Antisense Approach to Isoform-Specific Blockade of Acetylcholinesterase

2020 ◽  
pp. 570-591
Keyword(s):  
Antisense Approach

scholarly journals A multi-target antisense approach against PDE4 and PDE7 reduces smoke-induced lung inflammation in mice

2009 ◽  
Vol 10 (1) ◽  
Author(s):  
Marylène Fortin ◽  
Hélène D'Anjou ◽  
Marie-Ève Higgins ◽  
Jasmine Gougeon ◽  
Paméla Aubé ◽  
...  
Keyword(s):  
Lung Inflammation ◽  
Antisense Approach

Repression of Wnt-5a impairs DDR1 phosphorylation and modifies adhesion and migration of mammary cells

2001 ◽  
Vol 114 (11) ◽  
pp. 2043-2053 ◽  
Author(s):  
Marzieh Jönsson ◽  
Tommy Andersson
Keyword(s):  
Mammary Epithelial Cells ◽  
Human Mammary Epithelial Cell ◽  
Mammary Epithelial ◽  
Epithelial Cell Line ◽  
Collagen Binding ◽  
Collagen Matrices ◽  
Endogenous Expression ◽  
Human Mammary Epithelial ◽  
Antisense Approach ◽  
And Migration

The Wnt-5a gene encodes a secreted protein that controls several normal processes during embryogenesis and development of adult tissues by as yet unknown mechanisms. Endogenous expression of Wnt-5a mRNA is known to occur in both mouse and human mammary cell lines. To investigate the biological role of Wnt-5a in the human mammary epithelial cell line HB2, we used an antisense approach to repress endogenous expression of Wnt-5a protein. We also generated a cell population that constitutively overexpresses this protein. We found that overexpression of Wnt-5a protein enhanced cell-to-collagen binding and abolished hepatocyte growth factor-stimulated migration of HB2 transfectants through collagen matrices. Conversely, repression of Wnt-5a protein led to cell scattering, impaired cell-collagen interaction and enhanced cell motility. As we were searching for modified collagen receptors in antisense cells, we discovered that the collagen-binding discoidin domain receptor 1 (DDR1) failed to undergo phosphorylation. In reciprocal experiments, phosphorylation of DDR1 was consistently enabled by expression of Wnt-5a-HA protein in non-Wnt-5a-producing MCF-7 breast cancer cells. Activation of the Wnt/β-catenin signalling pathway did not influence or mimic the Wnt-5a-mediated effect on DDR1 phosphorylation. These data demonstrate that Wnt-5a protein participates in regulation of adhesion to and migration through collagen and is also a co-factor necessary for collagen-induced activation of DDR1 receptors in mammary epithelial cells.

Regulation of PAI-1 Concentration in Platelets by Systemic Administration of Antisense Oligonucleotides to Rats

2001 ◽  
Vol 85 (06) ◽  
pp. 1086-1089 ◽  
Author(s):  
Zofia Pawlowska ◽  
Ewa Chabielska ◽  
Anna Kobylanska ◽  
Anna Maciaszek ◽  
Maria Swiatkowska ◽  
...  
Keyword(s):  
Antisense Oligonucleotide ◽  
Antisense Oligonucleotides ◽  
Low Doses ◽  
Significant Delay ◽  
Occlusion Time ◽  
Antisense Approach ◽  
First Time ◽  
Pai 1 ◽  
Arterial Thrombolysis

SummaryIn this report we tested the effect of oligodeoxyribonucleotides antisense to PAI-1 mRNA administered into rats on PAI-1 concentration in platelets. Low doses of the antisense oligonucleotide (MPO-16R) reduced PAI-1 activity, both in rat blood plasma and platelet lysates by 20.5% and 28.7%, respectively. There was no change in platelet count after treatment with MPO-16R but treated platelets showed lower aggregability as compared with controls (37 ± 13% and 54 ± 12%, respectively). In an experimental model of rat arterial thrombosis, low doses of MPO-16R caused a significant delay in the occlusion time (31.8%). These data further support for the role of PAI-1 as a major determinant of arterial thrombolysis resistance and for the first time demonstrate the possibility of reduction of platelet PAI-1 concentration by antisense approach.

