Publications (2)9.11 Total impact
-
Article: Serotonin mechanisms in heart valve disease I: serotonin-induced up-regulation of transforming growth factor-beta1 via G-protein signal transduction in aortic valve interstitial cells.
[show abstract] [hide abstract]
ABSTRACT: Clinical disorders associated with increased serotonin [5-hydroxytryptamine (5-HT)] levels, such as carcinoid syndrome, and the use of serotonin agonists, such as fenfluoramine have been associated with a valvulopathy characterized by hyperplastic valvular and endocardial lesions with increased extracellular matrix. Furthermore, 5-HT has been demonstrated to up-regulate transforming growth factor (TGF)-beta in mesangial cells via G-protein signal transduction. We investigated the hypothesis that increased exposure of heart valve interstitial cells to 5-HT may result in increased TGF-beta1 expression and activity because of serotonin receptor-mediated signal transduction with activation of Galphaq, and subsequently up-regulation of phospholipase C. Thus, in the present study we performed a clinical-pathological investigation of retrieved carcinoid and normal valve cusps using immunohistochemical techniques to detect the presence of TGF-beta1 and other proteins associated with TGF-beta expression, including TGF-beta receptors I and II, latent TGF-beta-associated peptide (LAP), and alpha-smooth muscle actin. Carcinoid valve cusps demonstrated the unusual finding of widespread smooth muscle actin involving the interstitial cells in the periphery of carcinoid nodules; these same cells were also positive for LAP. Normal valve cusps were only focally positive for smooth muscle actin and LAP. In sheep aortic valve interstitial cell cultures 5-HT induced TGF-beta1 mRNA production and increased TGF-beta1 activity. 5-HT also increased collagen biosynthesis at the dosages studied. Furthermore, TGF-beta1 added to SAVIC cultures increased the production of sulfated glycan and hyaluronic acid. In addition, overexpression of Galphaq using an adenoviral expression vector for a constitutively active Galphaq mutant (Q209L-Galphaq) resulted in increased phospholipase C activity as well as up-regulation of TGF-beta expression and activity. These results strongly support the view that G-protein-related signal transduction is involved in 5-HT up-regulation of TGF-beta1. In conclusion, 5-HT-associated valve disease may be, in part, because of TGF-beta1 mechanisms.American Journal Of Pathology 01/2003; 161(6):2111-21. · 4.89 Impact Factor -
Article: Gene delivery to pig coronary arteries from stents carrying antibody-tethered adenovirus.
[show abstract] [hide abstract]
ABSTRACT: Deployment of coronary stents to relieve atherosclerotic obstruction has benefitted millions of patients. However, gene therapy to prevent in-stent restenosis, while promising in experimental studies, remains a challenge. Conventional strategies for viral vector administration utilize catheters that deliver infusions of viral suspensions, which result in suboptimal localization and potentially dangerous distal spread of vector. Stent-based gene delivery may circumvent this problem. We hypothesized that site-specific delivery of adenoviral gene vectors from a stent could be achieved through a mechanism involving anti-viral antibody tethering. Stents were formulated with a collagen coating. Anti-adenoviral monoclonal antibodies were covalently bound to the collagen surface. These antibodies enabled tethering of replication defective adenoviruses through highly specific antigen-antibody affinity. We report for the first time successful stent-based gene delivery using antibody-tethered adenovirus encoding green fluorescent protein (GFP), demonstrating efficient and highly localized gene delivery to arterial smooth muscle cells in both cell culture and pig coronary arteries. Overall arterial wall transduction efficiency in pigs was 5.9 +/- 1.1% of total cells. However, neointimal transduction was more than 17% of total cells in this region. Importantly, when specific antibody was used to tether adenovirus, no distal spread of vector was detectable by PCR, in either distal organs, or in the downstream segments of the stented arteries. Control adenovirus stents, with nonspecific antibody plus adenovirus, demonstrated only a few isolated foci of transduction, and poor site-specific transduction with distal spread of vector. We conclude that a vascular stent is a suitable platform for a localizable viral vector delivery system that also prevents systemic spread of vector. Gene delivery using stent-based anti-viral antibody tethering of vectors should be suitable for a wide array of single or multiple therapeutic gene strategies.Human Gene Therapy 03/2002; 13(3):443-54. · 4.22 Impact Factor
