Since its discovery, the study of the biological role of α-synuclein and its pathological implications has been the subject of increasing interest. The propensity to adopt different conformational ...states governing its aggregation and fibrillation makes this small 14-kDa cytosolic protein one of the main etiologic factors associated with degenerative disorders known as synucleinopathies. The structure, function, and toxicity of α-synuclein and the possibility of different therapeutic approaches to target the protein have been extensively investigated and reviewed. One intriguing characteristic of α-synuclein is the different ways in which it interacts with lipids. Though in-depth studies have been carried out in this field, the information they have produced is puzzling and the precise role of lipids in α-synuclein biology and pathology and
is still largely unknown. Here we provide an overview and discussion of the main findings relating to α-synuclein/lipid interaction and its involvement in the modulation of lipid metabolism and signaling.
Pathological α-synuclein (α-syn) overexpression and iron (Fe)-induced oxidative stress (OS) are involved in the death of dopaminergic neurons in Parkinson's disease (PD). We have previously ...characterized the role of triacylglycerol (TAG) formation in the neuronal response to Fe-induced OS. In this work we characterize the role of the α-syn variant A53T during Fe-induced injury and investigate whether lipid metabolism has implications for neuronal fate. To this end, we used the N27 dopaminergic neuronal cell line either untransfected (UT) or stably transfected with pcDNA3 vector (as a transfection control) or pcDNA-A53T-α-syn (A53T α-syn). The overexpression of A53T α-syn triggered an increase in TAG content mainly due to the activation of Acyl-CoA synthetase. Since fatty acid (FA) β-oxidation and phospholipid content did not change in A53T α-syn cells, the unique consequence of the increase in FA-CoA derivatives was their acylation in TAG moieties.
Control cells exposed to Fe-induced injury displayed increased OS markers and TAG content. Intriguingly, Fe exposure in A53T α-syn cells promoted a decrease in OS markers accompanied by α-syn aggregation and elevated TAG content.
We report here new evidence of a differential role played by A53T α-syn in neuronal lipid metabolism as related to the neuronal response to OS.
Effect of A53T α-syn overexpression and Fe exposure on neuronal lipid metabolism. A53T α-syn overexpression increased FAS expression levels, ACS activity and TAG content. A53T α-syn diminished ROS generation and lipid peroxidation induced by Fe-exposure. A53T α-syn cells exposed to Fe showed increased TAG content. The appearance of TAG in A53T α-syn cells could constitute a marker of neuronal injury during the early stages of PD. Display omitted
•A53T α-syn overexpression induced TAG and lipid droplet accumulation in dopaminergic neurons.•Dopaminergic neurons that overexpressed A53T α-syn displayed augmented Acyl-CoA synthetase activity.•Fe induced A53T α-syn aggregation and increased TAG content in neurons.•Blockage of TAG synthesis reduced A53T α-syn cell viability.
α-synuclein (α-syn) accumulation and aggregation is a common pathological factor found in synucleinopathies, a group of neurodegenerative disorders that includes Parkinson´s disease (PD). It has been ...proposed that lipid dyshomeostasis is responsible for the occurrence of PD-related processes, however, the precise role of lipids in the onset and progression of neurodegenerative disorders remains unclear. Our aim was to investigate the effect of α-syn overexpression on neutral lipid metabolism and how this impacts on neuronal fate. We found lipid droplet (LD) accumulation in cells overexpressing α-syn to be associated with a rise in triacylglycerol (TAG) and cholesteryl ester (CE) levels. α-syn overexpression promoted diacylglycerol acyltransferase 2 upregulation and acyl-CoA synthetase activation, triggering TAG buildup, that was accompanied by an increase in diacylglycerol acylation. Moreover, the CE increment was associated with higher activity of acyl-CoA:cholesterol acyltransferase. Interestingly, α-syn overexpression increased cholesterol lysosomal accumulation. We observed that sterol regulatory element-binding protein (SREBP)-1 and SREBP-2 were differentially regulated by α-syn overexpression. The latter gave rise to a reduction in SREBP-1 nuclear translocation and consequently in fatty acid synthase expression, whereas it produced an increase in SREBP-2 nuclear localization. Surprisingly, and despite increased cholesterol levels, SREBP-2 downstream genes related to cholesterolgenesis were not upregulated as expected. Notably, phospholipid (PL) levels were diminished in cells overexpressing α-syn. This decrease was related to the activation of phospholipase A2 (PLA2) with a concomitant imbalance of the PL deacylation-acylation cycle. Fatty acids released from PLs by iPLA2 and cPLA2 action were esterified into TAGs, thus promoting a biological response to α-syn overexpression with uncompromised cell viability. When the described steady-state was disturbed under conditions favoring higher levels of α-syn, the response was an enhanced LD accumulation, this imbalance ultimately leading to neuronal death.
Brain iron accumulation constitutes a pathognomonic indicator in several neurodegenerative disorders. Metal accumulation associated with dopaminergic neuronal death has been documented in Parkinson's ...disease. Through the use of in vivo and in vitro models, we demonstrated that lipid dysregulation manifests as a neuronal and glial response during iron overload.
