significant reduction in PTP1b levels in the presence of Compound No. 288267 compared to control cells (antibody available from Upstate Cell Signaling Solutions). 
     To further determine the action of LMW-PTPase antisense compounds on the insulin signaling pathway duplicate cell culture well comprising primary mouse hepatocytes were treated with 100 ng of either Compound No. 288267, Compound No. 288291, control Compound No. 141923, an antisense inhibitor of PTP1b or a combination of Control No. 288291 and the antisense inhibitor of PTP1b. A saline treated control was used as well. For each treatment group, one of the wells was incubated with 5 nM of insulin for 10 minutes. Following incubation the cell were analyzed for levels of phosphorylated IR-.beta., phosphoAkt, unphosphorylated Akt, PTP1b or LMW-PTPase using western blot techniques. These data show that IR-.beta. is phosphorylated in the presence of LMW-PTPase antisense compounds, but not in the presence of PTP-1b antisense compounds either alone or combined with the LMW-PTPase antisense compound. Western blot techniques are well known to those of ordinary skill in the art and antibodies are readily available from a number of commercial vendors (e.g., Upstate Cell Signaling Solutions, Charlottesville, Va.). (Harlow, E and Lane, D., Antibodies: A Laboratory Manual, (1988) Cold Spring Harbor Press). 
     EXAMPLE 12 
     Design of Oligomeric Compounds Targeting Human LMW-PTPase 
     A series of oligomeric compounds was designed to target different regions of human LMW-PTPase, using published sequences cited in Table 1. The compounds are shown in Table 12. All compounds in Table 12 are chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of 10 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′) by five-nucleotide “wings”. The wings are composed of 2′-O-(2-methoxyethyl) nucleotides, also known as 2′-MOE nucleotides. The internucleoside (backbone) linkages are phosphorothioate throughout the oligonucleotide. All cytidine residues are 5-methylcytidines. 
     
       
         
           
               
               
             
               
                 TABLE 12 
               
             
            
               
                   
               
               
                 Chimeric oligonucleotides having 2′-MOE wings and 
                   
               
               
                 deoxy gap targeting human LMW-PTPase 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 Target 
                   
                   
                   
                   
               
               
                 Compound 
                 SEQ ID 
                 Target 
                   
                 SEQ ID 
               
               
                 No. 
                 NO 
                 Site 
                 Sequence (5′to 3′) 
                 NO 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 105809 
                 1 
                 1 
                 ACCCCGTTCCGCACGCCCCC 
                 279 
                   
               
               
                   
               
               
                 105811 
                 1 
                 41 
                 TAGCCTGTTCCGCCATCTTC 
                 280 
               
               
                   
               
               
                 105812 
                 1 
                 241 
                 GCTCATGGGAATGCCGTGCC 
                 281 
               
               
                   
               
               
                 105813 
                 1 
                 271 
                 ATCTTCTTTGGTAATCTGCC 
                 282 
               
               
                   
               
               
                 105814 
                 1 
                 421 
                 ATAGGGATCTTCAATAATAA 
                 283 
               
               
                   
               
               
                 105815 
                 1 
                 451 
                 CACCGTCTCAAAGTCAGAGT 
                 284 
               
               
                   
               
               
                 105816 
                 1 
                 571 
                 CCGACTGAGAAATGCAGGAC 
                 285 
               
               
                   
               
               
                 105817 
                 1 
                 611 
                 AACAAAGAGCTGGCTTTGGG 
                 286 
               
               
                   
               
               
                 105818 
                 1 
                 671 
                 CAAACACAACTGATTTCCAT 
                 287 
               
               
                   
               
               
                 105819 
                 1 
                 701 
                 TGAATCAAACATTTTTATTG 
                 288 
               
               
                   
               
               
                 105820 
                 1 
                 781 
                 TTGTTCTACTATTTTTGTAA 
                 289 
               
               
                   
               
               
                 105821 
                 1 
                 811 
                 GTGAGGTTTTCCTTCATTGT 
                 290 
               
               
                   
               
               
                 105822 
                 1 
                 961 
                 ACTACTGTCAATCCACAAAA 
                 291 
               
               
                   
               
               
