Compositions, methods and kits relating to remodel

Details Compositions, methods and kits relating to remodel Compositions, methods and kits relating to remodel

2000-10-19

2003-10-07

Chan, Christina (Department: 1644)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

C536S023500, C435S455000, C435S252300, C435S320100

Reexamination Certificate

active

06630325

ABSTRACT:

BACKGROUND OF THE INVENTION
The present invention relates to identifying novel processes involved in mediating arterial remodeling.
Arterial stenosis with reduction in blood flow is a common problem in many vascular diseases. Several growth factors have been implicated in the mechanisms leading to vascular stenosis. For instance, fibroblast growth factor 2 (FGF-2) has been identified as an important factor in mediating proliferation of smooth muscle cells leading to intimal lesion formation. Furthermore, it has been demonstrated that arterial stenosis in response to angioplasty is largely due to negative remodeling as a result of adventitial fibrosis. As more fully set forth below, transforming growth factor beta (TGF-&bgr;) signaling has been demonstrated to play an important role in arterial stenosis in that, among other things, inhibition of TGF-&bgr; signaling using a soluble TGF-&bgr; receptor type II dramatically reduced lumen narrowing by decreasing negative remodeling and adventitial matrix deposition as well by decreasing neointima formation. These results indicate the crucial role of TGF-&bgr; signaling in arterial response to injury.
Vascular remodeling is a response of blood vessels to both physiological and pathological stimuli, leading to either vessel enlargement (positive remodeling) or shrinkage (negative remodeling). It has been demonstrated that neointimal proliferation or intimal mass following angioplasty shows little correlation with restenosis because of permanent changes in vascular geometry (Kakuta et al., 1994, Circulation 89:2809-2815; Nunes et al., 1995, Arterioscler. Thromb. Vasc. Biol. 15:156-165). Negative remodeling has been shown to account for most of the restenosis process (Mintz et al., 1993, Circulation 88:1-654), and is now generally considered the predominant cause of restenosis. A successful therapeutic approach to restenosis, therefore, would target negative vascular remodeling.
Several growth factors have been implicated in the mechanisms leading to vascular stenosis, such as fibroblast growth factor-2 (FGF-2) and transforming growth factor-&bgr; (TGF-&bgr;). Specifically, cellular responses involving TGF-&bgr; in the adventitia have gained increased attention for their potential involvement in adventitial remodeling (Wilcox et al., 1996, Int. J. Radiat. Oncol. Biol. Phys. 36:789-796; Wilcox and Scott, 1996, Int. J. Cardiol. 54S:S21-35; Shi et al., 1996, Circulation 93:340-348). There is evidence that proliferative events occurring in the adventitia contribute to vascular remodeling and restenosis in response to vascular injury (Wilcox et al., 1996, Int. J. Radiat. Oncol. Biol. Phys. 36:789-796; Wilcox et al., 1997, Ann. N.Y. Acad. Sci. 811:437-447; Scott et al., 1996, Circulation 93:2178-2187). There is now general agreement that TGF-&bgr; is a potential factor in the adventitial remodeling process (Shi et al., 1996, Arterioscler. Thromb. Vasc. Biol. 16:1298-1305).
Although it is known that the TGF-&bgr; family of cytokines can have a variety of effects on vascular cells, very little is known about the role of this family of cytokines in vascular remodeling. TGF-&bgr; affects many functions including proliferation of smooth muscle cells (SMC) (Halloran et al., 1995, Am. J. Surg. 170:193-197). It has been demonstrated that inhibition of SMC proliferation by TGF-&bgr; occurs via extension of the G2 phase of the cell cycle (Grainger et al., 1994, Biochem. J. 299:227-235). In contrast, it has also been shown that inhibition of SMC proliferation by TGF-&bgr;1 is due to arrest in the late G1 phase of the cell cycle (Reddy and Howe, 1993, J. Cell Physiol. 156:48-55). SMC derived from atherosclerotic lesions responded to TGF-&bgr;1 with an increase in proliferation, and lower levels of TGF-&bgr; receptor II (TGF-&bgr;RII) have been implicated in the lack of inhibition by TGF-&bgr; in these cells (McCaffrey et al., 1995, J. Clin. Invest. 96:2667-2675).
