Inhibition of TGF-β and VEGF expression in human endothelium cell lines with antisense-oligonucleotides: potential tools for gene therapy of chronic renal transplant rejection

Hallscheidt, Peter; Wallich, Reiner; Kauffmann, G. W.; Pomer, S.

doi:10.1016/s0041-1345(01)01926-1

Cited by 6 publications

(5 citation statements)

References 7 publications

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“…Inhibition of VEGF using antisense oligonucleotides has been reported in human endothelial cell lines (Hallscheidt et al 2001), human glioma cells (Zheng et al 2000) and retinopathy animal models (Robinson et al 1995;Garrett et al 2001). Robinson (1996) demonstrated that antisense oligonucleotides against VEGF inhibited retinal neovascularization in a murine model of proliferative retinopathy, suggesting their potential use in the treatment of retinal neovascular diseases.…”

Section: Discussionmentioning

confidence: 99%

Nanoparticle formulation enhances the delivery and activity of a vascular endothelial growth factor antisense oligonucleotide in human retinal pigment epithelial cells

Aukunuru

Ayalasomayajula

Kompella

2003

Journal of Pharmacy and Pharmacology

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The objective of this study was to investigate the delivery and activity of a vascular endothelial growth factor (VEGF) antisense oligonucleotide in a human retinal pigment epithelial cell line (ARPE-19) using a biodegradable nanoparticulate delivery system. A 19-mer antisense phosphorothioate oligonucleotide (PS-ODN) complementary to bases 6-24 relative to the translational start site of the VEGF mRNA, a sense PS-ODN and a mismatch PS-ODN were examined for the inhibition of secretion and mRNA expression of VEGF using an enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction, respectively. Nanoparticles of the antisense oligonucleotides were formulated using a poly(lactide-co-glycolide) (50:50) copolymer using a double emulsion solvent evaporation method. After preparing nanoparticles, drug loading, encapsulation efficiency and particle size were determined. The cells were exposed to either plain solution of oligonucleotide or nanoparticles of oligonucleotide from Day 3 through Day 6. Alternatively, the cells were incubated with PS-ODNs and lipofectin for 4 h on Day 4. In all studies, VEGF secretion and mRNA expression were determined on Day 6. The particle size, drug loading and encapsulation efficiency were 252 nm, 5.5% and 16.5%, respectively. The antisense PS-ODN inhibited VEGF mRNA and protein secretion when delivered using nanoparticles or lipofectin but not in its free form. This was consistent with the ability of nanoparticles and lipofectin to elevate the cellular uptake of the oligonucleotide by 4-fold and 13-fold, respectively. Neither mismatch nor sense oligonucleotides inhibited VEGF secretion. In conclusion, biodegradable nanoparticles enhance cellular delivery of a VEGF antisense oligonucleotide and inhibit VEGF secretion and mRNA expression in a human retinal pigment epithelial cell line.

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Section: Discussionmentioning

confidence: 99%

Nanoparticle formulation enhances the delivery and activity of a vascular endothelial growth factor antisense oligonucleotide in human retinal pigment epithelial cells

Aukunuru

Ayalasomayajula

Kompella

2003

Journal of Pharmacy and Pharmacology

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“…CRLR is expressed in endothelial cells and is a receptor for adrenomedullin, a potent vasodilator and permeability factor, 30 whereas IL-1␤ has been shown to induce VEGF expression in human endothelial cell lines. 31,32 IL-1␤ also mediates ischemic injury in the retina as a consequence of increased ocular pressure 33 and has been shown to be upregulated in the retina in early diabetes. 21 PAI2 is a fibrinolytic factor that is associated with endothelial proliferation and migration, 34 whereas the FGF family has been implicated in a number of biological activities including angiogenesis and mitogenesis.…”

Section: Discussionmentioning

confidence: 99%

Argon Laser Photocoagulation–Induced Modification of Gene Expression in the Retina

Wilson

Hobbs

Shen

et al. 2003

Invest. Ophthalmol. Vis. Sci.