                 105823 
                 1 
                 1051 
                 TCTTCCCTATCTTTTCAATA 
                 292 
               
               
                   
               
               
                 105824 
                 1 
                 1091 
                 GTATTGAAGGTGCCAACGAC 
                 293 
               
               
                   
               
               
                 105825 
                 1 
                 1171 
                 TAAGTTTGAGAGGGAAAGTG 
                 294 
               
               
                   
               
               
                 105826 
                 1 
                 1201 
                 CATACAAGTGTCCTTCTTTC 
                 295 
               
               
                   
               
               
                 105827 
                 1 
                 1291 
                 TTATTTTAAAAAATAAGGCA 
                 296 
               
               
                   
               
               
                 105828 
                 1 
                 1321 
                 AAATAATAACAGTTTTCCCA 
                 297 
               
               
                   
               
            
           
         
       
     
     EXAMPLE 13 
     Design of Oligomeric Compounds Targeting Mouse LMW-PTPase 
     A series of oligomeric compounds was designed to target different regions of mouse LMW-PTPase, using published sequences cited in Table 1. The compounds are shown in Table 13. All compounds in Table 13 are chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of 10 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′) by five-nucleotide “wings”. The wings are composed of 2′-O-(2-methoxyethyl) nucleotides, also known as 2′-MOE nucleotides. The internucleoside (backbone) linkages are phosphorothioate throughout the oligonucleotide. All cytidine residues are 5-methylcytidines. 
     
       
         
           
               
               
             
               
                 TABLE 13 
               
             
            
               
                   
               
               
                 Chimeric oligonucleotides having 2′-MOE wings and 
                   
               
               
                 deoxy gap targeting mouse LMW-PTiPase 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 Target 
                   
                   
                   
                   
               
               
                 Compound 
                 SEQ ID 
                 Target 
                   
                 SEQ ID 
               
               
                 No. 
                 NO 
                 Site 
                 Sequence (5′ to 3′) 
                 NO 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 349037 
                 11 
                 246 
                 AGGTTTAGTTAGTCTAAGAA 
                 298 
                   
               
               
                   
               
               
                 349038 
                 11 
                 248 
                 AGAGGTTTAGTTAGTCTAAG 
                 299 
               
               
                   
               
               
                 349039 
                 11 
                 250 
                 TCAGAGGTTTAGTTAGTCTA 
                 300 
               
               
                   
               
               
                 349040 
                 11 
                 254 
                 AAGGTCAGAGGTTTAGTTAG 
                 301 
               
               
                   
               
               
                 349041 
                 11 
                 256 
                 GCAAGGTCAGAGGTTTAGTT 
                 302 
               
               
                   
               
               
                 349042 
                 11 
                 258 
                 CCGCAAGGTCAGAGGTTTAG 
                 303 
               
               
                   
               
               
                 349043 
                 11 
                 296 
                 TTTGTTCCATATTTGCTTGT 
                 304 
               
               
                   
               
               
                 349044 
                 12 
                 307 
                 ATGGGTGACCGGCAAATGTT 
                 305 
               
               
                   
               
               
                 349045 
                 12 
                 308 
                 AATGGGTGACCGGCAAATGT 
                 306 
               
               
                   
               
               
                 349046 
                 12 
                 311 
                 TGCAATGGGTGACCGGCAAA 
                 307 
               
               
                   
               
               
                 349047 
                 12 
                 312 
                 CTGCAATGGGTGACCGGCAA 
                 308 
               
               
                   
               
               
                 349048 
                 12 
                 3i3 
                 TCTGCAATGGGTGACCGGCA 
                 309 
               
               
                   
               
               
                 349049 
                 12 
                 314 
                 TTCTGCAATGGGTGACCGGC 
                 310 
               
               
                   
               
               
                 349050 
                 12 
                 315 
                 CTTCTGCAATGGGTGACCGG 
                 311 
               
               
                   
               
               
                 349051 
                 12 
                 316 
                 GCTTCTGCAATGGGTGACCG 
                 312 
               
               
                   
               
               
                 3490521 
                 12 
                 318 
                 CTGCTTCTGCAATGGGTGAC 
                 313 
               
               
                   
               
               
