Regulation of androgen action by receptor gene inhibition.
Regulated functions of hormonal agents play a critical role in health and disease. Target cell responsiveness to a hormonal signal is a product of both cellular concentrations of the hormone ligand and the corresponding receptor protein. The major thrust of the drug design for treatment of endocrine-related problems, so far, has been directed to ligand derivatives. In certain cases, receptor regulation through antigene technology has much to offer with improvements in both target cell and hormonal specificity. Three different antigene approaches are currently being explored. The first approach is to inhibit the expression of the receptor gene by disrupting the DNA protein interaction at critical cis-elements by short triple helix-forming oligonucleotides. The second approach is to sequester and inactivate the receptor mRNA by the antisense mRNA produced in the target tissue directed by a heterologous tissue-specific promoter. The third approach is the tissue-specific expression of a catalytic ribozyme that binds to the specific receptor mRNA and selectively degrades it before its translation into the protein. In this study, we have characterized the promoter of the rat androgen receptor, and by progressive deletion from its 5' end have identified two critical cis-regulatory elements, one at the -960 to -940 region and the other at the -554 to -574 positions. The former is an activator while the latter is an inhibitor domain. The inhibitory domain is the binding site for the nuclear factor kappa B (NF-kappa B) and more specifically, the p50/p50 homodimer of this transcription factor family. We have also provided correlative data to show that under normal physiological conditions, the NF-kappa B functions as an antiandrogen during the age-dependent desensitization of the liver. In addition to the naturally functioning antiandrogenic influence of NF-kappa B, we have designed an artificial antiandrogenic agent, a triplex-forming oligonucleotide (TFO) directed to the -960/-940 activator domain of the rat androgen receptor gene promoter. This oligonucleotide at a TFO-to-promoter ratio of 500 is able to cause about 60% inhibition of rAR promoter function in transfected COS-1 cells. These results clearly demonstrate the feasibility of the antigene approach for effective inhibition of steroid hormone action.
Song
C S
CS
Department of Cellular & Structural Biology, University of Texas Health Science Center at San Antonio 78284, USA.
Jung
M H
MH
Supakar
P C
PC
Chen
S
S
Vellanoweth
R L
RL
Chatterjee
B
B
Roy
A K
AK
D000728:Androgens / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D011944:Receptors, Androgen / Q000235:genetics* / Q000378:metabolism
D000728:Androgens; D016328:NF-kappa B; D011944:Receptors, Androgen; D004247:DNA