Compositions and methods for treating and preventing neointimal stenosis
Methods for treating or preventing neointima stenosis are disclosed. The methods generally involve the use of a TGFβ inhibitor, a SMAD2 inhibitor, an FGF Receptor agonist, a Let-7 agonist, or a combination thereof, to inhibit endothelial-to-mesenchymal transition (Endo-MT) of vascular endothelial cells into smooth muscle cells (SMC) at sites of endothelial damage. The disclosed methods can therefore be used to prevent or inhibit neointimal stenosis or restenosis, e.g., after angioplasty, vascular graft, or stent. Also disclosed are methods for increasing the patency of biodegradable, synthetic vascular grafts using a composition that inhibits Endo-MT. A cell-free tissue engineered vascular graft (TEVG) produced by this method is also disclosed.
1. A cell-free polymeric vascular graft, scaffold, or stent comprising an effective amount of a TGFβ inhibitor, a SMAD2 inhibitor, an FGF receptor agonist, a Let-7 agonist, or a combination thereof, to increase the recruitment of host cells to the graft, scaffold, or stent, and promote endothelial cell growth but not smooth muscle cell proliferation, relative to the graft, scaffold, or stent without the TGFβ inhibitor, a SMAD2 inhibitor, an FGF receptor agonist, a Let-7 agonist, or a combination thereof.
2. The cell-free polymeric vascular graft, scaffold, or stent of claim 1 , comprising biodegradable or bioabsorbable polymers.
3. The cell-free polymeric vascular graft, scaffold, or stent of claim 2 , wherein the biodegradable or bioabsorbable polymers are selected from the group consisting of poly(lactic acid), poly(glycolic acid), polyanhydrides, poly(ortho)esters, polyesters, polyurethanes, poly(butic acid), poly(valeric acid), poly(caprolactone), poly(hydroxyalkanoates), and poly(lactide-co-caprolactone), or combinations, blends or co-polymers thereof.
4. The cell-free polymeric vascular graft, scaffold, or stent of claim 2 , wherein the biodegradable or bioabsorbable polymers are formed into a fiber-based mesh.
5. The cell-free polymeric vascular graft, scaffold, or stent of claim 4 wherein the fiber-based mesh is a non-woven mesh.
6. The cell-free polymeric vascular graft, scaffold, or stent of claim 2 , wherein the vascular graft further comprises a polymeric sealant.
7. The cell-free polymeric vascular graft, scaffold, or stent of claim 6 , wherein the polymeric sealant comprises a co-polymer of ε-caprolactone and L-lactide.
8. The cell-free polymeric vascular graft, scaffold, or stent of claim 1 , wherein the TGFβ inhibitor or SMAD2 inhibitor is selected from the group consisting of 4-[4-(1,3-Benzodioxol-5-yl)-5-(2-pyridinyl)-1H-imidazol-2-yl]-benzamide (SB431542), 2-(5-Chloro-2-fluorophenyl)-4-[(4-pyridyl)amino]pteridine (SB208), 6-[2-(1,1-Dimethylethyl)-5-(6-methyl-2-pyridinyl)-1H-imidazol-4-yl]quinoxaline (SB525334), 4-(5-(benzo[d][1,3]dioxol-5-yl)-4-(6-methylpyridin-2-yl)-1H-imidazol-2-yl)4yridin[2.2.2]octane-1-carboxamide (SM16), 4-(2-(6-methylpyridin-2-yl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)4yridine4-6-carboxamide (LY2157299), 3-((5-([1,2,4]triazolo[1,5-a]4yridine-6-yl)-4-(6-methylpyridin-2-yl)thiazol-2-ylamino)methyl)benzonitrile (EW-7203), and (2-butyl-4-chloro-1-{[2′-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl}-1H-imidazol-5-yl)methanol (Losartan).
9. The cell-free polymeric vascular graft, scaffold, or stent of claim 1 , wherein the Let-7 agonist is a polynucleotide encoding a Let-7 pri-miRNA, pre-miRNA, mature miRNA, or RNAi effective in silencing TGFβRI gene expression.
10. The cell-free polymeric vascular graft, scaffold, or stent of claim 9 , wherein the Let-7 agonist comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:102, 103, 104, 105, 105, 106, 107, 108, 109, 110, 111, 112, or 113, or a fragment or variant thereof capable of binding a let-7 target sequence and silence TGFβRI gene expression.
11. The cell-free polymeric vascular graft, scaffold, or stent of claim 1 , wherein the vascular graft, scaffold, or stent further comprises an active agent selected from the group consisting of anti-thrombogenic agents, anti-proliferative agents, anti-inflammatory agents, antiproliferative agents, anesthetic agents, anti-coagulants, cholesterol-lowering agents, vasodilating agents, and agents which interfere with endogenous vascoactive mechanisms.
12. The cell-free polymeric vascular graft, scaffold, or stent of claim 1 , wherein the TGFβ inhibitor, SMAD2 inhibitor, FGF receptor agonist, Let-7 agonist, or combination thereof, is dispersed throughout the vascular graft, scaffold, or stent.
13. The cell-free polymeric vascular graft, scaffold, or stent of claim 1 , wherein the TGFβ inhibitor, SMAD2 inhibitor, FGF receptor agonist, Let-7 agonist, or combination thereof, is encapsulated in the form of microspheres, nano spheres, microparticles and/or microcapsules that are seeded into the vascular graft, scaffold, or stent.
14. The cell-free polymeric vascular graft, scaffold, or stent of claim 1 , wherein the TGFβ inhibitor or the SMAD2 inhibitor comprises an antibody, an antibody fragment, an antigen-binding variable domain of an antibody, a soluble receptor, or a mutant ligand.
15. The cell-free polymeric vascular graft, scaffold, or stent of claim 1 , wherein the effective amount the TGFβ inhibitor, the SMAD2 inhibitor, the FGF receptor agonist, the Let-7 agonist, or the combination thereof, is an amount effective to inhibit endothelial to mesenchymal transition of vascular endothelial cells at the site of the vascular graft, scaffold, or stent implantation.
16. The cell-free polymeric vascular graft, scaffold, or stent of claim 1 , wherein the cell-free polymeric vascular graft, scaffold, or stent retains its patency for at least about two weeks following implantation.