                 3490531 
                 12 
                 320 
                 TACTGCTTCTGCAATGGGTG 
                 314 
               
               
                   
               
               
                 349054 
                 12 
                 322 
                 AATACTGCTTCTGCAATGGG 
                 315 
               
               
                   
               
               
                 349055 
                 12 
                 327 
                 TCCTGAATACTGCTTGTGCA 
                 316 
               
               
                   
               
               
                 349056 
                 12 
                 330 
                 GTTTGCTGAATACTGCTTCT 
                 317 
               
               
                   
               
               
                 349057 
                 12 
                 332 
                 GAGTTTCCTGAATACTGCTT 
                 318 
               
               
                   
               
               
                 349058 
                 12 
                 335 
                 TACCAGTTTCCTGAATACTG 
                 319 
               
               
                   
               
               
                 349059 
                 12 
                 337 
                 GTTACCAGTTTCCTGAATAC 
                 320 
               
               
                   
               
               
                 349060 
                 12 
                 339 
                 CAGTTACCAGTTTCCTGAAT 
                 321 
               
               
                   
               
               
                 349061 
                 11 
                 477 
                 TGTATGCTCGCTCCTGCTCT 
                 322 
               
               
                   
               
               
                 349062 
                 12 
                 503 
                 ATCGAATGTGGCAAAGTCTT 
                 323 
               
               
                   
               
               
                 349063 
                 11 
                 506 
                 CCGGCGCGCTCGCTCCGTCT 
                 324 
               
               
                   
               
               
                 349064 
                 12 
                 507 
                 TATAATCGAATGTGGCAAAG 
                 325 
               
               
                   
               
               
                 349065 
                 12 
                 509 
                 TATATAATCGAATGTGGCAA 
                 326 
               
               
                   
               
               
                 349066 
                 12 
                 512 
                 TAGTATATAATCGAATGTGG 
                 327 
               
               
                   
               
               
                 349067 
                 12 
                 513 
                 ATAGTATATAATCGAATGTG 
                 328 
               
               
                   
               
               
                 349068 
                 12 
                 515 
                 ACATAGTATATAATCGAATG 
                 329 
               
               
                   
               
               
                 349069 
                 12 
                 517 
                 ATACATAGTATATAATCGAA 
                 330 
               
               
                   
               
               
                 349070 
                 12 
                 531 
                 GATTGCTTTCATCCATACAT 
                 331 
               
               
                   
               
               
                 349071 
                 12 
                 533 
                 CAGATTGCTTTCATCCATAC 
                 332 
               
               
                   
               
               
                 349072 
                 12 
                 536 
                 TCTCAGATTGCTTTCATCCA 
                 333 
               
               
                   
               
               
                 349073 
                 11 
                 537 
                 TTGTCGCCTCCCGCGTCGTG 
                 334 
               
               
                   
               
               
                 349074 
                 12 
                 539 
                 ATCTCTCAGATTGCTTTGAT 
                 335 
               
               
                   
               
               
                 349075 
                 12 
                 541 
                 AGATCTCTCAGATTGCTTTC 
                 336 
               
               
                   
               
               
                 349076 
                 12 
                 543 
                 TGAGATCTCTCAGATTGCTT 
                 337 
               
               
                   
               
               
                 349077 
                 11 
                 574 
                 CTCCTGCCACATGTAGTCCG 
                 338 
               
               
                   
               
               
                 349078 
                 11 
                 643 
                 GTGCTCGGGCCCTTATTTTC 
                 339 
               
               
                   
               
               
                 349079 
                 12 
                 665 
                 CAGCTCGAAGTCAGAGTCAT 
                 340 
               
               
                   
               
               
                 349080 
                 12 
                 667 
                 ACCACCTCGAAGTCAGAGTC 
                 341 
               
               
                   
               
               
                 349081 
                 12 
                 669 
                 ACACCACCTCGAAGTCAGAG 
                 342 
               
               
                   
               
               
                 349082 
                 12 
                 670 
                 TACACCACCTCGAAGTCAGA 
                 343 
               
               
                   
               
               
                 349083 
                 12 
                 671 
                 GTACACCACCTCGAAGTCAG 
                 344 
               
               
                   
               
               
