Inhibition of vascular smooth muscle cell proliferation in vitro and in vivo by c-myc antisense oligodeoxynucleotides.

Abstract

Restenosis after angioplasty is due predominantly to accumulation of vascular smooth muscle cells (VSMCs). The resistance of restenosis to pharmacological treatment has prompted investigation of genes involved in VSMC proliferation. We have examined the effect on VSMC proliferation of blocking expression of the c-myc proto-oncogene with antisense oligodeoxynucleotides, both in vitro and in a rat carotid artery injury model of angioplasty restenosis. Antisense c-myc oligodeoxynucleotides reduced average cell levels of c-myc mRNA and protein by 50-55% and inhibited proliferation of VSMCs when mitogenically stimulated from quiescence or when proliferating logarithmically (IC5o = 10 ug/ml). Corresponding sense c-myc, two-basepair mismatch antisense c-myc, antisense a-actin or glyceraldehyde phosphate dehydrogenase oligodeoxynucleotides did not suppress c-myc expression or inhibit VSMC proliferation. Antisense c-myc inhibition was relieved by overexpression of an exogenous c-myc gene. After balloon catheter injury, peak cmyc mRNA expression occurred at 2 h. Antisense c-myc applied in a pluronic gel to the arterial adventitia reduced peak c-myc expression by 75% and significantly reduced neointimal formation at 14 d, compared with sense c-myc and gel application alone. We conclude that c-myc expression is required for VSMC proliferation in vitro and in the vessel wall. C-myc is a therefore a potential target for adjunctive therapy to reduce angioplasty restenosis. (J. Clin. Invest. 1994. 93:820-828.

Vascular Smooth Muscle Cell Proliferation and Regrowth after Mechanical Injury in Vitro Are. Egr-1/NGFI-A-Dependent

“…To determine the temporal pattern with which E11 associates with SMCs, the PS-ODN was 5Ј end-labeled with 32 P by kinase reaction and incubated with the cells for various times. 32 P-E11 was taken up most efficiently within the first 4 hours of exposure and optimally after 6 hours ( Figure 3).…”

High efficiency reporter gene transfection of vascular tissue in vitro and in vivo using a cationic lipid–DNA complex

“…remained constant at 30 to 50%, despite drug regimens Many protocols involving cationic liposomes for the which have been used in an attempt to control it. 1,2 The delivery of DNA, [16][17][18][19]21 or antisense oligonucleoexact cause of restenosis remains controversial, although tides 11,12,22 into cells and tissues have been described in the migration and overproliferation of vascular smooth vitro and in vivo. 23 Indeed, the protocols have progressed muscle cells (VSMCs) and simultaneous production of to the point where a number of human gene therapy extracellular matrix have been implicated.…”

Tissue-selective expression of dominant-negative proteins for the regulation of vascular smooth muscle cell proliferation

“…Antisense oligonucleotides targeted against c-myc and c-myb have been reported to result in reduced c-myc and c-myb protein levels and inhibition of proliferation of human and rat VSMCs in vitro. [16][17][18][19] Similarly, in vivo transfection studies utilizing antisense c-myc and c-myb oligonucleotides have reported reduced c-myc or c-myb expression and reduced neointimal thickening following vascular injury in a number of animal models. 16,[20][21][22] However, therapeutic application of antisense oligonucleotides is hampered by cellular uptake and stability, as well as by their lack of cell-and possibly target mRNA-specificity.…”

“…[16][17][18][19] Similarly, in vivo transfection studies utilizing antisense c-myc and c-myb oligonucleotides have reported reduced c-myc or c-myb expression and reduced neointimal thickening following vascular injury in a number of animal models. 16,[20][21][22] However, therapeutic application of antisense oligonucleotides is hampered by cellular uptake and stability, as well as by their lack of cell-and possibly target mRNA-specificity. [23][24][25][26] Consequently, antisense gene expression vectors containing the c-myc cDNA have been created and shown to reduce cmyc expression and cellular proliferation in a number of cell types.…”