scholarly journals Acetyl-CoA carboxylase inhibition alters tubulin acetylation and aggregation in thrombin-stimulated platelets

2021 ◽  
Vol 42 (Supplement_1) ◽  
Author(s):  
M Octave ◽  
L Pirotton ◽  
A Ginion ◽  
V Robaux ◽  
S Lepropre ◽  
...  
Keyword(s):  
Platelet Aggregation ◽  
Lipid Content ◽  
Thrombus Formation ◽  
Lipid Synthesis ◽  
Phospholipid Content ◽  
Acetate Incorporation ◽  
Acetyl Coa Carboxylase ◽  
Acetyl Coa ◽  
Tubulin Acetylation ◽  
Platelet Functions

Abstract Introduction Acetyl-CoA carboxylase (ACC), the first enzyme regulating lipid synthesis, promotes thrombus formation by increasing platelet phospholipid content. Inhibition of its activity decreases lipogenesis and increases the content in acetyl-CoA which can serve as a substrate for protein acetylation. This posttranslational modification plays a key role in the regulation of platelet aggregation, via tubulin acetylation. Purpose To demonstrate that ACC inhibition may affect platelet functions via an alteration of lipid content and/or tubulin acetylation. Methods Platelets were treated 2 hours with CP640.186, a pharmacological ACC inhibitor, prior to thrombin stimulation. Platelet functions were assessed by aggregometry and flow cytometry. Lipogenesis was measured via 14C-acetate incorporation into lipids. Lipidomics analysis was carried out on the commercial Lipidyzer platform. Protein phosphorylation and acetylation were evaluated by western blot. Results Treatment with CP640.186 drastically decreased platelet lipogenesis. However, the quantitative lipidomics analyses showed that preincubation with the compound did not affect global platelet lipid content. Interestingly, this short-term ACC inhibition was sufficient to increase tubulin acetylation level, at basal state and after thrombin stimulation. It was associated with an impaired platelet aggregation, in response to low thrombin concentration, while granules secretion was not affected. Mechanistically, we highlighted a decrease in Rac1 activity, associated with a reduced phosphorylation of its downstream effector PAK2. Surprisingly, actin cytoskeleton was not impacted but we evidenced a significant decrease in ROS production which could result from a decreased NOX2 activity. Conclusion Pharmacological ACC inhibition decreases platelet aggregation upon thrombin stimulation. The mechanism depends on increased tubulin acetylation, with subsequent alteration of the Rac1/PAK2/NOX2 signaling pathway FUNDunding Acknowledgement Type of funding sources: Other. Main funding source(s): Fonds pour la formation à la Recherche dans l'Industrie et l'Agriculture (FRIA)

scholarly journals Acetyl-CoA Carboxylase Inhibitor CP640.186 Increases Tubulin Acetylation and Impairs Thrombin-Induced Platelet Aggregation

2021 ◽  
Vol 22 (23) ◽  
pp. 13129
Author(s):  
Marie Octave ◽  
Laurence Pirotton ◽  
Audrey Ginion ◽  
Valentine Robaux ◽  
Sophie Lepropre ◽  
...  
Keyword(s):  
Platelet Aggregation ◽  
Actin Cytoskeleton ◽  
De Novo ◽  
Lipid Synthesis ◽  
Human Platelets ◽  
Acetyl Coa Carboxylase ◽  
Acetyl Coa ◽  
Signaling Systems ◽  
Tubulin Acetylation ◽  
The Impact