                 349084 
                 12 
                 672 
                 GGTACACCACCTCGAAGTCA 
                 345 
               
               
                   
               
               
                 349085 
                 12 
                 674 
                 CTGGTACACCACCTCGAAGT 
                 346 
               
               
                   
               
               
                 349086 
                 12 
                 675 
                 GCTGGTACACCACCTCGAAG 
                 347 
               
               
                   
               
               
                 349087 
                 12 
                 676 
                 TGCTGGTACACCACCTCGAA 
                 348 
               
               
                   
               
               
                 349088 
                 11 
                 676 
                 TACGACCGCGACGGCCGCGT 
                 349 
               
               
                   
               
               
                 349089 
                 12 
                 677 
                 TTGCTGGTACACCACCTCGA 
                 350 
               
               
                   
               
               
                 349090 
                 11 
                 745 
                 CGAAGGTGTTCGTGTTACTC 
                 351 
               
               
                   
               
               
                 349091 
                 11 
                 824 
                 GGGTTGGCGCGTCGTCGCTG 
                 352 
               
               
                   
               
               
                 349092 
                 11 
                 876 
                 GGGTGGTAGTGGCGGGTGGG 
                 353 
               
               
                   
               
               
                 349093 
                 11 
                 931 
                 GTGGCTGGGTGGTGTCGTGT 
                 354 
               
               
                   
               
            
           
         
       
     
     EXAMPLE 14 
     Antisense Inhibition of LMW-PTPase Expression In Vivo: Mouse Model of Diet-Induced Obesity 
     The C57BL/6 mouse strain is reported to be susceptible to hyperlipidemia-induced atherosclerotic plaque formation. When these mice are fed a high-fat diet, they develop diet-induced obesity. Accordingly these mice are a useful model for the investigation of obesity and treatments designed to treat this conditions. In a further embodiment of the present invention, the oligomeric compounds of the invention are tested in a model of diet-induced obesity. 
     Male C57BL/6 mice received a 60% fat diet for about 12-13 weeks, after which mice were subcutaneously injected with Compound No. 288267 (SEQ ID NO: 186), an antisense oligonucleotide targeted to LMW-PTPase, or the control compound Compound No. 141923 (SEQ ID NO: 274) at a dose of 25 mg/kg two times per week for 6 weeks. Each treatment group was comprised of about 8 to 10 animals. Saline-injected high-fat fed animals serve as a control. As an additional control, mice fed a normal chow diet were treated with saline alone. 
     Body weight and accumulated food intake were measured throughout the study for each treatment group. No significant alterations in accumulated food intake were observed for the animals fed a high-fat diet, regardless of the treatment. At the end of the study, body composition was assayed by MRI. Treatment with Compound No. 288267 caused about a 12% decrease in fat content of the high-fat fed mice over the treatment period. The fat content of high-fat fed animals treated with saline alone or with the control compound Compound No. 141923 did not decrease over the course of the study. Therefore, another embodiment of the present invention is a method of reducing adiposity in an animal by administering an oligomeric compound targeted to LMW-PTPase. 
     To assess the physiological effects resulting from inhibition of target mRNA, the diet-induced obese mice that received treatment were further evaluated at beginning of the study (week 0), during the 3rd week of treatment (week 3.5), and at the end of the treatment period (week 6) for plasma triglyceride and plasma cholesterol levels. Triglycerides and cholesterol are measured by routine clinical analyzer instruments (e.g. Olympus Clinical Analyzer, Melville, N.Y.). Average results for each treatment group are shown in Table 14 in mg/dL. 
     
       
         
           
               
             
               
                 TABLE 14 
               
             
            
               
                   
               
               
                 Effects of antis ense inhibition of LMW-PTPase on plasma lipid levels 
               
            
           
           
               
               
               
            
               
                   
                 Cholesterol 
                 Triglycerides 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                 Week 
                   
                 Week 
                 Week 
                   
               
               
                 Treatment 
                 Week 0 
                 3.5 
                 Week 6 
                 0 
                 3.5 
                 Week 6 
               
               
                   
               
               
                 Saline, high-fat fed 
                 176 
                 175 
                 184 
                 83 
                 65 
                 76 
               