In this study, we show that cholesterol content and triacylglycerol (TAG) hydrolysis were strongly elevated in mice midbrain. Lipid cacostasis was concomitant with the loss of dopaminergic neurons, astrogliosis and elevated expression of α-synuclein. Exacerbated lipid peroxidation and markers of ferroptosis were evident in the midbrain from mice challenged with iron overload. An imbalance in the activity of lipolytic and acylation enzymes was identified, favoring neutral lipid hydrolysis, and consequently reducing TAG and cholesteryl ester levels. Notably, these observed alterations were accompanied by motor impairment in iron-treated mice.
In addition, neuronal and glial cultures along with their secretomes were used to gain further insight into the mechanism underlying TAG hydrolysis and cholesterol accumulation as cellular responses to iron accumulation. We demonstrated that TAG hydrolysis in neurons is triggered by astrocyte secretomes. Moreover, we found that the ferroptosis inhibitor, ferrostatin-1, effectively prevents cholesterol accumulation both in neurons and astrocytes. Taken together, these results indicate that lipid disturbances occur in iron-overloaded mice as a consequence of iron-induced oxidative stress and depend on neuron-glia crosstalk. Our findings suggest that developing therapies aimed at restoring lipid homeostasis may lead to specific treatment for neurodegeneration associated with ferroptosis and brain iron accumulation.
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•Iron accumulation (IA) in mice midbrain triggers astrogliosis and movement disorders.•Midbrain IA leads to ferroptosis and dopaminergic cell death.•Acylation/lipolytic imbalance results in neutral lipid cacostasis during IA.•TAG hydrolysis induced by IA depends on neuro-glia crosstalk.•Cholesterol accumulation is retreated by inhibiting ferroptosis.
Alzheimer’s disease (AD) is a neurodegenerative disorder associated with memory impairment and cognitive deficit. Most of the drugs currently available for the treatment of AD are ...acetylcholinesterase (AChE) inhibitors. In a preliminary study, significant AChE inhibition was observed for the ethanolic extract of Grindelia ventanensis (IC50=0.79mg/mL). This result prompted us to isolate the active constituent, a normal labdane diterpenoid identified as 17-hydroxycativic acid (1), through a bioassay guided fractionation. Taking into account that 1 showed moderate inhibition of AChE (IC50=21.1μM), selectivity over butyrylcholinesterase (BChE) (IC50=171.1μM) and that it was easily obtained from the plant extract in a very good yield (0.15% w/w), we decided to prepare semisynthetic derivatives of this natural diterpenoid through simple structural modifications. A set of twenty new cativic acid derivatives (3–6) was prepared from 1 through transformations on the carboxylic group at C-15, introducing a C2–C6 linker and a tertiary amine group. They were tested for their inhibitory activity against AChE and BChE and some structure–activity relationships were outlined. The most active derivative was compound 3c, with an IC50 value of 3.2μM for AChE. Enzyme kinetic studies and docking modeling revealed that this inhibitor targeted both the catalytic active site and the peripheral anionic site of this enzyme. Furthermore, 3c showed significant inhibition of AChE activity in SH-SY5Y human neuroblastoma cells, and was non-cytotoxic.
We have previously shown that phospholipase D (PLD) pathways have a role in neuronal degeneration; in particular, we found that PLD activation is associated with synaptic injury induced by oxidative ...stress. In the present study, we investigated the effect of α-synuclein (α-syn) overexpression on PLD signaling. Wild Type (WT) α-syn was found to trigger the inhibition of PLD1 expression as well as a decrease in ERK1/2 phosphorylation and expression levels. Moreover, ERK1/2 subcellular localization was shown to be modulated by WT α-syn in a PLD1-dependent manner. Indeed, PLD1 inhibition was found to alter the neurofilament network and F-actin distribution regardless of the presence of WT α-syn. In line with this, neuroblastoma cells expressing WT α-syn exhibited a degenerative-like phenotype characterized by a marked reduction in neurofilament light subunit (NFL) expression and the rearrangement of the F-actin organization, compared with either the untransfected or the empty vector-transfected cells. The gain of function of PLD1 through the overexpression of its active form had the effect of restoring NFL expression in WT α-syn neurons. Taken together, our findings reveal an unforeseen role for α-syn in PLD regulation: PLD1 downregulation may constitute an early mechanism in the initial stages of WT α-syn-triggered neurodegeneration.
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•We studied the impact of WT α-syn overexpression on PLD1 signaling.•Overexpression of WT α-syn downregulates PLD1 and NFL expression.•ERK1/2 is downregulated and mainly located in nuclear fraction in WT α-syn neurons.•Pharmacological PLD1 inhibition triggers NFL loss and ERK1/2 nuclear localization in non-transfected neurons.•Overexpression of eGFP-PLD1 reestablishes NFL levels in WT α-syn neurons.