Acetyl-CoA carboxylase (ACC) is the first enzyme regulating de novo lipid synthesis via the carboxylation of acetyl-CoA into malonyl-CoA. The inhibition of its activity decreases lipogenesis and, in parallel, increases the acetyl-CoA content, which serves as a substrate for protein acetylation. Several findings support a role for acetylation signaling in coordinating signaling systems that drive platelet cytoskeletal changes and aggregation. Therefore, we investigated the impact of ACC inhibition on tubulin acetylation and platelet functions. Human platelets were incubated 2 h with CP640.186, a pharmacological ACC inhibitor, prior to thrombin stimulation. We have herein demonstrated that CP640.186 treatment does not affect overall platelet lipid content, yet it is associated with increased tubulin acetylation levels, both at the basal state and after thrombin stimulation. This resulted in impaired platelet aggregation. Similar results were obtained using human platelets that were pretreated with tubacin, an inhibitor of tubulin deacetylase HDAC6. In addition, both ACC and HDAC6 inhibitions block key platelet cytoskeleton signaling events, including Rac1 GTPase activation and the phosphorylation of its downstream effector, p21-activated kinase 2 (PAK2). However, neither CP640.186 nor tubacin affects thrombin-induced actin cytoskeleton remodeling, while ACC inhibition results in decreased thrombin-induced reactive oxygen species (ROS) production and extracellular signal-regulated kinase (ERK) phosphorylation. We conclude that when using washed human platelets, ACC inhibition limits tubulin deacetylation upon thrombin stimulation, which in turn impairs platelet aggregation. The mechanism involves a downregulation of the Rac1/PAK2 pathway, being independent of actin cytoskeleton.

scholarly journals Acetyl-CoA carboxylase inhibition alters tubulin acetylation and aggregation in thrombin-stimulated platelets

2021 ◽  
Vol 13 (2) ◽  
pp. 182-183
Author(s):  
M. Octave ◽  
L. Pirotton ◽  
A. Ginion ◽  
V. Robaux ◽  
S. Lepropre ◽  
...  
Keyword(s):  
Acetyl Coa Carboxylase ◽  
Acetyl Coa ◽  
Tubulin Acetylation

scholarly journals Doxorubicin contributes to thrombus formation and vascular injury by interfering with platelet function

2020 ◽  
Vol 319 (1) ◽  
pp. H133-H143 ◽  
Author(s):  
Haichen Lv ◽  
Ruopeng Tan ◽  
Jiawei Liao ◽  
Zhujing Hao ◽  
Xiaolei Yang ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Platelet Aggregation ◽  
Vascular Injury ◽  
Platelet Function ◽  
Thrombus Formation ◽  
Platelet Activity ◽  
Vascular Toxicity ◽  
Platelet Functions

Doxorubicin therapy in mice (antitumor dosage) markedly enhanced platelet functions measured as agonist-induced platelet aggregation, degranulation, and adhesion to endothelial cells, actions leading to thrombus formation and thrombosis-independent vascular injury. Clopidogrel treatment ameliorated thrombus formation and vascular toxicity induced by doxorubicin via inhibiting platelet activity.

scholarly journals The sensitivity of acetyl-coenzyme A carboxylase to citrate stimulation in a homogenate of rat liver containing subcellular particles

1970 ◽  
Vol 117 (2) ◽  
pp. 385-395 ◽  
Author(s):  
Jill Iliffe ◽  
N. B. Myant
Keyword(s):  
Fatty Acids ◽  
Rat Liver ◽  
Specific Radioactivity ◽  
Acid Synthesis ◽  
Acetate Incorporation ◽  
Acetyl Coa Carboxylase ◽  
Acetyl Coa ◽  
Carboxylase Activity ◽  
Highly Sensitive ◽  
Activated State

1. Although citrate is known to activate purified preparations of acetyl-CoA carboxylase, it had no stimulatory effect on the incorporation of [14C]acetate into long-chain fatty acids in a whole homogenate of rat liver (S0.7) under conditions in which the activity of acetyl-CoA carboxylase was rate-limiting for fatty acid synthesis. 2. The rate of incorporation of acetyl carbon into fatty acids was estimated in S0.7 preparations incubated with [14C]acetate, by measuring the specific radioactivity of the acetyl carbon of acetyl-CoA and the incorporation of 14C into fatty acids. These estimates were compared with estimates of acetyl-CoA carboxylase activity in the S0.7 preparation obtained by direct assay in conditions in which the enzyme was in the fully activated state. 3. In the absence of citrate, incorporation of acetyl carbon into fatty acids was about 75% of the value expected if the acetyl-CoA carboxylase in the S0.7 preparation were in the fully activated state. 4. Incorporation of acetyl carbon into fatty acids in the S0.7 preparation was stimulated by citrate, but the effect was many times less than the stimulation of [14C]acetate incorporation by citrate in particle-free preparations. 5. When the mitochondria and microsomes were removed from the S0.7 preparation, [14C]acetate incorporation into fatty acids fell to a negligible value and the preparation became highly sensitive to stimulation by citrate. 6. It is suggested that in the presence of mitochondria and microsomes, and in the intact liver cell, the degree of activation of acetyl-CoA carboxylase is such that citrate activation may not be of physiological significance.