               
                 Compound No. 
                 182 
                 174 
                 181 
                 78 
                 70 
                 59 
               
               
                 141923 
               
               
                 Compound No. 
                 176 
                 157 
                 136 
                 79 
                 60 
                 56 
               
               
                 288267 
               
               
                 Saline, normal diet 
                 82 
                 68 
                 68 
                 74 
                 76 
                 61 
               
               
                   
               
            
           
         
       
     
     As shown in Table 14, Treatment with Compound No. 288267 caused a decrease in plasma cholesterol in diet-induced obese mice. Therefore embodiments of the present invention include methods of lowering cholesterol in an animal by administering an oligomeric compound of the invention. 
     The effects of target inhibition on glucose and insulin metabolism were also evaluated in the diet-induced obese mice treated with the oligomeric compounds of the invention. Plasma glucose was measured at beginning of the study (week 0), during the 3rd week of treatment (week 3.5), and at the end of the treatment period (week 6). Plasma insulin was similarly measured at beginning of the study (week 0), during the 3rd week of treatment (week 3.5), and at the end of the treatment period (week 6). Glucose levels were measured using standard methods (for example, with a YSI glucose analyzer, YSI Scientific, Yellow Springs, Ohio) and insulin levels were measured using a commercially available kit (for example, an Alpco insulin-specific ELISA kit, Windham, N.H.). Hypoglycemia was not observed in animals treated with Compound No. 288267. Insulin levels are shown in Table 15 as a percentage of insulin levels measured for high-fat fed animals treated with saline alone. 
     
       
         
           
               
             
               
                 TABLE 15 
               
             
            
               
                   
               
               
                 Effects of antisense inhibition of LMW-PTPase 
               
               
                 on blood glucose and insulin levels 
               
            
           
           
               
               
               
            
               
                   
                 Insulin 
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 Treatment 
                 Week 0 
                 Week 3.5 
                 Week 6 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 Saline, high-fat 
                 100 
                 114 
                 179 
               
               
                   
                 fed 
               
               
                   
                 Compound No. 
                 100 
                 107 
                 164 
               
               
                   
                 141923 
               
               
                   
                 Compound No. 
                 100 
                 86 
                 100 
               
               
                   
                 288267 
               
               
                   
                 Saline, normal 
                 21 
                 86 
                 79 
               
               
                   
                 diet 
               
               
                   
                   
               
            
           
         
       
     
     As shown in Table 15, treatment with Compound No. 288267 prevented the increase in insulin levels over the course of the study observed in high-fat fed animals treated with saline or with the control compound Compound No. 141923. Therefore, another embodiment of the present invention is a method of improving insulin sensitivity in an animal by administering an oligomeric compound targeted to LMW-PTPase. In one embodiment, improved insulin sensitivity is indicated by a reduction in circulating insulin levels. 
     Glucose tolerance tests were also administered in fasted mice. During the fifth week of treatment, mice were fasted overnight and then received intraperitoneal injections of glucose at a dose of 1 g/kg, and the blood glucose were measured before the glucose challenge and at 30 minute intervals for up to 2 hours. Results are shown in Table 16 for each treatment group. 
     
       
         
           
               
             
               
                 TABLE 16 
               
             
            
               
                   
               
               
                 Effects of antisense inhibition of LMW-PTPase on glucose tolerance 
               
            
           
           
               
               
            
               
                   
                 Glucose (mg/dL) 
               
            
           
           
               
               
               
               
               
               
            
               
                 Treatment 
                 0 min. 
                 30 min. 
                 60 min. 
                 90 min. 
                 120 min. 
               