Further studies have established that TGF-&bgr;1 stimulates SMC proliferation in vitro. Low doses of TGF-&bgr;1 stimulated SMC proliferation via platelet-derived growth factor (PDGF)-amino acid (AA)-dependent and PDGF-AA-independent mechanisms, while higher doses of TGF-&bgr;1 were inhibitory (Battegay et al., 1990, Cell 63:515-524; Stouffer and Owens, 1994, J. Clin. Invest. 93:2048-2055). Bifunctional effects of TGF-&bgr;1 in migration assays with SMC were also demonstrated (Koyama et al., 1990, Biochem. Biophys. Res. Commun. 169:725-729; Mii et al., 1993, Surgery 114:464-470).
TGF-&bgr;1 also plays a role in intimal lesion formation as indicated by a 5-7 fold induction of TGF-&bgr;1 mRNA in the balloon-injured rat carotid artery, with elevated levels of TGF-&bgr;1 mRNA persisting for 2 weeks (Majesky et al., 1991, J. Clin. Invest. 88:904-910). During the 2 week period, elevated TGF-&bgr;1 mRNA levels correlated with increases in mRNA expression of fibronectin and alpha-2 (I) and alpha-1 (III) collagens. These studies also demonstrated that infusion of recombinant TGF-&bgr;1 caused an increase in intimal SMC proliferation in vivo (id.).
Among clinically significant findings regarding the role of TGF-&bgr; signaling in arterial response to injury, it has been demonstrated that TGF-&bgr;1 mRNA expression in restenotic lesions compared to primary atherosclerotic lesions is increased (Nikol et al., 1992, J. Clin. Invest. 90:1582-1592). In the rat balloon injury model, treatment with TGF-&bgr;1 antibodies caused a small but significant reduction in neointima formation (Wolf et al., 1994, J. Clin. Invest. 93:1172-1178). Overexpression of TGF-&bgr;1 in the rat carotid artery by adenoviral gene transfer led to transient neointima formation with cartilaginous metaplasia that almost completely resolved within 8 weeks (Shulick et al., 1998, Proc. Natl. Acad. Sci. USA 95:6983-6988). Without wishing to be bound by any particular theory, TGF-&bgr;1 may also effect vascular tone since the factor was found to suppress nitric oxide synthase expression (Perella et al., 1996, J. Biol. Chem. 271:13776-13780) while at the same time inducing the vasoconstrictor endothelin in SMC in vitro (Kurihara et al., 1989, Biochem. Biophys. Res. Commun. 159:1435-1440). Further, TGF-&bgr;1 has been implicated in anti-apoptotic effects in SMC (Herbert and Carmeliet, 1997, FEBS Lett. 413:401-404).
Studies examining the expression of TGF-&bgr; ligand and TGF-&bgr; receptor (TGF-&bgr;R) mRNAs using reverse transcriptase polymerase chain reaction (RT-PCR) analysis revealed that TGF-&bgr;1, TGF-&bgr;3, and TGF-&bgr;RII mRNA levels were increased in the media of the injured rat carotid artery (Ward et al., 1997, Arterioscler. Thromb. Vasc. Biol. 17:2461-2470) and expression of TGF-&bgr;2 and TGF-&bgr;3 were also reported in SMC of the lung vasculature (Khalil et al., 1996, Am. J. Respir. Cell Mol. Biol. 14:131-138; Pelton et al., 1991, Am. J. Respir. Cell Mol. Biol. 5:522-530). However, reduced levels of TGF-&bgr;RII were demonstrated in human atherosclerotic lesions (McCaffrey et al., 1995, J. Clin. Invest. 96:2667-2675). The three TGF-&bgr; ligands have overlapping functions and all of them induce expression of the alpha-1 (I), alpha-2 (I) and alpha-1 (III) chains of collagen (Bray et al., 1998, Hypertension 31:986-994).
The role of TGF-&bgr; isoforms in vascular repair processes was examined using a rat balloon catheter denudation model (Smith et al., 1999, Circ. Res. 84:1212-1222). Proliferating and quiescent SMC in denuded vessels expressed high levels of mRNA for TGF-&bgr;1, TGF-&bgr;2, and TGF-&bgr;3, and lower levels of TGF-&bgr;RII mRNA (Smith et al., 1999, Circ. Res. 84:1212-1222). The role of TGF-&bgr; signaling in the rat carotid artery balloon injury model was tested and it was shown that control vessels developed an extensive neointima and adventitial fibrosis with abundant collagen production. Vessels from animals injected with a recombinant soluble TGF-&bgr;RII (designated as “TGF-&bgr;R:Fc”) revealed only little neointima formation and much less collagen deposition in the adventitia. The adventitia also contained sign

Affiliated with

Also associated with

Rating

Compositions, methods and kits relating to remodel does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Compositions, methods and kits relating to remodel, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Compositions, methods and kits relating to remodel will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3157454