BAY i 7351, A New Platelet Inhibitor, and antithrombotic Compound

1979 ◽  
Author(s):  
F. Seuter ◽  
W.D. Busse ◽  
U. Hoerlein ◽  
H. Boeshagen ◽  
F. Hoffmeister ◽  
...  
Keyword(s):  
Platelet Aggregation ◽  
Jugular Vein ◽  
Ex Vivo ◽  
Thrombus Formation ◽  
Minimal Effective Dose ◽  
Platelet Inhibitor ◽  
Small Doses ◽  
Effective Doses ◽  
Platelet Functions

4,4′ -Dichloro-N ,N′ -bis [( 1 -methyl-4-piperidinyl) -methyl] -2,2′ di thiobisbenzamide = BAY i 7351 was tested in animal models (rat and rabbit) of traumatically induced thrombosis. After prophylactic administration of small doses of BAY i 7351 (0.3 mg/kg p.o.) to rats the thrombus formation is inhibited (p < 0.05) both in the arterial and the venous system by 79% and 57%, respectively. Further on thrombi already formed (20 and 24 hold) are reduced in weight. The minimal effective doses of these thrombolytic effects in rats are 2 mg/kg p.o. in the carotid artery, and 6 mg/ kg p.o. in the jugular vein, when administered in two single doses of 1 and 3 mg/ kg. The compound is not an anticoagulant or a fibrinolytic drug. It is an inhibitor of platelet aggregation - induced with various aggregating agents including ADP -, in vitro with minimal effective concentrations in the range of 1-10 μg/ml as well as ex vivo (minimal effective dose: 3-10 mg/ kg p.o., rat). Further investigations as to v arious platelet functions, influence on thromboxane and prostacyclin formation will be reported separately. BAY i 7351 is a compound with properties which are considered favourable for treatment of thromboembolic diseases.

Fatty-acid biosynthesis and acetyl-coa carboxylase as a target of diclofop, fenoxaprop and other aryloxy-phenoxy-propionic acid herbicides

1988 ◽  
Vol 43 (1-2) ◽  
pp. 47-54 ◽  
Author(s):  
Klaus Kobek ◽  
Manfred Focke ◽  
K. Lichtenthaler Botanisches
Keyword(s):  
Fatty Acid ◽  
Propionic Acid ◽  
Fatty Acid Biosynthesis ◽  
De Novo ◽  
Free Acid ◽  
Acetate Incorporation ◽  
Acetyl Coa Carboxylase ◽  
Acid Biosynthesis ◽  
Acetyl Coa ◽  
Acid Herbicides

The effect of the herbicides and aryloxy-phenoxy-propionic acid derivatives diclofop, fenoxaprop, fluazifop and haloxyfop and their ethyl, methyl or butyl esters on the de novo fatty-acid biosynthesis of isolated chloroplasts was investigated with intact chloroplasts isolated from sensitive grasses (Poaceae) and tolerant dicotyledonous plants (Pisum, Spinacia). The 4 herbicides (free-acid form) block the de novo fatty-acid biosynthesis ([2-14C]acetate incorporation into the total fatty-acid fraction) of the sensitive Avena chloroplasts in a dose-dependent manner. The I50- values (a 50% inhibition of the [14C]acetate incorporation) lie in the range of 10-7 to 2 x 10-6 ᴍ. The ethyl or methyl esters (diclofop, fenoxaprop, haloxyfop) and butyl ester (fluazifop) do not affect the de novo fatty-acid biosynthesis of isolated chloroplasts or only at a very high concentration of ca. 10-4 ᴍ. In contrast, the de novo fatty-acid biosynthesis of the tolerant dicotyledonous species (pea, spinach) is not affected by the 4 aryloxy-phenoxy-propionic acid herbicides. In an enzyme preparation isolated from chloroplasts of the herbicide-sensitive barley plants the de novo fatty-acid biosynthesis from [14C]acetate and [14C]acetyl-CoA is blocked by all 4 herbicides (free acids), whereas that of [14C]malonate and [14C]malonyl-CoA is not affected. This strongly suggests that the target of all 4 herbicides (free-acid form) is the acetyl-CoA carboxylase within the chloroplasts. The applied ester derivatives, in turn, which are ineffective in the isolated chloroplast test system, have equally little or no effect on the activity of the acetyl-CoA carboxylase. It is assumed that the acetyl-CoA carboxylase of the tolerant dicot plants investigated is modified in such a way that the 4 herbicides cannot bind to and affect the target