               
                   
               
               
                 Saline, high-fat 
                 110 
                 346 
                 264 
                 219 
                 194 
               
               
                 fed 
               
               
                 Compound No. 
                 112 
                 317 
                 243 
                 210 
                 185 
               
               
                 141923 
               
               
                 Compound No. 
                 118 
                 277 
                 210 
                 188 
                 168 
               
               
                 288267 
               
               
                 Saline, normal 
                 100 
                 249 
                 184 
                 151 
                 133 
               
               
                 diet 
               
               
                   
               
            
           
         
       
     
     A plot of the data in Table 16 as glucose level as a function of time allows for comparison of the area under the curve for each treatment group. These data reveal improved glucose tolerance in high-fat fed animals treated with Compound No. 288267. Therefore, another aspect of the present invention is a method of improving glucose tolerance in an animal by administering an oligomeric compound of the invention. 
     Insulin tolerance tests were also administered in fasted mice. During the fourth week of treatment, mice were fasted for approximately 4 to 5 hours, and then received intraperitoneal injections of insulin at a dose of 0.5 U/kg. Glucose levels were measured before the insulin challenge and at 30 minute intervals for up to 2 hours. Results are shown in Table 17. 
     
       
         
           
               
             
               
                 TABLE 17 
               
             
            
               
                   
               
               
                 Effects of antisense inhibition of LMW-PTPase on insulin tolerance 
               
            
           
           
               
               
            
               
                   
                 Glucose (m/dL) 
               
            
           
           
               
               
               
               
               
               
            
               
                 Treatment 
                 0 min. 
                 30 min. 
                 60 min. 
                 90 min. 
                 120 min. 
               
               
                   
               
               
                 Saline, high-fat 
                 218 
                 122 
                 123 
                 129 
                 154 
               
               
                 fed 
               
               
                 Compound No. 
                 208 
                 127 
                 108 
                 116 
                 145 
               
               
                 141923 
               
               
                 Compound No. 
                 172 
                 104 
                 87 
                 88 
                 117 
               
               
                 288267 
               
               
                   
               
            
           
         
       
     
     A plot of the data in Table 17 as glucose level as a function of time allows for comparison of the area under the curve for each treatment group. These data reveal improved insulin tolerance in high-fat fed animals treated with Compound No. 288267. Therefore, another aspect of the present invention is a method of improving insulin tolerance in an animal by administering an oligomeric compound of the invention. 
     EXAMPLE 15 
     Antisense Inhibition of LMW-PTPase Expression in a Mouse Model of Diet-Induced Obesity 
     Liver and fat tissues from C57BL/6 mice fed a high fat diet and treated as described in Example 14 were further evaluated at the end of the study for target reduction. Analysis of target reduction in tissues was performed as described herein. Treatment with Compound No. 288267 reduced LMW-PTPase gene expression by about 90% in liver and about 75% in fat (white adipose tissue), whereas treatment with Compound No. 141923 did not reduce LMW-PTPase expression in either tissue. Treatment with Compound No. 288267 also caused a significant reduction in hepatic glucose-6-phosphatase expression, suggesting a suppression of hepatic glucose output. Treatment with Compound No. 288267 did not have an effect on the expression of SHP2, SHPTP2 or PTEN expression. Similar results are seen with ob/ob mice. Therefore, another embodiment of the present invention is a method of reducing hepatic glucose output in an animal by administering an oligomeric compound of the invention. Another embodiment of the present invention is a method of modulating genes involved in glucose metabolism by administering an oligomeric compound of the invention. In one embodiment, the modulated gene is glucose-6-phosphatase. 
     Liver triglyceride levels were also assessed, using a commercially available kit (for example, using the Triglyceride GPO Assay from Roche Diagnostics, Indianapolis, Ind.). Liver triglyceride content was reduced with Compound No. 288267 treatment as compared to treatment with Compound No. 141923 (about 35 mg/g vs. about 56 mg/g tissue, respectively). 
     Hepatic steatosis may also be assessed by routine histological analysis of frozen liver tissue sections stained with oil red 0 stain, which is commonly used to visualize lipid deposits, and counterstained with hematoxylin and eosin, to visualize nuclei and cytoplasm, respectively. 
     Therefore, another embodiment of the present invention is a method of decreasing hepatic triglyceride accumulation in an animal by administering an oligomeric compound of the invention. Another embodiment of the present invention is a method of treating steatosis in an animal by administering an oligomeric compound of the invention. In one embodiment, the steatosis is steatohepatitis. In one embodiment, the steatosis is NASH. 
     Another embodiment of the present invention is a method of treating diabetes or metabolic syndrome comprising administering an oligomeric compound of the invention. In some embodiments, the oligomeric compound inhibits LMW-PTPase expression in the liver, in fat, or in both tissues.