Method for the inhibition of function of STAT transcription factors

#1. Field of the Invention

#The present invention relates to transcription factors, which induce the transcription of genes. More specifically, the present invention relates to a method of inhibiting transcription factors for use in the therapeutic treatment of disease associated with hyper-activated signal transduction pathways, particularly malignancy.

#2. Description of Related Art

#Other advantages of the present invention will be readily appreciated as the same becomes better understood by reference to the following detailed description when considered in connection with the accompanying drawings wherein:

#FIG. 1 shows the sequence of the sense strand of double-stranded DNA fragments that were used for gel mobility assays and for the inhibition of the activation of STAT5;

#FIG. 2 shows the constitutive activation of a STAT-like DNA-binding factor in Dami/HEL and Meg-01 cells; Nuclear extracts from each cell line cultured in the absence of cytokine (CTL) or in the presence of as much as 400 ng/ml TPO or 40 ng/ml IL-3, were incubated with [32P]-labeled, double-stranded IRF-1 GAS (SEQ ID NO. 2) oligonucleotide; The DNA-binding complex and unbound probes were separated electrophoretically on 5% non-denaturing polyacrylamide gels; The autoradiograph shows the STAT-like DNA-binding factor (DBF) and the nonspecific bands (NSB);

#Generally, the present invention relates to a method of inhibiting the function of transcription factors in viable cells, using specific, double-stranded oligonucleotides that contain the optimal nucleotide binding site(s) recognized by a given transcription factor. This method is applicable in vitro for inhibiting DNA binding and function of any transcription factor that targets a specific nucleotide sequence to which it binds on native, endogenous DNA to exert its transcriptional activation function. The method can also use a specific nucleotide sequence which represents a consensus sequence of the targeted DNA upon which the transcription factor acts. Some examples of such transcription factors include (but are not limited to) the STAT family (STATs 1, 2, 3, 4, 5a, 5b, and 6), fos/jun, NF kappa B, HIV-TAT, and the E2F family.

#The present invention provides a method of inhibiting the function of transcription factors as shown in the Example herein.

#Contrary to that used in the art, i.e. antisense oligonucleotides, DNA triplex-forming oligonucleotides, and ribozymes that are engineered to bind to specific DNA or RNA sequences, the present invention uses double-stranded oligonucleotides that contain, for example, the consensus STAT5 binding site, TTCNNNGAA, to competitively inhibit the ability of activated STAT to bind to its endogenous DNA targets.