175 EFFECT OF ACETYL-CoA CARBOXYLASE (ACC) INHIBITOR ON THE LIPID CONTENT AND NUCLEAR MATURATION OF CANINE OOCYTES

2014 ◽  
Vol 26 (1) ◽  
pp. 202 ◽  
Author(s):  
J. McGill ◽  
G. Reddy ◽  
L. Simon ◽  
G. Wirtu
Keyword(s):  
Lipid Content ◽  
In Vitro Maturation ◽  
Animal Health ◽  
Fluorescent Microscopy ◽  
Cumulus Cell ◽  
Nile Red ◽  
Acetyl Coa Carboxylase ◽  
Acetyl Coa ◽  
Nuclear Maturation

Compared with other domestic species, embryo technologies are least developed for the dog. This is mainly due to difficulties in producing mature oocytes in vitro. Canine oocytes contain exceptionally high amounts of lipid. High lipid content increases the chilling sensitivity of oocytes and embryos. Mechanical and chemical reductions of the lipid content have been used to improve the cryotolerance of oocytes. Additionally, chemical stimulation of lipid catabolism improved oocyte in vitro maturation (IVM) rates in other species (You et al. 2012 Theriogenology 78, 235–543). Acetyl-CoA carboxylase (ACC) is the rate-limiting enzyme in de novo lipogenesis and its expression has been reported in oocytes and embryos. In somatic cells, inhibition of ACC reduces lipogenesis and enhances β-oxidation. Our hypothesis is that treatment of oocytes with an inhibitor of ACC (CP640186, Pfizer Animal Health, New York, NY, USA) reduces lipid content and improves IVM rate of oocytes. Ovaries were collected from a spay clinic and sliced in HEPES-buffered TCM-199 to recover oocytes. In vitro maturation was conducted at 38.5°C, 5% CO2, and high humidity in TCM-199 supplemented with 1% fetal bovine serum, glutamine, sodium pyruvate, β-mercaptoethanol, oestradiol, epidermal growth factor, and antimicrobial agents (Songsasen et al. Mol. Reprod. Dev. 79, 186–196). During the first 19 to 21 h, the IVM media contained 4 concentrations of the inhibitor (0+DMSO, 0.02, 0.1, and 0.5 μM, designated as treatments 1, 2, 3, and 4, respectively) and then oocytes were transferred to a medium without the inhibitor and cultured for an additional 27 to 29 h. At the end of culture (total of 48 h), oocytes were denuded of cumulus layers, washed, fixed, and stained with Nile red (lipid) and Hoechst-33342 (chromatin), and then mounted on a microscope slide. Lipid content and chromatin status were evaluated using fluorescent microscopy (TRITC and DAPI filters, respectively). The relative lipid content was measured by the corrected total cell fluorescence (CTCF) using ImageJ software (http://rsbweb.nih.gov/ij/). Data on CTCF and proportions of chromatin status of oocytes were analysed using one-way ANOVA (SigmaPlot 11.0). The mean CTCF for each treatment was 5.5 × 109 (n = 51, 5.2 × 109 (n = 44), 4.5 × 109 (n = 31), and 4.8 × 109 (n = 34), respectively (P = 0.3; 4 replicates). At the highest dose, the agent induced relatively more cumulus cell layer expansion but inhibited their attachment to the dish; the latter effect was reversible because cumulus cells attached and proliferated after washing the oocytes of the agent. Metaphase II was rare (≤3.1%); however, the proportion of oocytes developing to ≥GVBD stage (Trt 1 14%, n = 37; Trt 2 41%, n = 56; Trt 3 5%, n = 22; Trt 4 11%, n = 43) was affected by treatments. Our preliminary data indicate that a low concentration of ACC inhibitor has a positive effect on the nuclear maturation of canine oocytes but the effect on lipid content as estimated by using Nile red fluorescence intensity appears to be minimal.

Sign in / Sign up

Export Citation Format

Share Document