The development of hybrids from natural products is a promising strategy for drug discovery. In cancer therapy, there is a need to discover novel agents that can induce apoptosis in cancer cells. To ...contribute to this field of interest, we investigated the effect of a synthetic hybrid from cativic acid and caffeic acid (
5
) on viability, proliferation, and apoptosis in human neuroblastoma cells (IMR-32). Three hybrids were prepared via Mitsunobu esterification from 17-hydroxycativic acid (
1
) and natural phenols. Cell viability was analyzed by MTT assay. SYTOX green and LDH leakage were used to determine the cytotoxic effect. Caspase-3 activity, cell cycle phases, and proliferation were analyzed in order to characterize the biological effects of hybrid
5
. The mitogen-activated protein kinase (MAPK) status was evaluated for elucidating the potential mechanisms involved in hybrid
5
effect. Hybrid
5
reduced the viability of IMR-32 cells in a time- and concentration-dependent manner (IC
50
= 18.0 ± 1.3 μM) as a result of its antiproliferative effect through changes in the cell cycle distribution and induction of apoptosis associated with activation of caspase-3. Exposure to
5
triggered ERK1/2 activation and nuclear translocation. Hybrid
5
also promoted an increase in nuclear localization of the transcription factor c-Jun. Inhibition of ERK1/2 and JNK potentiated
5
-induced inhibition of IMR-32 viability. Hybrid
5
displays cell growth inhibition by promoting cell cycle arrest and apoptosis, through ERK1/2 and JNK participation.
Alzheimer's disease (AD) is a neurodegenerative disorder associated with memory impairment and cognitive deficit. Most of the drugs currently available for the treatment of AD are ...acetylcholinesterase (AChE) inhibitors. Plants of the Amaryllidaceae family are known to synthesize alkaloids, which have shown AChE inhibitory activity. Habranthus tubispathus and H. jamesonii are two Amaryllidaceae that can be found growing wild to the southwest of Buenos Aires in Argentina. Acetyl- and butyrylcholinesterase inhibition was observed for the extracts obtained from bulbs of H. tubispathus and bulbs and aerial parts of H. jamesonii. The strongest cholinesterase inhibition was observed for the alkaloid extract obtained from the aerial parts for H. jamesonii (AChE IC50= 0.7 μg/mL; BChE IC50= 6.7 μg/mL). The AChE inhibition observed for H. jamesonii could be explained by the presence of galanthamine and sanguinine, two potent AChE inhibitors. The levels of lycorine and hippeastidine, moderate AChE inhibitors, observed in the bulbs of H. tubispathus could be responsible for the significant AChE inhibition observed. The alkaloids present in these Amaryllidaceae were identified by means of GC-MS analysis. In the case of H. tubispathus, hippeastidine and 3-O-demethylhippeastidine, were isolated and completely characterized by 1H and 13C NMR spectroscopy.
A bioassay‐guided phytochemical analysis of the ethanolic extract of Grindelia argentina Deble & Oliveira‐Deble (Asteraceae) allowed the isolation of a known flavone, hispidulin, and three new ...oleanane‐type saponins, 3‐O‐β‐D‐xylopyranosyl‐(1→3)‐β‐D‐glucopyranosyl‐2β,3β,16α,23‐tetrahydroxyolean‐12‐en‐28‐oic acid 28‐O‐β‐D‐xylopyranosyl‐(1→2)‐β‐D‐apiofuranosyl‐(1→3)‐β‐D‐xylopyranosyl‐(1→3)‐α‐L‐rhamnopyranosyl‐(1→2)‐α‐L‐arabinopyranosyl ester (2), 3‐O‐β‐D‐glucopyranosyl‐2β,3β,23‐trihydroxyolean‐12‐en‐28‐oic acid 28‐O‐β‐D‐xylopyranosyl‐(1→2)‐β‐D‐apiofuranosyl‐(1→3)‐β‐D‐xylopyranosyl‐(1→3)‐α‐L‐rhamnopyranosyl‐(1→2)‐α‐L‐arabinopyranosyl ester, (3) and 3‐O‐β‐D‐xylopyranosyl‐(1→3)‐β‐D‐glucopyranosyl‐2β,3β,23‐trihydroxyolean‐12‐en‐28‐oic acid 28‐O‐β‐D‐xylopyranosyl‐(1→2)‐β‐D‐apiofuranosyl‐(1→3)‐β‐D‐xylopyranosyl‐(1→3)‐α‐L‐rhamnopyranosyl‐(1→2)‐α‐L‐arabinopyranosyl ester (4), named grindeliosides A–C, respectively. Their structures were determined by extensive 1D‐ and 2D‐NMR experiments along with mass spectrometry and chemical evidence. The isolated compounds were evaluated for their inhibitory activities against LPS/IFN‐γ‐induced NO production in RAW 264.7 macrophages and for their cytotoxic activities against the human leukemic cell line CCRF‐CEM and MRC‐5 lung fibroblasts. Hispidulin markedly reduced LPS/IFN‐γ‐induced NO production (IC50 51.4 μM), while grindeliosides A–C were found to be cytotoxic, with grindelioside C being the most active against both CCRF‐CEM (IC50 4.2±0.1 μM) and MRC‐5 (IC50 4.5±0.1 μM) cell lines.
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