Peptide nucleic acid restores colistin susceptibility through modulation of MCR-1 expression in Escherichia coli

2020 ◽  
Author(s):  
Xiaoming Wang ◽  
Yao Wang ◽  
Zhuoren Ling ◽  
Chaoyang Zhang ◽  
Mingming Fu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Peptide Nucleic Acid ◽  
Promoter Activity ◽  
Sequence Variation ◽  
Promoter Sequence ◽  
E Coli ◽  
Expression In Escherichia Coli ◽  
Expression Host ◽  
Antisense Approach ◽  
Increased Susceptibility

Abstract Background Plasmid-mediated mechanisms of drug resistance accelerate the spread of polymyxin resistance, leaving clinicians with few or no antibacterial options for the treatment of infections caused by MDR bacteria, especially carbapenemase-producing strains. Objectives To evaluate the associations among promoter sequence variation, mcr-1 expression, host factors and levels of colistin resistance and to propose antisense agents such as peptide nucleic acids (PNAs) targeting mcr-1 as a tool to restore colistin susceptibility through modulation of MCR-1 expression in Escherichia coli. Methods A β-galactosidase assay was performed to study mcr-1 promoter activity. Quantitative real-time PCR and western blot assays were used to identify the expression level of MCR-1 in WT strains and transformants. Three PNAs targeting different regions of mcr-1 were designed and synthesized to determine whether they can effectively inhibit MCR-1 expression. MIC was measured to test colistin susceptibility in the presence or absence of PNA-1 in mcr-1-carrying E. coli. Results Variation in the mcr-1 promoter sequence and host species affect promoter activity, MCR-1 expression levels and colistin MICs. One PNA targeting the ribosome-binding site fully inhibited the expression of mcr-1 at a concentration of 4 μM, resulting in significantly increased susceptibility to colistin. The MIC90 of colistin decreased from 8 to 2 mg/L (P < 0.05) in the presence of 4 μM PNA. Conclusions These findings suggest that the antisense approach is a possible strategy to combat mcr-1-mediated resistance as well as other causes of emerging global resistance.

scholarly journals Hyperketonemia (Acetoacetate) Upregulates NADPH Oxidase 4 and Elevates Oxidative Stress, ICAM-1, and Monocyte Adhesivity in Endothelial Cells

2015 ◽  
Vol 35 (1) ◽  
pp. 364-373 ◽  
Author(s):  
Preeti Kanikarla-Marie ◽  
Sushil K. Jain
Keyword(s):  
Oxidative Stress ◽  
Endothelial Cells ◽  
High Glucose ◽  
Microvascular Dysfunction ◽  
New Drugs ◽  
Cellular Injury ◽  
Nadph Oxidase 4 ◽  
Antisense Approach ◽  
Cellular Dysfunction

Background/Aims: The incidence of developing microvascular dysfunction is significantly higher in type 1 diabetic (T1D) patients. Hyperketonemia (acetoacetate, β-hydroxybutyrate) is frequently found along with hyperglycemia in T1D. Whether hyperketonemia per se contributes to the excess oxidative stress and cellular injury observed in T1D is not known. Methods: HUVEC were treated with ketones in the presence or absence of high glucose for 24 h. NOX4 siRNA was used to specifically knockdown NOX4 expression in HUVEC. Results: Ketones alone or in combination with high glucose treatment cause a significant increase in oxidative stress, ICAM-1, and monocyte adhesivity to HUVEC. Using an antisense approach, we show that ketone induced increases in ROS, ICAM-1 expression, and monocyte adhesion in endothelial cells were prevented in NOX4 knockdown cells. Conclusion: This study reports that elevated levels of ketones upregulate NOX, contributing to increased oxidative stress, ICAM-1 levels, and cellular dysfunction. This provides a novel biochemical mechanism that elucidates the role of hyperketonemia in the excess cellular injury in T1D. New drugs targeting inhibition of NOX seems promising in preventing higher risk of complications associated with T1D.

scholarly journals Phosphoinositide-Specific Inositol Polyphosphate 5-Phosphatase IV Inhibits Inositide Trisphosphate Accumulation in Hypothalamus and Regulates Food Intake and Body Weight

Endocrinology ◽  
2006 ◽  
Vol 147 (11) ◽  
pp. 5385-5399 ◽  
Author(s):  
Daniela F. Bertelli ◽  
Eliana P. Araújo ◽  
Maristela Cesquini ◽  
Graziela R. Stoppa ◽  
Miriam Gasparotto-Contessotto ◽  
...  
Keyword(s):  
Body Weight ◽  
Food Intake ◽  
Insulin Receptor ◽  
Tyrosine Phosphorylation ◽  
Body Weight Loss ◽  
Pi3 Kinase ◽  
Inositol Polyphosphate ◽  
Intracellular Signals ◽  
Antisense Approach ◽  
Inositol Phosphatases