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“…Inhibition of constitutive AGT expression by blockade of autocrine TGF-1 in human lung myofibroblasts. Primary fibroblasts from fibrotic (HIPF) human lung were cultured in serum-free medium with and without TGF-1-neutralizing antibodies (anti-TGF ), isotype-matched control antibodies (NS IgG), or with antisense (AS) or scrambled control (SCR) oligonucleotides against TGF-1 mRNA [34]. After 24 hours, cells were harvested and subjected to realtime PCR for AGT and -actin.…”

Section: A Profibrotic Angiotensin-tgf-1 Auto-crine Loop In Human Lunmentioning

confidence: 99%

Angiotensin-TGF-β1 Crosstalk in Human Idiopathic Pulmonary Fibrosis:Autocrine Mechanisms in Myofibroblasts and Macrophages

Uhal

Kim

et al. 2007

CPD

135

126

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Angiotensin II (ANGII) has been identified as a proapoptotic and profibrotic factor in experimental lung fibrosis models, and patients with the ID/DD polymorphism of ANG converting enzyme (ACE), which confers higher levels of ACE, are predisposed to lung fibrosis (Hum. Pathol. 32:521-528, 2001). Previous work from this laboratory has shown that human lung myofibroblasts isolated from patients with Idiopathic Pulmonary Fibrosis (IPF) synthesize the ANGII precursor angiotensinogen (AGT) constitutively. In attempts to understand the mechanisms and consequences of constitutive AGT synthesis by myofibroblasts, we studied myofibroblast-rich primary cultures of lung fibroblasts from patients with IPF (HIPF isolates), primary fibroblasts from normal human lung (NLFs), the IMR90 and WI38 human lung fibroblasts cell lines, and paraffin sections of lung biopsies from patients with IPF. Compared to the normal NLF isolates, HIPF primary fibroblast isolates constitutively synthesized more AGT and TGF-beta1 mRNA, and released more AGT protein, ANGII and active TGF-beta1 protein into serum-free conditioned media (both p<0.01). Incubation of HIPF fibrotic isolates with the ANGII receptor antagonist saralasin reduced both TGF-beta1 mRNA and active protein, suggesting that the constitutive expression of AGT drives the higher expression of TGF-beta1 by the HIPF cells. Consistent with this premise, treatment of either the primary NLFs or the WI38 cell line with 10(-7) M ANGII increased both TGF-beta1 mRNA and soluble active TGF-beta1 protein. Moreover, induction of the myofibroblast transition in the IMR90 cell line with 2 ng/ml TGF-beta1 increased steady state AGT mRNA levels by realtime PCR (8-fold, p<0.01) and induced expression of an AGT promoter-luciferase reporter construct by over 10-fold (p<0.001). Antisense oligonucleotides against TGF-beta1 mRNA or TGF-beta neutralizing antibodies, when applied to the fibrotic HIPF cells in serum-free medium, significantly reduced AGT expression. In lung sections from IPF patient biopsies, immunoreactive AGT/ANGI proteins were detected in myofibroblasts, epithelial cells and presumptive alveolar macrophages. Together, these data support the existence of an angiotensin/TGF-beta1 "autocrine loop" in human lung myofibroblasts and also suggest ANG peptide expression by epithelia and macrophages in the IPF lung. These findings may explain the ability of ACE inhibitors and ANG receptor antagonists to block experimental lung fibrosis in animals, and support the need for evaluation of these agents for potential treatment of human IPF. This manuscript discusses the data described above and their implications regarding IPF pathogenesis.

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Inhibition of TGF-β and VEGF expression in human endothelium cell lines with antisense-oligonucleotides: potential tools for gene therapy of chronic renal transplant rejection

Cited by 6 publications

References 7 publications

Nanoparticle formulation enhances the delivery and activity of a vascular endothelial growth factor antisense oligonucleotide in human retinal pigment epithelial cells

Nanoparticle formulation enhances the delivery and activity of a vascular endothelial growth factor antisense oligonucleotide in human retinal pigment epithelial cells

Argon Laser Photocoagulation–Induced Modification of Gene Expression in the Retina

Angiotensin-TGF-β1 Crosstalk in Human Idiopathic Pulmonary Fibrosis:Autocrine Mechanisms in Myofibroblasts and Macrophages

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Inhibition of TGF-β and VEGF expression in human endothelium cell lines with antisense-oligonucleotides: potential tools for gene therapy of chronic renal transplant rejection

Cited by 6 publications

References 7 publications

Nanoparticle formulation enhances the delivery and activity of a vascular endothelial growth factor antisense oligonucleotide in human retinal pigment epithelial cells

Nanoparticle formulation enhances the delivery and activity of a vascular endothelial growth factor antisense oligonucleotide in human retinal pigment epithelial cells

Argon Laser Photocoagulation–Induced Modification of Gene Expression in the Retina

Angiotensin-TGF-&#946;1 Crosstalk in Human Idiopathic Pulmonary Fibrosis:Autocrine Mechanisms in Myofibroblasts and Macrophages

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Angiotensin-TGF-β1 Crosstalk in Human Idiopathic Pulmonary Fibrosis:Autocrine Mechanisms in Myofibroblasts and Macrophages