The enzyme phosphatidylinositol 3-kinase (PI3-kinase) exerts an important role in the transduction of the anorexigenic and thermogenic signals delivered by insulin and leptin to first-order neurons of the arcuate nucleus in the hypothalamus. The termination of the intracellular signals generated by the activation of PI3-kinase depends on the coordinated activity of specific inositol phosphatases. Here we show that phosphoinositide-specific inositol polyphosphate 5-phosphatase IV (5ptase IV) is highly expressed in neurons of the arcuate and lateral nuclei of the hypothalamus. Upon intracerebroventricular (ICV) treatment with insulin, 5ptase IV undergoes a time-dependent tyrosine phosphorylation, which follows the same patterns of canonical insulin signaling through the insulin receptor, insulin receptor substrate-2, and PI3-kinase. To evaluate the participation of 5ptase IV in insulin action in hypothalamus, we used a phosphorthioate-modified antisense oligonucleotide specific for this enzyme. The treatment of rats with this oligonucleotide for 4 d reduced the hypothalamic expression of 5ptase IV by approximately 80%. This was accompanied by an approximately 70% reduction of insulin-induced tyrosine phosphorylation of 5ptase IV and an increase in basal accumulation of phosphorylated inositols in the hypothalamus. Finally, inhibition of hypothalamic 5ptase IV expression by the antisense approach resulted in reduced daily food intake and body weight loss. Thus, 5ptase IV is a powerful regulator of signaling through PI3-kinase in hypothalamus and may become an interesting target for therapeutics of obesity and related disorders.

Involvement of protein kinase D in Fcγ-receptor activation of the NADPH oxidase in neutrophils

2002 ◽  
Vol 363 (1) ◽  
pp. 95-103 ◽  
Author(s):  
Jan K. DAVIDSON-MONCADA ◽  
Guillermo LOPEZ-LLUCH ◽  
Anthony W. SEGAL ◽  
Lodewijk V. DEKKER
Keyword(s):  
Protein Kinase ◽  
Nadph Oxidase ◽  
Receptor Activation ◽  
Protein Kinase D ◽  
Fcγ Receptor ◽  
Fcγ Receptors ◽  
Down Regulation ◽  
Antisense Approach ◽  
Nadph Oxidase Activation

Protein kinases involved in the activation of the NADPH oxidase by Fcγ receptors in neutrophils were studied. Of three different protein kinase C (PKC) inhibitors, Gö 6976 inhibited the NADPH oxidase completely, whereas bisindolylmaleimide I and Ro 31-8220 caused a 70–80% inhibition. Thus a Gö 6976-sensitive, bisindolylmaleimide I/Ro 31-8220-insensitive component contributes to NADPH oxidase activation induced by Fcγ receptors. Down-regulation of PKC isotypes resulted in inhibition of Fcγ-receptor-activated NADPH oxidase, but a down-regulation-insensitive component was still present. This component was sensitive to Gö 6976, but insensitive to Ro 31-8220. It has been shown previously that protein kinase D/PKC-μ (PKD) shows this same pharmacology in vitro. We show that PKD is present in neutrophils and that, in contrast with PKC isotypes, PKD is not down-regulated. Therefore PKD may participate in NADPH oxidase activation. To obtain direct evidence for this we adopted an antisense approach. Antisense PKD inhibited NADPH oxidase induced by Fcγ-receptor stimulation by 50% and the Ro 31-8220-insensitive component in the activation was inhibited by antisense PKD. In vitro kinase assays showed that PKD is activated by presenting IgG-opsonized particles to neutrophils. Furthermore, PKD localizes to the area of particle intake in the cell and phosphorylates two of the three cytosolic components of the NADPH oxidase, p40phox and p47phox. Taken together, these data indicate that Fcγ receptors engage PKD in the regulation of the NADPH oxidase.

Identification of REST-Regulated Genes and Pathways Using a REST-Targeted Antisense Approach

2013 ◽  
Vol 23 (6) ◽  
pp. 389-400 ◽  
Author(s):  
Yalda Sedaghat ◽  
Huynh-Hoa Bui ◽  
Curt Mazur ◽  
Brett P. Monia
Keyword(s):  
Antisense Approach
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